AI HOC NONG LAM TP.

HO CH MINH
KHOA MOI TRNG VA TAI NGUYEN
WX

TS. Le Quoc Tuan

Bai giang

VI SINH moi trng

Lu hanh noi bo -2009-

GII THIEU
Vi sinh vat hoc moi trng la nganh nghien cu cac oi vi sinh vat ton tai trong moi trng t nhien va nhan tao. Nguon goc cua cac nghien cu bat au t s quan sat cua Antony van Leeuwenhoek (1677). Van Leeuwenhoek a s dung mot knh hien vi e kham pha nhng g ma ong goi la nhng ong vat nho chung song va sinh san trong nc ma, nc gieng, nc bien va nc bang tan. Trong suot nhieu the ky, s hieu biet cua chung ta ve vi sinh vat moi trng da tren nhng quan sat chi tiet va cac th nghiem vi s giup cua knh hien vi va cac cong cu hoa sinh cung nh toan hoc hien ai. Nhieu nghien cu cua van Leeuwenhoek da vao viec kiem tra cac mau vat c at trong ong nghiem, trong chai thuy tinh tai nha ong. Hien tai, ngi ta cho rang ong a tao nen mot moi trng nhan tao, cung nh o ma ngay nay chung ta co nhng ky thuat nuoi cay vi sinh vat trong phong th nghiem e xac nh cau truc cung nh chc nang cua chung. Nh chung ta a biet vi sinh vat hien dien khap ni, trong at, trong nc, khong kh, trong c the sinh vat khac, ac biet chung co the ton tai trong nhng moi trng khac nghiet nhat. Chung ong mot vai tro quan trong trong vong tuan hoan vat chat. V the, chung c xem la la mot mac xch quan trong trong qua trnh chuyen hoa vat chat. Trong phan vi sinh vat moi trng nay, chung ta se tm hieu cac qua trnh chuyen hoa vat chat trong moi trng t nhien va nhan tao. Qua o, co the nam bat c cac quy luat chuyen hoa chat hu c va vo c bi vi sinh vat nham ieu khien va ap dung chung mot cach hieu trong cac cong trnh x ly chat thai. Nghien cu s tac ong tng ho gia cac c the vi sinh vat, gia vi sinh vat va moi trng (cac tac nhan ly hoa va sinh hoc) nham kiem soat s phat trien cua chung, va nham tang cao hieu qua x ly chat thai cua chung khi c ap dung. T o chung ta se co nhng hieu biet ung an ve vi sinh vat va tam quan trong cua chung trong moi trng t nhien cung nh nhan tao.

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Khoa Moi Trng va Tai Nguyen ai Hoc Nong Lam TP. HCM

MUC LUC
Chng 1. S phan bo cua vi sinh vat trong moi trng 1.1. Moi Trng at va s phan bo cua vi sinh vat trong at 1.1.1. Moi trng at 1.1.2. S phan bo cua vi sinh vat trong at va moi quan he gia cac nhom VSV 1.1.3. Moi quan he gia at, vi sinh vat va thc vat 1.2. Moi trng nc va s phan bo cua vi sinh vat trong nc 1.2.1. S phan bo cua vi sinh vat trong nc 1.3. Moi trng khong kh va s phan bo cua vi sinh vat trong khong kh Chng 2. Kha nang chuyen hoa cac hp chat trong moi trng t nhien cua VSV 2.1. Vong tuan hoan nitrogen trong t nhien 2.2. Qua trnh amon hoa 2.2.1. S amon hoa urea 2.2.2. S amon hoa protein 2.3. Qua trnh nitrate hoa 2.3.1. Giai oan nitrite hoa 2.3.2. Giai oan nitrate hoa 2.4. Qua trnh phan nitrate hoa 2.5. Qua trnh co nh nitrogen phan t 2.6. S chuyen hoa cac hp chat phosphore cua vi sinh vat 2.6.1. Vong tuan hoan phosphore trong t nhien 2.6.2. S phan giai phosphore hu c trong at do vi sinh vat 2.6.3. S phan giai phosphore vo c do vi sinh vat 2.7. S chuyen hoa cac hp chat lu huynh cua vi sinh vat 2.7.1. Vong tuan hoan lu huynh trong t nhien 2.7.2. S oxy hoa cac hp chat lu huynh 2.7.3. S kh cac hp chat lu huynh vo c do vi sinh vat Chng 3. Sinh trng va phat trien vi sinh vat 3.1. Mau ly thuyet ve sinh trng va phat trien cua vi khuan 3.2. Sinh trng va phat trien cua vi khuan trong ieu kien nuoi cay tnh ng cong sinh trng 3.2.1. Pha lag 3.2.2. Pha log 3.2.3. Pha on nh 3.2.4. Pha t vong 3.3. Sinh trng cua vi khuan trong qua trnh nuoi cay lien tuc 3.4. Lam ong bo s phan chia te bao 3.5. Cac phng phap xac nh sinh trng va phat trien cua vi khuan 3.5.1. Cac phng phap xac nh so lng te bao 3.5.2. Cac phng phap xac nh sinh khoi te bao Trang 1 1 1 4 5 5 7

8 8 8 9 10 10 10 11 12 12 12 13 14 15 15 15 16 17 19 19 20 23 24 25 27 29 29 30

Tac dung cua cac yeu to ben ngoai len sinh trng va phat trien cua vi khuan 3.6.1. C che tac dung cua cac yeu to ben ngoai len vi khuan 3.6.2. Cac yeu to vat ly 3.6.3. Cac yeu to hoa hoc 3.6.4. Cac yeu to sinh hoc Chng 4. Thanh phan vi sinh vat tham gia trong qua trnh x ly nc thai 4.1. Vi khuan 4.1.1. Cau truc te bao 4.1.2. ieu kien moi trng 4.1.3. S phat trien cua vi khuan 4.1.4. ong hoc cua qua trnh x ly sinh hoc 4.1.5. ng dung s phat trien cua vi khuan va hoat ong s dung chat nen trong x ly sinh hoc. 4.2. Nam 4.3. Tao Chng 5. X ly nc thai bang vi sinh vat 5.1. X ly nc thai bang vi sinh dnh bam trong moi trng hieu kh 5.1.1. C s ly thuyet cua phng phap 5.1.2. Be loc sinh hoc co vat lieu tiep xuc khong ngap nc 5.1.3. Be loc sinh hoc co lp vat lieu ngap trong nc 5.2. X ly nc thai bang vi sinh vat yem kh trong moi trng can l lng va moi trng vi sinh dnh bam 5.2.1. Cac qua trnh sinh hoc va phan loai cong trnh 5.2.2. Be x ly yem kh co lp can l lng 5.2.3. Be loc yem kh 5.2.4. anh gia qua trnh Chng 6. Cac qua trnh kh nitrogen bang vi sinh vat 6.1. S chuyen hoa amonia bang qua trnh nitrate hoa sinh hoc 6.1.1. Mo ta qua trnh 6.1.2. Phan loai cac qua trnh nitrate 6.1.3. S oxy hoa carbon va nitrate hoa giai oan n (s o phoi hp) 6.1.4. Nitrate hoa giai oan kep (s o tach biet) 6.2.1. Loai bo nitrogen bang nitrate hoa/phan nitrate hoa sinh hoc 6.2.2. Phan loai cac qua trnh nitrate hoa/phan nitrate hoa Chng 7. Kh phosphorus bang cac phng phap sinh hoc 7.1. Cac qua trnh kh phosphorus 7.1.1. Qua trnh A/O (kh phosphorus dong chnh) 7.1.2. Qua trnh PhoStrip (kh phosphorus dong phu) 7.1.3. Be phan ng me lien tuc 7.1.4. So sanh cac qua trnh kh phosphorus sinh hoc 7.2. Viec kh nitrogen va phosphorus ket hp bang cac phng phap x ly

3.6.

31 32 33 36 38 40 40 41 42 45 47 47 48 51 51 52 59 60 60 63 65 66 68 68 69 70 73 74 76 82 84 84 85 85 86

sinh hoc 7.2.1. Qua trnh A2/O 7.2.2. Qua trnh 5 giai oan 7.2.3. So sanh cac qua trnh kh nitrogen va phosphorus sinh hoc ket hp Chng 8. Cac he thong x ly t nhien va ng dung 8.1. Cac he thong x ly t nhien 8.1.1. S phat trien cua cac he thong x ly t nhien 8.1.2. Toc o cham 8.1.3. R nhanh 8.1.4. He chay tran be mat 8.1.5. at ngap nc 8.1.6. He thc vat thuy sinh bac cao 8.1.7. He nuoi trong thuy san 8.2. Nhng nghien cu c ban trong viec ng dung he thong x ly t nhien 8.2.1. Cac ac tnh cua nc thai va c che x ly 8.2.2. Nhng van e ve sc khoe cong ong Tai lieu tham khao

87 87 88 90 90 93 95 97 99 100 100 101 101 106 108

Chng 1 S PHAN BO CUA VI SINH VAT TRONG MOI TRNG

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1.1. Moi trng at va s phan bo cua vi sinh vat trong at

1.1.1. Moi trng at. at la mot moi trng thch hp nhat oi vi vi sinh vat, v the no la ni c tru rong rai nhat cua vi sinh vat, ca ve thanh phan cung nh so lng so vi cac moi trng khac. S d nh vat la do trong at co mot lng ln cac chat hu c. o la nguon thc an cho cac nhom vi sinh vat d dng (vi sinh vat phan huy cac chat carbon hu c, nhom vi sinh vat phan huy cac chat nitrogen hu c). Cac chat vo c co trong at cung la nguon dinh dng cho cac nhom vi sinh vat t dng (cac nhom phan huy cac chat vo c, chuyen hoa cac chat S, P, Fe) Cac chat dinh dng khong nhng tap trung nhieu tang at mat ma con phan tan xuong cac tang at sau. Bi vay, cac tang at khac nhau, s phan bo vi sinh vat khac nhau phu thuoc va ham lng cac chat dinh dng. Mc o thoang kh cua at cung la mot ieu kien anh hng en s phan bo cua vi sinh vat. Cac nhom hieu kh phat trien nhieu nhng ni co nong o oxy cao. Nhng ni yem kh, ham lng oxy thap thng phan bo nhieu loai vi sinh vat k kh. o am va nhiet o trong at cung anh hng en s phat trien cua vi sinh vat at. at vung nhiet i thng co o am 70-80% va nhiet o 200C 300C. o la nhiet o va o am thch hp vi a so vi sinh vat. Bi vay, trong moi gam at thng co hang chuc trieu en hang t te bao vi sinh vat bao gom nhieu nhom khac nhau ve v tr phan loai cung nh hoat tnh sinh ly, sinh hoa va sinh thai. 1.1.2. S phan bo cua vi sinh vat trong at va moi quan he gia cac nhom vi sinh vat. 1.1.2.1. S phan bo cua vi sinh vat trong at. Vi sinh vat la nhng c the nho be de dang phat tan nh gio va cac sinh vat khac. Bi vay chung co the di chuyen de dang en moi ni trong t nhien. Nhat la nhng vi sinh vat co bao t, bao t cua chung co kha nang song tiem sinh trong cac ieu kien kho khan. Khi gap ieu kien thuan li, chung lai phat trien va sinh soi. Tuy nhien, at la ni vi sinh vat ton tai nhieu nhat so vi cac moi trng khac. S phan bo cua vi sinh vat at con goi la khu he vi sinh at. Chung bao gom cac nhom co ac tnh sinh ly, sinh hoa va sinh thai rat khac nhau. Cac nhom vi sinh vat chnh c tru trong at bao gom: Vi khuan, Vi nam, Xa khuan, Virus, Tao, Nguyen sinh ong vat. Trong o vi khuan la nhom chiem nhieu nhat ve so lng. Chung bao gom vi khuan hieu kh, vi khuan k kh, vi khuan t dng, vi khuan d dngNeu chia theo cac nguon dinh dng th no lai co nhom t dng carbon, t dng amin, d dng amin, vi khuan co nh nitrogen
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Theo nhieu tai lieu th trung bnh trong at vi khuan chiem khoang 90% tong so. Xa khuan chiem khoang 8%, vi nam 1%, con lai 1% la tao, nguyen sinh ong vat. T le nay thay oi tuy theo cac loai at khac nhau cung nh khu vc a ly, tang at, thi vu, che o canh tac nhng at co ay u chat dinh dng, o thoang kh tot, nhiet o, o am va pH thch hp th vi sinh vat phat trien nhieu ve so lng va thanh phan. S phat trien cua vi sinh vat lai chnh la nhan to lam cho at them ph nhieu, mau m. S phan bo cua vi sinh vat trong at co the chia ra nh sau: * Phan bo theo chieu sau. Quan the vi sinh vat thng tap trung nhieu nhat tang canh tac. o la ni tap trung re cay, chat dinh dng, co cng o chieu sang, nhiet o, o am thch hp nhat. So lng vi sinh vat giam dan theo tang at, cang xuong sau cang t vi sinh vat. Rieng oi vi cac at bac mau, do hien tng ra troi, tang 0 20 cm t chat hu c hn tang 20 40 cm. Bi vay, tang nay so lng vi sinh vat nhieu hn tang tren. Sau o giam dan cac tang di. Thanh phan vi sinh vat cung thay oi theo tang at: vi khuan hieu kh, vi nam, xa khuan thng tap trung tang mat v tang nay co nhieu oxy. Cang xuong sau, cac nhom vi sinh vat hieu kh cang giam manh. Ngc lai, cac nhom vi khuan k kh nh vi khuan phan nitrate hoa phat trien manh o sau 20 40 cm. vung kh hau nhiet i nong am thng co qua trnh ra troi, xoi mon nen tang 0 20 cm de bien ong, tang 20 40 cm on nh hn. * Phan bo theo cac loai at. Cac loai at khac nhau co ieu kien dinh dng, o am, o thoang kh, pH khac nhau. Bi vay s phan bo cua vi sinh vat cung khac nhau. at lua nc, tnh trang ngap nc lau ngay lam anh hng en o thong kh, che o nhiet, chat dinh dngCh co mot lp mong tren khoang 0 3 cm la co qua trnh oxy hoa, tang di qua trnh kh oxy chiem u the. Bi vay, trong at lua nc cac loai vi sinh vat k kh phat trien manh. V du nh vi khuan amon hoa, vi khuan phan nitrate hoa. Ngc lai, cac loai vi sinh vat hieu kh nh vi khuan nitrate hoa, vi khuan co nh nitrogen, vi nam va xa khuan eu rat t. Ty le gia vi khuan hieu kh va yem kh luon nho hn 1. at trong hoa mau, khong kh lu thong tot, qua trnh oxy hoa chiem u the, bi vay cac loai vi sinh vat hieu kh phat trien manh, vi sinh vat yem phat trien yeu. Ty le gia vi khuan hieu kh va yem kh thng ln hn 1, co trng hp at ti 4 5. at giau dinh dng nh at phu sa song Hong, so lng vi sinh vat tong so rat cao. Ngc lai, vung at bac mau Ha Bac co so lng vi sinh vat t nhat. * Phan bo theo cay trong. oi vi tat ca cac loai cay trong, vung re cay la vung vi sinh vat phat trien manh hn so vi vung khong co re. S d nh the v re cay cung cap mot lng ln chat hu c khi no chet i. Khi con song, ban than re cay cung thng xuyen tiet ra cac chat hu c lam nguon dinh dng cho vi sinh vat. Re cay con lam cho at thoang kh, gi c o
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am. Tat ca nhng nhan to o lam cho so lng vi sinh vat vung re phat trien manh hn vung ngoai re. Tuy nhien, moi loai cay trong trong qua trnh song cua no thng tiet qua bo re nhng chat khac nhau. Bo re khi chet i cung co thanh phan cac chat khac nhau. Thanh phan va so lng cac chat hu c tiet ra t bo re quyet nh thanh phan va so lng vi sinh vat song trong vung re o. V du nh vung re cay ho au thng phan bo nhom vi khuan co nh nitrogen cong sinh con vung re lua la ni c tru cua cac nhom co nh nitrogen t do hoac hoi sinh v.v So lng va thanh phan vi sinh vat cung thay oi theo cac giai oan phat trien cua cay trong. at vung phu sa song Hong, so lng vi sinh vat at cc ai giai oan lua choi nhanh, e nhanh, giai oan nay la cay lua sinh trng manh. Bi vay thanh phan va so lng chat hu c tiet qua bo re cang ln o la nguon dinh dng cho vi sinh vat vung re. So lng vi sinh vat at cc tieu thi ky lua chn. Thanh phan vi sinh vat cung bien ong theo cac giai oan phat trien cua cay phu hp vi ham lng cac chat tiet qua bo re. 1.1.2.2. Moi quan he gia cac nhom vi sinh vat trong at. S phan bo cua vi sinh vat trong at vo cung phong phu ca ve so lng cung nh thanh phan. Trong qua trnh song chung nh the, chung co mot moi quan he tng ho vo cung chat che. Da vao tnh chat cua cac loai quan he gia cac nhom vi sinh vat, ngi ta chia ra lam 4 loai quan he: ky sinh, cong sinh, ho sinh va khang sinh. * Quan he ky sinh: Quan he ky sinh la hien tng vi sinh vat nay song ky sinh tren vi sinh vat khac, hoan toan an bam va gay hai cho vat chu. V du nh cac loai virus song ky sinh trong te bao vi khuan hoac mot vai loai vi khuan song ky sinh tren vi nam. Cac loai vi khuan co nh nitrogen cong sinh thng hay b mot loai thc khuan Rhizobium ky sinh, tren moi trng dch the co hien tng moi trng ang uc tr nen trong. Nguyen nhan la do thc khuan the xam nhap va lam tan tat ca cac te bao vi khuan goi la hien tng sinh tan. Khi nuoi cay vi khuan tren moi trng ac cung co hien tng nh vay. Cac thc khuan nay ton tai trong at trong cay ho au lam anh hng rat ln en qua trnh hnh thanh not san cay au. * Quan he cong sinh La quan he hai ben cung co li, ben nay khong the thieu ben kia trong qua trnh sinh song. vi sinh vat ngi ta t quan sat thay quan he cong sinh. Co mot so gia thiet cho rang: ty the c quan ho hap cua te bao vi nam chnh la mot vi khuan cong sinh vi vi nam. Gia thiet o da tren cau tao cua ty the co ca bo may DNA rieng biet, co the t sao chep nh mot c the oc lap. Gia thiet nay cha c cong nhan hoan toan. Lai co gia thiet cho rang: cac plasmid co trong vi nam va vi khuan chnh la s cong sinh gia virus va vi nam hay vi khuan o. V du nh cac plasmid mang gen khang thuoc a mang lai moi li cho vi khuan chu la khang c thuoc khang sinh v the ma hai ben cung co li va goi la quan he cong sinh.
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* Quan he ho sinh La quan he hai ben cung co li nhng khong nhat thiet phai co nhau mi song c nh quan he cong sinh. Quan he nay thng thay trong s song cua vi sinh vat vung re. V du nh moi quan he gia nam moc phan huy tinh bot thanh ng va nhng nhom vi khuan phan giai loai ng o. Moi quan he gia nhom vi khuan phan giai phosphore va nhom vi khuan phan giai protein cung la quan he ho sinh, trong o nhom th nhat cung cap P cho nhom th hai va nhom th hai cung cap N cho nhom th nhat. * Quan he khang sinh Quan he khang sinh la moi quan he oi khang lan nhau gia hai nhom vi sinh vat. Loai nay thng tieu diet loai kia hoac han che qua trnh song cua no. V du ien hnh la xa khuan khang sinh va nhom vi khuan man cam vi chat khang sinh do xa khuan sinh ra. Khi nuoi cay 2 nhom nay tren moi trng thach a, ta co the thay ro hien tng khang sinh: xung quanh ni xa khuan moc co mot vong vo khuan, tai o vi khuan khong moc c. Ngi ta can c vao ng knh cua vong vo khuan o ma anh gia kha nang khang sinh cua xa khuan. Tat ca cac moi quan he tren ay cua khu he vi sinh vat at tao nen nhng he sinh thai vo cung phong phu trong tng loai at. Chung lam nen o mau m cua at, thay oi tnh chat ly hoa cua at va t o anh hng en cay trong. 1.1.3. Moi quan he gia at, vi sinh vat va thc vat 1.1.3.1. Quan he gia at va vi sinh vat at. at co ket cau t nhng hat nho lien ket vi nhau thanh cau truc at. Co quan iem cho rang vi sinh vat ong vai tro gian tiep trong s lien ket cac hat at vi nhau. Hoat ong cua vi sinh vat, nhat la nhom hieu kh a hnh thanh nen mot thanh phan cua mun la acid humic. Cac muoi cua acid humic tac dung vi ion Ca2+ tao thanh mot chat deo gan ket nhng hat at vi nhau. Sau nay ngi ta a tm ra vai tro trc tiep cua vi sinh vat trong viec tao thanh ket cau at. Trong qua trnh phan giai chat hu c, nam moc va xa khuan phat trien mot he khuan ty kha ln trong at. Khi nam moc va xa khuan chet i, vi khuan phan giai chung tai thanh cac chat deo co kha nang ket dnh cac hat at vi nhau. Ban than vi khuan khi chet i va t phan huy cung tao thanh cac chat ket dnh. Ngoai ra lp dch nhay bao quanh cac vi khuan co vo nhay cung co kha nang ket dnh cac hat at vi nhau. Cac chat ket dnh tao thanh ket cau at con c goi la mun hoat tnh. Nh vay mun khong nhng la ni tch luy chat hu c lam nen o ph nhieu cua at ma con la nhan to tao nen ket cau at. * Tac ong cua phan bon en vi sinh vat at Khi ta bon phan vao at, phan tac dung nhanh hay cham en cay trong la nh hoat ong cua vi sinh vat. Vi sinh vat phan giai phan hu c thanh dang vo c cho cay trong hap thu, bien dang vo c kho tan thanh de tan. Ngc lai cac loai phan bon cung anh hng en sinh trng va phat trien cua vi sinh vat trong at.

* Tac ong cua che o nc oi vi vi sinh vat: ai a so cac loai vi khuan co ch eu phat trien manh o am 60 80%. o am qua thap hoac qua cao eu c che vi sinh vat. Ch co nam moc va xa khuan la co the phat trien c ieu kien kho. * Tac ong cua che o canh tac khac ti vi sinh vat: Ngoai cac che o phan bon, nc, lam at, che o canh tac khac cung co tac dung ro ret ti hoat ong cua vi sinh vat. V du nh che o luan canh cay trong. Moi loai cay trong eu co mot khu he vi sinh vat ac trng song trong vung re cua no. Bi vay luan canh cay trong lam cho khu he vi sinh vat at can oi va phong phu hn. Ngi ta thng luan canh cac loai cay trong khac vi cay ho au e tang cng ham lng am cho at. Cac lai thuoc hoa hoc tr sau, diet co gay tac ong co hai ti vi sinh vat cung nh he sinh thai at noi chung. Viec dung cac loai thuoc hoa hoc lam o nhiem moi trng at, tieu diet phan ln cac loai vi sinh vat va ong vat nguyen sinh trong at. Tat ca nhng bien phap canh tac co anh hng trc tiep va sau sac en s phat trien cua vi sinh vat trong at, t o anh hng en qua trnh hoat ong sinh hoc, cu the la s chuyen hoa cac chat hu c thanh vo c trong at, anh hng en qua trnh hnh thanh mun va ket cau at. 1.1.3.2. Moi quan he gia vi sinh vat va thc vat Moi loai cay eu co mot khu he vi sinh vat vung re ac trng cho cay o v re thc vat thng tiet ra mot lng ln cac chat hu c va vo c, cac chat sinh trng, thanh phan va so lng cua cac chat o khac nhau tuy loai cay. Nhng chat tiet cua re co anh hng quan trong en vi sinh vat vung re. Tren be mat cua re va lp at nam sat re cha nhieu chat dinh dng nen tap trung vi sinh vat vi so lng ln. Cang xa re so lng vi sinh vat giam cang giam i. Thanh phan vi sinh vat vung re khong nhng phu thuoc vao loai cay trong ma con phu thuoc vao thi ky phat trien cua cay. Vi sinh vat phan giai cellulose co rat t khi cay con non nhng khi cay gia th rat nhieu. ieu o chng to vi sinh vat khong nhng s dung cac chat tiet cua re ma con phan huy re khi re cay gia va chet i. Vi sinh vat song trong vung re co quan he mat thiet vi cay, chung s dung nhng chat tiet cua cay lam chat dinh dng, ong thi cung cap chat dinh dng cho cay qua qua trnh hoat ong phan giai cua mnh. Vi sinh vat con tiet ra cac vitamin va chat sinh trng co li oi vi cay trong. Ben canh o co rat nhieu vi sinh vat gay benh cho cay, co nhng loai c che s sinh trng cua cay, co nhng loai tan pha mua mang nghiem trong.

1.2.

Moi trng nc va s phan bo cua vi sinh vat trong nc.

1.2.1. S phan bo cua vi sinh vat trong moi trng nc. Vi sinh vat co mat khap ni trong cac nguon nc. S phan bo cua chung hoan toan khong ong nhat va rat khac nhau tuy thuoc vao ac trng cua tng loai moi trng. Cac yeu to moi trng quan trong quyet nh s phan bo cua vi sinh vat la o man, chat
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hu c, pH, nhiet o va anh sang. Nguon nhiem vi sinh vat cung rat quan trong v ngoai nhng nhom chuyen song nc ra con co nhng nhom nhiem t cac moi trng khac vao. moi trng nc ngot, ac biet la nhng ni luon co s nhiem khuan t at, hau het cac nhom vi sinh vat co trong at eu co mat trong nc, tuy nhien vi t le khac biet. Nc ngam va nc suoi thng ngheo vi sinh vat nhat do nhng ni nay ngheo chat dinh dng. Trong cac suoi co ham lng sat cao thng cha cac vi khuan nh Leptothrix ochracea. cac suoi cha lu huynh thng co nhom vi khuan lu huynh mau luc hoac mau ta. Nhng nhom nay eu thuoc loai t dng hoa nang va quang nang. nhng suoi nc nong thng ch ton tai cac nhom vi khuan a nhiet nh Leptothrix thermalis. ao, ho va song do ham lng chat dinh dng cao nen so lng va thanh phan vi sinh vat phong phu hn nhieu. Ngoai nhng vi sinh vat t dng con co nhieu nhom vi sinh vat d dng co kha nang phan huy cac chat hu c. Hau het cac nhom vi sinh vat trong at eu co mat ay. nhng ni b nhiem ban bi nc thai sinh hoat con co mat cac vi khuan ng ruot va cac vi sinh vat gay benh khac. Tuy nhng vi khuan nay ch song trong nc mot thi gian nhat nh nhng nguon nc thai lai c o vao thng xuyen nen luc nao chung cung co mat. ay chnh la nguon o nhiem vi sinh nguy hiem oi vi sc khoe con ngi. nhng thuy vc co nguon nc thai cong nghiep o vao th thanh phan vi sinh vat cung b anh hng theo cac hng khac nhau tuy thuoc vao tnh chat cua nc thai. S phan bo cua vi sinh vat trong thuy vc con phu thuoc vao cac tang nc khac nhau. tang mat nhieu anh sang thng co nhng nhom vi sinh vat t dng quang nang. Di ay ho giau chat hu c thng co cac nhom vi khuan d dng phan giai chat hu c. nhng tang ay co s phan huy chat hu c manh tieu thu nhieu oxy tao ra nhng vung khong co oxy hoa tan th ch co mat nhom k kh bat buoc. Co nhng vi sinh vat co kha nang chu man cao, nhng co nhng vi sinh vat ch co the song trong nc ngot. Cac vi sinh vat song trong moi trng nc man noi chung co kha nang s dung chat dinh dng co nong o thap. Chung phat trien cham hn nhieu so vi vi sinh vat at. Chung thng bam vao cac hat phu sa e song. Vi sinh vat bien thng thuoc nhom a lanh va chu c ap suat cao. Noi chung cac nhom vi sinh vat song cac nguon khac nhau rat a dang ve hnh thai cung nh hoat tnh sinh hoc. Chung tham gia vao viec chuyen hoa vat chat. trong moi trng nc cung co mat ay u cac nhom tham gia vao cac chu trnh chuyen hoa cac hp chat carbon, nitrogen va cac chat khoang khac. Moi quan he gia cac nhom vi nhau cung rat phc tap, quan he ky sinh, cong sinh, ho sinh, khang sinh nh trong moi trng at. Ngay nay cac nguon nc, ngay ca nc ngam va nc bien nhng mc o khac nhau a b o nhiem do cac nguon chat thai khac nhau. Do o khu he vi sinh vat b anh
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hng rat nhieu va do o kha nang t lam sach cac nguon nc do hoat ong phan giai cua vi sinh vat cung b anh hng.

1.3.

Moi trng khong kh va s phan bo cua vi sinh vat trong khong kh.

Moi trng khong kh khong phai la ong nhat, tuy tng vung khac nhau, moi trng kh rat khac nhau ve thanh phan cac loai kh. nhng vung khong kh trong lanh nh vung nui, ty le kh O2 thng cao. nhng vung khong kh b o nhiem, ty le cac kh oc nh H2S, SO2, CO2thng cao, nhat la thanh pho va cac khu cong nghiep. S phan bo cua vi sinh vat trong khong kh cung khac nhau tuy tng vung. Khong kh khong phai la moi trng song cua vi sinh vat. Tuy nhien trong khong kh co rat nhieu vi sinh vat ton tai. Nguon goc cua nhng vi sinh vat nay la t at, t nc, t con ngi, ong vat, thc vat, theo gio, theo bui phat tan i khap ni trong khong kh. Mot hat bui co the mang theo rat nhieu vi sinh vat, ac biet la nhng vi sinh vat co bao t co kha nang ton tai lau trong khong kh. Neu o la nhng vi sinh vat gay benh th o chnh la nguon gay benh co trong khong kh. Cac vi sinh vat gay benh ng ho hap co the ton tai lau trong khong kh. Khi ngi ht phai khong kh co nhiem khuan o se co kha nang nhiem benh. Nhng vi khuan gay benh r sat co the theo gio bay i va lay benh cho cac canh ong rat xa nguon benh.

Chng 2 KHA NANG CHUYEN HOA CAC HP CHAT TRONG MOI TRNG T NHIEN CUA VI SINH VAT
WX

2.1. Vong tuan hoan nitrogen trong t nhien

Trong cac moi trng t nhien, nitrogen ton tai cac dang khac nhau, t nitrogen phan t dang kh cho en cac hp chat hu c phc tap co trong c the ong vat, thc vat va con ngi. Trong c the vi sinh vat, nitrogen ton tai chu yeu di dang cac hp chat hu c nh protein, acid amin. Khi c the vi sinh vat chet i, lng nitrogen hu c nay ton tai trong at. Di tac ong cac nhom vi sinh vat hoai sinh, protein c phan giai thanh cac acid amin. Cac acid amin lai c mot nhom vi sinh vat phan giai thanh NH3 hoac NH4+ goi la nhom vi khuan amon hoa. Qua trnh nay goi la s khoang hoa chat hu c v qua o nitrogen hu c c chuyen thanh dang nitrogen khoang. Dang NH4+ se c chuyen hoa thanh dang NO3- nh nhom vi khuan nitrate hoa. Cac hp chat nitrate lai c chuyen hoa thanh nitrogen phan t, qua trnh nay goi la phan nitrate hoa c thc hien bi nhom vi khuan phan nitrate. Kh N2 se c co nh lai trong te bao vi khuan va te bao thc vat sau o c chuyen hoa thanh dang nitrogen hu c nh nhom vi khuan co nh nitrogen. Nh vay, vong tuan hoan nitrogen c khep kn. Trong hau het cac khau chuyen hoa cua vong tuan hoan eu co s tham gia cua cac nhom vi sinh vat khac nhau. Neu s hoat ong cua mot nhom nao o ngng lai, toan bo s chuyen hoa cua vong tuan hoan cung se b anh hng nghiem trong.

2.2. Qua trnh amon hoa

Trong thien nhien ton tai nhieu dang hp chat nitrogen hu c nh protein, acid amin, acid nucleic, ureaCac hp chat nay i vao at t nguon xac ong vat, thc vat, cac loai phan chuong, phan xanh, rac thai hu c. Thc vat khong the ong hoa c dang nitrogen hu c phc tap nh tren, no ch co the s dung c sau qua trnh amon hoa. Qua qua trnh amon hoa, cac dang nitrogen hu c c chuyen hoa thanh NH4+ hoac NH3. 2.2.1. S amon hoa urea. Urea co trong thanh phan nc tieu cua ngi va ong vat, chiem khoang 2.2% nc tieu. Urea cha ti 46.6% nitrogen, v the no la mot nguon dinh dng am tot oi vi cay trong. Tuy nhien, thc vat khong the ong hoa trc tiep urea ma phai qua qua trnh amon hoa. Qua trnh amon hoa urea chia lam 2 giai oan, giai oan au di tac dung cua enzyme urease tiet ra bi cac vi sinh vat, urea se b thuy phan tao thanh muoi carbonate amoni. Giai oan 2, carbonate amoni chuyen hoa thanh NH3, CO2 va H2O. CO(NH2)2 + 2 H2O (NH4)2CO3 (NH4)2CO3 2NH3 + CO2 + H2O
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Trong nc tieu con co acid uric, ton tai trong at mot thi gian acid uric se b phan giai thanh urea va acid tactronic. Sau o urea tiep tuc b phan giai thanh NH3. Nhom vi sinh vat phan giai urea va acid uric con co kha nang amon hoa cyanamid calci la mot loai phan bon hoa hoc. Chat nay sau khi i vao at cung b chuyen hoa thanh urea roi sau o qua qua trnh amon hoa c chuyen thanh NH3. CN-Nca + 2 H2O CN-NH2 + H2O CN-NH2 + Ca(OH)2 CO(NH2)

Nhieu loai vi khuan co kha nang amon hoa urea, chung eu tiet ra enzyme urease. Trong o co mot so loai co hoat tnh phan giai cao nh Planosarcina urea, Micrococcus urea, Bacillus amylovorum, Proteus vulgaris a so vi sinh vat phan giai urea thuoc nhom hieu kh hoac k kh khong bat buoc, chung a pH trung tnh hoac hi kiem. Bi vay khi s dung urea lam phan bon ngi ta thng ket hp vi bon voi hoac tro, ong thi lam thoang at. 2.2.2. S amon hoa protein. Protein la thanh phan quan trong cua te bao sinh vat, khi ong vat, thc vat chet i, nguon protein co trong te bao cua chung c tch luy trong at. Protein cha ti 15 17% nitrogen, nhng cay trong khong the hap thu trc tiep protein ma phai thong qua s phan huy cua vi sinh vat. Nhom vi sinh vat phan huy protein co kha nang tiet ra enzyme protease bao gom proteinase va peptidase. Di tac dung cua proteinase phan t protein se c phan giai thanh cac chuoi polypeptide va oligopeptide (cha t 3 5 acid amin). Sau o di tac dung cua enzyme peptidase cac polypeptide va oligopeptide se c phan giai thanh cac acid amin. Mot phan acid amin se c te bao vi sinh vat hap thu lam chat dinh dng. Phan khac se thong qua qua trnh kh amin tao thanh NH3 va nhieu san pham trung gian khac. S kh amin co the xay ra theo mot trong nhng phng thc sau: R-CH(NH2)COOH R-CH(NH2)COOH + H2O R-CH(NH2)COOH + O2 R=CHCOOH + NH3 R-CH2OH-COOH + CO2 + NH3 R-CO-COOH + NH3

Mot so acid amin b deamin hoa bi vi sinh vat nh enzyme deaminase, sau o tao ra san pham cuoi cung la amon, v du: Alamin + O2
Ala-deaminose

Piprevar + NH4

oi vi cac acid amin co vong nh triptophan, khi phan giai se tao thanh cac hp chat co mui thoi nh indon va scaton. Khi phan giai cac acid amin cha S nh methionin, cystein, vi sinh vat giai phong ra H2S, chat nay oc oi vi cay trong. Mot so hp chat amin sinh ra trong qua trnh amon hoa co tac dung oc oi vi ngi va ong vat. V du
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nh histamin, armatino chnh la nguyen nhan b nhiem oc thc an tht ca thiu thoi hoac tht hop e qua lau (o nhiem thc pham). Ty le C:N trong at rat quan trong oi vi nhom vi sinh vat phan huy protein. Neu nh ty le nay qua cao, trong at qua t am vi sinh vat se tranh chap thc an am oi vi cay trong, chung phan huy c bao nhieu la hap thu bay nhieu. Neu ty le C:N qua thap, am d tha, qua trnh phan huy se cham lai, cay trong khong co am khoang e hap thu. Nhieu cong trnh nghien cu a rut ra ty le C:N bang 20 la thch hp nhat cho qua trnh amon hoa protein, co li nhat oi vi cay trong. Nhieu vi sinh vat co kha nang amon hoa protein. Trong nhom vi khuan co Bacillus mycoides, B. mesentericus, B. subtilis, Pseudomonas fluorescens, Clostridium sporogenesXa khuan co Streptomyces griseusVi nam co Aspergillus oryzae, A. flavus, A. niger, Penicilium camemberti Ngoai protein va urea, nhieu loai vi sinh vat co kha nang amon hoa kitin. Kitin la thanh phan cua vo nhieu loai con trung, giap xac. Hang nam kitin c tch luy lai trong at vi mot lng khong nho, nhom vi sinh vat phan huy kitin co kha nang tiet enzyme kitinase va kitobiase phan huy phan t kitin thanh cac goc n phan t, sau o goc amin c amon hoa tao thanh NH3.

2.3. Qua trnh nitrate hoa.

Sau qua trnh amon hoa NH3 c hnh thanh, mot phan phan ng vi cac anion trong at tao thanh cac muoi amon. Mot phan muoi amon cung c cay trong hap thu, phan con lai c oxy hoa thanh dang nitrate goi la qua trnh nitrate hoa. Nhom vi sinh vat tien hanh qua trnh nay goi chung la nhom vi khuan nitrate hoa bao gom 2 nhom tien hanh qua 2 giai oan. 2.3.1. Giai oan nitrite hoa Qua trnh oxy hoa NH4+ tao thanh NO2- c tien hanh bi vi khuan nitrite hoa. Chung thuoc nhom vi sinh vat t dng hoa nang co kha nang oxy hoa NH4+ bang oxy khong kh vao tao ra nang lng. NH4+ + 3/2 O2 NO2- + H2O + 2 H + Q Nang lng c vi khuan s dung e ong hoa CO2 thanh carbon hu c. Enzyme xuc tac cho qua trnh nay la cac enzyme cua qua trnh ho hap hieu kh. Nhom vi khuan nitrite hoa bao gom 4 chi khac nhau: Nitrosomonas, Nitrosocystis, Nitrosolobus va Nitrosospira chung eu thuoc loai t dng bat buoc, khong co kha nang song tren moi trng thach. Bi vay phan lap chung rat kho, phai dung silicagen thay cho thach. 2.3.2. Giai oan nitrate hoa Qua trnh oxy hoa NO2- thanh NO3- c thc hien bi nhom vi khuan nitrate. Chung cung la nhng vi sinh vat t dng hoa nang co kha nang oxy hoa NO2- tao thanh nang lng. Nang lng nay c dung e ong hoa CO2 tao thanh ng.
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NO2- + O2

NO3- + Q

Nhom vi khuan tien hanh oxy hoa NO2- thanh NO3- bao gom 3 chi khac nhau: Nitrobacter, Nitrospira va Nitrococcus. Ngoai nhom vi khuan t dng hoa nang noi tren, trong at con co mot so loai vi sinh vat d dng cung tien hanh qua trnh nitrate hoa. o la cac loai vi khuan va xa khuan thuoc cac chi Pseudomonas, Corynebacterium, Streptomyces Qua trnh nitrate hoa la mot khau quan trong trong vong tuan hoan nitrogen, nhng oi vi nong nghiep no co nhieu ieu bat li. Dang am nitrate thng de b ra troi xuong cac tang sau, de b i vao qua trnh phan nitrate hoa tao thanh kh N2 lam cho at mat am. Anion NO3- thng ket hp vi ion H+ trong at tao thanh HNO3 la cho pH at giam xuong rat bat li cho cay trong. Hn na, lng NO3 d tha trong at c cay trong hap thu nhieu lam cho ham lng nitrate trong san phan lng thc, thc pham cao gay oc cho ngi va gia suc. Bi vay ngay nay ngi ta thng han che viec bon phan am hoa hoc co goc nitrate.

2.4. Qua trnh phan nitrate hoa

Cac hp chat am dang nitrate trong at rat de b kh bien thanh nitrogen phan t. Qua trnh nay goi la qua trnh phan nitrate hoa. No khac vi qua trnh oxy hoa nitrate tao thanh NH4+ con goi la qua trnh amon hoa nitrate. Co the phan biet c hai qua trnh tren qua s o sau. Amon hoa nitrate NH2OH NO3 NO2 NO N2O N2 NH3

Phan nitrate hoa Qua trnh amon hoa nitrate do mot so vi khuan d dng tien hanh trong ieu kien hieu kh co chc nang cung cap NH4+ cho te bao vi khuan e tong hp acid amin. Phan ng kh NO3- thanh N2 ch xay ra trong ieu kien k kh. NO3- la chat nhan ien t cuoi cung trong chuoi ho hap k kh, nang lng tao ra c dung e tong hp nen ATP. Nhom vi sinh vat thc hien qua trnh phan nitrate hoa phan bo rong rai trong at. Thuoc nhom t dng hoa nang co Thiobacillus denitrificans, Hydrogenomonas agilis Thuoc nhom d dng co Pseudomonas denitrificans, Micrococcus denitrificanas, Bacillus licheniformissong trong ieu kien k kh, trong nhng vung at ngap nc.

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oi vi nong nghiep qua trnh phan nitrate hoa la mot qua trnh bat li v no lam cho at mat am. Qua trnh nay xay ra manh trong ieu kien k kh. Oxy co tac dung c che cac enzyme xuc tac cho qua trnh kh nitrate, o la cac enzyme nitrate reductase va nitrite reductase. cac ruong lua nc ngi ta thng lam co xuc bun e han che qua trnh nay, ong thi bon am amon ch khong bon am nitrate. Trong cac moi trng t nhien ngoai qua trnh phan nitrate sinh hoc noi tren con co qua trnh phan nitrate hoa hoc thng xay ra khi pH<5.5. Cac qua trnh nay khong co s tham gia cua vi sinh vat NH4Cl + HNO3 N2 + HCl + H2O N2 + R-OH + H2O R-NH2 + HNO3

2.5. Qua trnh co nh nitrogen phan t (xem phan vi sinh vat hoc ai cng) 2.6. S chuyen hoa cac hp chat phosphore cua vi sinh vat.
2.6.1. Vong tuan hoan phosphore trong t nhien. Trong t nhien, P nam trong nhieu dang hp chat khac nhau. P hu c co trong c the ong thc vat, c tch luy trong at khi ong vat va thc vat chet i. Nhng hp chat phosphore hu c nay c vi sinh vat phan giai tao thanh cac hp chat phosphore vo c kho tan, mot so t c tao thanh dang de tan. Cac hp chat phosphore vo c kho tan co nguon goc t nhng quang thien nhien nh apatit, phosphorite, phosphate sat, phosphate nhomNhng hp chat nay rat kho hoa tan va cay trong khong the hap thu trc tiep c. Cay trong ch co the hap thu c khi chung c chuyen hoa thanh dang de tan. Qua trnh nay c thc hien mot phan quan trong la nh vi sinh vat phan huy phosphore vo c. Cac muoi cua acid phosphoric dang de tan c cay trong hap thu va van chuyen thanh cac hp chat phosphore hu c trong c the thc vat. ong vat va ngi s dung cac san pham thc vat lam thc an lai bien phosphore hu c cua thc vat thanh P hu c cua ong vat va ngi. Vong tuan hoan cua cac dang hp chat phosphore trong t nhien c dien ra. Vi sinh vat ong mot vai tro quan trong trong vong tuan hoan o. Neu nh thieu s hoat ong cua mot nhom vi sinh vat nao o th s chuyen hoa cua vong tuan hoan se b anh hng nghiem trong. Vong tuan hoan cua cac dang phosphore trong t nhien c bieu dien nh s o sau.

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P vo c de tan P vo c kho tan P hu c trong at

Phan P (Chat bai tiet)

P hu c thc vat

P hu c ong vat 2.6.2. S phan giai phosphore hu c trong at do vi sinh vat. Cac hp chat P hu c trong at co nguon goc t xac ong vat, thc vat, phan xanh, phan chuongHp chat phosphore hu c quan trong nhat c phan giai ra t te bao vi sinh vat la nucleotide. Nucleotide co trong thanh phan nhan te bao. Nh tac ong cua cac nhom vi sinh vat hoai sinh trong at, chat nay tach ra khoi thanh phan te bao va c phan giai thanh 2 phan protein va nuclein. Protein se i vao vong chuyen hoa cac hp chat nitrogen, nuclein se i vao vong chuyen hoa cac hp chat phosphore. S chuyen hoa cac hp chat phosphore hu c thanh muoi cua H3PO4 c thc hien bi nhom vi sinh vat phan huy phosphore hu c. Nhng vi sinh vat nay co kha nang tiet ra enzyme phosphatase e xuc tac cho qua trnh phan giai. Nhom vi sinh vat phan giai phosphore hu c c phat hien t nam 1911 do J. Stoklasa, ong a phan lap c 3 loai vi khuan co kha nang phan huy phosphore hu c eu thuoc giong Bacillus. Sau o ong nuoi cay nhng vi khuan nay trong moi trng ch co acid nucleic lam nguon P va N duy nhat va nhan thay lng phosphore c phan giai t 15 en 23%. Neu bo sung vao moi trng mot t (NH4)2SO4 th lng P c phan giai tang len. Sau o ngi ta a tm ra nhieu loai vi sinh vat co kha nang phan huy P hu c theo s o tong quat sau: Nucleoprotein nuclein a. nucleic H3PO4

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H3PO4 thng phan ng vi cac kim loai trong at tao thanh cac muoi phosphate kho tan nh Ca3(PO4)2, FePO4, AlPO4 Vi sinh vat phan giai P hu c chu yeu thuoc 2 chi Bacillus va Pseudomonas. Cac loai co kha nang phan giai manh la B. megatherium, B. mycoides va Pseudomonas sp. Ngay nay, ngi ta a phat hien thay mot so xa khuan va vi nam cung co kha nang phan giai phosphore hu c. 2.6.3. S phan giai phosphore vo c do vi sinh vat. Cac hp chat phosphore vo c c hnh thanh do qua trnh phan giai lan hu c (con goi la qua trnh khoang hoa lan hu c) phan ln la cac muoi phosphate kho tan. Cay trong khong the hap thu c nhng dang kho tan nay. Neu khong co cac qua trnh phan giai cac hp chat phosphore kho tan bien thanh dang de tan th ham lng phosphore tong so trong at dau co nhieu cung tr thanh vo dung. Ve c che cua qua trnh phan giai phosphore vo c do vi sinh vat cho en nay van con nhieu tranh cai. Nhng ai o so cac nha nghien cu eu cho rang s san sinh acid trong qua trnh song cua mot so nhom vi sinh vat a lam cho no co kha nang chuyen cac hp chat phosphore t dang kho tan thanh dang de tan. a so cac vi sinh vat phan giai phosphore vo c eu sinh CO2 trong qua trnh song, CO2 se phan ng vi H2O co trong moi trng tao thanh H2CO3. H2CO3 se phan ng vi phosphate kho tan tao thanh phosphate de tan theo phng trnh sau: Ca3(PO4)2 + 4 H2CO3 + H2O Dang kho tan Ca(H2PO4)2 + H2O + 2 Ca(HCO3)2 Dang de tan Dang de tan

Cac vi khuan nitrate hoa trong at cung co kha nang phan giai phosphore vo c do no co kha nang chuyen hoa NH3 thanh NO3-. NO3- se phan ng vi H+ tao thanh HNO3. Sau o HNO3 phan ng vi muoi phosphate kho tan tao thanh dang de tan. Ca3(PO4)2 + 4 HNO3 Ca(H2PO4)2 + 2 Ca(NO3)2 Cac vi khuan sulphate hoa cung co kha nang phan giai phosphate kho tan do s tao thanh H2SO4 trong qua trnh song. Ca3(PO4)2 + 2 H2SO4 Ca(H2PO4)2 + 2 CaSO4 Ngoai ra cac nhom vi sinh vat co kha nang tao thanh cac acid hu c trong qua trnh song cung co the lam cho dang phosphate kho tan chuyen thanh dang de tan. Tuyet ai a so cac vi sinh vat phan huy phosphore vo c trong qua trnh song eu lam giam pH cua moi trng. Tuy nhien, gan ay co mot vai tac gia a cong bo tm ra mot vai chung vi khuan phan giai phosphore ma trong qua trnh nuoi cay khong lam giam pH moi trng. Rat nhieu vi sinh vat co kha nang phan giai phosphore vo c, trong o nhom vi khuan c nghien cu nhieu hn ca. Cac loai co kha nang phan giai manh la Bacillus megatherium, B. butyricus, B. mycoides, Pseudomonas radiobacter, P. gracilis trong
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nhom vi nam th Aspergillus niger co kha nang phan giai manh nhat. Ngoai ra mot so xa khuan cung co kha nang phan giai phosphore vo c.

2.7. S chuyen hoa cac hp chat lu huynh cua vi sinh vat.

2.7.1. Vong tuan hoan lu huynh trong t nhien. Cung nh phosphore, lu huynh la mot trong nhng chat dinh dng quan trong cua cay trong. Trong at no thng dang cac hp chat muoi vo c nh CaSO4, Na2SO4, FeS2, Na2Smot so dang hu c. Trong c the sinh vat, S nam trong thanh phan cua cac acid amin chu lu huynh nh methionin, cystein va trong nhieu loai enzyme quan trong khac. Thc vat hut cac hp chat lu huynh vo c trong at chu yeu di dang SO42- va chuyen sang dang S hu c cua te bao. ong vat va ngi s dung thc vat lam thc an va cung bien S cua thc vat thanh S cua ong vat va ngi. Khi ong thc vat chet i e lai mot lng lu huynh hu c trong at. Nh s phan giai cua vi sinh vat, S hu c se c chuyen hoa thanh H2S. H2S va cac hp chat vo c khac co trong at se c oxy hoa bi cac nhom vi khuan t dng thanh S va SO42-, mot phan c tao thanh lu huynh hu c cua te bao vi sinh vat. SO42- lai c thc vat hap thu, c the vong chuyen hoa cac hp chat lu huynh dien ra lien tuc. Trong o cac nhom vi sinh vat ong mot vai tro quan trong khong the thieu c. 2.7.2. S oxy hoa cac hp chat lu huynh. 2.7.2.1. S oxy hoa cac hp chat lu huynh do vi khuan t dng hoa nang. Trong nhom vi khuan t dng hoa nang co mot so loai co kha nang oxy hoa cac hp chat lu huynh vo c nh thiosulphate, kh hydro sulfur va lu huynh nguyen chat thanh dang SO42- theo cac phng trnh sau: 2H2S + O2 2 S + 3 O2 + 2 H2O 5 Na2S2O3 + H2O + 4 O2 Na2S4O6 + O2 2 H2O + 2 S + Q 2 H2SO4 + Q 5 Na2SO4 + 2 S2 + H2SO4 + Q 3 Na2SO4 + 5 H2SO4 + Q

H2SO4 sinh ra lam pH at ha xuong (diet tr c benh thoi do Streptomyces gay ra va pH thap vi khuan cua mot so loai gay benh cho cay trong khong song c). Nang lng sinh ra trong qua trnh oxy hoa tren c vi sinh vat s dung e ong hoa CO2 tao thanh ng. ong thi mot so t hp chat dang S cung c ong hoa tao thanh S hu c cua te bao vi khuan. Cac loai vi khuan co kha nang oxy hoa cac hp chat lu huynh theo phng thc tren la Thiobacillus thioparus va Thiobacillus thioxidans. Ca hai loai nay eu song pH thap, thng la pH=3, oi khi pH=1 1.5 hai loai nay van co the phat trien. Nh ac iem nay ma ngi ta dung 2 loai vi khuan tren e lam tang o hoa tan cua quang apatide. Ngoai 2 loai vi khuan tren con co 2 loai vi khuan khac co kha nang oxy hoa cac hp chat lu huynh vo c, o la Thiobacillus denitrificans va Begiatra minima.
15

Thiobacillus denitrificans co kha nang va kh nitrate va oxy hoa lu huynh theo phng trnh sau: 5 S + 6 KNO3 + 2CaCO3 2 K2SO4 + CaSO4 + 2 CO2 + 3 N2 + Q Vi khuan Begiatra minima co the oxy hoa H2S hoac S. Trong ieu kien co nhieu H2S no se oxy hoa H2S tao thanh S tch luy trong te bao. Trong ieu kien thieu H2S cac hat S se c oxy hoa en khi S d tr het th vi khuan chet hoac trang thai tiem sinh. 2.7.2.2. S oxy hoa cac hp chat lu huynh do vi khuan t dng quang nang. Mot so nhom vi khuan t dng quang nang co kha nang oxy hoa H2S tao thanh H2S ong vai tro chat cho ien t trong qua trnh quang hp cua vi khuan. Cac vi khuan thuoc ho Thiodaceae thng oxy hoa H2S theo phng trnh sau: SO42-. anh sang CO2 + H2S + H2O sau: C6H12O6 + H2SO4 Cac vi khuan thuoc ho Chlorobacteriaceae thng oxy hoa H2S theo phng trnh anh sang CO2 + H2S + H2O C6H12O6 + S nhom vi khuan tren, S c hnh thanh khong tch luy trong c the ma ngoai moi trng. 2.7.3. S kh cac hp chat lu huynh vo c do vi sinh vat. Ngoai qua trnh oxy hoa, trong at con co qua trnh kh cac hp chat lu huynh vo c thanh H2S. Qua trnh nay con goi la qua trnh phan sulphate hoa. Qua trnh nay c tien hanh ieu kien k kh, nhng tang nc sau. Nhom vi sinh vat tien hanh qua trnh nay goi la nhom vi khuan phan sulphate hoa. C6H12O6 + 3 H2SO4 6 CO2 + 6 H2O + 3 H2S + Q ay chat hu c ong vai tro cung cap hydro trong qua trnh kh SO4 co the la ng hoac cac acid hu c hoac cac hp chat hu c khac. H2SO4 se b kh dan ti H2S. Qua trnh phan sulphate hoa dan en viec tch luy H2S trong moi trng lam o nhiem moi trng, anh hng en i song cua thc vat va ong vat trong moi trng o. Lua moc trong ieu kien yem kh co qua trnh phan sulphate hoa manh se b en re va anh hng xau en qua trnh sinh trng va phat trien cua chung.

16

Chng 3 SINH TRNG VA PHAT TRIEN VI SINH VAT

WX Sinh trng va phat trien la thuoc tnh c s cua sinh vat. Cung nh ong vat va thc vat, vi sinh vat cung sinh trng va phat trien. Sinh trng la s tang kch thc va khoi lng cua te bao, con phat trien (hoac sinh san) la s tang so lng te bao. Trong phan nay chung ta chu yeu e cap en sinh trng va phat trien cua vi khuan. Khi noi ve sinh trng va phat trien cua vi khuan tc la e cap en sinh trng va phat trien cua mot so lng ln te bao cua cung mot loai. Ro rang, viec nghien cu mot ca the te bao vi khuan qua nho la rat kho. Tuy nhien, s tang khoi lng te bao khong phai bao gi cung dien ra song song vi s tang sinh khoi. Chang han, khi chat dinh dung trong moi trng a can, vi khuan tuy con phan chia 1 - 2 lan nhng cho 2 -4 te bao nho hn te bao bnh thng; trong pha m au, sinh khoi vi khuan tang len, nhng so te bao khong thay oi, ngc lai, vao cuoi pha logarit kch thc te bao giam i nhng so te bao van con tang. V vay can phan biet cac thong so va hang so khac nhau khi xac nh so lng hoac khoi lng vi khuan. Khi xac nh so lng hay khoi lng cua vi khuan ta thng dung dch treo ong eu cua cac te bao trong moi trng dch the nao o ma xac nh nong o vi khuan (so te bao trong 1ml) hoac mat o vi khuan (mg/ml). T ket qua o cac ch so nay co the tnh c hang so toc o phan chia te bao (the hien bang so lan tang oi nong o vi khuan sau 1 gi) va ai lng ngc lai, tc thi gian the he (thi gian can cho so lng te bao trong mot quan the vi khuan tang gap oi). Trong ieu kien phong th nghiem thng ta theo doi anh hng cua cac yeu to moi trng len sinh trng va phat trien cua vi khuan nh cac phng phap nuoi cay thch hp. Tuy tnh chat thay oi trong he vi khuan - moi trng ta phan biet hai phng phap c ban nuoi cay vi khuan : nuoi cay tnh va nuoi cay lien tuc. Mot vi sinh vat trong ieu kien moi trng thch hp se khong ngng hap thu chat dinh dng va tien hanh trao oi chat. Neu qua trnh ong hoa ln hn qua trnh d hoa th tong so nguyen sinh chat (khoi lng, the tch, kch thuc) se khong ngng tang len. o la hien tng sinh trng cua ca the. Neu qua trnh sinh trng can bang th cac thanh phan cua te bao cung tang len theo nhng t le thch hp, khi at en mot mc o nhat nh th se hnh thanh s sinh san. Khi o so lng ca the tang len. Ca the ban au se phat trien dan len thanh mot quan the, s tiep tuc sinh trng cua ca the trong quan the tao ra s sinh trng cua quan the.

3.1. Mau ly thuyet ve sinh trng va phat trien cua vi khuan

Gia s trong mot bnh kn cha mot lng ln moi trng dinh dng, ta cay vao o mot te bao vi khuan. Neu thanh phan moi trng hoan toan phu hp vi nhu cau cua te bao, vi khuan se sinh trng, tang khoi lng va the tch, tong hp cac thanh phan cua
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te bao (thanh te bao, mang te bao chat, DNA, RNA, protein...) cho en khi kch thc ln gap oi. Roi vi khuan phan chia cho hai te bao. Hai te bao nay lai tiep tuc sinh trng va phan chia e cho 4 roi 8, 16... te bao. Neu so te bao ban au khong phai la 1 ma la No th sau n lan phan chia ta se co so te bao tong cong la N : N = No . 2 n (1) Cac gia tr N va No co the xac nh nh phong em hoac tnh so khuan lac moc tren moi trng ac. Con gia tr n (so the he) co the tnh nh logarit thap phan : logN = logNo + n.log2 T o : n=
1 . (logN - logNo) log 2

(2)

V du vi khuan phan chia n lan sau thi gian t, khoang thi gian gia hai lan phan chia lien tiep (hoac thi gian can cho viec tang oi so te bao) goi la thi gian the he va bieu th bang g : g=
t 2 t1 t = log2 n log N log No

(3)

ay t2 - t1 bieu th s sai khac gia thi gian au (t1) va thi gian cuoi (t2) tnh bang gi trong o so te bao c xac nh. Gia tr ao ngc cua thi gian the he hay la so lan phan chia sau mot n v thi gian (tc sau 1 gi) goi la hang so toc o phan chia C Hang so toc o phan chia phu thuoc vao mot so ieu kien : a. Loai vi khuan : V du : 370C, C = 3 oi vi E. coli va C = 0,07 oi vi Mycobacterium tuberculosis. b. Nhiet o nuoi cay : V du : E. coli nuoi cay trong nc tht :

Nhiet o (0C) 18 22 30 37 42 43

Thi gian the he (phut) 120 60 30 20 20 -

Hang so toc o phan chia C 0,5 1 2 3 3 0

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c. Moi trng nuoi cay Khi nuoi cay B. subtitis 370C, trong : nc tht moi trng khoang - glucose moi trng khoang - citrate moi trng khoang - glucose - citrate C=2 C = 0,8 C = 0,3 C = 1,2

oi vi mot so c chat a cho hang so toc o phan chia khong phu thuoc vao nong o trong mot gii han rong. Chang han, oi vi B. subtitis, khi nuoi cay trong moi trng cha glucose, du nong o glucose bang 0,3 hay 50g/l ta van co C = 0,8 ; C ch giam khi nong o ng vt ra ngoai cac gii han noi tren. Tuy nhien, khong phai bao gi sinh trng (tang sinh khoi vi khuan) cung dien ra song song vi sinh san (tang so lng vi khuan), trng hp nay ch gap trong pha logarit. V vay, khi nghien cu ong hoc trong cac qua trnh nuoi cay lien tuc ta thng theo doi sinh trng va sinh san cua quan the vi khuan bang mot tieu chuan khac. Cung co the bieu th bang so lng te bao nhng pho bien hn la bieu th bang sinh khoi vi khuan, bang chat kho hay bang mat o quang hoc ... (can chu y rang mat o quang hoc t le vi so te bao).

3.2. Sinh trng va phat trien cua vi khuan trong ieu kien nuoi cay tnh. ng cong sinh trng
Neu theo doi sinh trng va phat trien cua vi khuan trong ieu kien phong th nghiem ta nhan thay so lng chung tang len rat nhanh. ieu nay de hieu, v vi ieu kien thch hp thi gian t the he (hay noi ung hn, thi gian tang oi) cua nhieu loai vi khuan ch vao khoang 30 phut. Ro rang cac qua trnh sinh tong hp cung nh cac qua trnh d hoa (nh ho hap) nham cung cap nguyen lieu va nang lng cho cac phan ng tong hp a dien ra trong te bao vi toc o rat nhanh, hoan toan khong thay cac sinh vat khac. D nhien hien tng noi tren khong bao gi xay ra v sinh trng va sinh san cua vi khuan trong mot he thong ong kn ch sau mot thi gian nhat nh, v nhieu nguyen nhan khac nhau, se b ngng lai. Phng phap nuoi cay ma trong suot thi gian o ta khong them vao chat dinh dng cung khong loai bo i cac san pham cuoi cung cua trao oi chat goi la nuoi cay tnh. S sinh trng trong mot he thong nh vay tuan theo nhng quy luat bat buoc khong nhng oi vi cac c the n bao ma ca oi vi cac c the a bao. 3.2.1. Pha lag Pha nay tnh t luc bat au cay en khi vi khuan at c toc o sinh trng cc ai. Trong pha lag vi khuan cha phan chia (ngha la cha co kha nang sinh san) nhng the tch va khoi lng te bao tang len ro ret do qua trnh tong hp cac chat, trc het la cac cao phan t (protein, enzyme, acid nucleic...) dien ra manh me. Mot so enzyme can
19

cho qua trnh tong hp thuoc cac endoenzyme loai proteinase, amylase va cac enzyme nam trong qua trnh chuyen hoa glucide, eu c hnh thanh trong pha nay. o dai cua pha lag phu thuoc trc het vao tuoi cua ong giong va thanh pham moi trng. Chang han, pha lag se khong co neu ta dung ong giong gom cac te bao ang pha sinh trng logarit va cay chung vao cung mot moi trng di nhng ieu kien nuoi cay nh nhau. Trai lai neu ta cay cac te bao pha on nh hoac cac bao t vao cung mot moi trng di nhng ieu kien nuoi cay nh nhau, pha lag van co. Thng thng te bao cang gia th pha lag cang dai. Ro rang nguyen nhan cua pha lag la s khac biet gia cac te bao pha on nh (hoac bao t) vi cac te bao ang sinh trng logarit. Trong pha lag dien ra viec xay dng lai cac te bao ngh thanh cac te bao sinh trng logarit. Nhng ngay khi dung cac te bao ang sinh trng logarit ma cay vao moi trng mi khac vi moi trng trc ay ta van thay pha lag. Nguyen nhan cua pha lag trong trng hp nay chnh la s thch ng cua vi khuan vi ieu kien nuoi cay mi. S thch ng o co lien quan vi s tong hp cac enzyme mi ma trc ay te bao cha can.. Cac enzyme mi nay c tong hp nh s cam ng vi cua cac c chat mi. Viec tm hieu o dai thi gian cua pha lag la can thiet trong viec phan oan ac tnh cua vi khuan va tnh chat cua moi trng. Nh ta a biet, pha lag bieu th s thch nghi cua vi khuan vi mot moi trng a cho. Trang thai sinh ly cua vi khuan cang xa vi sinh trng logarit trong moi trng mi th pha lag cang dai. Nhng yeu to th hai anh hng en o dai thi gian cua pha lag la toc o cua cac qua trnh dien ra trong te bao, trc het la toc o tong hp cac enzyme thch ng mi. Co rat nhieu yeu to anh hng en pha lag. ang chu y nhat la 3 yeu to sau ay: Tuoi giong cay : Tuoi cua quan the giong cay tc la chung ang giai oan sinh trng nao, co anh hng rat ro en pha lag. Thc nghiem chng minh neu giong cay giai oan pha lag (hay pha ch so) th pha lag se ngan. Ngc lai neu giong cay pha t vong th pha lag vat nuoi se keo dai. Lng cay giong : noi chung lng cay giong nhieu th pha lag ngan va ngc lai. Trong cong nghiep len men ty le cay giong thng mc 1/10. Thanh phan moi trng : Moi trng co thanh phan dinh dng phong phu (thng la moi trng co c chat thien nhien) th cho pha lag ngan. Trong cong nghiep len men thanh phan cua moi trng len men thng tranh sai khac nhieu so vi thanh phan cua moi trng nhan giong.

3.2.2. Pha log Trong pha nay vi khuan sinh trng va phat trien theo luy tha, ngha la sinh khoi va so lng te bao tang theo phng trnh N = No . 2ct hay X = Xo . ct. kch thc cua te
20

bao, thanh phan hoa hoc, hoat tnh sinh ly ... noi chung khong thay oi theo thi gian. Te bao trang thai ong hoc va c coi nh la nhng te bao tieu chuan. Neu ta lay truc tung la so te bao, truc hoanh la thi gian ta se co ng bieu dien la ng cong I.

Mot o th nh vay ro rang khong co li cho viec tnh toan. Cho nen ngi ta thng lay truc tung la logarit cua so te bao va ng bieu dien sinh trng theo luy tha cua vi khuan se la ng thang. V pha sinh trng theo luy tha cua vi khuan c bieu dien bang s phu thuoc theo ng thang gia thi gian va logarit cua so te bao nen pha nay c goi la pha logarit. Hn na, ngi ta thng dung logarit c so 2 la thch hp
21

hn ca v s thay oi mot n v cua log2 tren truc tung chnh la s tang oi so lng vi khuan va thi gian can e tang mot n v cua log2 lai la thi gian the he. Ba thong so quan trong cua pha log la thi gian the he (hoac thi gian tang oi) g, hang so toc o phan chia c va hang so toc o sinh trng . Neu xac nh thi gian the he g theo so te bao bang phng phap noi tren ta se nhan c gia tr trung bnh. Tren thc te, trong mot quan the vi khuan luon luon co mot so te bao khong co kha nang phan chia. V vay cac te bao phan chia manh thng co thi gian the he nho hn gia tr trung bnh tm thay. Cac hang so c va co the tnh c t phng trnh :
=
log 2 X 2 log 2 X 1 log 2 e(t 2 t1 )

Cac hang so nay thay oi tuy theo hang loat yeu to moi trng va ieu kien nuoi cay. Tuy nhien trong ieu kien th nghiem co the ieu chnh sao cho toc o sinh trng cua vi khuan ch man cam, ngha la ch phu thuoc vao mot yeu to, con cac yeu toc khac cua moi trng khong co anh hng g. Trong trng hp nh vay yeu to a cho la yeu to han che toc o sinh trng. Monod (1942) la ngi au tien a chng minh rang neu tat ca cac chat dinh dng eu nong o d tha tr mot chat han che th hang so toc o phan chia (hoac sinh trng) se la ham so cua nong o chat dinh dng han che nay. Chat dinh dng han che co the la ng (saccharose, glucose) acid amin (tryptophan, arginin), chat vo c (phosphate, CO2). Sinh trng theo luy tha cua mot vi khuan co thi gian the he la 20 phut Thi gian (phut ) 0 20 40 60 80 100 So lan phan chia 0 1 2 3 4 5 So te bao bieu th bang so so hoc log2 0 1 1 2 2 4 3 8 4 16 5 32

log10 0,000 0,301 0,602 0,903 1,204 1,505

V vay Monod a neu len mot cach tng t moi quan he gia cac hang so c va vi nong o chat dinh dng han che qua cac phng trnh :
c = c max [S ] K s + [S ]

va

= max

[S ] K s [S ]

ay cmax va max lan lt la hang so toc o phan chia va hang so toc o sinh trng cc ai, Ks la hang so bao hoa va [S] la nong o chat dinh dng han che. Giong
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nh hang so Michaelis km, hang so bao hoa Ks bieu th ai lc cua te bao oi vi chat dinh dng : Gia tr cua Ks cang nho th ai lc cang ln. o la nong o cua chat dinh dng han che ma o at c na gia tr cc ai, tc = thap. Mot so hang so bao hoa
1

max

. Gia tr cua Ks thng rat

Vi sinh vat E. coli E. coli E. coli E. coli E. coli E. coli M. tuberculosis A. aerogenes B. megatherium E. coli

Chat dinh dng han che glucose mannitol lactose glucose tryptophan tryptophan glucose phosphate O2 O2 O2

Ks 4mg/l 2mg/l 20mg/l 7,5mg/l 0,2 en 1,0 g/l 0,4 g/l 3, 9g/l 0,6mmol/l 3,1 . 10-8 mol/l 6,0 . 107 mol/l 2,2 . 10-8 mol/l

3.2.3. Pha on nh Trong pha nay quan the vi khuan trang thai can bang ong hoc, so te bao mi sinh ra bang so te bao cu chet i. Ket qua: so te bao va ca sinh khoi khong tang cung khong giam. Toc o sinh trng bay gi phu thuoc vao nong o c chat. Cho nen khi giam nong o c chat (trc khi c chat b can hoan toan) toc o sinh trng cua vi khuan cung giam. Do o viec chuyen t pha lag sang pha on nh dien ra dan dan. Nguyen nhan ton tai cua pha on nh la do s tch luy cac san pham oc cua trao oi chat (cac loai ru, acid hu c) va s can chat dinh dng (thng la chat dinh dng co nong o thap nhat). Nguyen nhan th nhat rat phc tap va kho phan tch, nguyen nhan th hai a c nghien cu ky hn. S tang sinh khoi tong cong ty le thuan vi nong o ban au cua chat dinh dng han che. G=K.c ay G la o tang sinh khoi tong cong, c la nong o ban au cua chat dinh dng han che. K la hang so hieu suat : K=G/c

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Hang so hieu suat K thng c bieu th bang so mg chat kho oi vi 1mg chat dinh dng. oi vi cac ng, K thng dao ong khoang t 0,20 en 0,30 ngha la t 100mg ng c tao thanh 20 - 30mg khoi lng kho cua te bao. Can chu y rang ngay trong pha on nh co the dien ra cac qua trnh nh s dung chat dinh dng, phan huy mot phan ribosome. Nhng te bao man cam se chet trc, nhng te bao khac con ton tai mot thi gian cho ti khi nao viec oxy hoa cac chat d tr hoac cac protein cua te bao a can. Lng sinh khoi at c trong pha on nh goi la hieu suat hoac san lng. San lng phu thuoc vao tnh chat va so lng cac chat dinh dng s dung va vao ieu kien nuoi cay. o la s sai khac gia so lng vi khuan cc ai va khoi lng vi khuan ban au. X = Xmax - Xo ai lng nay c bieu th bang gam. 3.2.4. Pha t vong Trong pha nay so lng te bao song co kha nang song giam theo luy tha (mac du so lng te bao tong cong co the khong giam). oi khi cac te bao t phan nh cac enzyme cua ban than. cac vi khuan sinh bao t th phc tap hn do qua trnh hnh thanh bao t. Thc ra cha co mot quy luat chung cho pha t vong. S chet cua te bao co the nhanh hay cham, co lien quan en s t phan hay khong t phan. Do sc song ln, bao t b chet cham nhat (trong nhng ieu kien thch hp nh kho va nhiet o thap bao t co the duy tr c kha nang song hang tram nam). Nguyen nhan cua pha t vong cha that ro rang, nhng co lien quan vi ieu kien bat li cua moi trng. Trong trng hp moi trng tch luy cac acid (Escherichia, Lactobacillus) nguyen nhan cua s chet te bao tng oi de hieu. Nong o chat dinh dng thap di mc can thiet, se lam giam hoat tnh trao oi chat, phan huy dan dan cac chat d tr va cuoi cung dan en s chet cua hang loat te bao. Ngoai ac tnh cua ban than chung vi khuan, tnh chat cua cac san pham trao oi chat tch luy cung anh hng en tien trnh cua pha t vong. Mot so enzyme the hien hoat tnh xuc tac cc ai trong pha t vong nh deaminase, decarboxylase, cac amilase va proteinase ngoai bao. Ngoai chc nang xuc tac mot so qua trnh tong hp nhng enzyme noi tren chu yeu xuc tac cac qua trnh phan giai. Toc o t vong cua te bao co lien quan trc tiep en thc tien vi sinh vat hoc va ky thuat, o la van e bao quan cac chung vi sinh vat quan trong ve mat ly thuyet (cac chung va cac bien chung ac biet) va ky thuat (cac chung sinh chat khang sinh, acid amin, vitamine... vi san lng cao). Ngoai kha nang song ta con can bao quan ca cac ac tnh di truyen cua vi khuan. Co nhieu phng phap bao quan khac nhau, nhng tat ca eu nham lam giam trao oi chat en toi thieu chu yeu bang cach giam nhiet o va o am. Sau ay la mot so phng phap thng dung trong viec bao quan vi sinh vat :

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a. Cay chuyen thng xuyen tren thach nghieng hoac trch sau vao thach. Sau khi a sinh trng, vi khuan c gi trong tu lanh + 40C. Phng phap nay n gian nhat va thng c dung, nhng kem hieu qua nhat. b. Bao quan di dau vo trung : dau parafin va ngan can moi trng kho va lam giam trao oi chat do can tr s xam nhap cua oxy. c. Bao quan trong cat hoac at set vo trung : Do cau truc ly - hoa cat va at set eu la nhng vat chat tot mang cac te bao vi sinh vat, chu yeu la cac bao t. Sau khi lam kho khong kh cat (hoac at set) cung vi vi khuan co the bao quan te bao rat lau. d. ong kho ; la phng phap hoan thien va co hieu qua nhat. Vi khuan c tron vi moi trng thch hp (sa, huyet thanh,...) roi lam lanh va lam kho nh bang kho. Sau may nam bao quan te bao van gi c kha nang song ma khong b bien oi ve di truyen. e. Bao quan trong glycerin (10%) va gi trong tu lanh sau (-600C hay -800C) : ay la phng phap rat thch hp nhng can mua c loai ong nha chu nhiet (khi kh trung).

3.3. Sinh trng cua vi khuan trong qua trnh nuoi cay lien tuc
Trong phng phap nuoi cay tnh noi tren, cac ieu kien moi trng luon luon thay oi theo thi gian, mat o vi khuan tang len con nong o c chat giam xuong. Vi khuan phai sinh trng va phat trien theo mot so pha nhat nh, sinh khoi at c khong cao. Tuy nhien trong nhieu nghien cu va thc tien san xuat ta can cung cap cho vi sinh vat nhng ieu kien on nh e trong mot thi gian dai chung co the van sinh trng trong pha log. D nhien mot mc o nao o co the cay chuyen te bao nhieu lan (qua nhng khoang thi gian ngan) vao moi trng dinh dng mi . Nhng n gian hn, ngi ta a lien tuc moi trng dinh dng mi vao bnh nuoi cay vi khuan ong thi loai khoi bnh mot lng tng ng dch vi khuan. ay chnh la c s cua phng phap nuoi cay lien tuc trong chemostat va turbidostat. Gia s ta co mot bnh nuoi cay trong o vi khuan ang sinh trng phat trien, cho chay lien tuc vao bnh moi trng mi co thanh phan khong oi. The tch bnh nuoi cay c gi khong oi, ngha la dong moi trng i vao c bu bi dong moi trng i ra ci cung toc o. Ta goi the tch bnh la v (lt) la toc o vao cua moi trng dinh dng, toc o dong i vao la f (lt/gi). Do o toc o pha loang (con goi la he so pha loang) D la f/v. ai lng D bieu th s thay oi the tch sau 1 gi. Neu vi khuan khong sinh trng va phat trien, chung se b rut khoi bnh nuoi cay vi toc o :
v = dX = D. X dt
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ay X la sinh khoi te bao (g/l).

Toc o sinh trng cua quan the vi khuan trong bnh cho bi phng trnh :
v+ = dX = X dt

toc o thay oi cuoi cung (tang hoac giam) mat o vi khuan trong nuoi cay lien tuc la s sai khac gia toc o tang va toc o giam v : v = v+ - v- =
dX dt

= ( - D)X

Neu < D gia tr v = dX/dt co gia tr dng, ngha la mat o vi khuan trong bnh tang, ngc lai neu < D, v se co gia tr am va mat o vi khuan trong bnh giam. Trong trng hp ac biet = D ta co v = 0, ngha la mat o te bao khong tang khong giam theo thi gian, quan the vi khuan trong trang thai can bang ong hoc. Neu bnh th nghiem co thiet b duy tr sao cho luon luon bang D ta se thu c quan the vi khuan sinh trng va phat trien theo luy tha thng xuyen mot mat o te bao khong oi va khong phu thuoc vao thi gian. Trong trng hp nh vay khong nhng kch thc trung bnh cua te bao, trang thai sinh ly cua chung ma ca moi trng nuoi cay eu khong oi va khong phu thuoc vao thi gian. ieu nay, mot mat tao ieu kien cho viec nghien cu sinh trng va sinh ly cua te bao vi khuan, mat khac cai thien qua trnh san xuat vi sinh vat quy mo cong nghiep. Chemostat va turbidostat la hai thiet b nuoi cay trong o ta co the duy tr c ieu kien = D Chemostat gom mot bnh nuoi cay, dung dch dinh dng t mot bnh d tr chay vao ay vi mot toc o khong oi. Nh thong kh va khuay c hoc bnh nuoi cay c cung cap ay u oxy va bao am viec phan bo nhanh va ong eu cac chat dinh dng trong dong moi trng i vao. Theo mc o vao cua moi trng ma dch vi khuan c rut khoi bnh mot cach thch hp. Sinh trng cua vi khuan trong chemostat c ieu chnh bi nong o c chat. Chat dinh dng han che nay co the la mot thanh phan chu yeu nao o cua moi trng. Pho bien hn ca la nguon carbon va nang lng, nhng thng ngi ta cung chon nguon nitrogen, phosphore, lu huynh hoac Mg2+. Nh a noi tren, la ham so cua nong o chat dinh dng han che. = max Ngha la co vo so hang so t = 0 en = max (hang so toc o sinh trng cc ai at c khi bao hoa c chat) tuy theo nong o chat dinh dng han che sao cho mat o vi khuan trong bnh khong b giam. Nh the se co mot toc o pha loang Dm, o sinh khoi vi khuan at c cc ai, ngha la Dm = max. Trai vi chemostat hoat ong cua turbidostat (t ch turbidity = o uc) da vao viec duy tr mat o (hoac o uc) vi khuan khong oi. Te bao quang ien o o uc ieu chnh moi trng i vao bnh qua mot he thong rle. Trong bnh nuoi cay tat ca chat dinh dng eu nong o d tha va vi khuan sinh trng vi toc o gan cc ai.
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Neu mat o te bao tang qua gia tr can thiet th he so pha loang se tang va phan te bao tha b loai ra ngoai (v D > max). Ngc lai neu mat o te bao giam xuong di mc chon la he so D lai giam va mat o te bao se tang (v max > D). ket qua cua s ieu chnh la duy tr cho he so pha loang D = max. Ro rang ve mat ky thuat phng phap turbidostat phc tap hn chemostat. Tom lai, co nhng sai khac chnh gia nuoi cay tnh va nuoi cay lien tuc: Nuoi cay tnh c xem nh la he thong ong, quan the te bao sinh trng trong o phai trai qua cac pha m au logarit, on nh va t vong. Moi pha sinh trng c ac trng bi nhng ieu kien nhat nh. Viec ieu khien t ong kho thc hien. Nuoi cay lien tuc, trai lai, la he thong m co khuynh hng dan en viec thiet lap mot can bang ong hoc. Yeu to thi gian ay, trong pham vi nhat nh, b loai tr. Te bao c cung cap nhng ieu kien moi trng khong oi, nh viec ieu chnh t ong.

3.4. Lam ong bo s phan chia te bao

Bnh thng quan the vi khuan ang sinh trng va phan chia la mot hon hp te bao cac pha sinh trng rat khac nhau cua s phan chia te bao, ngha la vao luc bat ky, trong moi trng nuoi cay ta co the ong thi gap cac te bao va phan chia xong, sap phan chia hoac nhng giai oan ke tiep. Tuy nhien, muon nghien cu cac qua trnh trao oi chat trong suot chu ky phan chia te bao ta can co mot quan the vi khuan trong o cac te bao phan chia ong thi (ong bo). Co the lam ong bo s phan chia te bao bang mot phng phap nhan tao nh thay oi sinh ly te bao hoac da vao viec chon loc c hoc cac te bao co cung kch thc. Phng phap sinh ly lam ong bo s phan chia te bao pho bien la phng phap gay choang nhiet. Nh ta biet, trong nhng ieu kien nuoi cay bnh thng nhiet o thch hp ( vi khuan la 300C, 370C) cac qua trnh tong hp phan ln cac cao phan t (DNA, RNA, protein) dien ra song song va trang thai can bang. Bang cach giam nhiet o ta se pha huy can bang nay. Trong khi tong hp DNA con tiep tuc mot thi gian th viec tong hp RNA va protein b ngng. Ket qua la s phan chia te bao cung b ngng. Neu phuc hoi lai nhiet o thch hp ban au th ch sau mot thi gian lag ngan, te bao se bat au phan chia nhng bay gi phan chia ong bo tc la phan ln te bao eu phan chia trong mot khoang thi gian tng oi ngan. Sau khi nang nhiet o qua trnh tong hp RNA va protein c phuc hoi con ham lng DNA van mc ban au tng ng vi chang ket thuc nhan oi cua nhiem sac the. Tuy nhien, s phan chia ong bo, khong phai la trang thai ben vng , ch sau vai the he tnh ong bo lai b pha huy va quan the vi khuan lai la mot hon hp te bao cac pha phan chia khac nhau. Phng phap nang cao nhiet o a c ng dung au tien vi ong vat nguyen sinh (Tetrahymena). Viec phan tch sinh ly cac te bao phan chia ong bo chng minh rang do s pha huy th t bnh thng cua s phan chia nhiem sac the va s phan chia te bao ma gay
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nen hien tng sinh trng mat can bang o la nguyen nhan cua s phan chia ong bo. Nh ta biet ieu kien e m au viec phan chia te bao la s ket thuc qua trnh nhan oi nhiem sac the. Mat khac, mot khi a trong trang thai nhan oi nhiem sac the se oc lap vi cac ieu kien sinh trng va chu trnh nhan oi se hoan thanh ngay khi tong hp protein b km ham bi cloramphenicol hoac bi thieu mot acid amin chu yeu. Tren c s cua nhng ket qua nay ta co the hieu c nguyen tac lam ong bo khong phai ch bang gay choang nhiet ma noi chung bang moi tac ong khac en trang thai sinh ly bnh thng cua te bao. Nhiet o thap lam ngng tong hp RNA va protein nhng viec nhan oi nhiem sac the van c hoan thanh mac du vi toc o cham. Do tong hp RNA va protein b km ham ma s phan chia te bao va sinh trng cung b ngng. Bi v de hieu rang, tong hp protein la can khong nhng cho viec m au mot chu trnh nhan oi mi cua DNA ma ca cho viec tong hp cac enzyme va cac cau truc can thiet oi vi s hnh thanh vach ngan ngang. Con sinh trng b ngng th co the hieu rang v protein la thanh phan chu yeu cua te bao va sinh trng oi hoi co nhng enzyme thng xuyen hoat ong khong nhng tham gia vao cac qua trnh sinh tong hp ma ca vao cac qua trnh phan gia (qua trnh cung cap nang lng va cac tien chat cho viec tong hp). Nguyen nhan trc tiep cua s phan chia ong bo sau khi phuc hoi nhiet o thch hp la trang thai giong nhau cua s nhan oi nhiem sac the. Vao cuoi luc gay choang lanh tat ca vi khuan eu cha mot nhiem sac the a nhan oi, bat ke te bao a giai oan nao cua s nhan oi khi ha nhiet o. Viec phuc hoi nhiet o dan en viec hoat ong ong thi cua cac qua trnh ke tiep s nhan oi : phan ly nhiem sac the va hnh thanh cac vach ngang. Mot phng phap sinh ly khac lam ong bo s phan chia te bao la thay oi thanh phan cua moi trng. Neu vao mot luc nao o ta loai khoi moi trng nuoi cay mot chat dinh dng hoac mot chat trao oi chu yeu nao o vi khuan se sinh trng mat can bang : sau khi phuc hoi nong o chat nay, te bao se phan chia ong bo. Chang han, co the gay oi timin trong thi gian ngan cac bien chng E. coli tr dng oi vi timin hoac gay oi mot so acid amin can thiet cho sinh tong hp cac protein cua te bao. Ve nguyen tac lam ong bo s phan chia te bao bang gay oi acid amin ta co the giai thch nh sau: Trong trng hp nay tong hp RNA va protein b ngng ngay nhng qua trnh nhan oi nhiem sac the, neu a m au, van co the dien ra cho en khi hoan thanh. Nguyen nhan ay lai la s sinh trng mat can bang trong thi gian oi acid amin. V bc m au mot chu trnh nhan oi mi lien quan chat che en tong hp protein nen nhiem sac the cua tat ca vi khuan sau khi a nhan oi th ngng lai. Viec them acid amin se phuc hoi tong hp RNA va protein, do o cung kch thch phan ng m au chu trnh nhan oi mi sau khi cac nhiem sac the nhan oi a c phan ly. Nh the lam ong bo s phan chia te bao la hau qua cua viec lam ong bo s phan chia nhiem sac the. Nguyen tac lam ong bo bang gay oi timin, t quan iem tong hp DNA, van cha c giai thch. Viec loai bo timin cung ong thi lam ngng tong hp DNA, ngha
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la cac te bao trong quan the cha nhiem sac the cac giai oan khac nhau cua s nhan oi. Khoang thi gian gay oi khong c qua gii han 30 - 45 phut, neu khong sau o te bao se chet v thieu timin. ay la qua trnh khong thuan nghch, khong the loai bo bang cach them ay u timin. Phng phap chon loc c hoc xuat phat t thc te cac te bao ca the vao thi ky trc khi phan chia eu tang the tch. Trong mot quan the vi khuan sinh trng di nhng ieu kien bnh thng, nhng te bao nho nhat chnh la nhng te bao va phan chia xong. Neu chon loc rieng nhng te bao nay ta se thu c cac vi khuan ma sau khi nuoi cay tiep se phan chia ong bo. Ngi ta a dung cac mang loc co kch thc lo tng ng vi cac vi khuan nho nhat, ket qua cho kha tot. Ngoai ra cung co the chon loc bang cach ly tam cac te bao. Y ngha cua viec lam ong bo s phan chia te bao la cho no cho ta kha nang nghien cu mot quan the te bao ong nhat, ngha la trong o cac ac tnh hnh thai va sinh ly cua mot te bao tng ng vi cua ca quan the. Nh o ta co the chng minh c rang cac thanh phan cua te bao vi khuan khong c tong hp vi toc o nh nhau vao thi gian gia hai lan phan chia. Chang han, trong qua trnh phan chia te bao Azotobacter vinelandii ngng tong hp DNA nhng tang cng tong hp cac acid amin t do. Trai lai, ham lng RNA va protein tang nh nhau trong pha nay.

3.5. Cac phng phap xac nh sinh trng va phat trien cua vi khuan
So lng te bao va sinh khoi mac du co moi lien he theo ty le thuan y = kx nhng moi lien he nay ch ung trong ieu kien th nghiem rat han che, khong co y ngha pho bien. ieu nay chnh la do kch thc va khoi lng cua tng vi khuan thay oi tuy theo loai va chung, trong qua trnh sinh trng th phu thuoc vao thi gian ( cac pha cua ng cong sinh trng), vao toc o sinh trng, thanh phan moi trng nuoi cay va vao ieu kien nuoi cay. V vay trong viec theo doi sinh trng va phat trien cua vi khuan can phan biet cac phng phap xac nh so lng te bao va sinh khoi. 3.5.1. Cac phng phap xac nh so lng te bao Trong mot quan the vi khuan khong phai moi te bao eu co kha nang song. Nhng vi khuan goi la song phai tao thanh khuan lac tren (hoac trong) moi trng thach va phat trien trong moi trng dch the. Do o tuy theo muc ch th nghiem ta co the xac nh so lng te bao tong cong (ca te bao song va chet) hoac so lng te bao song. e xac nh so lng te bao tong cong ngi ta thng dung phng phap em te bao trc tiep di knh hien vi nh cac phong em (phong em Neubauer, Thom hoac Petrof - Hauser). Neu chieu day cua lp dch cha vi khuan la 0,02mm va canh hnh vuong la 0,05mm (the tch 5.10-8cm3) th muon xac nh so lng te bao trong 1ml phai nhan so tm thay vi 2.107. Can chu y la sau khi te bao vao phong em phai i mot thi gian e vi khuan lang va nam trong mot mat phang quang hoc. Neu te bao chuyen ong tch cc phai dung formaldehyde giet chet trc.
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Co the dung ky thuat nhuom ac biet e phan biet te bao song va te bao chet. Do te bao song co mang sinh chat hoat ong, khong tham thuoc nhuom nen hai loai te bao bat mau khong giong nhau. Chang han o congo ch nhuom mau te bao chet con te bao song khong mau. Tuy nhien, ket qua nhuom mau phu thuoc vao nhieu yeu to : loai vi khuan, o pha loang cua thuoc nhuom, pH...V du, cac te bao song cua C. acetobutylicum bat mau xanh va cac te bao chet bat mau o khi dung Acridin orange vi nong o 1 : 10.000. Ket qua se ngc lai neu dung Acridin orange 1 : 5.000. Trong trng hp xac nh so lng te bao song ngi ta thng em so khuan lac tao thanh bi cac vi khuan song trong ieu kien sinh trng thuan li. Dch treo vi khuan c pha loang va mot the tch nhat nh c a len moi trng thach trong hop petri. Sau khi nuoi cay ngi ta em so khuan lac moc len. Neu gia du rang moi te bao tao thanh mot khuan lac th em so khuan lac ta se em c so te bao song. Tat nhien ieu nay khong ung oi vi cac vi khuan moc thanh chuoi, thanh oi hay thanh am hoac oi vi cac dch treo vi khuan khong ong nhat. Ngoai ra cung can chon o pha loang vi khuan thch hp sao cho so khuan lac khong t (kem chnh xac), khong nhieu (khuan lac co the b st vao nhau va 2 te bao co the cho 1 khuan lac). Thng tren mot hop thach so khuan lac t 40 - 400 la thch hp. So khuan lac cung rat phu thuoc vao thanh phan moi trng. Te bao moc thanh khuan lac tren moi trng nay khong nhat thiet cung cho xuat hien khuan lac tren moi trng khac. Vi mot so loai vi khuan kho phat trien tren moi trng thach ngi ta co the kien tra so lng bang phng phap pha loang lien tiep sau o cay t moi o pha loang 1ml vao tng ong ng moi trng ch th (neu co 1 vi khuan a vao cung u cho phan ng dng tnh sau khi nuoi cay). Phng phap nay goi la phng phap so lng co kha nang nhat (MPN, Most Probable Number). Co the cay vao 5 ong va lay so lieu 3 o pha loang cuoi (co phan ng dng tnh) roi tra bang e tm ra so lng gan ung mat o vi khuan. Con co the loc chat dch qua mang loc vi khuan roi dat mang loc len a moi trng thach e kiem tra so khuan lac se moc va suy ra mat o vi sinh vat trong chat dch. 3.5.2. Cac phng phap xac nh sinh khoi te bao Viec chon phng phap e xac nh sinh khoi vi khuan tuy thuoc vao muc ch nghien cu. Chang han, muon anh gia san lng te bao th ta can sinh khoi ti hoac kho sau khi a ly tam te bao; muon xac nh cng o trao oi chat hay hoat tnh men, ngi ta lai tnh ham lng protein hoac nitrogen. Nh vay, trong thc te co the s dung cac phng phap trc tiep hoac gian tiep. Cac phng phap trc tiep gom co : 1. Xac nh sinh khoi ti hoac sinh khoi kho (phng phap nay kem chnh xac)

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2. Xac nh ham lng nitrogen tong so (phng phap micro - Kjehdal) va phng phap vi khuan tan xac nh NH3 hay ham lng carbon tong so (theo Van SlikeFolch). Cac phng phap nay cho o chnh xac cao. 3. Trong thc te hang ngay ngi ta thng xac nh ham lng protein cua vi khuan. Co the dung phng phap biure cai tien hoac phng phap so mau khac. Cac phng phap vi lng da vao viec o so lng cac thanh phan ac trng cua protein nh tirosine, triptophan (theo Lowry hoac Folin-Ciocalteu) cung cho ket qua tot. Co the noi xac nh ham lng protein trong sinh khoi la phng phap thch hp nhat v mot mat protein la thanh phan chu yeu cua chat kho, mat khac o la nhng thanh phan hoat ong trong sinh khoi (hau het protein cua te bao la enzyme) Cac phng phap gian tiep e o sinh khoi vi khuan: 1. o o uc cua dch treo te bao. ay la phng phap rat thuan li. Trong thc te ta thng o mat o quang hoc cua dch treo (dch huyen phu). Trong mot so trng hp ngi ta cung xac nh s khuech tan anh sang. Tuy nhien s phu thuoc theo ng thang gia hai ch so nay vi sinh khoi vi khuan ch thay trong vung cac gia tr rat thap cua mat o te bao. V s khuech tan anh sang phu thuoc vao ng knh, hnh dang va ch so chiet quang cua cac hat khuech tan nen thnh thoang can kiem tra lai tng quan gia cac ai lng quang hoc va cac ch so o nh sinh khoi kho, ham lng nitrogen hoac ham lng carbon. Cung can chu y rang oi vi kh cung mot sinh khoi vi khuan trong dch treo nhng co the co mat o quang hoc khac nhau. Chang han mat o quang hoc oi vi 1mg/ml chat kho cua E.coli trong pha log la 1,54 con trong pha on nh la 2,38. gia tr cua mat o quang hoc (O.D)/mg chat kho cung thay oi theo toc o sinh trng . 2. o cac ch so cng o trao oi chat nh hap thu O2, tao thanh CO2 hay acid, v cac ch so nay lien quan trc tiep vi sinh trng. D nhien ta ch dung cac phng phap nay trong trng hp khong s dung cac phng phap khac, v du khi mat o sinh khoi rat nho. e o cac ch so noi tren co the dung cac phng phap chuan o, ien hoa...

3.6.

Tac dung cua cac yeu to ben ngoai len sinh trng va phat trien cua vi khuan

Sinh trng va trao oi chat cua vi khuan lien quan chat che vi cac ieu kien cua moi trng ben ngoai. Cac ieu kien nay bao gom hang loat cac yeu to khac nhau tac ong qua lai vi nhau. a so cac yeu to o eu co mot ac tnh tac dung chung bieu hien 3 iem hoat ong : toi thieu, toi thch va cc ai. Vi tac dung toi thieu cua yeu to moi trng vi khuan bat au sinh trng va m au cac qua trnh trao oi chat, vi tac dung toi thch vi khuan sinh trng, vi tac dung

31

cc ai va bieu hien hoat tnh trao oi chat, trao oi nang lng ln nhat, vi tac dung cc ai vi khuan ngng sinh trng va thng chet. Anh hng cua cac yeu to moi trng len vi khuan co the la thuan li hoac bat li. Anh hng bat li se dan en tac dung c khuan hoac diet khuan. Do tac dung c khuan cua yeu to moi trng, te bao ngng phan chia, neu loai bo yeu to nay khoi moi trng vi khuan lai tiep tuc sinh trng va phat trien. Khi co mat chat diet khuan, trai lai vi khuan ngng sinh trng, phat trien va chet nhanh chong. S chet cua te bao thng khong xay ra ngay mot luc trong ca quan the ma dien ra dan dan, co the bieu dien bang ng cong t vong logarit. Mot so yeu to, chu yeu la cac hoa chat co the hien tac dung c khuan hoac diet khuan tuy nong o. Tac dung khang khuan cua cac yeu to ben ngoai chu anh hng cua mot so ieu kien nh tnh chat va cng o tac dung cua ban than yeu to, ac tnh cua c the va tnh chat cua moi trng. 3.6.1. C che tac dung cua cac yeu to ben ngoai len vi khuan Cac yeu to cua moi trng ben ngoai tac dung len te bao vi khuan thuoc 3 loai : yeu to vat ly (o am, nhiet o...), yeu to hoa hoc (pH moi trng, the oxy hoa kh...) va yeu to sinh hoc (chat khang sinh). Du la yeu to nao nhng khi a tac dung bat li len te bao th thng trc het gay ton hai en cac cau truc quan trong cho s song cua te bao.. Nhng ton hai o dan en pha huy chc phan hoat ong cua cac cau truc va lam te bao chet.. Tac dung co hai cua cac yeu to ben ngoai te bao vi khuan the hien chu yeu nhng bien oi sau : 3.6.1.1. Pha huy thanh te bao. Mot so chat nh lisozyme (cha trong la lach, bach cau, long trang trng,...) co kha nang phan huy thanh te bao vi khuan dan en tao thanh cac nguyen lap chu yeu vi khuan G+ va cac cau lap vi khuan GTrong s co mat cua penicillin, cac cau lap c hnh thanh va de phan huy. vi khuan G-, do tac dung cua chat khang sinh nay, te bao tao thanh cac dang hnh cau man cam vi ap suat tham thau tng t vi cac dang L cua vi khuan. 3.6.1.2. Bien oi tnh tham cua mang te bao chat Mot so chat khong nhat thiet phai xam nhap te bao nhng van gay tac dung khang khuan. Do tac dung len mot hoac mot so chc nang sinh ly cua mang te bao chat nay lam vi khuan mat kha nang sinh san. Tac dung khang khuan cua cac chat oxy hoa va cac chat kh (H2O2, cac halogien...) la do anh hng cua chung len cac thanh phan cua mang te bao chat. Cung co the xep vao nhom cac hp chat nay, cac ru, phenol, cac chat tay ra tong hp va mot so khang sinh.
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3.6.1.3. Thay oi ac tnh keo cua nguyen sinh chat Cac yeu to vat ly cung nh hoa hoc eu co the gay nen tac dung nay. Chang han, nhiet o cao lam bien tnh protein va lam chung ong tu. 3.6.1.4. Km ham hoat tnh Mot so chat tac ong vao cac he thong sinh nang lng cua te bao, cyanide km ham enzyme cytochrome - oxydase, fluoride ngan can qua trnh ng phan. Cac chat oxy hoa manh (H2O2, cac halogien) pha huy cac he thong te bao lam ton hai en chc nang trao oi chat.... 3.6.1.5. Huy hoai cac qua trnh tong hp Trong s co mat cua mot so chat tng t ve mat cau truc vi cac chat trao oi t nhien, goi la chat antimetabolite qua trnh sinh tong hp co the b c che. C che tac dung cua cac chat antimetabolie khong giong nhau. Mot so gan vi trung tam hoat ong cua enzyme nhng khong tham gia vao phan ng khien enzyme mat hoat tnh phan huy c chat; mot so khac co the tham gia vao phan ng enzyme va c lap vao san pham cua phan ng nhng sau o khong c s dung trong trao oi chat vi cung mc o nh trong trng hp cua c chat thc. 3.6.2. Cac yeu to vat ly 3.6.2.1. o am Hau het cac qua trnh song cua vi khuan co lien quan en moi trng nc do o o am la mot yeu to quan trong cua moi trng. a so vi khuan thuoc cac sinh vat a nc ngha la chung can nc dang t do, de hap thu. Ch mot so xa khuan co the xep vao bon a kho v chung s dung c ca nc hydroscopic gan tren be mat cac hat at dang cac phan t. Khi thieu nc se xay ra hien tng loai nc khoi te bao vi khuan, trao oi chat b giam va te bao chet. So vi te bao dinh dng, cac bao t chu ng c kho han hn rat nhieu. Neu lam lanh te bao ong thi lam kho trong chan khong ta co the bao quan c sc song cua te bao trong mot thi gian dai. ay la nguyen tac cua phng phap lam ong kho vi khuan. 3.6.2.2. Nhiet o Hoat ong trao oi chat cua vi khuan co the coi la ket qua cua cac phan ng hoa hoc. V cac phan ng nay phu thuoc chat che vao nhiet o nen yeu to nhiet o ro rang anh hng sau sac en cac qua trnh song cua te bao. Te bao thu c nhiet chu yeu t moi trng ben ngoai, mot phan cung do c the thai ra do ket qua cua hoat ong trao oi chat. Hoat ong cua vi sinh vat gii han trong moi trng cha nc co the hap thu. Vung nay cua nc nam t 20 en khoang 1000 goi la vung sinh ong hoc. Hau het te bao sinh dng cua vi sinh vat b chet nhiet o cao protein b bien tnh, mot hoac hang loat enzyme b bat hoat. Cac enzyme ho hap ac biet la cac enzyme trong chu trnh Krebs rat man cam vi nhiet o. S chet cua vi khuan nhiet o cao cung co the con la hau qua cua s bat hoat hoa RNA va s pha hoai mang te bao chat.
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Nhiet o thap (di vung sinh ong hoc) co the lam bat hoat qua trnh van chuyen cac chat hoa tan qua mang te bao chat do thay oi cau hnh khong gian cua mot so permease cha trong mang hoac anh hng en viec hnh thanh va tieu thu ATP can cho qua trnh van chuyen chu ong cac chat dinh dng. Vi khuan thng chu ng c nhiet o thap. nhiet o di iem bang hoac thap hn chung khong the hien hoat ong trao oi chat ro ret. Nhiet o thap co the coi la yeu to c khuan neu lam lanh qua nhanh. Neu lam lanh trong chan khong cac tinh the bang se thang hoa. o la phng phap ong kho e bao quan vi sinh vat. Gii han gia nhiet o cc tieu va nhiet o cc ai la vung nhiet sinh trng cua vi sinh vat. Gii han nay rat khac nhau gia cac loai vi khuan : tng oi rong cac vi khuan hoai sinh, nhng rat hep cac vi khuan gay benh. Tuy theo quan he vi vung nhiet co the chia vi khuan thanh mot so nhom : a. Vi khuan a lanh : sinh trng tot nhat nhiet o di 200C, thng gap trong nc bien, cac ho sau va suoi nc lanh. Hoat tnh trao oi chat cac vi khuan nay thap. Trong ieu kien phong th nghiem nhieu vi khuan a lanh de dang thch ng vi nhiet o cao hn. b. Vi khuan a am: chiem a so, can nhiet o trong khoang 200C - 400C. Ngoai cac dang hoai sinh ta con gap cac loai ky sinh gay benh cho ngi va ong vat. c. Vi khuan a nong : sinh trng tot nhat 550C. Nhiet o sinh trng cc ai cua cac vi khuan a nong dao ong gia 750C va 800C. Cac vi khuan a nong gom chu yeu la cac xa khuan, cac vi khuan sinh bao t, thanh tao va nam moc. Thng gap chung trong suoi nc nong, trong phan u. Ap lc, ap suat tham thau va ap suat thuy tnh co the anh hng en cau truc cua te bao vi khuan. Mang te bao chat cua vi khuan la ban tham do cac hien tng tham thau va viec ieu chnh tham ap qua cac he thong permease eu co lien quan en mang. Trong moi trng u trng te bao mat kha nang rut nc va cac chat dinh dng hoa tan bao quanh : te bao chu trang thai kho sinh ly, b co sinh chat va co the b chet neu keo dai. Trong thc te ngi ta ng dung tac dung co sinh chat cua cac nong o muoi cao (10 15%) hoac ng cao (50 - 80%) e bao quan thc pham (muoi da, ca, p tht ca) hoa qua (lam mt) v a so vi sinh vat rat man cam vi tham ap cao cua moi trng. Ngc lai, khi vi khuan vao dung dch nhc trng nc se xam nhap te bao, ap lc ben trong se tang len. Tuy nhien do co thanh te bao cng vi khuan khong xay ra hien tng v sinh chat nh te bao thc vat. a so vi khuan sinh trng tot trong moi trng cha t hn 2% muoi, nong o cao hn co hai cho te bao. Nhng cung co mot so vi khuan lai sinh trng tot nhat trong moi trng cha ti 30% muoi. Ta goi la cac vi khuan a muoi. Trong hoat ong song cua mnh vi khuan thng chu anh hng cua nhng thay oi ap lc thuy tnh. nhiet o bnh thng ap lc cao co the lam cham hoac lam mat
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kha nang di ong, lam ngng sinh trng, lam yeu ong lc va lam thay oi trao oi chat nhng khong lam chet vi khuan. Tuy nhien, nhieu vi khuan ay bien va cac mo dau co the chu ap suat thuy tnh ti 200 300 atm. Ta goi o la cac vi khuan a ap. Ve c che tac dung cua ap suat thuy tnh cao len vi khuan van cha co y kien dt khoat. Co le do ap suat cao ma the tch te bao b giam, o nht cua noi thay te bao tang len, t o dan en lam bat hoat mot so enzyme, nhat la cac enzyme nam trong qua trnh phan chia te bao va lam giam toc o hoac lam ngng cac phan ng sinh hoa. Cung co the, do ap lc thuy tnh cao ma chc nang cua mang te bao chat b ton thng. Am thanh. Song am thanh, ac biet trong vung sieu am (tren 20kHz), co anh hng ln en sinh trng cua vi khuan. Cac te bao sinh dng b chet nhanh chong, te bao non man cam hn nhieu so vi te bao gia. Man cam nhat oi vi tac dung cua sieu am la cac vi khuan hnh si, t man cam hn la cac trc khuan va co sc e khang cao nhat la cac cau khuan. ac biet, sieu am hau nh khong anh hng g len bao t vi khuan va cac vi khuan khang acid. Sc cang be mat. Khi sinh trng trong moi trng dch the vi khuan chu anh hng cua sc cang be mat cua moi trng. a so cac moi trng dch the dung trong phong th nghiem co sc cang be mat khoang 0,57 - 0,63mN/cm. Nhng thay oi manh me sc cang be mat co the lam ngng sinh trng va lam te bao chet. Khi sc cang be mat thap, cac thanh phan cua te bao chat b tach khoi te bao. ieu nay chng to mang te bao chat b ton thng. Cac chat nang cao sc cang be mat chu yeu la cac muoi vo c. Cac chat lam giam sc cang be mat chu yeu la cac acid beo, alchohol va cac chat khac vi chuoi carbon dai, thang va thm. Cac chat nay c goi la cac chat co hoat tnh be mat. Tac dung cua chung the hien trong viec lam thay oi cac ac tnh be mat cua vi khuan., trc het la nang cao tnh tham cua te bao. Trong thc te ngi ta s dung hien tng nay trong viec nuoi cay cac vi khuan khang acid. Sc cang be mat thap con ngan can vi khuan gan vao be mat cng, tranh cho chung khoi canh tranh sinh trng. Cac tia bc xa. Anh sang co the gay ra nhng bien oi hoa hoc va do o nhng ton thng sinh hoc neu c te bao hap thu. Mc o gay hai tuy thuoc vao mc nang lng trong lng t anh sang c hap thu va mc nang lng trong lng t lai phu thuoc gian tiep vao chieu dai song cua tia chieu. Cac lng t bc xa gay nen nhng bien oi hoa hoc cua cac phan t va nguyen t co chieu dai song khoang 10.000 . Bao gom anh sang mat tri, ti t ngoai, tia X, tia gamma va tia vu tru. Cac tia vu tru tia gamma va tia X co nang lng rat ln. Khi c vat chat hap thu chung co the ban ra cac electron t cac nguyen t cua vat chat o. V vay cac tia nay c goi la cac bc xa ion hoa. Anh sang mat tri. La nguon tia chieu t nhien nhat co tac dung pha huy te bao vi khuan (ngoai le : cac vi khuan quang hp lai s dung anh sang mat tri lam nguon nang lng). Tac dung nay b yeu i neu te bao cha sac to hay cac vo nhay. So vi cac bc xa ion hoa th tia t ngoai (10 - 300nm) co nang lng nho hn. Khi b vat chat hap phu, tia t ngoai khong gay nen hien tng ion hoa nhng kch thch cac phan t, ngha la chuyen cac ien t en mot nang lng cao hn. Di anh hng cua
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tia t ngoai vi khuan b chet hoac b ot bien tuy theo loai vi khuan va lieu lng chieu, bao t cua moc co sc e khang cao. ieu ang chu y la nhng h hai do tia t ngoai gay nen trong te bao phan nao co the ao ngc. Neu sau khi chieu tia t ngoai ta lai cho vi khuan chu tac dung cua anh sang ban ngay th nhieu vi khuan co kha nang song sot va tiep tuc sinh trng, phan chia. Hien tng nay c goi la quang tai hoat. Tia sang mat tri tuy co cha mot phan tia t ngoai nhng phan ln nhng tia nay b kh quyen (ozon, may...) gia lai. V vay anh nang co tac dung diet khuan nho hn so vi tia t ngoai dung trong phong th nghiem. Hien nay cac bc xa ion hoa c ng dung trong viec kh trung thc pham, dc pham... va e gay ot bien vi sinh vat. 3.6.3. Cac yeu to hoa hoc Trong so cac yeu to hoa hoc anh hng en te bao trc het phai ke nong o ion hydrro (pH), the oxy hoa kh (Eh) cua moi trng, cac chat sat trung va cac chat hoa tr lieu. 3.6.3.1. Anh hng cua pH moi trng pH cua moi trng co y ngha quyet nh oi vi sinh trng cua nhieu vi sinh vat. Cac ion H+ va OH- la hai ion hoat ong ln nhat trong tat ca cac ion, nhng bien oi du nho trong nong o cua chung cung co anh hng manh me. Cho nen viec xac nh thch hp ban au va viec duy tr pH can thiet trong thi gian sinh trng cua te bao la rat quan trong. Cac gia tr pH (cc tieu, toi thch, cc ai) can cho sinh trng va sinh san cua vi khuan tng ng vi cac gia tr pH can cho hoat ong cua nhieu enzyme. Gii han pH hoat ong oi vi vi sinh vat trong khoang 4 - 10. a so vi khuan sinh trng tot nhat pH trung bnh (7,0). Cac vi khuan nitrate hoa, vi khuan not san, xa khuan, vi khuan phan giai urea lai a moi trng hi kiem. Mot so vi khuan chu acid nh vi khuan lactic, Acetobacter... co the sinh trng pH<1. nam si va nam men a pH acid (pH : 4 - 6) pH cua moi trng khong nhng anh hng manh me en sinh trng ma con tac ong sau sac en cac qua trnh trao oi chat. Mang te bao chat cua vi sinh vat tng oi t tham oi vi cac ion H+ va OH-. V vay, mac du pH cua moi trng ben ngoai dao ong trong gii han rong, nong o cua 2 ion noi tren trong te bao chat noi chung van on nh. Anh hng cua pH moi trng len hoat ong cua vi sinh vat co the la do ket qua tac ong qua lai gia ion H+ va enzyme cha trong mang te bao chat va thanh te bao. e duy tr pH thch hp oi vi vi sinh vat trong thi gian nuoi cay, nhat la oi vi cac vi khuan sinh acid nhng lai khong chu c acid (Lactobacillus, Enterobacteriacea, nhieu Pseudomonas), ngi ta thng them vao moi trng chat em hoac dung cac c chat oxy hoa hoan toan. Dung dch em thng dung la muoi cua cac acid yeu (phosphate, acetate, carbonate).
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3.6.3.2.

The oxy hoa kh (Eh)

e bieu th mc o thoang kh cua moi trng ngi ta dung ai lng rH. Theo nh ngha : rH = -log (H2), ay (H2) la ap lc cua hydro trong kh quyen. Dung dch nc bao hoa hydrro co rH = 0, bao hoa oxy co rH = 41. Thang t 0 en 41 xac nh mc o thoang kh cua moi trng. pH co anh hng en gia tr rH cua moi trng, s phu thuoc nay c bieu th bi phng trnh : rH =
Eh 0.03

+ 2pH (Eh = the oxy hoa kh tnh ra volt)

Cac vi sinh vat k kh bat buoc co the sinh san nhng gia tr rH rat thap (khong qua 8 - 10), cac vi sinh vat hieu kh hoac k kh tuy tien gii han kha rong rH t 0 en 30 con cac vi sinh vat hieu kh bat buoc rH t 10 en 30. Cac gia tr rH > 30 khong co li cho s sinh san cua cac vi sinh vat hieu kh bat buoc. Cac vi khuan k kh co the sinh trng gia tr Eh kha thap. bang cach them mot so chat nhat nh vao moi trng co the giam gia tr Eh. Oxy co vai tro het sc quan trong trong hoat ong song cua vi sinh vat. Tuy thuoc vao nhu cau oi vi oxy ma ngi ta chia vi sinh vat thanh cac nhom sau: Hieu kh bat buoc : Thuoc nhom nay la cac vi sinh vat ch co the sinh trng c khi co mat oxy phan t (O2). Chung co chuoi ho hap hoan chnh, dung O2 lam the nhan hydro cuoi cung. Trong te bao co cha enzyme SOD (superocide dismutase) va peroxidase. Tuyet ai a so vi nam va so ong vi khuan thuoc nhom nay. Hieu kh khong bat buoc Thuoc nhom nay la cac vi sinh vat co the sinh trng c ca trong ieu kien co oxy lan trong ieu kien khong co oxy. Trong te bao co cha SOD va peroxidase. Co oxy chung sinh trng tot hn. Phan ln nam men va nhieu vi khuan thuoc nhom nay. Co the ke en cac loai nh Saccharomyces cerevisiae, E. coli ... Vi hieu kh : Thuoc nhom nay la cac vi sinh vat ch co the sinh trng c ieu kien ap lc oxy rat thap (khoang 0.01 0.03atm). Chung cung thong qua chuoi ho hap va dung oxy lam the nhan hydro cuoi cung. Co the ke en cac loai nh Vibrio cholerae, Hydrogenomonas spp. ... K kh chu dng : o la nhng vi khuan k kh nhng lai ton tai c khi co mat oxy. Chung khong s dung oxy, khong co chuoi ho hap nhng s co mat cua oxy khong co hai oi vi chung. Trong te bao co SOD, peroxidase nhng thieu hydrogenperoxidase. Thuoc nhom nay co the ke en Streptococcus lactis, Lactobacillus lactis... K kh :
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Vi cac vi sinh vat thuoc nhom nay s co mat cua oxy phan t la co hai. Chung khong sinh trng c tren moi trng ac hoac ban ac khi e trong khong kh hay trong khong kh co cha 10% CO2. Chung ch sinh trng c lp dch the sau, ni khong co oxy, qua trnh len men, qua trnh phosphoryl hoa quang hp, qua trnh methane. Trong te bao cua cac vi sinh vat nay khong co SOD, cytochromoxydase, phan ln khong co hydrogen peroxidase. Co the ke en rat nhieu loai trong cac chi Clostridium, Fusobacterrium, Bifidobacterium ... 3.6.3.3. Cac chat diet khuan (sat trung) Cac chat diet khuan thng dung nhat la phenol va cac hp chat cua phenol, cac alcohol, halogen, kim loai nang, H2O2, ... Hien nay ngoai viec dung mot so hoa chat e sat trung da, cac dung cu dc pham, thc pham, nc uong... ngi ta con dung cac dung dch nh nc brom 1%, HgCl2 0,1%, con, AgNO3 0,05%, hypochloride calci (1% Cl2) e sat trung be mat hat. Hat c ngam trong cac dung dch tng ng khoang 5 - 30 phut, trc o can x ly hat bang xa phong va cac chat co hoat tnh be mat khac e am bao cho be mat hat tham hoan toan, x ly xong phai ra hat nhieu lan bang nc vo trung. a so cac chat dung lam thuoc nhuom vi khuan eu la chat km ham chung. 3.6.3.4. Cac chat hoa tr lieu o la cac chat hoa hoc co tac dung oc len vi khuan nhng khong gay hai cho c the bac cao (khac vi cac chat sat trung). Noi chung thanh phan hoa hoc cua cac chat nay a c biet, do o ta co the ieu chnh bang con ng nhan tao. C che tac dung cua cac chat hoa tr lieu ro rang da vao s tng t ve cau truc cua cac chat nay vi cac hp chat ma vi khuan can e tao thanh cac coemzyme, protein va cac acid nucleic. Cac chat hoa tr lieu canh tranh v tr gan vi cac hp chat o tren phan t enzyme va km ham nhieu phan ng sinh hoa quan trong. a so cac chat hoa tr lieu c s dung e ieu tr cac benh khac nhau. Cac chat hoa tr lieu quan trong nhat la cac sulfonamide dan xuat t acid paminosulfonic. o la nhng chat oi khang cua acid p-aminobenzoic (APAB). Do canh tranh v tr gan tren phan t enzyme vi PAB cac sulfonamide km ham viec tao thanh acid folic la tien chat cua coenzyme tham gia vao qua trnh tong hp mot so acid amin va purin. 3.6.4. Cac yeu to sinh hoc Trong so cac yeu to sinh hoc co anh hng co hai len cac qua trnh song cua vi khuan can ke en cac khang the va cac chat khang sinh. Khang the la cac globulin trong mau ong vat, co kha nang lien ket ac hieu vi khang nguyen a kch thch sinh ra no, khang the nay con c goi la khang the ac hieu. Khang the c tm thay chu yeu trong huyet thanh cua ong vat, v vay huyet thanh cha khang the ac hieu khang nguyen c goi la khang huyet thanh.

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Khang the cung co the c tm thay trong cac dch khac cua c the, nh sa. Nhng khang the co san trong sa hay huyet tng cua ngi hay ong vat t trc khi co s tiep xuc vi khang nguyen c goi la khang the t nhien hay khang the khong ac hieu.

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Chng 4 THANH PHAN VI SINH VAT THAM GIA TRONG QUA TRNH X LY NC THAI
WX Vi sinh vat la mot thanh phan qua trong trong qua trnh x ly nc thai sinh hoc. Ve mat cau truc va chc nang, vi sinh vat c xep vao loai sinh vat co nhan va tien nhan. Bon tien nhan gom (eubacteria, archaebacteria) la quan trong nhat trong x ly sinh hoc va c hieu n gian la vi khan. Bon co nhan bao gom thc vat, ong vat va nguyen sinh vat. Bon co nhan quan trong trong qua trnh x ly sinh hoc bao gom nam, ong vat nguyen sinh va trung banh xe, tao.

4.1.

Vi khuan (bacteria)

La cac sinh vat tien nhan n bao. Hnh thc sinh san chu yeu la nhan oi mac du co mot so loai sinh san hu tnh hoac bang cach nay choi. Cho du co hang ngan loai vi khuan khac nhau nhng dang tong quat ch co ba loai sau: spherical, cylindrical va helical. Kch thc chung bien oi. Mot vai kch thc cua vi khuan nh sau : ng knh t 0.5 - 1.0m oi vi spherical, rong t 0.5-1m va dai t 1.5-3m oi vi cylindrical va rong t 0.5-5m dai t 6-15m oi vi helical. 4.1.1. Cau truc te bao Co cau truc te bao n gian (hnh 4.1) ben trong te bao goi la te bao chat, cha protein, carbonhydrate va cac chat hu c phc tap. Vung dch bao cha ribonucleic acid (RNA), co vai tro chu yeu trong viec sinh tong hp protein. Cung ben trong te bao chat co vung cha deoxyribonucleic acid (DNA), DNA cha tat ca nhng thong tin can thiet cho s san xuat tat ca cac cau thanh cua te bao va co the c goi la vat lieu di truyen cua te bao. Thanh phan te bao: kiem tra tren mot so lng ln cac vi khuan khac nhau ngi ta cho rang chung cha khoang 80% la nc va 20% la chat kho trong o co 90% la chat hu c va 10% la vo c. Thanh phan c ban cua te bao vi khuan nh sau:
Nguyen to
Carbon (C) Oxygen (O) Nitrogen (N) Hydrogen (H) Phosphorus (P) Sulfur (S) Potasium (K) Sodium (Na) Calcium (Ca) Magnesium (Mg) Chlorine (Cl) Sat (Fe) Cac chat khac

Phan tram trong lng kho Khoang Trung bnh

45-55 16-22 12-16 7-10 2-5 0.8-1.5 0.8-1.5 0.5-2.0 0.4-0.7 0.4-0.7 0.4-0.7 0.1-0.4 0.2-0.5 50 20 14 8 3 1 1 1 0.5 0.5 0.5 0.2 0.3

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Hnh 4.1: Cau truc te bao di knh hien vi ien t Cong thc c c tnh cho chat hu c n gian nhat la C5H7O2N, theo cong thc nay th co khoang 53% ve trong lng c the la carbon. Cong thc C60H87O23N12P co the c s dung khi co tnh en phosphorus. Thanh phan vo c bao gom P2O5 (50%), SO3 (15%), Na2O (11%), CaO (9), MgO (8), K2O (6%) va Fe2O3 (1%). Bi v tat ca cac n chat va hp chat eu co nguon goc t moi trng, nen s thieu hut mot trong nhng chat nay trong chat nen se la nhan to han che va trong mot vai trng hp co the tac ong en s phat trien cua vi khuan. 4.1.2. ieu kien moi trng Cac ieu kien cua moi trng ve nhiet o va pH co anh hng quan trong len s ton tai va phat trien cua vi khuan. Noi mot cach tong quat, s phat trien toi u ch xay ra trong mot gii han hep cua nhiet o va pH, cho du vi khuan co the ton tai trong mot khoang rong ve gii han cua hai yeu to nay. Nhiet o di nhiet o toi u thng co anh hng y ngha len s phat trien hn la nhiet o tren nhiet o toi u. Toc o phat trien cua vi khuan se tang len gap oi khi nhiet o tang len 100C cho en khi at en nhiet o toi u. pH cua moi trng cung la nhan to anh hng chu yeu en s phat trien cua vi khuan. Hau het vi khuan khong the thch ng c pH>9.5 hoac pH<4.0. pH toi u cho s phat trien cua vi khuan nam trong khoang 6.5-7.5.

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4.1.3. S phat trien cua vi khuan Viec kiem soat moi trng co hieu qua trong x ly sinh hoc da vao s hieu biet cac quy tac c ban nham che ng s phat trien cua vi sinh vat. Sau ay ban luan ve s phat trien cua vi khuan, cac vi sinh vat quan trong trong x ly sinh hoc. S phat trien cua vi khuan trong nuoi cay thuan khiet. Nh a e cap trc ay, vi khuan co the sinh san bang cach t nhan oi, sinh san gii tnh hoac sinh san bang nay mam, nhng chu yeu chung sinh san bang cach nhan oi (tc la t mot te bao ban au sinh ra hai te bao mi). Thi gian cho moi mot qua trnh nhan oi te bao c goi la thi gian sinh san, co the thay oi t vai ngay cho en t hn 20 phut. V du, neu thi gian gia hai the he la 30 phut, mot ca the vi khuan se nhan len thanh 16.777.216 ca the sau 12 gi. ieu nay c tnh theo ly thuyet, oi vi vi khuan se khong tiep tuc phan chia bi v mot so gii han cua nhan to moi trng nh nong o dinh dng, pH, nhiet o va tham ch ca kch thc cua he thong. S phat trien qua tng giai oan theo so lng vi khuan Hnh thc phat trien tong quat cua vi khuan trong nuoi cay c mo ta qua o th, qua o th ta thay s phat trien cua vi khuan trai qua 4 giai oan, moi mot giai oan tng ng vi moi so lng vi khuan khac nhau. (hnh 4.2)

Hnh 4.2. ng cong bieu dien cac giai oan phat trien cua vi khuan Ve so lng theo thang logarit (1) Giai oan phat trien cham (pha lag): trong suot thi ky nay vi khuan phai tiet vao moi trng chat khang the nham thch ng vi moi trng mi cua chung va bat au phan chia.

42

(2) Giai oan tang trng ve so lng theo Logarit (pha log) : suot thi gian nay vi khuan nhan oi vi mot toc o xac nh bi thi gian sinh san va kha nang thu nhan va ong hoa thc an cua chung (toc o tang trng theo phan tram la khong oi). (3) Giai oan phat trien on nh : Trong giai oan nay quan the vi khuan trong trang thai on nh. Ly do trc tien cua hien tng nay la: cac te bao a s dung can kiet chat nen hoac chat dinh dng can thiet cho s phat trien; s phat trien cua te bao mi can bang vi s chet cua te bao cu. (4) Giai oan vi khuan t chet : Suot giai oan nay, toc o chet cua vi khuan vt qua s san sinh ra te bao mi. Toc o chet thng la mot chc nang cua quan the song va cac ac tnh cua moi trng. Trong mot vai trng hp, tr so logarit cua so lng te bao chet tng ng vi tr so cua so lng te bao sinh ra gian oan sinh trng nhng co dau ngc lai. S phat trien ve mat so sinh khoi. S phat trien c thao luan ay trong moi tng quan cua nhng thay oi ve sinh khoi cua vi sinh vat theo thi gian. S phat trien nay bao gom 4 giai oan. (hnh 4.3).

Gia tr log so lng te bao vi sinh vat

Pha tang trng

Pha on nh Pha Chet

Pha thch nghi

Thi gian (gi)

Hnh 4.3. ng bieu dien s tang sinh khoi cua vi sinh vat
(1) Giai oan tang trng cham. Mot lan na vi khuan oi hoi phai thch ng vi moi trng dinh dng cua chung. Giai oan tang trng cham oi vi sinh khoi khong khong keo dai nh giai oan tang trng cham oi vi so lng bi v sinh khoi bat au tang trc khi s phan chia te bao xay ra (te bao ln len roi mi phan chia).

43

(2) Giai oan tang sinh khoi theo logarit : luon co tha thc an xung quanh vi sinh vat, va toc o trao oi chat va tang trng cua vi khuan ch phu thuoc vao kha nang x ly chat nen cua chung. (3) Giai oan tang trng cham dan: toc o tang trng sinh khoi cua vi khuan giam bi v s can kiet dan chat dinh dng. (4) Giai oan ho hap noi bao: vi khuan bat buoc thc hien qua trnh trao oi chat bang chnh cac nguyen sinh chat co trong te bao bi v nong o cac chat dinh dng cap cho te bao a b can kiet. Suot giai oan xay ra hien tng giam dan sinh khoi trong khi o cac chat dinh dng con lai trong te bao chet khuech tan ra ngoai e nuoi dng nhng te bao con con song. Phat trien trong moi trng hon hp Cac qua trnh phat trien tren lien quan en mot quan the vi sinh vat. Hau het cac qua trnh x ly sinh hoa xay ra trong moi trng hon hp gom nhieu chung loai vi sinh tac ong len moi trng va co tac ong tng ho lan nhau. Moi loai vi sinh vat trong he thong co mot ng cong sinh trng va phat trien rieng cua no, v tr va cac dang ng cong tang trng theo thi gian phu thuoc vao thc an va chat dinh dng co san va phu thuoc vao cac nhan to moi trng nh nhiet o, pH, va ca ieu kien k kh hoac hieu kh. S bien oi ve u the vi sinh vat theo thi gian trong qua trnh on nh hieu kh chat thai hu c c mo ta hnh 4.4. Co nhieu loai vi sinh vat ong vai tro quan trong trong qua trnh on nh chat hu c co trong nc thai. Khi thiet ke hoac phan tch qua trnh x ly sinh hoc, nha thiet ke nen ngh en cac giai oan cua mot he thong sinh thai hoac quan xa nh c mo ta hnh 4.4.

Hnh 4.4. S phat trien cua vi sinh vat trong nc thai

44

4.1.4. ong hoc cua qua trnh x ly sinh hoc. e am bao cho qua trnh x ly sinh hoc dien ra co hieu qua th phai tao c cac ieu kien moi trng nh pH, nhiet o, chat dinh dng, thi gian...tot nhat cho he vi sinh. Khi cac ieu tren c bao am qua trnh x ly dien ra nh sau: Tang trng te bao: ca hai trng hp nuoi cay theo tng me hay nuoi cay trong cac be co dong chay lien tuc, nc thai trong cac be nay phai c khuay tron mot cach lien tuc va hoan chnh. Toc o tang trng cac te bao vi sinh co the bieu dien bang cong thc sau: rt = X Trong o : rt : toc o tang trng cua vi khuan. Khoi lng/n v the tch trong mot n v thi gian (g/m3.s). : toc o tang trng rieng. 1/thi gian = 1/s. X : nong o sinh vat trong be hay nong o bun hoat tnh (g/m3=mg/l) Chat nen - gii han tang trng Trong trng hp nuoi cay theo me neu chat nen va chat dinh dng can thiet cho s tang trng ch co vi so lng han che th cac chat nay se c dung en can kiet va qua trnh tang trng ngng lai. trng hp nuoi cay trong be co dong cap chat nen va chat dinh dng lien tuc th anh hng cua viec giam bt dan chat nen va chat dinh dng co the
= m
S Ks + S

bieu dien bang phng trnh Monod e xuat (1942, 1949) Trong o: : toc o tang trng rieng (1/s) m : toc o tang trng rieng cc ai (1/s) S : nong o chat nen trong nc thai thi iem s tang trng b han che. Ks : hang so ban toc o, the hien anh hng cua nong o chat nen thi iem toc o tang trng bang toc o cc ai (g/m3, mg/l) S tang trng te bao va s dung chat nen. Trong ca hai trng hp nuoi cay theo me va nuoi cay trong be co dong chay lien tuc, mot phan chat nen c chuyen thanh cac te bao mi, mot phan c oxy hoa thanh chat vo c va hu c on nh. Bi v so te bao mi sinh ra lai hap thu chat nen va sinh san tiep nen co the thiet lap quan he gia toc o tang trng va lng chat nen c s dung theo phng trnh sau: rt = -Yrd
45

Trong o: rt : toc o tang trng cua te bao (g/m3.s) Y : he so nang suat s dung chat nen cc ai (mg/mg) (la ty so gia khoi lng te bao va khoi lng chat nen c tieu thu o trong mot thi gian nhat nh giai oan tang trng Logarit). rd : toc o s dung chat nen (g/m3.s) Anh hng cua ho hap noi bao. Trong cac cong trnh x ly nc thai khong phai tat ca cac te bao vi sinh vat eu co tuoi nh nhau va eu giai oan sinh trng Logarit ma co mot so ang giai oan chet va giai oan sinh trng cham. Khi tnh toan toc o tang trng cua te bao phai tnh toan to hp hien tng nay, e tnh toan gia thiet rang: s giam khoi lng cua cac te bao do chet va tang trng cham ty le vi nong o vi sinh vat co trong nc thai va goi s giam nay la do phan huy noi bao. Qua trnh ho hap noi bao co the bieu dien n gian bang phan ng sau: C5H7O2N + 5O2 113 160 1 1.42 5CO2 + 2H2O + NH3 + Q

T phng trnh tren cho thay : neu tat ca cac te bao b oxy hoa hoan toan th lng COD cua cac te bao bang 1.42 lan nong o te bao. rd (do phan huy noi bao) = -Kd.X Trong o Kd : he so phan huy noi bao (1/s) X : nong o te bao (nong o bun hoat tnh) (g/m3)
rt =
'

m XS
(K s + S )

Kd X

Ket hp vi tang trng noi bao, toc o tang trng thc te cua te bao : Hay rt = -Yrd - KdX. Trong o : rt : toc o tang trng thc cua vi khuan (1/s)
' = m
S Kd Ks + S

Toc o tang trng rieng thc se la : Toc o tang sinh khoi bun hoat tnh se la :
'

yb =

rt rd

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Anh hng cua nhiet o Nhiet o nc co anh hng rat ln en toc o cua phan ng sinh hoa trong qua trnh x ly nc thai. Nhiet o khong ch anh hng en hoat ong chuyen hoa cua vi sinh vat ma con tac ong ln en qua trnh hap thu kh oxy vao nc thai va qua trnh lang cac bong can vi sinh vat be lang t 2. Anh hng cua nhiet o en toc o phan ng sinh hoa trong qua trnh x ly nc thai c bieu dien bang cong thc : rT = r20(T-20) Trong o : rT : toc o phan ng T0C r20 : toc o phan ng 200C : he so hoat ong do nhiet o T : nhiet o nc o bang 0C Gia tr trong qua trnh x ly sinh hoc dao ong t 1.02-1.09 thng lay 1.04 4.1.5. ng dung s phat trien cua vi khuan va hoat ong s dung chat nen trong x ly sinh hoc. Trc khi thao luan ve cac qua trnh phat trien rieng biet c s dung cho x ly chat thai, th viec ap dung cac ong thai phat trien sinh hoc va s dung chat nen se c giai thch. Muc ch ay la mo ta (1) s phat trien cua vi sinh vat va s can bang chat nen va (2) d oan c nong o cac vi sinh vat va chat nen au ra. Trong phan thao luan nay, qua trnh x ly hieu kh thc hien trong mot phan ng tron hoan chnh khong co s hoi lu. Qua nghien cu cho thay qua trnh tren giong nh qua trnh bun hoat tnh khong co s hoi lu.

4.2.

Nam (fungi)

Nam co the c xem la nguyen sinh vat a te bao khong quang hp va d dng. Nam thng c phan loai da vao hnh thc sinh san. Chung sinh san hu tnh hoac vo tnh bang cach nhan oi, nay choi hoac tao thanh bao t. Moc hay nam that thng co hnh dang si goi la mycelium, men cung la nam nhng khong the tao thanh mycelium va thng khong phai la dang te bao. Hau het cac loai nam eu co the song trong ieu kien thieu kh. Chung co kha nang phat trien trong ieu kien o am thap va co the thch ng c trong moi trng pH thap. pH toi u cho s phat trien cua hau het cac loai nam la 5.6 va khoang bien ong t 2-9. Nam cung co nhu cau nitrogen thap, nhu cau cua chung ch khoang bang so vi vi khuan. Kha nang cua nam co the ton tai trong ieu kien pH va nhu cau nitrogen thap, kha nang phan huy cellusose gap oi, lam cho chung tr nen quan trong trong x ly sinh hoc cac chat thai cong nghiep va cac chat thai hu c dang ran.
47

4.3.

Tao (algae)

Tao la bon thc vat a bao hay n bao, t dng va quang hp c. Chung co tam quan trong trong cac qua trnh x ly sinh hoc bi hai ly do. Trong thuy vc chung co kha nang tao ra oxygen qua qua trnh quang hp la s song cua he sinh thai moi trng nc. oi vi ao hieu kh hoac ao oxy hoa k kh khong bat buoc hoat ong hieu qua, th tao la can thiet cho viec cung cap oxygen cho vi khuan d dng hieu kh. Tao thng song troi noi va phan phoi trong ca khoi nc, cung co loai tao bam (bam vao nhng vat chm trong nc) hoac song ay (tao thanh lp tren be mat cua nen ay). Nhng loai hien dien chnh trong x ly sinh hoc gom: Tao lam (cyanophyta) gan giong vi vi khuan. Tao luc (chlorophyta) Tao nau (chrysophyta) Tao mat (euglena) Moi tng quan sinh hoc gia tao va vi khuan trong x ly nc thai the hien mo hnh di ay.
S dung cho muc ch khac Chat thai hu c (cha C, N, P) Oxygen Sinh khoi tao

Vi khuan (C68H95O27N14)

Vi tao (C106H181O45N16P)

Nc thai a x ly

Sinh khoi vi khuan

CO2, NH4+, PO43Dinh dng khoang

Bc xa anh sang mat tri

CO2 kh quyen

Hnh 4.5. Moi tng quan gia tao va vi khuan trong x ly nc thai (theo W. J. Oswald, 1977)
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Vi khuan Nc thai + O2 bun vi khuan + nc thai a c x ly Oxy trong trng hp nay c cung cap bi thc vat phu du va trao oi nc be mat va khong kh. Thc vat + anh sang sinh khoi thc vat + O2 Muoi khoang + CO2 Chlorophylle cha trong vi tao cho phep chung s dung anh sang mat tri nh la nguon nang lng: o la cai c ban cua qua trnh quang hp. Tao phat trien di anh sang ong thi trch lay CO2 va chat khoang trong nc e lam nguon thc an cho c the roi thai oxy vao moi trng. Qua trnh quang hp c mo ta bi phng trnh sau: CO2 + 2H2O (CH2O)x + O2 + H2O

(CH2O)x c xem la chat hu c cua tao, ket qua phan tch tao cho ra cong thc phc tap nh sau: Tao = C106H181O45N16P. Va phng trnh c viet lai Anh sang 160CO2 + 182H2O + 16NH4+ + HPO42C106H181O45N16P + 118O2 + 117H2O + 14H+

Phng trnh nay la bang chng cho s co mat can thiet cua muoi dinh dng trong s san sinh tao. Can chu trong en s can bang v trong can bang nay so mol O2 c giai phong ra t hn so mol CO2 c tieu thu. S tang pH do CO2 chuyen dch can bang nh sau: Ca2+ + (HCO3-)2 CaCO3 + CO2 + H2O

Vao ban em, qua trnh quang hp b ngng lai va s ho hap cua tao gop phan vao viec giam ham lng oxy hoa tan (CH2O)x + O2 CO2 + H2O

49

Tao cung co vai tro quan trong trong cac qua trnh x ly sinh hoc nhng van e ngan can s phat trien vt mc cua tao trong cac loai nc tiep nhan (receiving waters) phai c quan tam en. Tao neu khong c s dung het bi bon ong vat phu du thng lang xuong ay, ieu o cho thay co mot s o nhiem xay ra nhng khong ang ke trong nen ay cua cac ao x ly. Nc thai sau khi c x ly o vao moi trng tiep nhan v vay ma co cha vi tao dang phu du cho thay s o nhiem hu c trc tiep (ham lng chat hu c l lng co the at en 0.2kg/m3). ieu o can phai co mot qua trnh thu hoi va lam tang gia tr cua tao khi thai ra. Mot vai ky thuat lang loc bang cac phng phap vat ly (ly tam, loc qua mang, lam kho, ong tu bang dong ien, tuyen noi bang ien...) qua ton kem v vay hoan toan khong the chap nhan c trong viec x ly sinh hoc. Phng phap kha thi c thc hien bang cach on nh va keo dai chuoi thc an nhng ao cuoi la nhng ao nuoi ca s dung tao lam nguon thc an. Trong tat ca cac trng hp, neu co s quan ly chnh xac (suc kh nhan tao trong cac ao, thu hoach ca thng xuyen...) ngi ta cho rang luc o co mot s km ham ang ke lng tao c thai ra moi trng tiep nhan. Mat khac, s hien dien cua cac sinh vat song (ca, tom, ong vat phu du...) cac ao cuoi la vat ch th sinh hoc co gia tr cho biet chat lng nc a c x ly Ngay nay, do phai tap trung vao viec ao thai chat dinh dng trong cac qua trnh x ly. Mot vai nha khoa hoc bien ho s loai bo nitrogen t cac chat thai, nhng ngi khac th e ngh s loai bo phosphorus va mot so khac lai khuyen loai bo ca hai. S la chon khach quan cho viec x ly tac ong en kieu cua qua trnh x ly c la chon.

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Chng 5

X LY NC THAI BANG VI SINH VAT

WX

5.1. X ly nc thai bang vi sinh dnh bam trong moi trng hieu kh (attached growth treatment process)
5.1.1. C s ly thuyet cua phng phap 5.1.1.1. Mo ta qua trnh hnh thanh sinh vat dnh bam tren vat lieu Phan ln vi khuan co kha nang sinh trng va phat trien tren be mat vat ran, khi co u o am va thc an la cac hp chat hu c, muoi khoang va oxy. Vi khuan dnh bam vao be mat vat ran bang chat gelatin do chung tiet ra va chung co the de dang di chuyen trong lp gelatin dnh bam nay. au tien vi khuan c tru tap trung mot khu vc, sau o mang vi sinh (biofilm) khong ngng phat trien, phu kn toan bo vat ran bang mot lp n bao. Chat dinh dng (hp chat hu c, muoi khoang) va oxy co trong nc thai can x ly tao ieu kien cho lp vi khuan nay phat trien. Sau mot thi gian, s phan lp hnh thanh: lp ngoai cung la lp hieu kh, c oxy khuyech tan tham nhap, lp trong la lp yem kh khong co oxy. Be day cua hai lp nay phu thuoc vao loai vat lieu nang (vat lieu loc), cng o gio va nc qua lp loc. Be day lp hoat tnh hieu kh thng khoang 300-400m. 5.1.1.2. Hieu qua va phan loai. Vi khuan trong mang vi sinh dnh bam hoat ong co hieu qua cao hn vi khuan trong moi trng hat can l lng. Qua trnh x ly bang vi sinh dnh bam hieu kh trong cac be loc sinh hoc ang c dung hien nay co the phan lam hai loai: Loai co vat lieu tiep xuc khong ngap nc Loai co vat lieu tiep xuc ngap trong nc.

51

5.1.2. Be loc sinh hoc co vat lieu tiep xuc khong ngap nc. 5.1.2.1. Cau tao Be loc sinh hoc nho giot Trong be loc, cac lp vat lieu co o rong va dien tch mat tiep xuc trong mot n v the tch ln nhat trong ieu kien co the. Nc thai c he thong phan phoi phun thanh giot eu khap tren be mat cua lp vat lieu. Nc sau khi cham lp vat lieu chia thanh cac hat nho chay thanh mang mong qua khe vat lieu i xuong di. Trong thi gian chay nh vay nc thai tiep xuc vi mang nhay gelatin bam quanh vat lieu loc. Sau mot thi gian lp nay day len ngan can oxy cua khong kh tham vao trong lp mang nhay. Do khong co oxy, tai lp trong cua lp mang nhay sat vi be mat cng cua vat lieu loc, vi khuan yem kh phat trien tao ra san pham phan huy yem kh cuoi cung la kh methan va CO2 lam troc lp mang ra khoi vat cng roi b nc cuon xuong pha di. Mat khac khi chat nen khong con khuyech tan ti lp ben trong na, cac vi sinh vat co trong lp a hieu kh se chet va t tieu i. Do vay xuat hien nhng khoang trong te bao cho cac vi sinh hieu kh va k kh khac. Khi chat nen that s can kiet, viec tieu huy cac te bao con lai lam cho mang sinh hoc b tach ri tng vung ra khoi be mat. Tren mat vat lieu loc hnh thanh lp mang mi. S tach mang sinh hoc c tao ra va thuc ay bi dong nc chay qua be mat. Hien tng nay c lap i lap lai tuan hoan va nc thai c lam sach BOD va cac chat dinh dng. e tranh hien tng tac nghen trong he thong phun, trong khe rong lp vat lieu, trc be sinh hoc nho giot, phai thiet ke song chan, li chan, be lang t 1. Nc sau be loc sinh hoc co nhieu bun l lng do cac mang sinh hoc troc ra nen phai c x ly tiep bang be lang t 2.

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Cau tao be loc sinh hoc nho giot (xem hnh 5.1)

53

a. Vat lieu loc. Vat lieu loc tot nhat la vat lieu co dien tch be mat tiep xuc trong mot n v the tch ln, o ben cao theo thi gian, khong b tat nghen va gia re. Tuy thuoc vao ieu kien ng dung co the chon : than a, a cuc, cuoi soi ln, a ong co kch thc trung bnh 60-100mm, neu kch thc vat lieu nho hn se lam giam o rong, gay tac nghen cuc bo. Neu kch thc ln hn th dien tch mat tiep xuc b giam nhieu, lam giam hieu qua x ly. Chieu cao lp vat lieu t 1.2-2.5m. Nhng thap nien gan ay, do ky thuat san xuat nha PVC phat trien, nhng tam nha uc ln song, gap nep va cac dang khac nhau cua qua cau nha a c dung lam lp vat lieu loc (xem hnh 5.1). Do vat lieu loc nhe, de lap at va thao d nen chieu cao be loc sinh hoc a c tang len t 6-9m goi la thap loc sinh hoc (xem hnh 5.2) tang chieu cao lam giam dien tch mat bang cua be loc sinh hoc.

b. He thong phan phoi nc. He thong phan phoi nc lam bang dan ong nha t quay a c a vao tieu chuan thiet ke be loc sinh hoc v co cau tao n gian, lam viec on nh, de quan ly. He thong gom ong ng dan nc vao c at tam be, nh ong lap khp quay hnh cau a nc ra 2 hoac 3 ong nhanh at nam ngang song song vi ban knh be (xem hnh 5-1) tren ong nhanh lap voi phun hoac lo phun nc xuong be mat be loc. Cac tia nc phun ra cung tren mot pha, vuong goc va ngc vi chieu quay cua ong nhanh. ong lng cua cac tia nc bien thanh lc lam cho dan ong nhanh quay quanh truc. Toc o quay thay oi theo lu lng nc, thng toc o quay khoang 1 vong trong 10 phut (tuy theo ieu kien thi tiet va muc ch x
54

ly ma lng nc a vao khac nhau nen toc o quay se khac nhau). Khi gian phun co nh phai bo tr eu lo phun nc tren toan dien tch be, phai co thung cha lap xiphong hoat ong t ong e cap nc cho dan phun theo tng me lien tiep. Khoang cach t be mat cua lp vat lieu en voi phun t 0.2-0.3m e lay khong kh va e cho cac tia nc phun ra v eu thanh giot nho tren toan dien tch be. c. San va thu nc. San va thu nc trong be loc sinh hoc lam hai nhiem vu: (1) Thu eu nc co cac manh v cua mang sinh hoc b troc ra chay t tren xuong e dan sang be lang t 2. (2) Phan phoi eu gio vao be loc e duy tr moi trng hieu kh trong cac khe rong. San lam bang tam be tong, sanh nung hay tam nha tang cng bang si thuy tinh co khoan lo hoac khe cho nc va kh i qua, ong thi c lp vat lieu loc. Khoang cach t san phan phoi en ay be thng 0.6-0.8m. ay be co o doc 1-2% ve mang trung tam. Xung quanh tng va o cao gia ay va san phan phoi at cac ca thong gio, tong dien tch cac ca so thong gio bang 20% dien tch san phan phoi. Gio i vao be loc theo dong oi lu cua khong kh c tao ra do chenh lech nhiet o gia nc thai va khong kh quanh be. Neu nhiet o nc thap hn nhiet o khong kh, th khong kh trong cac khe rong cua lp loc lanh hn khong kh ben ngoai, gio se i t tren mat be loc xuong ay be roi thoat ra qua cac ca thong gio. Ngc lai, neu nhiet o nc cao hn nhiet o khong kh, gio se i t di len. Khi nhiet o nc va khong kh bang nhau, khong co gio i vao be loc, v vay nhng be dien tch va chieu cao ln nen bo tr cac quat thong gio. 5.1.2.2. Phan loai be loc sinh hoc nho giot. Be loc sinh hoc nho giot c phan loai theo tai trong thuy lc hoac tai trong cac chat hu c, be co tai trong thap, be co tai trong cao. Theo bang 5.1.

55

Bang 5.1. Phan biet tai trong cac be loc sinh hoc nho giot. (cac ch tieu thiet ke)
Thong so Chieu cao lp vat lieu Loai vat lieu Tai trong BOD Toc o tai thuy He so tuan hoan Toc o tai thuy tren be mat cua be lang t 2 Hieu qua kh BOD sau be loc va be lang t 2 kg BOD5/m3.ngay m3/m2.ngay R=Qc/Q m3/m2.ngay % n v o m Tai trong thap 1-3 a cuc, than cuc, a ong, cuoi ln 0.08-0.4 1-4.1 Tuy chon 0-1 25 80-90 Tai trong cao 0.9-2.4 (a) 6-8 (nha tam) a cuc, tan cuc, soi ln, tam nha uc, cau nha. 0.4-1.6 4.1-40.7 0.5-2 16 65-85

Ghi chu: Toc o tai thuy neu trong bang la ty so cua lu lng nc x ly Q (m3/ngay) cong vi lu lng tuan hoan Qc (m3/ngay) (neu co) chia cho dien tch be mat cua be loc. Be loc sinh hoc nho giot tai trong thap quan ly n gian, hieu qua x ly on nh ngay ca khi nguon nc co chat lng dao ong ln, hieu qua x lycua be loc phu thuoc vao che o ti nc tc la phu thuoc vao vong quay cua thiet b ti, hay the tch thung o va tch nc roi lay i bang xi phong, thi gian gian oan khoang 5phut. 5.1.2.3. Tuan hoan nc. oi vi be loc cao tai muon tang hieu qua x ly phai tuan hoan nc lai e tang thi gian tiep xuc cua nc thai vi vi sinh dnh bam va giam tai trong hu c. Khi tuan hoan lai nc, tai trong thuy lc tang len, ay manh qua trnh tach mang vi sinh vat cu va hnh thanh mang mi tren be mat vat lieu, lam giam hien tng tac nghen trong cac lo rong cua lp vat lieu, tang lu lng trong he phan phoi e am bao toc o quay cua dan ong. Noi chung, lp loc co lng nc vao ln, can co s tuan hoan. Trong trng hp nay lng nc cap can bao am sao cho co s ong hoa cac vi khuan nhieu mc o khac nhau. S t vet cua vat lieu lp loc ma tren nhng vat lieu nay ch ton tai mot lp mang hoat tnh mong se tao ra s trao oi nhanh chong va thuc ay manh me ngay tai lp loc s phan huy cac chat te bao a c tao nen. Viec khoang hoa (on nh) c am nhan bi cac thiet b khac trong he thong nh cac be lang s cap va th cap. Co the ap dung mot trong 3 s o sau: (hnh 5.3) - S o a: be loc sinh hoc cao tai, chieu cao lp loc t 0.9-2m. Tuan hoan nc lien tuc, nc tuan hoan lay t sau be lang t 2 hoac co the sau be loc, a ve trc be lang t 1. Bun lang be lang t 2 cung a ve be lang t 1 e tang cng qua trnh keo tu trong be lang t 1. Bun xa ra t ay be lang t 1 a i x ly tiep.
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- S o b: nc tuan hoan lay t sau be loc a ve trc be loc, s o nay co the ap dung cho ca be loc tai trong thap va be loc tai trong cao. Khi can nitrate hoa (chuyen NH4+ thanh NO3-) triet e nen dung s o nay cho be tai trong thap. - S o c: c s dung khi thiet ke dan phan phoi la dan phun ma lien tuc at cao hn lp loc 0.5-0.6m, cac giot ma nho trai eu khap dien tch loc. Khi ap dung be loc sinh hoc e x ly nc thai can quan tam en moi trng e loai tr kha nang sinh san cua ruoi, muoi, giun san, oc, reu...

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Hnh 5.3.

58

5.1.2.4. Be lang t 2. Be lang t 2 at sau be loc sinh hoc lam nhiem vu lang gan ra khoi nc nhng bong can do cac vay mang sinh hoc troc ra. Khac vi be lang t 2 at sau be aerotank, be lang t 2 at sau be loc sinh hoc ch co tuan hoan nc ma khong can hoan bun trc tiep vao be loc sinh hoc, neu tuan hoan bun phai a vao au be lang t 1, cho nen nong o bun trong nc i vao be lang thng nho hn 500mg/l khong xay ra hien tng lang han che. Tnh toan be lang t 2 nay giong nh tnh toan be lang t 1, ch khac la trong tai be mat be lang tnh vi lu lng nc x ly cong them lu lng tuan hoan. Tai trong be mat thng lay t 16-25m3/m2.ngay. 5.1.3. Be loc sinh hoc co lp vat lieu ngap trong nc T au nhng nam 1990 en nay, cac nha khoa hoc trong lnh vc x ly nc thai a nghien cu va ap dung thanh cong vao san suat cong nghe loc sinh hoc co lp vat lieu loc ngap trong nc. My, Phap, Uc cong nghe nay a ap dung e x ly nc thai sinh hoat va cong nghiep thc pham cong suat 40.000m3/ngay, a vao van hanh t 1994. Viet Nam cung a ap dung thanh cong cong nghe nay t nam 1995 e x ly nc thai Benh vien a khoa Khanh Hoa, Ninh Thuan, Long Xuyen. 5.1.3.1. Cau tao (xem hnh 5.4)

1. 2. 3. 4. 5.

Mang phan phoi nc thai sau khi qua be lang 1 vao cac be. Dan ong co khoan lo phan phoi nc vao va thu nc xa ra. Ong xa nc ra loc. Mang thu nc loc Ong dan nc a loc sang be loc t 2 hoac vao be tiep xuc kh trung nc.
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Ong dan va dan ong phan phoi kh Hop ngan nc tr lai may gio. Ong dan gio t may nen ti Hat vat lieu loc noi polystyrene ng knh 2-5mm. Dien tch be mat 700-800m2/m3 vat lieu. 10. Li chan Inox co mat li 1.5x1.5mm co the thay bang san gan chup loc, co khe h 1.5mm at ngc. 11. Khoang trong e lp vat lieu loc gian n khi ra = chieu day lp loc. 12. Chieu cao lp nc e ra loc thng t 1.2-1.4m. 5.1.3.2. Quy trnh van hanh. Nc thai a qua be lang t 1 c bm len mang phan phoi 1, theo dan ong 2 phan phoi eu tren dien tch ay be, nc c tron eu vi khong kh cap t ngoai vao qua dan ong phan phoi 6. Hon hp kh nc i cung chieu t di len qua lp vat lieu loc. Trong lp vat lieu loc xay ra qua trnh kh BOD va chuyen hoa NH4+ thanh NO3-, lp vat lieu loc co kha nang gi lai can l lng. Nc trong c thu vao mang 4 theo ong 5 i ra ngoai. Neu muon kh BOD, NO3- va P, nen loc t hai bac tr len, bac loc cuoi, dan phan phoi kh at vao gia lp vat lieu loc cao o sao cho lp vat lieu loc nam di dan phan phoi kh co u the tch la vung thieu kh e kh NO3- va P. o chenh lech mc nc gia cac be loc lam viec noi tiep H=0.5m. Be loc sinh hoc dung vat lieu noi co kha nang gi c trong khe rong cac vay troc ra cua mang sinh hoc bam quanh hat, nen mac du cng o gio ln, nhng ham lng can l lng trong nc ra khoi be loc eu 20mg/l. Do o khong can thiet ke be lang t 2. Tom lai viec x ly nc thai bang vi sinh dnh bam co hieu qua cao, trong nhieu trng hp co the at en 95% viec loai bo BOD ra khoi nc thai, ac biet la he thong loc cao ap co cong suat lam viec rat ln, nhng dien tch mat bang chiem t cho nen no rat thuan tien trong cong viec lap at thiet ke he thong cung nh de dang van hanh va kiem soat thng xuyen. 5.2. X ly nc thai bang vi sinh yem kh trong moi trng can l lng va moi trng vi sinh dnh bam. 5.2.1. Cac qua trnh sinh hoc va phan loai cong trnh. 5.2.1.1. Cac qua trnh sinh hoc. X ly sinh hoc vi sinh yem kh la qua trnh phan huy cac chat hu c, vo c co trong nc thai khi khong co oxy. Quy trnh nay c ap dung t trc en nay e x ly on nh can va x ly nc thai cong nghiep co nong o BOD, COD cao. Mi nam tr lai ay do
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6. 7. 8. 9.

cong nghe sinh hoc phat trien, quy trnh x ly bang vi sinh yem kh c ap dung e x ly nc thai sinh hoat va nc thai cong nghiep co nong o BOD tng t. Khi nong o BOD trong nc thai ln hn 500mg/l ap dung quy trnh x ly 2 bac. Bac 1 x ly yem kh, bac 2 x ly hieu kh. Qua trnh chuyen hoa cac hp chat hu c trong nc thai bang vi sinh yem kh xay ra theo 3 bc: (1) Mot nhom vi sinh t nhien co trong nc thai thuy phan cac hp chat hu c phc tap va lipid thanh cac chat hu c n gian co trong lng nhe nh monosaccharide, amino acid e tao ra nguon thc an va nang lng cho vi sinh hoat ong. (2) Nhom vi khuan tao ra men acid bien oi cac hp chat hu c n gian thanh cac acid hu c thng la acid acetic. Nhom vi khuan nay thng c goi la nhom acidogen hay acid former. Trong so cac vi khuan c phan lap t be k kh bao gom Clostridium spp., Peptococcus anaerobus, Bifidobacterium spp., Staphylococcus va Escherichia coli. (3) Nhom vi khuan tao methane chuyen hoa hydro va acid acetic thanh kh methane va carbonic. Nhom vi khuan nay c goi la vi khuan sinh methane hay methanogen hay methane former. Bon vi khuan nay thng c tm thay trong da day cua ong vat va trong chat nen hu c lay t song va ho. Cac chi vi khuan chu yeu c xac nh bao gom nhom vi khuan hnh que (Methanobacterium, Methanobacillus) va nhom vi khuan hnh cau (Methanococcus, Methanosarcina). Vai tro quan trong cua chung la tieu thu hydro va acid acetic, chung tang trng rat cham va qua trnh x ly yem kh chat thai c thc hien khi kh methane va carbonic thoat ra khoi hon hp. ieu quan trong can nh la vi khuan sinh methane co the s dung mot so gii han cac chat nen e tao thanh kh methane. Ngay nay, ngi ta biet rang vi khuan sinh methane co the s dung cac chat nen sau: CO2+H2, formate, acetate, methanol, methylamin va CO. Phan ng chuyen hoa sinh nang lng ien hnh lien quan en cac phc hp sau: 4 H2 + CO2 4HCOOH CH3COOH 4CH3OH 4(CH3)3N + H2O CH4 + 2H2O CH4 + 3 CO2 + 2H2O CH4 + CO2 3CH4 + CO2 + 2H2O 9CH4 + 3CO2 + 6H2O + 4NH3

e duy tr s on nh cua cac qua trnh x ly yem kh, phai duy tr c tnh trang can bang ong cua qua trnh theo 3 bc a neu tren. Muon vay phai : - Khong co oxy - Khong co ham lng qua mc cua kim loai nang - Gia tr ph cua hon hp t 6.6 en 7.6.
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Phai duy tr o kiem u khoang 1000-1500mg/l lam dung dch em e ngan can ph giam xuong di 6.2 Nhiet o cua hon hp (nc thai) t 27-380c. Phai co u chat dinh dng theo ty le cod:n:p = 350:5:1 va nong o thap cua cac kim loai sat, nikel, ong ....

5.2.1.2. Phan loai qua trnh. Theo nguyen tac hoat ong va cau tao cong trnh cac be phan ng x ly bun va x ly nc thai tren thc te va trong cac tai lieu x ly nc thai qua trnh x ly c phan ra nh sau: Be phan ng yem kh tiep xuc: nc thai cha x ly c tron eu va lien tuc vi bun yem kh tuan hoan lai trong be kn. Be phan ng vi sinh yem kh dnh bam co tren cac tam phang at trong be, co dong nc t di len, hoac t tren xuong. Be phan ng vi sinh yem kh dnh bam tren cac hat cat l lng trong nc do van toc nc i t di len lam gian n lp cat. Be phan ng co dong nc x ly i t di len qua lp can l lng (UASB-Upflow Anaerobic Sludge Blanket) Be phan ng co dong nc i qua lp can l lng va loc tiep qua lp vat lieu loc co nh (upflow sludge blanket/fixed bed). Ho x ly yem kh.

Trong phan nay ch gii thieu be UASB va be loc yem kh la hai loai cong trnh a c xay dng va van hanh co hieu qua nc ta.

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5.2.2. Be x ly yem kh co lp can l lng (UASB). 5.2.2.1. Cau tao (Xem hnh 5.5)

Hnh 5.5. S o cau tao va nguyen tac hoat ong cua be phan ng yem kh UASB 1. Be ieu hoa lu lng va tram bm nc thai. 2. Bo phan o va ieu chnh pH. 3. nh lng chat dinh dng N, P neu can. 4. Ong dan va dan ong phan phoi eu nc thai trong be. 5. The tch vung phan ng yem kh. 6. Ca tuan hoan lai can lang. 7. Tam chan kh. 8. Ca dan hon hp bun nc sau khi a tach kh vao ngan lang. 9. The tch vung bun lang. 10. Mang thu nc. 11. Ong dan hon hp kh methane. 12. Ong dan nc sang be x ly hieu kh (t 2). 13. Thung cha kh. 14. Ong dan kh ot. 15. Ong xa bun d tha.
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5.2.2.2. Quy trnh hoat ong. Nc thai sau khi ieu chnh pH theo ong dan vao he thong phan phoi am bao phan phoi eu nc tren dien tch ay be. Nc thai i t di len vi van toc V=0.6-0.9m/h. Hon hp bun yem kh trong be hap phu chat hu c hoa tan trong nc thai, phan huy va chuyen hoa chung thanh kh (khoang 70-80% la methane, 20-30% la carbonic). Bot kh sinh ra bam vao hat bun can noi len tren lam xao tron va gay ra dong tuan hoan cuc bo trong lp can l lng, khi hat can noi len tren va phai tam chan (7) v ra, kh thoat len tren, can ri xuong di. Hon hp bun nc a tach het kh ra ca (8) vao ngan lang. Nc thai trong ngan lang tach bun lang xuong ay qua ca (6) tuan hoan lai vung phan ng yem kh. Nc trong dang len tren c thu vao mang (10) theo ong (12) dan sang be lam sach hieu kh (lam sach t 2). Kh biogas c gian ong (11) thu ve bnh cha (13) roi theo ong dan kh ot (14) i ra ngoai. 5.2.2.3. Phan bo bun trong be. Bun trong be la sinh khoi ong vai tro quyet nh trong viec phan huy va chuyen hoa chat hu c, bun c chia thanh 2 vung ro ret trong be phan ng. chieu cao khoang be tnh t ay len, lp bun hnh thanh do cac hat can keo tu nong o t 5-7%, tren lp nay la lp bun l lng nong o t 1000-3000mg/l gom cac bong can chuyen ong gia lp bun ay va bun tuan hoan t ngan lang ri xuong. Tren mat tiep giap vi pha kh, nong o bun trong nc be nhat. Nong o cao cua bun hoat tnh trong be lam viec vi tai lng chat hu c cao. e hnh thanh khoi bun hoat tnh u nong o, lam viec co hieu qua oi hoi thi gian van hanh khi ong t 3-4 thang. Neu cay vi khuan tao acid va vi khuan tao men methane trc vi nong o thch hp va van hanh vi che o thuy lc cong suat thiet ke, thi gian khi ong co the rut xuong t 2-3 tuan. Can d tha nh ky xa ra ngoai. Lng can d tha ch bang 0.15-0.2 ham lng COD, tc bang can sinh ra so vi khi x ly hieu kh. Can xa ra on nh co the a trc tiep en thiet b lam kho. 5.2.2.4. Qua trnh lang Hon hp vi sinh yem kh phan huy chat hu c trong be tnh trang tron lan gia 3 pha: kh, nc, bun. e a nc ra khoi be, trc het phai tach kh ra khoi hon hp bang cac tam tach kh at nghieng so vi phng ngang 550 (kch thc va cach bo tr xem hnh 4-1). Sau khi tach kh, hon hp bun nc chay theo ca (8) vi van toc 9-10m/h vao ngan lang (9). The tch ngan lang tnh theo thi gian lu nc 1 gi. Can ri xuong ay hnh non cua ngan lang chay qua khe (6) tr lai ngan phan huy yem kh (5). Tong chieu cao ngan lang 2m, chieu cao phan lang 1 m.

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5.2.2.5. Ch tieu thiet ke. Khi thiet ke be phan ng yem kh UASB va be loc yem kh co the tham khao so lieu cho trong bang 5.1, e chon thong so thiet ke thch hp. Bang 5.2. So lieu ky thuat t ket qua van hanh be UASB va be loc yem kh
Nguon nc thai Nc thai sinh hoat Nha may ru, men ru Che bien bot khoai tay Che bien sa Nha may hoa chat hu c tong hp Che bien rau va hoa qua Giay cac loai Che bien hai san Ham lng COD au vao (mg/l) 500-800 20.000 4.500-7.000 3.000-3.400 18.000 8.300 7.700 2.300-3.000 Thi gian lu nc trong be 4-10 5-10 5-10 5-10 5-10 5-10 5-10 5-10 Tai lng COD (kgCOD/m3.ngay) 4-10 14-15 8-9 12 7-9 18 12 8-10 Hieu qua kh COD (%) 70-25 60 75-80 80 90 55 80 75-80

5.2.3. Be loc yem kh. Be loc yem kh la cot loc dung vat lieu loc noi thng la polyspirene, co ng knh hat t 3-5 mm, chieu day 2m. Cau tao be loc (xem hnh 5.6).

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Nc thai i vao be c phan phoi eu theo dien tch ay be. Dong nc i t di len tiep xuc vi khoi bun l lng lp loc roi tiep xuc vi khoi hat loc co vi khuan yem kh dnh bam. Chat hu c hoa tan trong nc thai c hap thu va phan huy, bun can c gi lai trong khe rong cua lp loc, sau thi gian 2-3 thang xa bun d mot lan. Nc i qua lp loc c tach kh roi chay vao mang thu theo ong dan a sang x ly hieu kh. 5.2.4. anh gia qua trnh. Co nhieu iem thuan li va khong thuan li trong viec x ly k kh cac hp chat hu c so vi cac qua trnh x ly hieu kh, van e chnh la toc o phat trien cua cac vi khuan sinh methane qua thap. Toc o phat trien thap oi hoi phai co thi gian lu nc dai trong be phan huy e qua trnh on nh chat thai xay ra. Tuy nhien, s phat trien cham c ghi nhan
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cho thay ch co mot phan chat hu c c s dung cho viec tong hp nen te bao mi. Vi vi khuan sinh methane th hau het cac chat thai hu c c chuyen hoa thanh kh methane la loai san pham hu dung dung lam nhien lieu. Bi v toc o phat trien cua te bao thap va s chuyen hoa cac hp chat hu c thanh kh methane va CO2, ket qua la chat ran hu c c on nh hp ly. Sau khi phi kho va tach nc, bun co the c s dung lam phan bon cho at. Nhiet o cao can thiet e at en x ly hoan toan thng c xem la bat tien nhat cua qua trnh x ly k kh; tuy nhien, nhiet o cao ch can thiet khi thi gian lu te bao u lau khong the at c tai mot nhiet o thap. Trong he thong x ly k kh th thi gian lu te bao vi sinh trong be phan ng bang thi gian lu nc cua dch tron trong be phan ng. Bi v nhiet o van hanh tang cao, thi gian lu te bao thap nhat giam xuong ang ke. Cho nen, cung cap nhiet cho be phan ng lam giam khong ch thi gian lu te bao can thiet e at en x ly hoan toan ma con thi gian lu nc, v the the tch be phan ng nho hn co the c s dung

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Chng 6 CAC QUA TRNH KH NITROGEN BANG VI SINH VAT

WX

6.1. S chuyen hoa ammonia bang qua trnh nitrate hoa sinh hoc (conversion of ammonia by biological nitrification)
Qua trnh ma trong o nitrogen cua nc thai cha x ly lan lt c chuyen hoa e tao thanh nitrate di tac dung cua vi sinh vat c goi la qua trnh nitrate hoa sinh hoc. Nc thai a c x ly khi c thai ra oi hoi thoa man cac yeu cau cua moi trng nc tiep nhan ni can co s giam thieu nhu cau oxy nitrogen (oxy can thiet e oxy hoa cac hp chat co cha nitrogen) hoac ni can thiet phai co s giam thieu kha nang gay oc cua ammonia. Trong phan nay, qua trnh nitrate hoa sinh hoc c trnh bay. Cac qua trnh x ly c phan loai va mo ta, ng dung, xem xet va so sanh ky lng. 6.1.1. Mo ta qua trnh Nitrate hoa la mot qua trnh t dng (v du, nang lng cho s phat trien cua vi khuan c lay t s oxy hoa cac hp chat co cha nitrogen, au tien la ammonia). Trai vi qua trnh d dng, cac vi khuan nitrate hoa s dung CO2 (carbon vo c) hn la carbon hu c e tong hp te bao. Qua trnh nitrate hoa ammonium la mot qua trnh gom hai bc lien quan en hai chi vi sinh vat, Nitrosomonas va Nitrobacter. Bc th nhat, ammonium c chuyen thanh nitrite; tiep o nitrite c chuyen thanh nitrate. Qua trnh chuyen hoa c mo ta nh sau: Bc 1: NH4+ + 3/2 O2 Bc 2: NO2- + O2 NO3- (2) NO2- + 2H+ + H2O (1)

Phng trnh 1 va 2 va nhng phan ng tao ra nang lng. Nitrosomonas va Nitrobacter s dung nang lng c sinh ra t nhng phan ng o cho s phat trien va duy tr te bao. Toan bo phan ng c trnh bay qua phng trnh sau: NH4+ + 2O2 NO3- + 2H+ + H2O

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Vi nang lng thu c, mot vai ion ammonium c tch luy trong te bao. Phan ng tong hp sinh khoi co the c the hien nh sau: 4CO2 + HCO3- + NH4+ + H2O C5H7O2N + 5O2

Cong thc hoa hoc C5H7O2N c s dung cho te bao vi khuan c tong hp Toan bo phan ng oxy hoa va tong hp co the c the hien nh sau: NH4++1.83O2+1.98HCO30.021C5H7O2N+0.98NO3-+1.041H2O + 1.88H2CO3

6.1.2. Phan loai cac qua trnh nitrate hoa Cac qua trnh nitrate hoa co the c phan loai theo mc o khac biet cua s oxy hoa carbon lien quan en nitrate hoa. S oxy hoa carbon va nitrate hoa co the xay ra trong mot phan ng n, lien quan en giai oan n. Trong qua trnh nitrate hoa tach biet, s oxy hoa carbon va nitrate hoa xay ra cac phan ng khac nhau. Phan ng tang cng chat l lng hoac tang cng chat ket dnh co the c s dung cho ca he thong phan ng n va phan ng tach biet. Mo hnh phan ng n va phan ng tach biet c mo ta hnh 6.1.

Hnh 6.1. Qua trnh oxy hoa carbon va nitrate hoa tang cng chat l lng : (a) s o loc sinh hoc tach biet, (b) s o phoi hp.
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Sinh vat nitrate hoa hien dien trong hau het cac qua trnh x ly sinh hoc, nhng so lng cua chung thng gii han. Kha nang cua cac qua trnh bun hoat tnh e nitrate hoa lien quan en t so BOD5/TKN (total Kjeldahl nitrogen - tong nitrogen tnh theo phng phap phan tch Kjeldahl). V ty so BOD5/TKN nam trong khoang t 1-3 tng xng vi cac gia tr gap phai trong cac he thong nitrate hoa tach biet. Ty so sinh vat nitrate hoa c anh gia bien ong t 0.21 tai BOD5/TKN = 1 en 0.083 tai BOD5/TKN=3 (xem bang 6.1). Trong hau het qua qua trnh bun hoat tnh truyen thong, ty so sinh vat nitrate hoa thng thap hn 0.083. ieu o cho thay rang khi BOD5/TKN >5, qua trnh co the c xem nh la mot qua trnh ket hp gia oxy hoa carbon va nitrate hoa, va khi t so nay <3, co the c xep vao qua trnh nitrate hoa tach biet. Bang 6.1. Moi tng quan gia phan so sinh vat nitrate hoa va ty so BOD5/TKN
Ty so BOD5/TKN 0.5 1 2 3 4 Phan so sinh vat nitrate hoa 0.35 0.21 0.12 0.083 0.064 Ty so BOD5/TKN 5 6 7 8 9 Phan so sinh vat nitrate hoa 0.054 0.043 0.037 0.033 0.029

6.1.3. S oxy hoa carbon va nitrate hoa giai oan n (s o phoi hp). Qua trnh nitrate hoa co the c thc hien trong he bun hoat tnh tang cng chat l lng. Cac qua trnh thng c s dung la chay truyen thong (conventional plug-flow), tron hoan chnh (complete-mix), suc kh tang cng (extended aeration) va nhieu dang sa oi cua mng oxy hoa (oxidation ditch). e at c nitrate hoa, oi hoi co s duy tr cac ieu kien co the cho s phat trien cua cac sinh vat nitrate hoa. V du, trong vung kh hau am ap, s nitrate hoa tang cng co the c gia tang nh s gia tang thi gian ton tai cua cac te bao trong he thong x ly va s cung cap kh. Ky thuat nay thng c s dung e at c nitrate hoa theo mua. Hai qua trnh tang cng ket dnh, co the c s dung cho qua trnh nitrate hoa va oxy hoa carbon ket hp, la loc nho giot va tiep xuc sinh hoc. Vi cac qua trnh tang cng chat l lng co the c tang cng nh viec ieu chnh cac thong so ieu khien. Nitrate hoa thng co the c thc hien bi viec lam giam toc o tai hoat c ap dung. Qua trnh tang cng chat l lng. ong thai cua qua trnh nitrate hoa c nh ngha ro rang nhat cho he thong tang cng chat l lng. ieu o cho thay cac bieu thc cua ong lc hoc c dung trong qua trnh tang cng chat l lng hieu kh co the c ap dung cho qua trnh nitrate hoa. T kinh nghiem va nhng nghien cu trong phong th nghiem, cho thay rang nhng nhan to sau co anh hng len qua trnh nitrate hoa : nong o ammonia va nitrite, t so
70

BOD5/TKN, nong o oxy hoa tan, nhiet o va pH. Anh hng cua nhng bien so len qua trnh nitrate hoa va viec tiep can e giai thch chung c trnh bay bang 5-2. Cac nhan to ong anh hng cua ong nang len qua trnh nitrate hoa tang cng chat l lng la kch c cua moi trng va o sau cua be. Viec ng dung nhng tiep can ve ong nang cho nhng phan tch qua trnh nitrate hoa trong mot phan ng phc hp hoan toan lien quan en cac bc sau: (1) La chon mot nhan to an toan e duy tr hoat o nh, suot ngay va thi gian tai hoat ngan. (2) La chon nong o oxy hoa tan thap nhat. DO thap nhat mc 2.0mg/l la thch hp e tranh viec giam hieu qua cua DO len toc o nitrate hoa. (3) Xac nh pH trong qua trnh van hanh. pH bien ong t 7.0-9.0 la thch hp. Moi mot mg/l NH4-N b oxy hoa co the gay nen s phan huy 7.14mg/l kiem (bieu hien qua CaCO3). (4) anh gia toc o phat trien cc ai cua cac vi khuan nitrate hoa khong anh hng en s thay oi cua nhiet o, DO va pH. (5) Xac nh thi gian ton tai t nhat cua te bao da vao toc o phat trien c xac nh trong bc (4). (6) Xac nh thi gian ton tai cua te bao theo d kien bang cach s dung nhan to an toan c xac nh bc (1). (7) Xac nh nong o nitrogen cua nc thai. (8) Xac nh thi gian gi nc e at en nong o nitrogen can thiet cua nc thai. (9) Xac nh toc o s dung chat hu tai ni ma qua trnh nitrate hoa - oxy hoa giai oan n c s dung.

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Bang 6.2. Anh hng cua nhng thay oi moi trng va qua trnh hoat ong len qua trnh nitrate hoa tang cng l lng.
Thong so moi trng Nong o NH4+ va NO2Mo ta anh hng Nong o NH4+ va NO2- anh hng en toc o tang trng rieng cc ai cua nitrosomonas va Nitrobacter. Toc o tang trng cua nitrobacter ln hn rat nhieu so vi nitrosomonas. Va toc o tang trng chung cua chung trong qua trnh la:

= m

N Ks + N

Ty so BOD/TKN

Nong o oxy hoa tan

: toc o tang trng rieng (1/s) m : toc o tang trng rieng cc ai (1/s) S : nong o chat nen trong nc thai thi iem tang trng b han che Ks : hang so ban toc o Lay m = 0.45 ngay -1 150C So phan tram cua cac hp chat hu c b nitrate hoa trong qua trnh kh BOD chu anh hng cua ty so BOD/TKN. Bieu th bang: FN=[0.16(NH3 b kh)]/[0.6(BOD5 b kh + 0.16 (NH3 b kh)] Mc o DO anh hng en toc o phat trien ac biet m cua cac sinh vat nitrate hoa. Anh hng o co the c mo hnh hoa vi moi tng quan sau:

'm = m

Da vao cac thong tin gii han co the lay Ko2=1.3mg/l Nhiet o (0C) Nhiet o anh hng rat ln en qua trnh nitrate hoa

DO K O2 + DO

= m e 0.098(T 15)
pH Toc o cc ai cua nitrate hoa xay ra trong khoang pH t 7.2 en 9.0. oi vi he thong nitrate hoa oxy hoa carbon anh hng cua pH co the c tnh theo cong thc

= m [1 0.833(7.2 pH )]

Van e chnh lien quan en phan tch nay la viec xac nh thi gian ton tai toi thieu cua te bao trong moi trng khac nghiet nhat va viec s dung nhan to an toan. S tiep can nay la can thiet giong nh cai c s dung trong viec thiet ke qua trnh bun hoat tnh tang cng chat l lng trong phan ng tron hoan toan. Cac qua trnh tang cng dnh bam (attached-growth processes). Qua trnh tang cng chat dnh bam la loc nho giot va be tiep xuc sinh hoc. Ngay nay, hau het viec tiep can e mo ta s van hanh cua cac qua trnh tang cng dnh bam la s dung cac nhan to tai hoat. oi vi loc nho giot s dung moi trng a, hoat tai chat hu c se anh hng en hieu suat cua qua trnh nitrate hoa v mang vi khuan chiem u the bi vi khuan d dng luc hoat tai chat hu c cao. e at c hieu qua nitrate hoa cao, s hoat tai chat hu c c duy tr trong khoang bien ong trong khoang 0.8-2m3/m2.phut. Bi v he loc s dung hop cha plastic co be mat tiep xuc ln hn (va so lng vi khuan hoat ong nhieu hn), hoat tai
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cao chat hu c co the c thc hien trong khi va at c nitrate hoa toi u. He loc tot khac co moi trng plastic la s thong gio tot hn, ieu nay cho phep viec luan chuyen oxygen cao hn. So sanh gia hai moi trng a va plactic cho thay rang moi trng plactic vi 80% dien tch tiep xuc se co the nitrate hoa khoang hn 60% ammonia tren mot n v the tch trong mot he thong ket hp nitrate hoa oxy hoa carbon. Trong he tiep xuc sinh hoc, lng ammonia co the b oxy hoa tuy thuoc vao be mat tiep xuc. Nitrate hoa ang ke se khong xay ra trong he tiep xuc sinh hoc cho en khi nong o BOD5 giam xuong 15mg/l hoac thap hn. 6.1.4. Nitrate hoa giai oan kep (s o tach biet). Ca hai qua trnh tang cng chat ran l lng va ket dnh eu c s dung trong nitrate hoa giai oan kep, moi qua trnh (oxy hoa carbon va nitrate hoa) co the c thc hien mot cach oc lap e at hieu qua cao nhat. Anh hng cua oc tnh co the giam xuong bi v chat hu c b phan huy sinh hoc, ma chnh no co the gay oc en cac vi khuan nitrate hoa, c loai bo trong giai oan oxy hoa carbon. Mc o loai bo carbon hu c trong giai oan oxy hoa carbon se anh hng en s la chon va thc hien qua trnh nitrate hoa. Carbon hu c thap trong nc thai co the la thuan li oi vi cac phan ng tang cng ket lang bi v mc o thap nh the khoi can phai lam sach sau qua trnh nitrate hoa. Trong cac phan ng nitrate hoa tang cng chat l lng, carbon hu c thap trong nc thai co the gay nen s mat can bang gia chat ran mat i t nen ay cua ao va chat ran c tong hp trong phan ng. S mat can bang nay thng can thiet e gia tang BOD trong phan ng nitrate hoa e duy tr gia tr chat ran sinh hoc trong he thong nitrate hoa. Tang cng chat l lng. Qua trnh nitrate hoa tang cng chat l lng giai oan kep giong nh trong thiet ke qua trnh bun hoat tnh. Khi nong o ammonia rat thap c mong i, cac phan ng tron hoan toan va chay doc (plug-flow) la hp ly. oi vi nitrate hoa giai oan ket hp, viec xac nh toc o nitrate hoa la mot tien trnh thng c s dung. Toc o o at thc nghiem c xem la co gia tr hn la toc o ly thuyet bi v s kho khan e at en s chia nho nitrate hoa cua dung dch phc hp. Toc o nitrate hoa tang khi nhiet o tang. Gia tr BOD5/TKN quan trong trong qua trnh nitrate hoa, vi toc o nitrate hoa tang khi ty so nay tang. Toc o nitrate hoa cung b anh hng bi gia tr pH cua hon hp dung dch va lam the nao e nh hng pH toi u cho qua trnh nitrate hoa.

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Tang cng dnh bam. Co hai loai khac nhau cua qua trnh tang cng dnh bam c s dung thng xuyen cho qua trnh nitrate hoa ket hp: loc nho giot va be tiep xuc sinh hoc. Loc nho giot co the c s dung cho nitrate hoa kep tiep theo sau qua trnh tang cng chat l lng s dung cho oxy hoa carbon. S ket hp pho bien hn cua qua trnh nay la viec s dung he thong loc nho giot hai giai oan: giai oan mot c s dung cho oxy hoa carbon va giai oan hai la nitrate hoa. Thap loc s dung chat mang plastic ac biet thch hp cho nitrate hoa bi v dien tch be mat ln. Thap loc nen c thiet ke e viec thong kh c tien hanh de dang, neu c oi hoi. Khi s dung cho qua trnh nitrate hoa sinh hoc, be tiep xuc sinh hoc c thiet ke thng da tren nong o ammonia hn la the tch nc thai hay nong o TKN. Cac ng dung ket hp oxy hoa carbon va nitrate hoa nh la mot tiep can co the dan en viec oi hoi dien tch be mat trong phan ng nitrate hoa.

6.2. Loai bo nitrogen bang nitrate hoa/phan nitrate hoa sinh hoc
Trong tat ca cac phng phap c s dung cho viec loai bo nitrogen, th nitrate hoa/ phan nitrate hoa sinh hoc la tot nhat v nhng ly do sau: (1) hieu suat loai bo cao, (2) tnh on nh va o chnh xac cua qua trnh cao, (3) de ieu khien, (4) dien tch at yeu cau thap, va gia thanh hp ly. Viec loai bo nitrogen bang qua trnh nitrate hoa/phan nitrate hoa sinh hoc la mot qua trnh gom hai bc. Bc th nhat, ammonia c chuyen hoa hieu kh thanh nitrate (nitrate hoa). Bc th 2, nitrate c chuyen hoa thanh kh nitrogen (phan nitrate hoa). 6.2.1. Mo ta qua trnh Viec loai bo nitrate bi s chuyen hoa thanh kh nitrogen co the c thc hien bang cac qua trnh sinh hoc trong ieu kien thieu kh. Viec lam giam NO3--N c thc hien thong qua ong hoa va d hoa. Trong qua trnh ong hoa lam giam nitrate, NO3--N c chuyen hoa thanh ammonia cho viec s dung cua te bao trong tong hp sinh hoc va xay ra khi NO3--N ch la mot dang nitrogen duy nhat co san. Trong qua trnh d hoa lam giam nitrate, kh nitrogen c hnh thanh t NO3--N: ay la ket qua cua qua trnh phan nitrate hoa trong nc thai. Trong hau het cac he thong nitrate hoa/phan nitrate hoa, nc thai c nitrate hoa phai cha u ham lng carbon (carbon hu c) e cung cap nguon nang lng cho viec chuyen hoa nitrate thanh kh nitrogen bi vi khuan. Nhu cau carbon co the c cung cap t cac nguon ben trong nh nc thai va nguyen lieu te bao hoac t ben ngoai (v du nh methanol).

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Bang 6.3. S so sanh viec la chon qua trnh nitrate hoa

Loai he thong Nitrate hoa oxy hoa carbon ket hp Tang cng chat l lng Thuan li Khong thuan li

Viec x ly ket hp carbon va ammonia trong mot giai oan n; ammonia nc thai thap; de kiem soat s on nh dung dch tron nh vao ty so BOD5/TKN cao. X ly ket hp carbon va ammonia trong mot giai oan n; viec on nh khong can lien ket vi viec lam sach th cap v cac sinh vat a c bam vao gia bam.

Tang dnh

cng

ket

Khong co s bao ve oi vi cac oc chat; s on nh trung bnh; viec on nh phai c lien ket vi ng dung cua viec lam sach cap hai e thu hoi sinh khoi; be phan ng yeu cau phai ln trong thi tiet lanh. Khong co s bao ve chong lai cac oc chat; ch on nh trung bnh; ammonia nc thai thng 1-3mg/l; khong ng dung c trong mua lanh trong tat ca moi trng hp.

Nitrate hoa giai oan kep Tang cng chat l lng

Co s bao ve tot chong lai hau het cac oc chat; hoat ong on nh; ammonia thap la co the c

Tang dnh

cng

ket

Co s bao ve tot chong lai vi hau het cac oc chat; hoat ong on nh; viec on nh khong can lien ket he lam sach th cap v cac sinh vat c ket dnh vao chat mang.

Nc thai oi hoi phai co s kiem soat can than khi t so BOD5/TKN thap; s on nh cua hoat ong phai lien ket vi hoat ong cua viec lam sach th cap e thu hoi sinh khoi; oi hoi nhieu hang muc x ly hn so vi nitrate hoa oxy hoa carbon ket hp. Ammonia nc thai thng thap 13mg/l; oi hoi nhieu hang muc x ly hn so vi nitrate hoa oxy hoa carbon ket hp.

Toc o nitrate hoa co the c mo ta bi cong thc sau:

= U

1 . 09

( T 20 )

(1 DO )

U1: toc o nitrate hoa tong so U0: toc o nitrate hoa xac nh. kg NO3--N/kg chat ran.d T: nhiet o nc thai. 0C. DO: oxy hoa tan trong nc thai. mg/l. Gia tr DO trong cong thc tren cho thay toc o nitrogen giam en tiem can khong khi nong o oxy hoa tan tien en 1mg/l. Toc o nitrate hoa xac nh tuy thuoc vao nguon carbon c cho bang 6.4.

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Bang 6.4. Cac toc o nitrate hoa ien hnh tuy theo nguon carbon.
Nguon carbon Methanol Methanol Nc thai Trao oi chat noi sinh Toc o nitrate hoa, U0, kg NO3--N/kg chat ran.ngay 0.21 - 0.32 0.12 - 0.90 0.03 - 0.11 0.017 - 0.048 Nhiet o, 0C 25 20 15 - 27 12 - 20

6.2.2. Phan loai cac qua trnh nitrate hoa/phan nitrate hoa. Cac qua trnh phan nitrate hoa c nh ngha nh la qua trnh tang cng l lng thieu kh va tang cng ket dnh thieu kh. Trong thao luan sau nay, viec phan loai da vao qua trnh nitrate hoa c thc hien nh the nao (1) trong cac he thong nitrate hoa/phan nitrate hoa oxy hoa carbon ket hp s dung nguon carbon noi tai va ben ngoai hoac (2) trong cac phan ng kep s dung methanol hoac mot nguon carbon hu c hp ly ben ngoai nao o. Nh a ghi chu trc ay, cac he thong ket hp thng lien quan en cac he thong bun n va cac he thong nitrate hoa/phan nitrate hoa s dung cac phan ng kep thng lien quan en cac he thong bun kep. Nen nh rang bun sinh ra trong he thong bun kep thng co tnh chat khac nhau. 6.2.2.1. He thong nitrate hoa/phan nitrate hoa ket hp (he thong bun n). Bi v gia thanh cua cac nguon carbon ben ngoai cao, cac qua trnh phat trien trong o cac bc nitrate hoa/phan nitrate hoa oxy hoa carbon thng c ket hp trong mot qua trnh n, nen s dung nguon carbon sinh ra t nhien trong nc thai. S thuan li hien nhien cua cac qua trnh nay bao gom (1) s giam the tch kh cung cap cho nitrate hoa va kh BOD5, (2) bo i viec cung cap nguon carbon hu c phu troi (v du nh methanol) cho qua trnh nitrate hoa, va (3) loai tr c he thong lam sach trung gian va hoi lu bun trong he thong nitrate hoa/phan nitrate hoa mot giai oan. Hau het cac he thong eu co the loai bo c t 60 en 80% tong nitrogen; toc o loai bo t 85 en 95%. Trong cac he thong ket hp nay, carbon trong nc thai va nguon carbon con lai sau khi phan huy te bao cua sinh vat c s dung e thc hien qua trnh nitrate hoa. oi vi qua trnh nitrate hoa, mot chuoi luan phien cac giai oan thieu kh va hieu kh (khong co on nh trung gian) c s dung. Cac vung thieu kh co the c tien hanh trong kenh oxy hoa bi viec kiem soat cac mc o oxy gen cung cap. Cac qua trnh phan ng theo me lien tuc ac biet cung c chap nhan e cung cap cac thi ky hieu kh va thieu kh suot chu ky hoat ong. Toc o nitrate hoa cc ai cho nc thai trong he thong giai oan n bien ong t 0.075 en 0.115 kg NO3--N/kg chat ran. ngay 200C trong phan ng thieu kh di ieu kien khong han che carbon. Toc o nitrate hoa trong he thong bun n khoang chng bang mot

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na toc o nitrate hoa cua he thong bun kep. S dung nguon carbon noi sinh, toc o nitrate hoa bien ong t 0.017 en 0.048 kg NO3--N/kg chat ran.ngay. 6.2.2.2. Qua trnh bon giai oan. Qua trnh bon giai oan (hnh 6.2 b) s dung nguon carbon trong nc thai va nguon carbon t s phan huy noi sinh e tien hanh nitrate hoa. Cac vung phan ng kep c s dung cho nitrate hoa oxy hoa carbon va phan nitrate hoa thieu kh. Nc thai au tien i vao vung phan nitrate hoa thieu kh va c tron vi dung dch c quay vong t khoang nitrate hoa oxy hoa carbon pha sau. Carbon hien dien trong nc thai c s dung e phan nitrate hoa, nitrate c quay vong lai. Bi v chat hu c cao, nen qua trnh phan nitrate hoa dien ra nhanh. Ammonia trong nc thai i qua khong thay oi qua ao thieu kh th nhat e c nitrate hoa trc khi qua ao hieu kh th nhat. Dch tron a c nitrate hoa t ao hieu kh th nhat bang qua vung thieu kh th hai, ni ay qua trnh phan nitrate hoa phu troi xay ra s dung nguon carbon noi sinh. Vung hieu kh th hai thng nho hn va c s dung e lay het kh nitrogen trc khi lam sach. Ammonia thoat ra t bun trong vung thieu kh th hai cung c nitrate hoa trong vung hieu kh cuoi cung. Nhng s bien oi cua he thong co the dung cho qua trnh loai bo nitrogen va phosphorus ket hp.

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Mng oxy hoa. Mng oxy hoa c s dung e tien hanh nitrate hoa va phan nitrate hoa (hnh 6.2 a). Trong kenh oxy hoa, dch tron chay vong theo kenh, c dan i va suc kh bi cac thiet b suc kh. oi vi nhng ng dung nitrate hoa/phan nitrate hoa, mot vung hieu kh c at ngay cuoi dong cua may suc, va vung k kh c thiet ke au dong cua may suc. Bang cach dan dong nc thai tai ngay au dong cua vung thieu kh, mot t nguon carbon cua nc thai c s dung cho phan nitrate hoa. Nc thai t be phan ng c lay t au cuoi cua vung hieu kh s dung cho viec lam sach. Bi v he thong ch co vung thieu kh, viec loai bo nitrogen thap hn so vi qua trnh bon giai oan. Thiet ke qua trnh cho he thong nitrate hoa/phan nitrate hoa ket hp. Cac thu tuc thiet ke cho he thong nitrate hoa/phan nitrate hoa tang cng l lng ket hp bien ong da tren loai qua trnh c s dung. Phng phap khuech ai e xac nh thi gian lu thieu kh, hieu kh va ty so quay vong c trnh bay di ay.
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Tha nhan qua trnh nitrate hoa hoan toan NO3--N quay vong en giai oan thieu kh va khong chu y en s ong hoa nitrogen, ty so quay vong oi hoi (dch tron + bun hoan lu) c cho bi cong thc
R =

(NH

+ 4

(NO

0 3

(NH
N

R: ty so hoan lu toan bo (dung dch tron + bun hoan lu) (NH4+-N)o; (NH4--N)e: ammonium au vao va au ra. mg/l. (NO3--N)e: nitrate au ra. mg/l. He thong phan nitrate hoa giai oan kep. Vao au nhng nam 1970, hau het nhng tiep can c chap nhan oi vi qua trnh phan nitrate hoa sinh hoc la s cong them vao cua mot he thong sinh hoc kep s dung methanol nh la nguon carbon e loai bo nitrate. Bi v qua trnh nitrate hoa/phan nitrate hoa oxy hoa carbon xay ra trong cac phan ng tach biet, nen bun sinh ra rieng biet trong moi phan ng, v the ten he thong bun tach biet c s dung thng xuyen. Bi v nguon carbon di dang methanol c them vao nhieu hn nhu cau oi hoi se c o di dang BOD, nen s chu y can than phai c danh het cho cong viec thiet ke va hoat ong cua nhan to nay cua he thong. Khi s dung methanol nh la mot nguon carbon th qua trnh phan nitrate hoa giai oan kep co the c mo ta nh sau. Phan ng nang lng co the c trnh bay qua phng trnh: Phan ng nang lng, bc 1: 6NO3- + 2CH3OH 6NO2- + 2CO2 + 4H2O

Phan ng nang lng, bc 2: 6NO3- + 2CH3OH 3N2 + 3CO2 + 3H2O + 6OH-

Phan ng nang lng toan phan: 6NO3- + 5CH3OH 5CO2 + 3N2 + 7H2O + 6OH-

Mot phan ng tong hp c a ra bi McCarry nh sau: 3NO3- + 14CH3OH + CO2 + 3H+ 3C5H7O2N + H2O

Trong thc nghiem, 25 en 30% lng methanol yeu cau cap nang lng cho viec tong hp. Nhng nghien cu c ban trong phong th nghiem, cong thc sau c a ra e mo ta phan ng loai nitrate toan phan.
79

Viec loai bo nitrate hoan toan NO3- + 1.08CH3OH + H+ 0.065C5H7O5N + 0.47N2 + 0.76CO2 + 2.44H2O

Neu tat ca nitrogen di dang nitrate, th nhu cau methanol tong so co the c xac nh bi cong thc tren. Tuy nhien, nc thai c x ly c nitrate hoa co the cha mot vai nitrite va DO. Ni ma nitrate, nitrite va DO hien dien, th nhu cau methanol co the c tnh toan theo kinh nghiem bi cong thc. Cm = 2.47N0 + 1.53N1 + 0.87D0 Trong o: Cm : nong o methanol yeu cau (mg/l). N0 : nong o nitrate ban au (mg/l) N1 : nong o nitrite ban au (mg/l) D0 : nong o oxy hoa tan ban au (mg/l) Tang cng chat l lng.Viec thet ke he thong phan nitrate hoa tang cng chat l lng giong vi viec thiet ke he thong bun hoat tnh c s dung e loai carbon hu c. Ca hai phan ng tron hoan toan va chay doc (plug - flow) c s dung. Bi v kh nitrogen giai phong suot qua trnh phan nitrate hoa thng bam vao chat ran sinh hoc, v the bc giai phong nitrogen bao gom gia thiet b phan ng va lang nen ay c s dung e tach chat ran sinh hoc. Viec loai bo kh nitrogen bam vao co the c thc hien hoac la trong kenh suc kh ma kenh nay c s dung e noi cac thiet b phan ng sinh hoc va on nh hoac trong be tach ma trong o chat ran c suc kh trong mot khoang thi gian ngan (5 en 10 phut). Chat ran l lng dch tron de bay hi trong cac phan ng nitrate hoa c tao thanh bi cac sinh vat co trach nhiem trong viec chuyen hoa carbon hu c (BOD) va qua trnh nitrate hoa. Toan bo chat ran l lng cua dch tron trong phan ng nitrate hoa thng cao hn t 50 en 100% chat ran l lng cua dch tron de bay hi va co the bao gom chat ket tua hoa hoc neu ket tua phen c s dung cho viec loai bo phosphorus. Trong cac phan ng phan nitrate hoa, chat ran l lng de bay hi cua dch tron c quan sat khoang 40 en 70% chat rang l lng dch tron. Anh hng cua nhng bien oi moi trng va van hanh chnh len qua trnh phan nitrate hoa giai oan kep la nong o nitrate, nong o carbon, nhiet o va pH.

80

Tang cng ket dnh. Mot so cac qua trnh phan nitrate hoa tang cng ket dnh, nhieu cai oc quyen, c phat trien. Cac qua trnh tang cng dnh bam chu yeu c mo ta bang 6-5. Cac be phan ng nen dung dch (fluidized-bed reactors) va he tiep xuc sinh hoc quay (rotating biological contactors - RBCs) thng c s dung nhat. Trong be phan ng nen dung dch, nc thai c x ly bang qua hng len tren mot lp nguyen lieu co hat mn, nh cat, vi toc o va phai en moi trng long. S dung moi hoa gia tang can thiet be mat xac nh va cho phep nong o sinh khoi cao trong be phan ng. Be phan ng can phai co khoang trong nho va n gian e thao tac. Bang 6-5. Mo ta cac he thong phan nitrate hoa tang cng ket dnh.
Phan loai Be phan ng nen ong kn ay kh (Gas-filled) Mo ta Toc o loai thai ien hnh 200C (kg/m3.d) 1.6 1.8

ay chat long (liquid-filled)

Be phan ng nen chat long Moi trng co o xop cao, cat mn Moi trng co o xop cao, carbon hoat tnh Be tiep xuc sinh hoc

Be c boc kn va lap ay bi kh nitrogen, ieu nay loai bo s can thiet phai nhan chm moi trng e duy tr cac ieu kien thieu kh. Be phan ng nen ong kn c lap ay chat long vi moi trng co o xop cao va thap, oi hoi phai co viec ra sach moi trng e kiem soat sinh khoi. o xop thay oi bang cach tang o am ac cua moi trng va toc o chay.

0.1 0.13

12 - 16 4.8 6.0

Be tiep xuc giong vi x ly hieu kh ngoai tr moi trng c nhan chm

Viec ng dung RBCs cho phan nitrate hoa giong nh ng dung cho cac x ly hieu kh, ngoai tr moi trng hoan toan chm e tranh viec oxy hoa dung dch. Cac tiep can hau nh c s dung trong viec anh gia s thc hien cua qua trnh phan nitrogen tang cng ket dnh lien quan en viec s dung cac thong so toc o loai thai. Cho dau mot vai d lieu ng dung c bao cao bang 6-3, cac bc cua he thong phai c ieu khien e xac nh ong thai cua qua trnh khi chung c xem xet e s dung.

81

Chng 7

KH PHOSPHORUS BANG CAC PHNG PHAP SINH HOC

WX

7.1. Cac qua trnh kh phosphorus

Trong nhng nam gan ay, mot so lng ln cac qua trnh kh phosphorus bang ky thuat sinh hoc c phat trien e thay the cho viec x ly hoa hoc. Nh a c mo ta phan trc, phosphorus c loai thai trong x ly sinh hoc bang viec hap thu orthophosphate, polyphosphate va hp chat phosphorus hu c vao trong te bao. Tong lng c loai thai tuy thuoc vao lng chat ran thc c sinh ra. Thanh phan phosphorus trong te bao vao khoang 1/5 ham lng nitrogen: thanh phan phosphorus thc te co the bien ong t 1/7 en 1/3 gia tr nitrogen tuy thuoc vao cac ieu kien xac nh cua moi trng. Tnh trung bnh, ham lng phosphorus c loai thai suot x ly th cap co the bien ong t 10 en 30% lng nc thai. Bang cach s dung mot trong cac qua trnh kh, viec kh at c ang ke co the vt ra khoi khoang bien ong nay. Nhan to quan trong trong viec kh phosphorus bang sinh hoc la vai tro cua vi sinh vat c luan phien trong cac ieu kien k kh va hieu kh. Nh c thao luan tiet trc cac ieu kien luan phien tap trung vao vi sinh vat e viec ong hoa phosphorus cua chung vt hn mc bnh thng. Phosphorus khong ch c s dung e duy tr te bao, tong hp va van chuyen nang lng ma con c tch luy cho viec s dung sau nay bi vi sinh vat. Nc thai cha phosphorus vt troi hoac la c lam sach hoac la c kh qua he thong phu e giai phong ham lng vt troi o. Bang chng cua viec luan phien t ieu kien k kh sang hieu kh co the c thc hien trong qua trnh x ly chnh, hoac dong chnh (mainstream) hoac dong bun hoi lu, hoac dong phu (sidestream) (xem hnh 7-1). Mot vai qua trnh x ly sinh hoc ien hnh c s dung cho viec kh phosphorus se c mo ta trong phan nay. Cac qua trnh o la (1) qua trnh A/O (Anearobic/Oxic stages) dung cho viec kh phosphorus dong chnh, (2) qua trnh PhoStrip (phosphorus strip - lam sach phosphorus) c s dung cho kh phosphorus dong phu, va (3) phan ng me lien tuc (sequencing batch reactor - SBR). SBR c dung cho x ly dong chay nho hn va cung co the tao nen s linh hoat cho qua trnh van hanh, ieu nay cho phep viec kh nitrogen co the ket hp them vao viec kh phosphorus. Thong tin ve thiet ke ien hnh cho cac qua trnh c trnh bay bang 7-1.

82

83

Bang 7-1. Thong tin thiet ke ien hnh cho cac qua trnh kh phosphorus ban phng phap sinh hoc.
Thong so thiet ke Ty so gia thc an va vi sinh vat (F/M) Thi gian lu chat ran Chat ran l lng trong bun long Thi gian lu nc Vung k kh Vung hieu kh Bun hoat tnh hoi lu Tuan hoan noi tai n v Kg BOD/kg chat nen.ngay Ngay Mg/l Gi A/O 0.2 - 0.7 2 - 25 2000 - 4000 0.5 - 1.5 1-3 25 - 40 Qua trnh PhoStrip 0.1 - 0.5 10 - 30 600 - 5000 8 - 12 4 - 10 20 - 50 10 - 20 Phan ng me lien tuc 0.15 - 0.5

2000 - 3000 1.8 - 3 1-4

% chat thai % chat thai

7.1.1. Qua trnh A/O (kh phosphorus dong chnh) Qua trnh A/O c s dung e oxy hoa carbon vi kh phosphorus ket hp t nc thai. Qua trnh A/O la mot he thong tang cng chat l lng bun n, he thong nay ket hp hai phan k kh va hieu kh theo th t (hnh 7-1 a). Viec cung cap co the c tien hanh cho qua trnh nitrate hoa bang keo dai thi gian lu nc trong giai oan hieu kh. Bun on nh c hoi lu cho au vao cua be phan ng va tron vi nc thai ang i vao. Di ieu kien k kh, phosphorus cha trong nc thai va sinh khoi cua te bao c giai phong di dang phosphate hoa tan. Viec lam giam mot t BOD cung xay ra trong giai oan nay. Roi phosphorus c ong hoa trong sinh khoi te bao vung hieu kh. Phosphorus c loai thai t dong dung dch trong bun hoat tnh. Nong o phosphorus au ra tuy thuoc chu yeu vao ty so BOD va phosphorus trong nc thai a c x ly. Theo bao cao th ty so nay vt qua 10/1, gia tr phosphorus hoa tan au ra co the at c 1mg/l hoac thap hn. Trong trng hp ty so BOD/phosphorus thap hn 10/1, cac muoi kim loai co the c them vao qua trnh e at en nong o phosphorus thap nhat. 7.1.2. Qua trnh PhoStrip (kh phosphorus dong phu) Trong qua trnh PhoStrip, mot phan cua bun hoat tnh hoi lu t qua trnh x ly sinh hoc c lam lech hng qua be lam sach phosphorus k kh (hnh 7-1 b), thi gian lu nc trong be lam sach nay bien ong ien hnh t 8 - 12 gi. Phosphorus c giai phong trong be lam sach i ra ben ngoai be trong chat noi tren mat, va bun hoat tnh ngheo phosphorus c a tr lai be hieu kh. Chat noi tren mat giau phosphorus c x ly vi voi hoac chat keo tu khac trong be tach va c ay vao cac be lang s cap hoac be ket bong/ gan loc e tach chat ran. Phosphorus c loai bo t he thong bang ket tua hoa hoc. He thong PhoStrip va cac he thong bun hoat tnh co lien quan khac co kha nang lam giam lng phosphorus au ra en gia tr nho hn 1.5mg/l trc khi loc.
84

7.1.3. Be phan ng me lien tuc (SBR). Be phan ng me lien tuc co the c ng dung e at en viec ket hp bat ky qua trnh oxy hoa carbon, giam nitrogen, va loai thai phosphorus nao. Qua trnh lam giam cac cau thanh cua nc thai co the c thc hien vi viec bo sung hoac khong bo sung them hoa chat bang cach thay oi s van hanh cua cac be phan ng. Phosphorus co the c loai thai bi viec bo sung them cac chat keo tu hoac bang sinh hoc khong bo sung them chat keo tu. Trong mo hnh 7-2, s giai phong phosphorus va ong hoa BOD se xay ra trong pha tron k kh, vi s ong hoa tiep sau trong pha tron hieu kh. Viec chnh sa thi gian phan ng lam cho qua trnh nitrate hoa va loai thai nitrogen co the cung c thc hien. Thi gian quay vong co the bien ong t 3 en 24 gi. Nguon carbon trong pha thieu kh c oi hoi e giup qua trnh phan nitrate hoa - hoac nguon carbon ben ngoai hoac s ho hap noi sinh cua sinh khoi ton tai trong nc thai.

Loc

Khuay k kh

Khuay hieu kh

Khuay thieu kh

O nh

lang

Hnh 7-2. Phan ng theo tng me lien tuc ng dung cho kh carbon, nitrogen va phosphorus. 7.1.4. So sanh cac qua trnh kh phosphorus sinh hoc. Mot s so sanh tong quat cua cac qua trnh kh phosphorus sinh hoc khac nhau c trnh bay qua bang 7-2. Da tren nhng thao luan trc ay, cac qua trnh sinh hoc mang lai nhieu thuan li cho viec loai bo cac thanh phan dinh dng trong viec x ly. V nhu cau loai bo chat dinh dng gia tang, cac viec sa cha oi vi cac qua trnh nay se luon luon phat trien va ngay cang co nhieu loai he thong x ly c s dung. Bi v viec ap dung thanh cong cua nhieu qua trnh tuy thuoc vao ieu kien xac nh cua a phng, cac khu x ly th iem phai c quan tam en e phat trien d lieu van hanh va cac thong so thiet ke.

85

Bang 7-2. S thuan li va bat li cua cac qua trnh loai thai phosphorus sinh hoc.
Qua trnh A/O Thuan li - S van hanh n gian so vi cac qua trnh khac. - Bun thai co thanh phan phosphorus cao (3-5%) va co gia tr lam phan bon. - Thi gian lu nc ngan. - Mc o lam giam hieu qua cua viec kh phosphorus co the chap nhan c, qua trnh co le at en viec nitrate hoa hoan toan. - Co the ket hp de dang vao trong cac he thong x ly bun hoat tnh ang ton tai. - Qua trnh linh hoat : qua trnh kh phosphorus khong b ieu khien bi ty so BOD/phosphorus. - Mot vai khu x ly US. - t s dung hoa chat hn so vi s ket tua hoa hoc dong chnh. - Co the at en nong o orthophosphate thap hn 1.5mg/l. - Qua trnh rat linh hoat cho viec ket hp viec kh nitrogen va phosphorus. - Qua trnh n gian e van hanh - Chat ran dch tron khong the c ra sach bi s dang nc Khong thuan li - Khong co kha nang at en mc o cao cua viec kh phosphorus va nitrogen ong thi. - Trong ieu kien thi tiet lanh, qua trnh van hanh khong bao am. - oi hoi ty so BOD/P cao. - Thi gian lu te bao hieu kh giam, th oi hoi phai co thiet b cung cap oxy vi toc o rat cao. - S linh ong kiem soat qua trnh b gii han. - Yeu cau them voi e ket tua phosphorus. - Yeu cau oxy hoa tan cao hn cua dch tron e ngan can s giai phong phosphorus trong be loc cuoi cung. - oi hoi phai co them be cho viec lam sach. - Can voi la mot van e cho viec duy tr.

PhoStrip

Phan ng tng me lien tuc

- Ch thch hp vi nhng dong chay nho. - oi hoi phai co cac n v tha. - Chat lng nc au ra tuy thuoc vao thiet b lang. - Thong so thiet ke han che.

7.2. Viec kh nitrogen va phosphorus ket hp bang cac phng phap x ly sinh hoc.
Mot so cac qua trnh sinh hoc a va ang phat trien cho viec kh ket hp nitrogen va phosphorus. Nhieu qua trnh cua phng phap nay la oc quyen va s dung mot dang cua qua trnh bun hoat tnh, nhng ket hp cac vung k kh, thieu kh va hieu kh hoac nhng ngan e tien hanh loai thai nitrogen va phosphorus. Mot vai qua trnh xuat phat t viec loai thai phosphorus sau o c ket hp vao cac he thong loai thai phosphorus va nitrogen. Hau het thng s dung cac qua trnh cho viec loai thai phosphorus va nitrogen ket hp la (1) qua trnh A2/O, (2) qua trnh Bardenpho 5 giai oan, nhng qua trnh nay se c mo ta trong phan nay. Hai qua trnh tren c mo hnh hoa hnh 7-3. Nhng thong tin thiet ke ien hnh c gii thieu bang 7-3. Phan ng theo me lien tuc, c mo ta phan au, cung c s dung cho viec kh nitrogen va phosphorus ket hp.

86

Bang 7-3. Cac thong so thiet ke cho qua trnh kh nitrogen va phosphorus ket hp
Thong so Ty so thc an / chat nen Thi gian lu chat ran Chat ran l lng trong dch Thi gian lu nc Vung k kh Vung thieu kh -1 Vung hieu kh -1 Vung thieu kh -2 Vung k kh -2 Bun hoat tnh hoi lu n v Kg BOD/kg chat nen.ngay Ngay Mg/l Gi A2/O 0.15-0.25 Qua trnh 5 giai oan 0.1-0.2 10-40 2.000-4.000 1-2 2-4 4-12 2-4 0.5-1 50-100

4-7 3.000-5.000 0.5-1.5 0.5-1 3.5-6

% nc thai

20-50

7.2.1. Qua trnh A2/O. Qua trnh A2/O la mot sa oi cua qua trnh A/O va cung cap them mot vung thieu kh cho s sa oi nay (hnh 7-2 ben tren tren). Thi gian lu nc trong vung thieu kh khoang chng 1 gi. Vung thieu kh thieu hut oxy hoa tan, nhng oxy hoa hp di dang nitrate va nitrite c a vao bi dch tron hoi lu t phan hieu kh. Nong o phosphorus trong nc thai au ra thng nho hn 2mg/l co the c mong i khong can phai qua loc, nhng neu qua loc th nong o phosphorus co the giam xuong thap hn 1.5mg/l. 7.2.2. Qua trnh 5 giai oan Qua trnh 5 giai oan co the c sa oi cho viec kh ket hp nitrogen va phosphorus. Viec sa oi qua trnh 5 giai oan ket hp them mot giai oan th 5 (giai oan k kh) dung cho viec kh phosphorus (xem hnh 7-3 ben di). Phng phap lien tuc va hoi lu khac vi qua trnh A2/O. He thong 5 giai oan cung cap cac giai oan k kh, thieu kh va hieu kh e kh phosphorus, nitrogen va carbon. Giai oan thieu kh th 2 c cung cap cho qua trnh nitrate hoa phu them s dung nitrate c sinh ra trong giai oan hieu kh nh la chat nhan ien t va carbon hu c noi sinh nh la chat cho ien t. Giai oan hieu kh cuoi cung c s dung e lam sach kh nitrogen t dung dch va lam giam toi thieu s giai phong phosphorus trong thiet b lam sach cuoi cung. Dch tron t vung hieu kh th nhat c hoi lu tr lai vung thieu kh.

87

7.2.3. So sanh cac qua trnh kh nitrogen va phosphorus sinh hoc ket hp. Viec so sanh cac qua trnh kh nitrogen va phosphorus sinh hoc ket hp c gii thieu bang 7-4. Cac thuan li c chia se bi tat ca cac qua trnh nay la so lng bun hoat tnh c sinh ra so vi viec san xuat bun t cac he thong bun hoat tnh truyen thong, va can t hoac khong can thiet co cac chat hoa hoc cho viec kh phosphorus. Mot vai qua trnh c sa oi co the c s dung ch e kh nitrogen hoac phosphorus.

88

Bang 7-4. Nhng thuan li va bat li cua qua trnh kh nitrogen va phosphorus ket hp.
Qua trnh A2/O Nhng thuan li Bun thai co thanh phan phosphorus kha cao (3-5%) va co gia tr lam phan bon. Cung cap kha nang phan nitrate hoa tot hn A/O Sinh ra bun hoat t nhat trong tat ca cac he thong loai phosphorus. Bun thai co thanh phan phosphorus kha cao va co gia tr lam phan bon. Tong nitrogen giam en mc o thap hn hau het cac qua trnh khac. Kiem quay tr lai he thong, bang cach nay lam giam hoac loai tr s can thiet cho viec bo sung hoa chat. c s dung rong rai Nam phi va cac d lieu quan trong co san. Khong thuan li Kho van hanh di thi tiet lanh. Phc tap hn A/O Tuan hoan noi ln tang nang lng cung cap va yeu cau duy tr. Kinh nghiem gii han US. oi hoi cung cap them hoa chat. oi hoi the tch be phan ng hn qua trnh A/O. S lap at ban au giam kha nang cua qua trnh e loai bo nitrogen va phosphorus. Yeu cau ty so BOD/P cao. Anh hng cua nhiet o len qua trnh khong ro rang.

Qua trnh 5 giai oan

89

Chng 8 CAC HE THONG X LY T NHIEN VA NG DUNG

WX

8.1. Cac he thong x ly t nhien

Trong moi trng t nhien, cac qua trnh vat ly, hoa hoc, sinh hoc xay ra khi nc at, he thc vat, vi sinh vat va kh quyen tng tac vi nhau. He thong x ly t nhien c thiet ke nham tan dung li the cua cac qua trnh nay e cung cap cho cac qua trnh x ly nc thai. Cac qua trnh lien quan trong he thong t nhien bao gom nhieu qua trnh c s dung trong cac he thong x ly c hoc hoac ban c hoc - lang ong chat nen, loc, trao oi kh, hap thu, trao oi ion, ket tua hoa hoc, oxy hoa kh (oxydation/reduction) hoa hoc, chuyen hoa (conversion) va bien thoai (degradation) sinh hoc - ac biet hn na la cac he thong sinh hoc nh quang hp, quang oxy hoa, ong hoa thc vat. Trong he thong t nhien, cac qua trnh xay ra vi toc o t nhien va co khuynh hng xay ra ong thi trong mot phan ng he sinh thai gian n, trai vi he thong c hoc ma trong o cac qua trnh xay ra tuan t trong cac be phan ng rieng biet vi toc o tang cao lien quan en viec cung cap nang lng. He thong x ly t nhien trong phan nay bao gom : (1) he thong x ly bang at - toc o cham (slow-rate), r nhanh (rapid infiltration), chay tran be mat (overland flow) va (2) he thong thuy sinh vat - at ngap nc t nhien (natural wetland) va he thong x ly bang thc vat thuy sinh (floating aquatic plant). Chu e chnh yeu trong trong nay la: (1) s phat trien cua cac he thong x ly bang at ai, (2) cac van e ang quan tam c ban trong cac he thong t nhien, (3) he thong toc o cham, (4) he thong tham nhanh, (5) he thong chay tran be mat, (6) cac he thong at ngap nc nhan tao, va (7) cac he thong thc vat thuy sinh. Viec ap dung cho nc thai c thao luan chng 9. 8.1.1. S phat trien cua cac he thong x ly t nhien Mot khai quat ve cac he thong x ly t nhien c cung cap trong chng nay. Lch s ng dung, cac tnh chat va muc tieu cua cac he thong c s dung trong hien tai se c trnh bay. 8.1.1.1. Cac he thong x ly t nhien My. Viec s dung cac he thong x ly t nhien bang at ai US a hnh thanh t nhng thap nien 1880. chau Au, canh ong thai (c s dung sm hn) a tr nen pho bien nh la mot bc tien e kiem soat o nhiem nc. Trong na au cua the ky 20, nhng he thong nay nhn chung a c thay the bi hoac la he thong x ly bang thc vat hoac bang cac
90

canh ong c quan ly ni ma nc thai a qua x ly c s dung cho viec san xuat nong san, vung ti tieu hoac khu vc lam sach nc ngam. Nhng he thong x ly mi bang at ai nay a co khuynh hng phat trien chiem u the pha Tay US, ni ma gia tr cua nc thai c xem nh la mot li the. So lng nhng vung US ang s dung viec x ly t nhien gia tang t 304 nam 1940 en 571 (phuc vu cho 6.6 trieu dan) nam 1972, nhng tong so nay ch cho thay mot phan tram nho trong 15.000 vung dan c s dung he thong x ly nay. Theo Clean Water Act vao nam 1972, viec au t vao cac he thong x ly bang at ai a c lam song lai nh la mot ket qua nhan manh rang nhng khu x ly nay c at lam ni s dung lai nc thai, quay vong chat dinh dng va s dung nc thai cho phat trien nong nghiep. S ho tr tai chnh bi Act a thuc ay nhng nghien cu rong rai va phat trien cong nghe he thong x ly t nhien, dan en s chap nhan no trong cac lanh vc cua ky thuat x ly nc thai nh la mot ky thuat quan ly se c xem la tng ng vi bat ky hnh thc x ly nao. Nhng phat trien gan ay nhat trong cong nghe he thong x ly t nhien la s dung cac vung at ngap nc vi thc vat noi va he thong thuy sinh vi thc vat l lng. Li ch cua viec x ly nc thai bang vung at ngap nc phat trien ket qua la mot hnh thc x ly mi ket vung at ngap nc vi thc vat thuy sinh va he thong x ly t nhien. Thc vat noi c s dung ban au e phat trien hnh thc x ly bang ho sinh hoc co truyen va cac ao on nh, nhng xa hn na la s phat trien cua ng dung nay a at c ket qua trong cong nghe oc ao cua he thong thuy sinh. 8.1.1.2. Cac tnh chat va muc tieu cua he thong x ly t nhien. Nhng ac tnh vat ly, muc tieu thiet ke, va kha nang x ly cua cac loai hnh khac nhau cua he thong t nhien c mo ta va so sanh trong phan nay. S so sanh cac ac tnh cua khu x ly, cac ac tnh thiet ke ien hnh, va so lng mong i cua nc thai a c x ly t cac loai hnh chu yeu cua cac he thong x ly t nhien se c gii thieu bang 8-1, 8-2 va 8-3. Tat ca cac hnh thc cua he thong x ly t nhien a c gii thieu trc ay qua mot vai dang tien x ly c hoc. oi vi nc thai, mot he lang nho la can thiet e loai bo cac chat ran tho co the gay can tr cho he thong phan phoi va tao ra ieu phien toai khong ang co. ieu can thiet e lam cho he tien x ly vt qua mot vai mc nho se tuy thuoc vao muc tieu cua he thong va nhng oi hoi thng xuyen. Kha nang cua he thong t nhien dung cho x ly nc thai la han che, cac he thong phai c thiet ke va quan ly phu hp vi kha nang cua he thong o. Cac chi tiet cua viec nh gia v tr, x ly ban au, va thiet ke qua trnh cho moi loai he thong c thao luan trong nhng phan sau.

Bang 8-1. So sanh cac ac iem cua v tr oi vi he thong x ly t nhien.

91

ac iem ieu kien kh hau

Toc o cham Can lu tr trong mua ong va suot thi gian tuyet ri 0.6 - 1m (t nhat)

R nhanh Khong (co the van hanh trong mua ong) 3m (o sau t hn co the chap nhan nhng ni co he thong thoat nc ngam Khong thanh van e, o doc qua mc th oi hoi nhieu cong sc hn. Nhanh (cat, cat mun)

o sau en nc ngam

Chay tran be mat Can lu tr trong mua ong va suot thi gian tuyet ri Khong co van e

at ngap nc Co the khong can lu tr trong thi tiet lanh Khong co van e

Thc vat thuy sinh Co the can lu tr trong thi tiet lanh Khong co van e

o doc

<15% oi vi at trong trot, <40% oi vi at rng Toc o trung bnh en nhanh

1 - 8%

Thng <5%

Thng <5%

o tham cua at

Thap (set, phu sa va at vi chan khong tham)

Thap en trung bnh

Thap en trung bnh

Bang 8-2. So sanh cac ac tnh thiet ke cua cac he thong x ly t nhien.
ac tnh Toc o cham (loai 1) Toc o cham (loai 2) R nhanh Chay tran mat at at nc ngap Thc vat thuy sinh

Ky thuat ap dung

Toc o tai thuy hang nam (m/nam) Dien tch yeu cau (ha/103m3/ngay) X ly tien ng dung thap nhat c cung cap Cach thc nc thai c ap dung

Phun nc hoac be mat 1.7-6.1

Phun nc hoac be mat 0.61-2.0

Luon luon be mat 6.1-91.4

Phun nc hoac be mat 7.3-56.7

Phun nc hoac be mat 5.5-18.3

Be mat

5.5-18.3

6-21.4 Tien x ly Lang nen ay Thoat hi nc va tham vao trong at

18.2-58.8 Tien x ly Lang nen ay Thoat hi nc va tham vao trong at

0.4-6.0 Tien x ly Lang nen ay Chu yeu la tham vao trong at

0.6-4.8 Sang rac

2.0-6.6 Tien x ly Lang nen ay Thoat hi nc va tham vao trong at

2.0-6.6 Tien x ly Lang nen ay Thoat hi nc mot t

Can cho thc vat

Yeu cau

Yeu cau

Khong bat buoc

Chay be mat va tham bay hi vi tham vao trong at mot t Yeu cau

Yeu cau

Yeu cau

Ghi chu: lang nen ay tuy thuoc vao viec s dung nc thai va loai cay trong.
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Bang 8-3. So sanh chat lng nc a c x ly t cac he thong toc o cham, r nhanh va chay tran mat at.
Gia tr, mg/l R nhanh Trung bnh Cc ai 2 <5 2 <5 0.5 <2 10 <20 1 <5

Thanh phan BOD SS NH4+-N Tong N (N) Tong P (P)

Toc o cham Trung bnh Cc ai <2 <5 <1 <5 <0.5 <2 3 <8 <0.1 <0.3

Chay tran mat at Trung bnh Cc ai 10 <15 15 <25 1 <3 5 <8 4 <6

8.1.2. Toc o cham (slow-rate) : X ly toc o cham la qua trnh x ly t nhien chiem u the ngay nay, lien quan en viec s dung nc thai cho cac vung at hoa mau dung cho viec x ly nc thai va cho nhu cau phat trien cua thc vat. Nc thai c s dung hoac la qua qua trnh bay hi hoac la tham vao trong at (hnh 8-1). Moi dong chay tren be mat c tap trung lai va s dung cho he thong, qua trnh x ly xay ra khi nc thai tham qua lp at. Trong hau het cac trng hp, qua qua trnh tham nc se i vao tang nc ngam, nhng trong mot vai trng hp, nc a qua x ly co the c tiep nhan bi he thong nc mat hoac vao trong he thong nc gieng. Toc o x ly ma he thong nay at c tren mot n v dien tch (toc o tai thuy - hydraulic loading rate), s la chon va quan ly thc vat la muc tieu cua viec thiet ke he thong va ieu kien cua vung at se c ban en phan sau. He toc o cham thng c xep vao loai 1 hoac loai 2 tuy thuoc vao muc tieu thiet ke. Mot he thong toc o cham c xem la loai 1 khi muc tieu chnh la x ly nc thai va toc o tai thuy la khong kiem soat c bi nhng nhu cau nc cho thc vat ch khong phai thong so thiet ke han che - kha nang tham cua at hoac tai lng cac thanh phan nc thai. Loai 2, c thiet ke vi muc tieu tai s dung nc thai qua viec san xuat nong pham hoac ti tieu cho khu giai tr (landscape irrigation) thng c noi en nh la mot he thong ti tieu nc thai hoac ti tieu ong ruong. Nc thai co the c dung cho at nong trai hoac vung cay xanh (bao gom ca at rng) bi cac phng thc phan tan a dang (hnh 8.1). Cac chu ky ng dung khong thng xuyen, ien hnh t 4-10 ngay, c dung e duy tr ieu kien hieu kh trong lp at. He toc o cham ket hp vi s co mat cua thc vat va he thong sinh thai at a lam cho he toc o cham mot tiem nang x ly ln nhat cua he thong x ly t nhien (bang 8-3).

93

Hnh 8.1. Cac phng thc phan tan nc thai

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8.1.3. R nhanh (rapid infiltration). Trong he thong r nhanh, nc thai a c x ly s bo c s dung cho mot thi han khong thng xuyen trong cac ao phan tan, nh hnh 8-3. X ly nc thai bi he thong ti toc o cao cung c tien hanh. Thc vat thng khong c cung cap cho cac ao loc nhng can thiet cho viec ng dung cac thiet b phun. Bi v toc o tai hoat thng cao, nen s thoat hi t va hau het nc thai tham qua lp at ni ma qua trnh x ly xay ra. Muc tieu thiet ke cho he loc nhanh bao gom (1) x ly e tai tao nguon nc ngam e tang nguon nc cap hoac bao ve s xam nhap cua nc man (saltwater intrusion), (2) x ly e tai tao nguon nc cua tui nc ngam nong hoac tai thu hoi nc bm, va (3) x ly e tai thiet dong chay va lay lai nguon nc mat. (xem hnh 8-2). Tiem nang cua he r nhanh thap hn he toc o cham bi v kha nang gi nc thap hn va cao hn ve toc o tai hoat thuy lc (xem bang 83).

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Hnh 8-2. Cac phng thc r nhanh (a) con ng tai nc, (b) con ng phuc hoi nc bang he thong thoat nc ngam, (c) con ng phuc hoi nc bang s dung he thong gieng.

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8.1.4. He chay tran be mat (overland-flow) Trong he chay tran be mat, nc thai a qua tien x ly c phan phoi doc theo cac ng doc xuong theo cac luong cay ieu nay cho phep nc thai co the chay tran be mat t cac luong en b e tiep nhan pha di (xem hnh 8-3). He chay tran be mat thng c s dung nhng vung co lp at be mat hoac lp at ben di khong tham tot, v the phng thc x ly nay c ng dung cho s bien ong ln ve kha nang tham cua at bi v at be mat co khuynh hng b ong cng theo thi gian. Qua trnh tham qua at la mot phng thc thuy lc yeu va hau het nc thai c tap trung theo cac dong chay be mat, mot phan nc se b mat i qua qua trnh boc hi, lng nc b boc hi bien ong theo thi gian trong nam va theo thi tiet cua vung o. Cac he thong nay c ng dung luan phien tuy theo tng mua va tuy thuoc vao muc tieu x ly. S phan phoi nc thai co the c tien hanh bang cac dung cu phan tan nc di ap suat cao, phun thanh dang sng ap suat thap hoac phan tan be mat bang cac ong dan.

97

Hnh 8-4.

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8.1.5. at ngap nc (wetland) at ngap nc la mot vung at co nc vi o sau cua nc nho hn 0.6m, thch hp cho s phat trien cua thc vat nho len be mat (emergent plant) nh uoi meo (cattail), co nen (bulrush), lau say (reed) va lach (sedge) (hnh 8-4). Thc vat tao nen be mat cho s bam vao cua vi khuan tao nen mot mang loc sinh vat, giup ch cho qua trnh loc va hap thu cac thanh phan dinh dng trong nc thai, trao oi oxygen trong cot nc va kiem soat toc o phat trien cua tao bang cach han che s xuyen qua cua anh sang mat tri. Ca hai loai at ngap nc t nhien (natural wetland) va at ngap nc nhan tao (constructed wetland) eu c s dung cho x ly nc thai, cho du viec s dung at ngap nc t nhien nhn chung b han che trong viec lam sach hoac x ly nc thai a c x ly th cap hoac a c x ly cap tien. 8.1.5.1. at ngap nc t nhien (natural wetland) T quan iem ieu chnh, cac vung at ngap nc c xem la ni tiep nhan nc. Ket qua la nc xa vao vung at ngap nc, trong hau het cac trng hp oi hoi phai co s ieu tiet, ma nhng oi hoi nay c quy nh ro rang la x ly th cap hoac x ly cap tien. Hn na, muc tieu chnh la khi xa vao vung at ngap nc t nhien se lam tang cng s phat trien cac quan the sinh vat ton tai trong o. Viec chnh sa cac vung at ngap nc t nhien nham tang cng kha nang x ly thng rat de b pha v oi vi he thong sinh thai t nhien va noi mot cach tong quat se khong at c. 8.1.5.2. at ngap nc nhan tao (constructed wetland) at ngap nc nhan tao thng co ay u cac kha nang x ly cua at ngap nc t nhien nhng khong ket hp vi viec xa vao he thong sinh thai t nhien. Hai loai at ngap nc nhan tao c phat trien cho viec x ly nc thai (1) he thong be mat nc t do (free water surface - FWS) va he thong chay di be mat (subsurface flow system - SFS). Khi c s dung e x ly nc mc o th cap hoac mc o cap tien, he thong FWS thng bao gom cac ao hoac kenh song song vi at khong tham ben di hoac chan di be mat (subsurface barrier), thc vat nho len mat nc va nc can 0.1 - 0.6m. Nc thai tien x ly thng c dung lien tuc cho nhng he thong nh the nay va qua trnh x ly xay ra khi nc chay cham qua than va re cua thc vat. He thong FWS co the cung c thiet ke vi muc ch tao ra nhng quan the hoang da mi hoac tang cng cac vung at ngap nc t nhien ang ton tai ben canh thc vat bac cao thuy sinh noi. Nhng he thong nh the nay bao gom s ket hp cua thc vat, vung nc m va cac ban ao cung vi s phat trien cua thc vat cung cap thc an cho cac quan the ong vat trong o. He thong chay di mat c thiet ke vi muc ch cho mc o x ly cap hai hoac x ly cap tien. Nhng he thong x ly nay
99

cung c goi la vung re (root zone) bao gom cac kenh vi lp ay khong tham c lap ay bi cac lp a va cat e nang thc vat. (hnh 8.5).

Hnh 8.5. Mat cat ngang cua he thong chay di mat (SFS) ien hnh 8.1.6. He thc vat thuy sinh bac cao (floating aquatic plant) He thong thc vat thuy sinh bac cao giong nh he thong FWS ngoai tr cac loai thc vat ay la thc vat noi nh beo Nhat ban, beo tam (hnh 8-4). o sau cua nc sau hn he thong at ngap nc dao ong t 0.5 - 1.8 m. S suc kh bo sung c s dung vi he thong thc vat noi e lam tang kha nang x ly va duy tr ieu kien hieu kh can thiet cho viec kiem soat sinh hoc s phat trien cua muoi. Ca hai he thong beo Nhat ban va beo tam c s dung e loai bo tao t nc thai cua ho sinh hoc va ao on nh, trong khi o he thong beo Nhat ban c thiet ke e cung cap cho mc o x ly cap hai va cap tien. Cac qua trnh tai thuy thng xuyen va oi hoi cac vung ac biet cho he thong thc vat noi giong vi he thong at ngap nc. (xem bang 8-2). 8.1.7. He nuoi trong thuy san (aquaculture) Nuoi trong thuy san la s phat trien ca va cac loai thuy san khac trong viec san suat nguon thc pham. Nc thai c s dung trong nhieu phng thc nuoi trong khap the gii. Tuy nhien, trong hau het cac trng hp, viec san xuat sinh khoi la muc tieu au tien cua he thong va bat ky hnh thc x ly ch la li tc phu them. Hau het hieu qua x ly at c trong he thong nuoi thuy san eu c quy cho vi khuan bam vao thc vat thuy sinh. Co t bang chng cho thay ca ong vai tro trc tiep trong x ly. Viec ket hp x ly t nhien vi nuoi trong thuy san trong viec ng dung oc lap oi hoi phai nghien cu nhieu hn. ac biet la nhng nguy hiem oi vi sc khoe khi s phat trien cua thuy sinh vat trong nc thai khong c kiem soat.
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8.2. Nhng nghien cu c ban trong viec ap dung he thong x ly t nhien. Biet c cac ac tnh cua nc thai, cac c che x ly, nhng anh hng en sc khoe cong ong va nhng yeu cau thng xuyen la nen tang cho viec thiet ke thanh cong va ng dung cac he thong x ly t nhien. 8.2.1. Cac ac tnh cua nc thai va c che x ly Nh a mo ta trong phan gii thieu, x ly nc thai trong he thong t nhien c thc hien bi cac qua trnh vat ly, hoa hoc va sinh hoc t nhien ma cac qua trnh nay xay ra trong he thong sinh thai at - nc - thc vat. He thong t nhien co kha nang loai bo t nhat mot vai thanh phan u the hay khong u the cua nc thai chung bao gom cac chat c xem la chat gay o nhiem - chat ran l lng, chat hu c, nitrogen, phosphorus, vi lng, vi lng hu c va vi sinh vat (bang 8-4). Cac qua trnh c ban chu trach nhiem loai bo cac thanh phan c mo ta trong phan nay. Bang 8-1. Cac thanh phan ien hnh cua nc thai sinh hoat cha x ly.
Chat gay o nhiem Chat ran tong (TS) Chat ran hoa tan tong (TDS) Co nh Bay hi Chat ran l lng (SS) Co nh Bay hi Chat ran co the on nh c BOD5 (200C) Tong Carbon hu c (TOC) COD Tong Nitrogen (tnh theo N) Hu c Ammonia Nitrite Nitrate Tong phosphorus (tnh theo P) Hu c Vo c Chloride Sulfate Kiem (tnh theo CaCO3) Chat beo Tong coliform Cac hp chat hu c bay hi (VOCs) n v mg/l mg/l mg/l mg/l mg/l mg/l mg/l ml/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l mg/l no/100ml g/l Nong o Thap 350 250 145 105 100 20 80 5 110 80 250 20 8 12 0 0 4 1 3 30 20 50 50 106-107 <100 Trung bnh 720 500 300 200 220 55 165 10 220 160 500 40 15 25 0 0 8 3 5 50 30 100 100 107-108 100-400 Cao 1200 850 525 325 350 75 275 20 400 290 1000 85 35 50 0 0 15 5 10 100 50 200 150 107-109 >400

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Chat ran l lng. Trong cac he thong ma tnh chat cua nc chay tren mat at - he chay tran, at ngap nc, va thc vat thuy sinh - chat ran l lng trong nc thai c loai bo mot phan nh s lang nen ay, tang cng bi qua trnh chay vi toc o rat cham va mot phan bi s loc cua thc vat. Viec loai bo chat ran cung xay ra tai be mat tiep xuc cua at. Trong cac he thong ma nc thai chay tren be mat at - he chay cham, he r nhanh, va SFS chat ran l lng au tien c loai bo bi s loc qua at hoac moi trng di lp at mat, v vay qua trnh lang nen ay co the ang ke trong cac ao r nhanh suot thi gian s dung. Trong he toc o cham va r nhanh, hau het chat ran b loai bo ngay lp at mat. V the, co y kien cho rang cac chat ran l lng co the lam ong cng lp at loc be mat cua he thong. Do o, cac he thong phai c thiet ke va s dung e lam giam thieu s bien mat kha nang loc cua at. Chat hu c. Chat hu c phan huy c co trong nc thai di dang hoa tan hoac l lng, thng c loai bo bang vi sinh vat phan huy. Vi sinh vat co trach nhiem phan huy thng lien ket vi mang loc, phat trien tren be mat cua cac hat at, thc vat va cac gia bam (litter). Nhn chung, cac he thong t nhien c thiet ke va hoat ong nham duy tr ieu kien hieu kh e qua trnh phan huy dien ra mot cach toi u va hoan toan hn phan giai k kh. Kha nang cua he thong x ly t nhien la lam giam chat hu c trong ieu kien hieu kh nhng b gii han bi s van chuyen oxygen vao trong he thong t kh quyen. V the, he thong phai c thiet ke sao cho tai lng BOD t hn so vi toc o van chuyen oxygen vao he thong. Nitrogen. Viec chuyen hoa va loai bo nitrogen trong cac he thong t nhien lien quan en mot loai cac qua trnh phc tap va cac phan ng c mo ta trong hnh 9-1. Cac c che lien quan trong viec loai bo nitrogen t nc thai tuy thuoc vao dang nitrogen - nitrate, ammonia, hoac nitrogen hu c. Nitrogen thng di dang ammonia hoac nitrogen hu c. Nitrogen hu c. Lien ket vi chat ran l lng trong nc thai, c loc bang lang nen ay va loc, nh c mo ta tren, hu c dang ran, co the c hap thu bi mun at, bao gom mot so rat ln cac phan t hu c phc tap cha cacbohydrate, protein, lignin. Mot vai nitrogen hu c b thuy phan thanh amino acid hoa tan ma cac amino acid nay co the tiep tuc c phan huy e giai phong NH4+. Ammonia (NH4+). Co the i theo vai con ng chuyen hoa trong he thong x ly t nhien. Ammonia hoa tan co the c loai bo bi qua trnh bay hi (volatilization) trc tiep i vao kh quyen dang kh NH3, cach nay ch chiem t le t (<10%), ngoai tr trng hp trong ao on nh ni ma nc thai c lu tr trong thi gian dai va pH cao lam tang cng ang ke qua trnh bay hi ammonia. Hau het anh hng va s chuyen hoa ammonia trong nc thai ch la tam thi qua cac phan ng trao oi ion trong at va cac chat hu c mang ien. Ammonia co the hap thu bi thc vat va vi sinh vat hoac c chuyen hoa thanh
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nitrate qua qua trnh nitrate hoa sinh hoc trong ieu kien hieu kh. Bi v kha nang hap thu ammonia cua he thong t nhien la gii han, nen qua trnh nitrate hoa la can thiet e giai phong ammonia. Chu trnh giai phong - hap thu nay ac biet quan trong trong he chay tran be mat ni ma s hap thu b han che doc theo be mat cac ng doc. Nitrate. c xem la mot ion tieu cc, khong c gi lai bi cac phan ng trao oi nhng van con dang dung dch va c chuyen hoa trong qua trnh tham. Neu chung khong c loai bo bi qua trnh ong hoa thc vat hoac kh nitrate hoa, nitrate se i sau va tham vao trong lp nc ngam. oi vi cac he thong the hien tnh tham thau cua nc nh he toc o cham, loc nhanh, va ng dung nc thai, nitrate trong qua trnh loc co the gay nguy hiem cho sc khoe cong ong. V the, nhng he thong nay phai c thiet ke va ap dung e tang cng mc o can thiet cua viec loai bo nitrogen e bao ve nguon nc ngam. Nitrate co the c hap thu bi thc vat, nhng s ong hoa ch xay ra vung xung quanh re trong suot thi ky phat trien cua thc vat. e at c s loai bo nitrogen t he thong bi ong hoa thc vat, thc vat can phai c thu hoach nh ky va loai ra khoi he thong. Neu thc vat con lai trong he thong, nitrogen trong thc vat se quay tr lai va i vao he thong di dang nitrogen hu c. ong hoa thc vat va thu hoach chung la c che loai bo nitrogen chu yeu trong he thong toc o cham. Kh nitrate sinh hoc (biological denitrification). Nitrate cung se c loai bo bi qua trnh kh nitrate sinh hoc va lan lt giai phong cac kh NO va N2 vao trong kh quyen. Kh nitrate sinh hoc la c che loai bo nitrogen chu yeu trong he chay tran mat at, r nhanh va thuy sinh. Kh nitrate c thc hien bi vi khuan tuy nghi (facultative bacteria) trong ieu kien thieu kh. Khong can thiet cho toan bo he thong la thieu kh cho kh nitrate hoa xay ra. Kh nitrate xay ra trong tieu vung thieu kh gan vi vung hieu kh. Tuy nhien, e phan ng kh nitrate hoa at toi a, th cac ieu kien oi hoi phai at c toi u. Trong ieu kien thieu kh t so C/N la can thiet quan trong e hoan thanh phan ng kh nitrate hoa. T so C/N t nhat la 2:1 (da vao TOC va total N) la can thiet e at en kh nitrate hoa hoan toan trong he thong t nhien. Carbon t xac ba thc vat co the c xem nh la nguon carbon, ac biet trong cac he thuy sinh, nhng trong he toc o cao nh chay tran mat at, r nhanh, nguon carbon phai bao gom ca nc thai c cung cap. V the s loai bo nitrogen toi a khong the at c trong nhieu he thong vi nc thai th cap co t so C/N thng di 1:1.

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Ma
NH3 NO3NH3 NO3-

Nc thai Nitrogen hu c NH3 Kh nitrate hoa N2 S bay hi NH3

Phan bon vo c NH3 NO3N2

Xac ba thc vat, phan xanh Nitrogen hu c protein Protein cua thc vat

Co nh nitrogen NO3Phan huy Nitrate hoa NH3 NH3 Nitrate hoa Proteins Phan huy NH3 Nitrate hoa NO3NH3 NO3-

S hap thu NO3NO3-

S hap thu

Trong at

Qua trnh loc

Nc ngam

Hnh 8.6. S chuyen hoa nitrogen trong he thong x ly t nhien

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Phosphorus. Cac qua trnh loai bo phosphorus trong he x ly t nhien la s ket tua va hap thu hoa hoc, cho du thc vat hap thu mot lng ln. Phosphorus ton tai chu yeu di dang orthophosphate c hap thu bi at set va mot vai phan nho at hu c trong tang at. Ket tua hoa hoc bi calcium (pH trung tnh) va sat hoac nhom (pH acid) xay ra toc o cham hn qua trnh hap thu, nhng tam quan trong th ngang bang. Phosphorus c hap thu co the b gi lai t va ben vng oi vi s loc qua at. Cho du kha nang hap thu phosphorus cua at la gii han, nhng no van kha ln oi vi at cat. Sau 88 nam cua he loc nhanh dung cho nc thai sinh hoat cha x ly tai Calumet, Michigan, nong o phosphorus trong nc ngam van thap (0.1-0.4mg/l). Tuy nhien, ap dung mot thi gian dai lam cho phosphorus hoa tan trong at tang len tren lp be mat (0.3m), cho thay rang lp nay tr nen bao hoa vi phosphorus. Mc o loai bo phosphorus co the at c bi he thong x ly t nhien tuy thuoc vao mc o tiep xuc cua nc va at nh he chay tran va he thuy sinh se gii han tiem nang loai thai phosphorus. Vi lng. Loai bo vi lng (chu yeu la kim loai) xay ra chu yeu qua hap thu (bao gom ca hap thu va cac phan ng ket tua) va bi s ong hoa cua thc vat oi vi mot vai kim loai. Kim loai c gi lai trong at hoac trong nen ay cua he thuy sinh. Kha nang gi lai cac kim loai cua hau het cac loai at va nen ay noi chung la rat cao, ac biet pH>6.5. Trong ieu kien pH thap va k kh, mot vai kim loai hoa tan manh hn va co the giai phong vao trong dch at. Loai bo kim loai bien ong gia cac he thong, tuy thuoc vao nong o nc thai va ieu kien vung at. Nhng hieu qua cua viec loai bo a c nghien cu vi hau het cac kim loai noi chung bien ong trong khoang 80-95%. Hieu qua thap hn co the oi vi at ngap nc FWS va he thong thc vat thuy sinh da vao s tiep xuc gii han gia nc vi at va nen ay cung nh ieu kien k kh cua nen ay. Chat hu c vi lng (trace organics). hp chat hu c vi lng c loai bo t nc thai qua qua trnh bay hi va hap thu, bi cac qua trnh phan giai sinh hoc va quang hoa. Noi cach tong quat, cac he t nhien co kha nang loai bo cac hp chat hu c vi lng. Tuy nhien theo cac c s d lieu hien nay qua t e d oan kha nang loai bo cac hp chat hu c co cau tao n gian. Nhng ket qua ien hnh oi vi viec loai bo cac chat hu c co the at c hieu suat t 85-99.99% oi vi tat ca cac he thong. Vi sinh vat. C che loai bo vi sinh vat (vi khuan va ky sinh trung - nguyen sinh ong vat va giun san) pho bien oi vi hau het cac he x ly t nhien bao gom : chet, loc nc, lang nen ay, phong xa, say kho va hap thu. Virus c loai bo ien hnh bi hap thu va t chet. He toc o cham va r nhanh ca hai eu co dong chay nc thai qua lp at nen co kha nang loai bo hoan toan vi sinh vat cua nc thai qua qua trnh loc. Trong moi trng at mn thng s dung he toc o cham, viec loai bo hoan toan co the at c khoang chng 1.5 m di chuyen cua nc. oi hoi phai co khoang cach di chuyen xa hn trong at e at c s loai bo trong he r nhanh, va khoang cach nay con tuy thuoc vao o tham cua at va toc o
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tai thuy lc. Tat ca cac hnh thc khac cua he x ly t nhien co the lam giam nong o vi sinh vat tuy theo o ln cua tng qua trnh x ly, nhng noi mot cach tong quat, neu nh khong at en s loai bo toi a th khong the xoa i moi hnh thc chung nga cac mam benh xuat phat t cac khu x ly t nhien. 8.2.2. Nhng van e ve sc khoe cong ong (public health issues) Yeu to sc khoe cong ong lien quan en viec x ly bang at ai bao gom : (1) mam vi khuan va s truyen benh co the co en cac dang sinh vat cao hn, gom ca con ngi, (2) hoa chat co the i vao nc ngam va gay nguy hiem cho sc khoe neu nc nay c dung cho hoat ong song va (3) chat lng mua mang cua cac vung at c ti tieu bang nc thai. Cac mam vi khuan (bacteriological agents). S song cua cac vi khuan va virus gay benh ngay trong nhng giot nc ban ra, tren va trong at va anh hng len ngi tiep nhan. ieu quan trong la tat ca nhng tiep xuc gia cac mam benh trong at qua nc thai va s nhiem benh ong vat va ngi eu phai co s chung nga dai lau va phc tap. Nhng cau hoi c at ra la nhng ton tai co lien quan ve viec phong nga co c thc hien hay khong? Cac may phun sng, c s dung trong x ly nc thai, phun ra nc di dang sng mu ma nhng hat sng nay co the di chuyen theo gio va vo cung nho ve kch thc cung nh khoi lng (co ng knh khoang 0.01-50m). chnh cac hat nay xuat phat t nc thai khong c kh trung se cha mot lng ln cac vi khuan va virus hoat ong. Tuy nhien trng hp phat tan bang cac hat sng co cha vi khuan nay ch chiem en 0.3% nc thai. Nhng nghien cu ve s di chuyen cua cac hat sng co cha vi khuan xuat phat t may phun sng la c s e s dung nc thai cha c x ly hoac cha kh trung. Cho du vi khuan di chuyen xa hn cac hat sng mang no t nc thai cha c kh trung, khoang cach bien ong ln nhat t 30-200m, ieu o cho thay vi khuan di chuyen theo gio se tang len theo s gia tang cua o am va toc o gio cung vi s giam nhiet o va tia t ngoai. S can thiet phai co cac vung em hoac kh trung e giam thieu rui ro cho sc khoe cong ong phai c at ra trong tat ca cac trng hp (1) mc o tiep can cua cong ong dan c vi khu x ly, (2) kch c cua khu x ly, (3) cung cap cac vung em hoac trong cay va (4) ac trng cua kh hau trong vung. Yeu cau ve vung em thng c anh gia thng xuyen bi cac nha chc trach, cach 15-60m t ng i, dai phan cach vung va cac khu dan c. S luan phien oi vi cac vung em bao gom trong cay, s dung may phun e phun nc xuong di, hoac theo mot quy ao vi toc o cham va ngng hoat ong cac may phun, hoac lam giam thieu toi a qua trnh phun trong khu x ly trong suot thi gian gio manh.
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Chat lng nc ngam (groudwater quality): cac he thong (chu yeu la toc o cham va r nhanh) ni ma mot phan nc thai tham qua va i vao trong mach nc ngam ma lng nc ngam nay phuc vu hoac co the phuc vu tiem tang nh nc cap phai c thiet ke va quan ly e duy tr chat lng nc ngam theo tieu chuan nc uong cua US.EPA. Bi v nitrate la nguyen nhan gay benh thieu mau tre em, nong o cua no trong nc uong c gii han bi tieu chuan nc uong (primary drinking water standard) la 10mg/l. S loai bo nitrogen nhat thiet phai at c qua tien x ly va x ly t nhien e duy tr chuan mc nay. Vi lng ap dung cho he x ly t nhien khong gay nguy hiem cho chat lng nc ngam bi v vi lng thng c loai bo qua qua trnh hap thu va ket tua hoa hoc khi chung tham qua ch mot vai chuc centimeter at, ngay ca trong he r nhanh vi toc o tai thuy cao. Trong nhieu nghien cu ve anh hng lau dai cua viec ng dung nc thai, thay rang khong co s gia tang nong o kim loai trong at lp tren cua at tren mc bnh thng oi vi at nong nghiep. Loai bo vi khuan t nc thai qua at mn kha hoan toan: no co the c m rong trong at co hat ln, at cat s dung cho he r nhanh. Ket cau at a co the lam cho vi khuan vt qua vai chuc met at t khu x ly vao ben trong tang at. Tnh trang nay co the tranh c bi viec anh gia ky lng ket cau a chat ban au khi la chon v tr ap dung. Chat lng vu mua (crop quality). Vi lng c gi lai trong at va nen ay cua he thong x ly t nhien va co san cho s ong hoa cua thc vat. T quan iem ve sc khoe cong ong, kim loai chu yeu co lien quan la cadmium. Cadmium co the tch luy trong thc vat mc o gay oc cho ngi va ong vat, va nhng mc o nay ben di nong o gay oc cho thc vat (phytotoxic). Ket qua cadmium la mot trong nhng thanh to gii han trong viec xac nh toc o tai hoat chat thai tren vung at nong nghiep. oi vi hau het cac ap dung, viec tch luy cadmium se khong phai la mot van e ang c tranh cai. Viec giam sat tai mot khu x ly Melbourn, Uc, ni ang tiep nhan mot lng nc thai trong 76 nam, cho thay rang khong co s gia tang trong viec tch luy cadmium trong thc vat khi so sanh vi s phat trien cua thc vat trong mot khu khong tiep nhan nc thai. Nhng kim loai khac hoac khong c ong hoa bi thc vat (nh ch) hoac gay oc cho thc vat nong o thap cho thay nguy c gay oc cho chuoi thc an (nh kem, ong, nickel).

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TAI LIEU THAM KHAO 1. Kieu Hu Anh, 1999. Giao Trnh Vi Sinh Vat Cong Nghiep. Nxb KH va KT. 2. Le Huy Ba, 2000. Moi Trng. Nxb ai Hoc Quoc Gia TP Ho Ch Minh. 3. Nguyen Lan Dung, Nguyen nh Quyen, Pham Van Ty, 2000. Vi Sinh Vat Hoc. Nxb Giao Duc. 4. Nguyen Thanh at, Mai Th Hang, 2001. Sinh Hoc Vi Sinh Vat. Nxb Giao Duc. 5. Tang Van oan, 2001. Ky Thuat Moi Trng. Nxb Giao Duc. 6. Hoang Hue, 1996. X Ly Nc Thai. Nxb Xay dng. 7. Trnh Xuan Lai, 2000. Tnh Toan Thiet Ke Cac Cong Trnh X Ly Nc Thai. Nxb Xay Dng. 8. Mai nh Yen, 1990. C S Sinh Thai Hoc. H Tong hp Ha noi. 9. Cuc Moi Trng, Vien Moi Trng va Tai nguyen, 1998. Cong Nghe Moi Trng. Nxb Nong nghiep. 10. Trung Tam ao Tao Nganh Nc va Moi Trng, 1999. So Tay X Ly Nc, tap I, II. Nxb Xay Dng. 11. Lacher W, 1983. Sinh Thai Hoc Thc Vat . Le Trong Cuc dch. Nxb H va THCN. 225tr. 12. Odum E. P, 1978. C s sinh thai hoc. Ban dch t tieng Anh cua Pham Bnh Quyen. Nxb H va THCN. 423tr. 13. Anthony F. Gaudy, J. Elizabeth T. Gaudy, 1980. Microbiology for Environmental Scientists and Engineers. Printed in United State of America. 14. Bowen H. J. M, 1966, Trace metals in biochemistry. Academic Press, New York. 15. Christion J. H, Ronald L. C., Guy R. Knudsen, Linda D. S, 2002. Manual of Environmental Microbiology. Printed in the United States of America. 16. Edwards P, 1980. Food potential of aquatic macrophytes. ICLARM Studies and Reviews, No. 5, P. 51. 17. Melcalt & Eddy. Inc, 1991. Wastewater Engineering, Treatment, Disposal and Reuse. Mc Graw-Hill Inter. Ed. Printed in Singapore. 18. Gabriel Bitton, 1999. Wastewater Microbiology. Printed in United State of America. 19. Gordon M.S; Chapman D.J; Kawasaki L.Y. 1982. Aquaculture approaches to recycling of dissolved nutrients in secondarily treated domestic wastewaters : IVconclusions, design and oporational consideration for artificial food chains. Vol. 16, No. 1, P. 67-71. 20. Masanori Fujita et al, 1999. Nutrient removal and starch production through cultivation of Wolffia arrhiza. Vol. 87, No. 2, P. 194-198. 21. US Environmental Protection Agency, 1978. Municipal wastewater aquaculture.
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22. US Environmental Protection Agency, 1981. Process Designe Manual for Land Applicaton of Municipal Sludges. 23. Tarifeno-Silva, E.; Kawasaki, L.Y.; Yu, D.P, 1982. Aquaculture approaches to recycling of dissolved nutrients in secondarily treated domestic wastewaters. IIIUptake of dissolved heavy metals by artificial food chains, vol. 16, no. 1, p51-57. 24. Water Pollution Control Federation, 1983. Nutrient Control, Manual of Pratice. Washington DC. 25. Yves Pietrasanta et Daniel Bondon, 1994. Le Lagunage Ecologique.

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