A STUDY OF HYDRODYNAMIC CAVITATION AS A METHOD OF WATER TREATMENT Candace Jane Botha ‘A thesis submitted in partial flftment of the requirements of the degree of Master of Science Hl in Engineering inthe Department of Chemical Engineering University of Nata Pollution Research Group Department of Chemica! Engineering University of Natal Durban September 1993Declaration of Candidate 1, Candace Jane Bota, declare tht ules indicated, tis dissertation is my own work and that it has not been submitted for a degeee at another Univesity or Institution. September 1993,Abstract With che quality of water declining de to pressures From a growing population and industy, the increasing demand on water resources and the coatoversie regarding the use of commonly zed disinfectants in water treatment proceses, its necestry to research new water treatment Droceses. A potential process is the Cav-ox process which involves a combination of ultraviolet irradiation, addition of hydrogen peronide and hydrodynamic cavitation, Cavitation isthe formation and colipse of vapour cavities in 2ligu. ‘The collapse oF implosion of these cavities creates high localised presures and temperatures. Cavitation can te induced ultrasonically, hydrodynamically, optically and on a particle evel, Hydrodynamic cavitation ‘occurs when the flow conditions create pressure variations in fluid, Ultrasonic cavitation involves the transmission of sound waves through a fluid resting in alteraate compresion and rarefeaction of a fluid due to pressure fluctuations, creating vapour cavities. Tn water the implosion of the vapour cavities results inthe Formation of hydroxy radicals, Ulirasonic cavitation can be used asa means of cell iuption snd homogenisation and hasbeen considered 8 8 method of disinfection when combined with ultraviolet radiation. As the effocis of ‘ultraviolet irradiation and hydrogen peroxide on microorzanisms are well known, the object ofthis investigation was to determine the potential of hydroyamic cavitation (generated in 1 none) forthe treatment of potable water. Hydrodynamic cavitation can frequently be enerated in existing sytems without the need for adsitional power supely. Initially a hydrodynamic cavitation rig was designed, built and evaluated in terms of nozzle flow characteristics, sound emission, cavitation number, hydrogen peroxide production and. bactericidal effect. The cavitation numbers ranged from 0,5 10 0,03 and the sound pressure levels reached 82 dB at 4 000 He. At the same frequency, 58 dB of sound was emitted by the ultrasonic bath. Hydrogen peroxide production by the cavitation rig was below the etection limit of 0,06 ma/t whereas 0,1 mg/t of hydrogen peroxide was produced by the Ultrasonic bath, The effect of hydrodynamic cavitation on rw water bacteria was determined by pumping raw water, obtained from Wiggins Water Treatment Plant, through the cavitation fig. Sundard microbiological techniques were wed to monitor any changes inthe number of viable colony forming units over varios periods and intensities of hydrodynamic cavitation Control experiments were undertaken in which raw water samples were ieeaed a an ultrasonic bath, Hydrodyeamic cavitation was found to te inappropriate for the inactivation oF raw water bacteria for water treatment a5 no reduction in colony forming units was detected. However scanning electron micrographs indicated that clumps of bacteria in matter became dispersed ‘when hydrodyamically cavitated. Significant inactivation of aw water bacteria (1,6 to? togreduction units) oocured after treatment in the ultrasonic bath. was further concluded that due wo differences in scale and residence time, the fesults ‘rom hydrodynamic cavitation were ot directly comparable with thote obtained from ultrasouic cavitation, Experiments combining hydrodynamic cavitation and ultavole: radiation were undertaken in order to atsess whether the dispersal of bacterial clumps would enkance bacteria activation by ultraviolet radiation. Te was concluded that there was mp significant enbancement of inactivation compared to only ultraviolet radiation. UY alone reduced the population from 1,5 t0 22 tog unite, andthe combination procestes resulted in similar log reductions (between Vand 2 log units). The effect woold be more pronounced in water containing a higher concentration of suspended ageresatesAcknowledgements | would ike to thank “The Water Research Commission for the funding of the proie: Diaree Diamond Sales for supplying the Cay-ox rg. “The Foundation for Research Development for Financial assistance during the course of this Prof C.A Buckley for hie supervision and advice, De FG, Neytell de Wilde or support, advise and encouragement throughout the project. Mr I Kaiter ofthe Department of Mechanical Engineering for asistance with the design and construction of the hydrodynamic cavitation vg Dr BS. Martincigh ofthe Department of Chemistry for the wie ofthe ultraviolet lamps and ‘help withthe hydrogen peroxide determination. Prof A.D. Broadhurst of the Department of Electronic Engineering forthe advice on the measurement and interpretation of sound pressure loves ‘The Electron Microscope Unit of the University of Pietermaritzburg forthe assistance with the scanning electron microscopy (Me 1 Bailey and Ms J, Abbott of Umgeai Water forthe help with microbiological techniques. Ms Sue Clemmett For the advice on photochemistry Ms B, Duvel ofthe Department of Chemical Engineering forthe ssistance with chemical analyses, “The workshop staff of the Department of Chemical Engineering forthe construction of evipment. Mre M. Calder of the CSIR for microbiological advice, Dr S, Hareson of the University of Cape Town for information on experiences with hydrodyaamis eavitaton. ‘Mom, Dad, Grant, Natalie and Lindsey for their encouragement and never-ending belie in ‘And Finally 2 special thanks 10 my husband Iva for his constant encouragement, help andNomenclature Symbol Explanation Coss sectional aca of pipe (mn?) Coss sectional area of ovfce (m8) Speed of sound (as) Concentration of disinfectant (8/8) Speed of light (0/3) Discharge coefficient (-) cont ime of disinfectant (3) Death rate () [Number of surviving orgavisms (-) Original number of viable organisms (-) Sound pressure (Pa) Threshold of hearing (iP) ‘Local fluid state pressure (Pa) Sutorated vapour presure a Fi tamperanir (Pa) Volumes flow rate (m9) Rasivs of reverberation Time (8) ‘Volume () Power (W) Cavitation number (-) Local fui velocity (0/2) Fluid deasity (kg/m)aor DwtF ec xTu Pps SEM SPL spc HM TCs wno wwrr Advanced Oxidation Process Drinking Water Health Effects Task Force Heterotophic Plae Count Nephelometsic Turbidity Unit Hydroxy radical Phosphate Buffered Saline Scanning Elector Microscopy Sound Pressure Level Standard Plte Count ‘Trinalomethane Dose of virus rasired to cause $0 infection in tse Utraviotet Radiation World Health Organisation Wigsins Water Tatment PlantGlossary Adenosine triphosphate (tr) ‘Aerobe Acar Antibody Autoclave Autotroh Bacills actericldal Bacteriophage (ohate) Carcinogente Chemolithotop Chemotronh Coliform bacteria ‘A compound containing purine, a pentose, and three phosphate groups; a major carrer of phosphate and enersy in iological systems, ‘A microorganism capable of growing or metabolising in the presence of Free oxyeen ‘A polysaccharide obtained from Celli (ed alga) and other seaweeds use as 2 sliitying agent in culture media |A microorganism capable of growing or metabolsing inthe absence of free oxygen; may be facultative or obligate (will, Dersh in the presence of Free oxigen. ‘A substance of microbisl origin tat Kill of inhibits certain other microorganisms, ‘An apparatus for steising by seam under pressre ‘A microorganism that requires only carbon dioxide and other Inorganic nutrients for growth [Any rod-shaped tacerium, or mote specifically the genus Bacilus Capable of Kiling bacteria. specified time (usualy les than ene how). ‘A buactril virus, ie. a vtus that infects bacteria [A cancer-producing substance. ‘An organism that obtains energy though chemical oxidation and vsesinorgani or mineral sutstances as electron donors, ‘Aa organism that obtains energy through chemical oxidations 138 oppo to photosynthesis, All aerobic and facutaively anserabic, gram negative, nonsporeforming bail that profuse acid and gas from the fermentation of stoveCtoplasm Deoxyribonuctec x (ova) Death Disinfect Disinfection Escherichia colt Becarytie cell Eatrophie Facultative (asacrobe) Fluorescence microscopy Gram stain Gram-negative [A macroscopically visible growth of microorganisms on 2 solid culture medium, Any growth, population, or cultivation of microorganisms, ‘The protoplatm of a cell exclusive of she mucteus. Single of double-stranded macromolecular chain of. nucleotides, the sequence of which determines the genetic code. Loss ofthe ability wo reproduce ( inactivation), ‘An agent that destroys the vegetative cells of infectious ‘microorganisms Destruction (or removal) of organisms capable of causing intestion. Faecal coliforms which tet iadole- postive at 44,5 °C Indication of faecal contamination A type of cell which has @ true noses be, a wel defined nuclear membrane Refers to waler which i high in strents and hence has excessive plant and algal growth, ‘Microorganisms that grow under either aerobi or anacsobic conditions, Coliform bacteria that produce gut From lactose in special buffered broth incubated at 44,5 °C, Indicator of Faecal The immediate emision of visible ight from a substance Which has absorbed radiation of ¢ shorter wavelength from mother source, ‘Microscopy in which she microorganisms are stained with a ‘iyorescent dye and observed by llumination with ulrevolet Tight {A diferent stain bated on the inability f certain bacteria 10 be readily decolourised with akobol once they have been stained witha combination of crystal violet and iodine. Bacteria which are readily decolourised with sicohol ia Grams method and will then pice up the colour of theGram-positive Heteroroph “Hleterotrophie Plate Count Inactivation Inoculation Mono Most Probable Number (pny Mutagenic substance Mutation Obtigate Organetroph Oxidation Pathogeate Peptidoglycan Photolithotrooh Bacteria which are not readily decoloursed with alcoko, thas fetaining the volt colour ofthe cxystalviolet-iodine dye comple. “The effect of toxins onthe liver ‘A microorganism that requires carbon in the organic form, ie. CO, cannot be used asa sole carbon source ‘An enumeration technique which selects for nerobic heterotrophic microoreantims. Alo referred to as Standard Plate Count ‘Loss ofthe ability to reproduce (ef death). ‘The invoduetion of microorganisms, infective material, of serum into culture media ving animal or pl Material (microorganisms, wnat] used ia the inocuaton, Within a cet {A disaccharide consisting of glucse and galactose, {A microorganism that requiesoaly carbon dioxide and other Inorganic nutrients for growh ‘A very specific antibody eg 1 a specific drug. Statistical estimate ofa bacterial popvltion through the use of éilution and moltine tube inoclatins. [An agent capable of exusing mutation [A genetic change, when tranemited to offepring gives se to [A effect onthe nervous system by x substance [Necessary oF required ©, oblignt arabes grow in the presence of free oxysen [A microorganisns that requites carbon in the organic form. [A chemical reaction in which a sabstance loses electrons or a hydrogen atom or gains an oxygen atom, arable of producing disease |A unique polymer (ound in the cll walls of procaryons Consists largely of alternating units of N-acetsiglucosmine and N-acetyimuramic ac, f0 which short peptide chins are attached to form a course molecular network, ‘An organism tha cbiains energy through photosynthesis and can utilise carbon dioxide ais sole source of earbonPhysicochemical Procaryote ell Raw water Ritonsceic acid (nay Standard late Count Steite Ste Strain Streptococel Seblethat Yeast Rofers tothe physical (8. pH, temperature) and chemical (ea, nitrate, mercury) characteris of water. ‘A type of call Icking a nuclear membrane and having other features which ditinguieh it from evcarytie calls, eg, Water in an untreated sae {A single or double-stranded macromolecular chain of nucleotide, the sequence of which specifies the order of Amino acids in polypeptide syntheis. There are three (spe riboromal RNA, mestenger RNA and transfer RNA. ‘A type of resting cll characteris by its resistance to ‘unfavourable environmental condton®, All microorganisms which product visible colonies on non-selective medivm. Indicator of the general ‘microbiological quality of the water Used for monitoring efficiency of water treatment processes. ree from al living organisms (ineluding vrases) ‘The proces of destroying or removing al lving organisms (and viruses) ‘A group of organisms within a species or variety, characteriad by rome particular quality (eg colonial morphology, toxin production. Spherical eels (oss) occurring in long or short chins The concentration or dose of a toxic substace below the ‘hreshold which causes death ‘An organism that grows a tmperstre of 45 10 $0 °C All bacteria which produce colones with a metalic sheen on im-Endo agar. Indication of general sanitary quality since many bacterin are of faced! origin [Any number of poironous substances produced by certain sicroorganisns, plants and anim, ‘A type of Fungus that grows primarily a single cells, reproducing by budding.Table of Contents AnsTRACT ACKNOWLEDGEMENTS. NOMENCLATURE, GLossaRY (CHAPTER 1 : INTRODUCTION 12 13 uM Water in Southern Africa Microbiological ater Quality Project Objective ‘Thesis Outline (CHAPTER 2 : RAW WATER BACTERIA AND DISINFECTION 24 22 ‘Bacteria inthe Aquatic Environment 2.11 Health Aspects 2.41.2. Water Analysis 2.13. Problems with Analyses 2.14 Microbiological Standards Disinfection of Water 2.24 Theory of Disinfection 2.22. Chlorine 22.2 Effect on Bacteria 2222 By-product Formation 223. Chlorine Dioxide 223.1 Effect on Bacteria 2.24 Chloramines 2.241 Effect on Bacteria 2242. By-prodvet Formation23 24 225 Ozone 22.5.1 Effect on Bacteria 22.52 By-product Formation 226. Ultraviolet Radiation 22.6.1 Effect on Bact 22.62 By-product Formation 2.7 Ultrasound 22.2.1 Effect on Bacteria 22.1.2 By-product Formatior Synergism 2.3. Ouonation and Ultraviolet Radiation 2.32. Ultrasound and Ozone 233 Ozone and Hydrogen Peroxle 2.34 Ultrsound and Chemical Disinfection 235, Ultrasound and Ultavotet Radiation 2.34 Hydrogen Peroxide and Uitrviolet Radiation 23.7 Ozone, Hydrogen Peroxide and Ultraviolet Radiation 2.38 Hydrogen Peroxide, Ultraviclet Radiation snd Hydrodynamic Cavitation Conctesion| (CHAPTER 3 : CAVITATION 32 33 a4 as “Theory af Cavitation Hydrodynamic Cavitation Uteasonie Cavitation Cavitation Phenomena 341 Sound 34.2. Chemical Reactions 3.43) Light Bactericidal Effect of Cav 3.5. Ultmionie Cavitation 3.52 Hydrodynamic Cavitation 353. Cell Disruption Methods 220 22 22 223 224 an 228 2.28 229 2.29 23 23 2s 235 2.36 an a436 Conclosions (CHAPTER 4 : EXPERIMENTAL RESULTS : HYDRODYNAMIC CAVITATION 4.1 The Hydrodynamic Cavitation Rig 42 Nozae Characteristics 421 Noctle Characteristic Curves 422. Cavitation Number 423, Energy Disipation 4.24 Sound Pressure Levels 43. Chemical Chartteristion 431 Hydrogen Poroxise Calibration Curve | 42 Hydrodynamic Cavitation and Hydrogen 44 Biological Effects of Hydrodynamic Cavitation 441 Heterotrophic Plate Count | 4412. Results of Heterotrophie Piste Counts 443. Dispersion of Agarepates in Raw Water 444. Scanning Electron Microscopy 45° Summary (CHAPTER § ; EXPERIMENTAL RESULTS : ULTRASONIC CAVITATION 5.1 Physical Configuration 5.11 Sound Presure Levels 52 Chemical Characterization 5.2.1 Hydrogen Peroxide Determination 53 Biological Effecs of Uteasound 53.1 Heterotrophie Plate Counts 532. Results of Heteretrophie Plate Count: 533. Scanning Electron Microscopy 54 Summary (CHAPTER 6 : PROCESS COMBINATION : CAVITATION AND UV 61 Chemical Actinometry 6.1.1 Preparation of Solutions 327 416 416 416 420 4a 423 sa 51 52 ss 56 56 56 8 59 ot 62 626a 63 6.12 Procedure 6.13. Results Ultraviolet Radiation 621 Results ‘Uttasound and UV Radiation 631 Resuks 64 Hydrodynamic Cavitation and UV Radiation 65 cHarrer oa 12 aa 1s 16 CHAPTER & APPENDIX B. APPENDIX E APPENDIX F APPENDIX 641 Results summary 6.5. UV Irradiation 652. Ultrasound and UV Radiation 653 Hydeodynamic Cavitation and UY Ratiaton DISCUSSION AND CONCLUSIONS, Raw Water ad Heterotophic Plate Count Analsis Hydrodynamic Cavitation Uneaonie Cavitation Contrast of Ultasonle and Hydrodynamic Cavitation Combination of Proceses RECOMMENDATIONS, HYDRAULIC CHARACTERISTICS OF SOUND PRESSURE LEVEL DATA ETEROTROPHIC PLATE COUNT DATA UMGENI WATER EXPERIMENTAL PROCEDURES [ENERGY INPUT INTO ULTRASONIC BATH CALCULATION OF POWER OUTPUT OF HBO LAMP 63 63 4 6-4 6-5 ou ou ot mM mM 12 1s 1List of Figures Figure 24 22 23 24 26 a 32 33 aM 35 36 Schematic outine of thesis Distribution diagram for chlorine in water Emission spectrum of the low pressure mercury ae lamp Emission spectrum of the high pressure mercury ate imp ‘A typical death curve for ultraviolet disinfection of secondary effluents Effect of ultrasound and ozone on Staphtococeusaweus and E. colt ‘Treatment of water with a combination of ultasound and ozone Phase diagram of a substance Bubble formation Schematic representation of cas Curves showing areas of cavitation for 82-inch tll valve ‘A sound pressre level meter with a etave filter set Effect of distance on sound meatured in a Free Feld environment and ia a room Process instrumentation diagram of cavitation vig Feed tank, pumps, valves, pressure gauges and transducer [At about 10 m above the free surface was the cavitaion Schematic digeam of cavitation nozzle Hydraulic characteristics of the cavitation nozae Cavitation momber bated on Pau for different inlet eressures Cavitation number based on Py, for different inlet pressures Data from Figure 45 reploted to nica cavitation numbers COperting conditions at which photos were taken Lines of diferent inlet pressures showing eneray disiption (Octave band Frequency spectrum for low intensity of cavitation 2s 2342 43 as aa aut 49 sa s2 53 SA 55 56 sa sa 59 61 62 63 64 6s 66 63 68 69 ‘Octave band frequency spectrum for medium level of cavitation (Octave band frequency spectrum For the highest level of cavitation Hydrogen peroxide calibration curve ‘The effect of low intensity of cavitation on colony formi The effect of medivm intensity cavitation on colony forming “The effect of high intensity on colony forming units in rw water ‘The effect of high intensity on colony forming units in raw water CFU/me for @ 12 b run of cavitation at condition C Schematic diagram showing the temperatere contol stem (Octave band freqvency spectrum For the ultrasonic bath “The conversion of sound measurements from the reverberant sound field tothe free field enviroament Comparison of the octave frequency spectrum for the ultrasonic bath and for the hydrodynamic cavitation rig in condition C Increate in temperature of the slteasoaic bath Log NIN, for rw water sonicated for 2h in the oltrsonic bath Log N/N, for rw water sonicated for 2h in the ultnsonic bath Raw water sonicated for 2 inthe ultrasonic bath Differences in shapes of death curves asa result of the aitferences in starting CFU/m¢ values Death curve for raw water bacteria iradiaed with fight at 254 nay with an inonsty of 0,2 mW em? Sonicated (10 min) raw water Followed by i 254 nm and an intensity of 0,2 mW/em? ation with UY at Sonicated raw water (I min) followed by UV ieradiason (6 min) Sonicaed raw water (1 iin) fotlowed by UV ieradiation (6 min) Sonicated raw water (1 min) followed by UV itadiaion (6 min) or 3 min followed by UV ieraciation Raw water eavtated for 3 min followed by UY iracation Raw water cavitated for 3 min followed by U Raw water cavitated for 3 min ata Jow intensity, fotowed by UV ieadiation “3 43 aa 419 420 st 52 ss 54 35 51 6s 66 66 ot 69 69TL: A modification of Figure 45 showing the postions of the ing condition wed in this project 7-3 ion number relative tothe oper List of Tables | 22 Diseases transmitted by Faecally contaminated water 25 | 2.6; Comparative sensitivity of microorgs 2:26 7: Results of the effect of ultasound of various Frequorces and durations as reported in selected references 16List of Plates Plate a a2 a Photographs 2 1, and of hydrodynamic cavitation Photographs e and f of hydrodynamic cav Scanning electron micrographs of non-cavtated and hydrodynamical Scanning electron micrographs of 1on-soncated and pncated 49 2m staChapter 1 INTRODUCTION Water 1s essential to tf 10 social development and to economic progress, 1s vale 8 well norecaied in undeveloped communities, where many individuals experience the tedium of Dearing heary burdens of water, possibly of dubious gual. oer fone disares to mee tet | ‘basic neds, Yet m developed societies with «higher quality of life the convenient and reliable | ‘availability of rlaively cheap, purified water seems often tobe taken for zranted. Shealé ‘those who ore responsibie forts supply ait in ther mission, he quality of Ue wll deteriorate land development willbe Seriausly impaired. (Departent of Water Affairs, 1986) 11 WATER IN SOUTHERN AFRICA Water isa public commodity its not physically protected and is subject tothe actions of water users and water polluters countrywide. South Aftica has an annual rainfall of approximately 497 mm, The eastern and southern coastlines receive most ofthe water leaving large ateas of thecountry receiv | less than 200 my of rainfall anaually, The uneven distribution of water over the ‘country results in difficulty and high costs associated with the distribution of the Surface runoff is the source of most of the available water with a volume of 53 500 milion m?/a (Department of Water Affairs, 1986). The exiting dams have 4 1oal capacity of 90% ofthe Total mean annual runoff. Omer seurces of aalible ‘water are pround water, effluent eure and unconventional sources suchas desalination, rainfall augmentation, reduction of evaporation from storage ares, the exploitation of icebergs and the abstaction of water from the atmosphere (Deparment of Water Alar, 1986) ‘The quality of water is declining asa result of several factors (Department of Water Altaits, 1986) (Salinaton astociated with the intensive wie and reuse of wat, eutrophication, trent whichis accelerated by industry and agriculture (i) Eutrophication isthe discharge of soluble nutrients such as phosphates and trogen into the water which results in the excessive grovth of algae which then has detrimental effects onthe water system.(ii) Pollution of the water by agrochemicals, leachate from mine dumps and andi sites, and metal from industry and other Sours. Efforts most be made to reduce the factors which cause a destine in water quali | addition 10 this, 2 balance berweon water supply and water demand must be maintained through effective management (Department of Water Affairs, 1986) Measures to maintain the telance have been suggested by the Department of Water [Affairs (1986), These include the efficient storage of water to case net veld; the reduction of evaporative loses from dams as approximately 27 ofthe water delivered to dame hasbeen lott due to evaporation; the potential of underground storage of water and the augmentation of water from ground water sources to be researched; the reservation of catchments by effective management; the hydrological planning of catchment development the contol of Heading the dalination of sea water ab an fea of esearch; the reuse of water, particularly by industry, savings in urban water use and the prevention of pollution, both industrial and sewage, ‘South Africas popsltion is increasing and the effect of increasing rral populations living slong the water course it intensfying. There ral communitos lack adequate sanitation, which leads the dichargeof sewage int watercourses and resulis in polluson. “The contamination of this water rele inthe occurence of disease associated with a oor quality of life, The pollted water, sored in the impoundments, becomes more Aifficutt and expensive 19 teat, The Department of Water Affair and Forestry recognises the problem and deals with policy decisions in a draft report on Water Supply snd Sanitation for Developing Communities (1991) where itis stated: the provision of water supply and sanitation services to developing communes is st to become a ey issur inthe management of ‘he water resources of Sout Africa. It alo considers the reasons forthe need for water of an acceptable quality The Department accepts that formal and informal communities warrant & supply of potable water for base human needs, and a sanitation srvce forthe disposal of waste Ina manner which does mot pose a helth risk tothe commuenty or which s detrimental {forthe envionment “Theres doubt as fo the quantity of water constituting abasic supply with the minimum vantity necessary to meet health and hygienie needs being 10 t0 15 € per day, A basic water supply which ie tuficient to mainain acceptable living standards is ‘estimated tobe between 20 and 25 £ per person per day, The Department recommends 4 quantity of not loss than 20 £ per day {nthe soltion tothe problems of increasing water demind and declining water quality, to avenver of action should be taken12 (The more difficult action involves the mamgement of catchments watercourses, This includes the improvement inthe living standard of the communitesin thearess. Thisaction woul result in improved living standards ‘of peopl, less pollution of water, an improved water quality, a redvetion in disease and an improved quality of life. Mor effective control of emissions by industry and the contol of pollution from sgicultare would also have to be achieved. (ii) Revearch into innovative methods of water veatment process. As the distribution of water to distant rural communties is cot, technology can tied fo develop water treatment facilities in cote proximity to the rural communities thus resulting in improved water quality and availability. ‘MICROBIOLOGICAL WATER QUALITY Pollution by sewage, an indicator of poor hygiene and lack of adequate sanitation facilites esults in elevated levels of bacterial pollutants in water. Keown drinking water pathogens include stains of Salmonela, Shigella, enterooxigenic Escherichia uli, Vidrio cholerae, Yersinia enterocolitica and Camiplebacer fe. These organisms ‘may cause diveates ranging from mild gastroentaritis 0 severe and potentially fatal dysentery, cholera or typhoid (World Health Oreansaten, vol 2, 1984). Is important that any new water treatment techaolgies ate effective in the disinfection of such (Chemical dsinfectants are currently used fr the production of potable water however, srowing awareness of posible toxic by-preduct formation has resulted in concerns reparding the heath effects of continued ute of chemical disinfectants. Chlorine is known to produce toxic residuals such as trialomethaes, the consumption of which ‘may result in carcinogenic, mutagenic, developmental and reproductive effects as well 1 neurotoxicity ané hepatoicty (Drinking Watee Hedth Effects Task Force, 1985). ‘The by-products of other agents such a8 chlorine dioxde, chloramines and ozone are ko currently being researched. The sfety of continued use of the conventional disinfection agents is questioned, resulting in researcs to develop alternative water tweatment proceses. Research on the use of witraviolttdisifecton systems, the wse of ozone and the development of innovative technologes is presently occuring. The potential ofthe use of technologies suchas cross-flow mirofiltation and hydrodyramic cavitation are being investigated at the University of Nata ‘The Cav-ox process introduced by Water Group Inc (USA) isa technology designed ‘o combine hydrodynamic cavitation (generated in a nce) and viraviolet (UV) light radiation withthe addition of small amount of hydrogen peroxide (Neytzell-de Wilde and Chety, 1990).‘Ulavotet radiation (UV) (Caiens, 1991) and hydrogen peroxide (Clarke and Knowles, 192) ace successful agents in the inactivation of bacteria. The efficacy of hydrodynamic cavitation for disinfection is uncertain, Consequently the bactericital tffect of hydrodynamic cavitation, produced by a ca this stad, ion nozzle, was selected or Perry (1973) defines cavitation as: the formation and collate of vapour cavities it & ‘loving liquid. A vapour cavity con form enyhere in @ flowing lau where the local pressures reduced to tat ofthe guid vapour pretsur othe temperanse of tke flowing liquid. Hydrodynamic cavitation cure by travelling cavitation, where bubbles form in the liquid and travel with the liquid at they expand and subiequentty collapse (CVoung, 1989). 1¢ has toon predicted that implosion (collapse) of wltrasoncally produced cavitation bubbles can produce instantaneout localised temperatures of $5 000 K and presares ranging from I G00 ate (Susie, 198; Mason, 1990) to 10 atm (Noltingh and Neppiea, 1950 in Hughes and Nybors, 1952). Cavitation is produced by ultrasound when the amplitude of the pressure variations (as a result of sound waves) is large enough to reduce the local presure to, or below the vapour pressure, As a result, minute bubbles and cavities form inthe sonicated Tiguid (Young, 1989). Ultrasonic treatment ata frequency of 26 ki is known to rede two Forms of eavitation, ie, trtsient cavitation and stable cwvitation. During transient cavitation the bubbles expand to 2 to 3 times their resonant size before imploding violently (Riese etal, 1990), Transient cavitation conotes a relatively ‘lent ctvity (bubble collapse) in which areas of high temperature and presure ‘ocur in very short (in the order of microseconds} bursts at highly localised locations in the toniates medium, These implotons may be accompanied by lcalised shock saver and/or the generation of highly gestive chemical species. In contrast, stable cavitation (the ess violet form) is associated with vibrating gastous bodies which Ihave been reported to have radii at intensities of 1 MHz and 50 kl of 4 ym and 0 yom eespactivaly (Bistrot a, 1000). The nature ofthis Foem of eavitaton consists of gaseous body that romaine sabilited within and, because ofthe ultrasonic field, silts or pulstes, When such volumetric oscillations are extablisted, the liguilike ‘mediam immediately adjacent to the gae bubble flows or stteams (termed Iicteetreaming), Mierostreaming har been shown t produce stessts sufficient 10 isrut cell membraaes (Schecba eta, 1991). The method of destruction isnot yet known although several authors has suggest mechanical sree, shock waves and shear stresses as posible mechasiams. ‘Hydrodynamic cavitation withthe formation an collapse of vapour cavities has the potential to produce shock waves, free radicals, turbulence and shear stresses which may inactivate bacteria in a similar manne to wltrasonic cavitation.13 14 PROJECT OBJECTIVE “The objective ofthis project was to determin te potential of hydrodynamic cavitation in water treatment (the production of potable water). To achleve this goal it was necessary to design a cavitation rig using a perspex repia of the nozae supplied with the Cav-on system, The effect of cavitation on raw vater bacteria was determined by enumeration of viable bacteria (using the Heterotophic plate count techniaue) before and after cavitation, As ultrasound generated in an ultrasonic bath produces cavitation, it was wsed as a compariton, The primary sim of the investigation was Tro determine the efficacy of hydrodynamic cavitation as @ means of inactivation of raw water bacteria. This was achieved quanttaively (By enumeration) and ‘ualatively (by seaming eleciron microscopy). The secondary aims of the project invelved the Comparison of atrasonc cavitation and hydvodynomic cavitation in terms of the Sound emited, the production of hydrogen peronide and the effect on raw water bacteria, Based on the results obtained forthe efficiency of hyérodynamic cavitation alone at 8 potential technique for bacterial inactivation, it was Secided to Undertake a preliminary investigation ta determine whether hydrodynamic cavitation had potential for we ix water traument when combined with uraviolet radiation, sing ultrasonic cavitation asa comparison. ‘THESIS OUTLINE Figure 1.1 shows the outliae ofthe thesis ‘Chapter 1 desls with the introduction to the projec: and highlights the need for tneatment of water in Southern Afric. This chao also indicates the niche for innovative water treatment technologies. Chapter 2 details the health aspects relating to water quality nd the conventional water disinfection technologies, highlighting the benefits and possible problems with the processes. The microbiological rechnigues ‘of water analysis are introduced and a varaty of techniques are reviewed, “The theory of cavitation produced both ulzasoncaly and hydrodynamical is presented in Chapter 3. This chapter describes phenomena of cavitation and the effect ofWATER QUALITY AND AVAILABILITY IN SA. (Chapter 1) RAW WATER BACTERIA AND DISINFECTION {Chapter 2) CAVITATION (Chapter 3) EXPERIMENTAL SECTIONS HYDRODYNAMIC CAVITATION ULTRASONIC CAVITATION (Chapter 4) (Chapter 5) puYSicat CHEMICAL powoicat _pHSICa, SHEMCAL. BIOLOGICAL Rig" Hydrogen * Death * Dimensions * Hydrogen Death tape Peromide” curves. gp pereride carves production , production + Curves sem Se PROCESS COMBINATION (Chapter 6) + ultraviolet radiation alone + Hydrodynamic cavitation and ultraviolet radiation * Ultrasound and ultraviolet radiation DISCUSSION AND CONCLUSIONS ‘Chapter 7) RECOMMENDATIONS {Chapter 8) FIGURE 1.1: Schematle outline of thesis (Chapter 4 cossttues the fist experimental sector, desing, with hydrodyas cavitation. ‘The physical configuration of the hydrodyenmc cavitation cgi described long wih the experiments on the hydraulic characteris of the system, The tigvas characterisedin terms of sound emitted (in sound pressure levels, SPL), hydrogen peroxide production and the effect on the vibiity of maw water bacteria. Scansog electron microscopy (SEM) provided a qualitative asessment of the effect of hydrodynamic cavitation. “The experiments involving ukraiound are described in Chapter $. ‘This workinvoves the characterisation of ultrasonic cavitation in terms of the sound emitted, hydrogen etonide production and the effect on the viability of aw water bacteria. Scanning lectton microscopy provided a qualitative indicatior of the effect of ultrasonic Since hydrodynamic cavitation was found to disperse aggregates in raw water, making the bacteria more accesible to the action of disinfectcn agents, Chapter 6 describes the combination of hydrodynamie cavitation with ultawilet radiation. Chapter 7 contains the discussion and conclusions while the recommendations are presented in ChaptesChapter 2 RAW WATER BACTERIA AND DISINFECTION Conventional water disinfection practices are reviewed and axed in terms of by-product formation. Alternative methods of bacterial inactivation ae popular research topics. Sever methods are described including vanced oxidation involving the production of hydroxyl radicals and the combination of several procesies such a wltraviolt radiation, hydrogen peroxide fand ozone. Innovative, non chemical means of disinfection, su2h as the Cav-ox process, re introduced, The current microbiological standards ae reported. The chapter concludes by highlighting the need for innovative, non-chemical water treatment technology. 2. BACTERIA AND Mi CROORGANISMS IN THE AQUATIC ENVIRONMENT “Most water contains sufficient nutrients to suppor a vaety of microorganisms. Some of the nateraly occurring species ate harmles while crrtain species can cause ilnss and death when ingested by humans. Unicelilar organisms are elsified according to interna strvcture ofthe cells. Protss are distinguished from plant and ania Kingdoms by the fact that pros exist 35 autonomously synthetic, unicellular organisms. Eukaryotes havea true nucleus because te nucleus ofeach cel it enclosed within a well-det ned nuclear membrane and is separated from the cytoplasm. Prokaryotes (bacteria) posess a nucleus which is not enclosed or separated from the cytoplasm although the nuclear materia is agarexated Bacteria are protected from adverse conditions by ¢ bacterial membrane. This membrane isthe target of many disinfection agents and tof relevance the inactivation of bacteria by means of ell disruption, a potential effect of hydrodynamic cavitation, Alt cells are bound by a surface membrane known 2 he cytoplasmic membrane, The ‘ytoplaamic membrane ofall organisms is composed ofa lipid layer in which proteins sueinserted. The lipids ate amphoteric (they conan bo hydrophobic and hydrophilic regions) and are oriented so thatthe hydrophobic portions of the molecule Hie within the membrane, from which water is excluded, while ‘he hydrophilic portions are in contact with the water of the aqueous phase on both sides, The ioserted proteins have hydrophobic amino acid residues buried within the membrane, and hydrophilic residues exposed on one or both ses,1 typical bacteria the rigidity nd strength of cell walls are de msily to strong Fires compored of heteropolymers generally called peptidogyeans or mucopeptides. The Deptidosleans consist of alternating units of N-acetylglucosamide and [N-acetylmaramicscid with -1-4linkages. A short pepe which varies in composition butalways contains a minimum of three amino acids alanine, glutamic aid, and either Aiaminopimelic acid or the structurally related amino acid lysine is Kinked to the ruramic acid The structural rigtity of the peptidoglycan is achieved by eros-inkng of the polymers. The type and extent of the cro inkage varies among different species (Frobisher et al, 1974). For example, che peotidogyeans of gram-posive ‘bacteria are more extensively crosslinked than those foand in gram-negative bacteria (Feobisher otal, 1974), The coll walls of gram-negative bacteria are thin (10 f0 15 nm) and make up twenty percent of the dry weight. Gram-positive bacterial cell walls are thicker (25 to 30 nm thick) and make up twenty to Forty percent of the dry weight. ‘The cytoplasmic membrane of prokaryotic css fa dint end sopaablestructre ‘Bacterial cell membranes consist of three layers. The two outer layers are electron dense and about 2,5 am thick, The middle layer, about $ nm thick is less electron ‘dense, The cell membrane ts theee functions: the first is to maintain a favourable Intracellslar osmotic presture as itis an osmotic barier which is impermeable to ‘molecules larger than glycerl, The second function the housing of @ sumber of sctive transport systems, The third functions i to provie asite for many ofthe key ‘enzymic reactions involved in energy metaboism (Frobisher eta, 1974). Most impounded water, regardles of source, contain sffcient nutrients 1 support the growth of various organisms. Algae, for example, require only mineral nutrients from the water and energy from the sunlight. Many bacteria, which may be photolithic for chemolithic, are able to grow in environments wits low nutrient concentrations. (Once the autotrophs flourish, a succession of heterorophs emerge to decompose the rgnnie material of the dond astatrphisslle and other organic matter (Frobisher. 1974). In lakes and rivers fre from sewage polltion the nuient levels are lower than in polluted streams. The number of tactria floating fee inthe water away feom zones Of nutrition, atthe bottom and edges, i limited, These bacteria may include soil saprophytes capable of growing with a small amount of organic and mineral sutstances in olution in the water. ‘The folowing species are representative ofthese bacteria Micrococcus, Flesobacterim, Chromobacteriam, Baciths, Proteus, Pteadomonas and Leptspira, The sil harbours psychropili (can grow at °C) bacteria such as CChromabacerim, Aeromonas, Micrococcus, Protas, Vitro, Clostridium to name a few. Species of Clostridium and other anaerobes, strict and facultative are often found in ‘decaying organic mater atthe tottom ofa lake, dam a river. Sulphur bacteria and sram-negatve, strictly anaerobic sulphate-reducing species such as Desulfovitvio and esl fomaculum are Foundaaa Hea “Microorganisms are the major cause of less associated food and water consumption, The World Health Organisation estimated that in 1980 25 000 pertons per day died from the consumption of contamiaated watr (Word Health Organisation, 1979 in Drinking Water Health Effects Task Force (Drinking Water Health Effects “Task Focce, 1989), Te has been noted that over the lat fiteen years, am average of two new enteric viruses (transmitted via potable wate} have been discovered each year (Gerba, 1984 in Drinking Water Health Effects Tsk Force, 1989). The most common and widespread danger sociated with drinking water is contamination with bacteria, This ean occur either ively or indirectly by sewage, wastes or human and animal excrement. The transfer of enteric diseases can orcur From faecal polltion, i the causal agents are present and living (World Health Organisation, vol 2, 1984). ‘Outbreak of dizense agents i water can occur fr several reasons, presented by Watkins and Cameron (1991) (Many microorganisms, particulary viruses and intestinal parasites, have a lower susceptibility to chlorine disinfection than faecal indicators such a8 . cl. (Reliance upon indicator testing, particularly whore disinfection is wed to mest water quality standards, may give a false sens of security ii) Some microorganisms may be protected from the action of chlorine by oranie times or even asacation with fee Eving amoekae (is) Sudden unexpected contamination ofa water parce may render an effective treatment ineffective. () The integrity of the distribution sytem may be ia question with posible ingress of contaminasts occuring through serve storage reservoirs, fractures for fitings on water maine ana Durst pipes (oi) Many pathogens are difficult to detect and the techriques may be low in recovery efficiency, time conteming, expensive and not conducive to routine (oii) Certain groups within the community serviced by the distribution system may bbe more suceptibie to infection the very young or very old White and Brandley (1972) in Drinking Water Healt Effects Task Foree, (1989) classify water-related disease orzanisms as (i) Water-borne : pathogens originating in Faecal material and are transmitted by drinking water i) Water-washed: orehnisms originating in farce and ansmitted through inadequate sanitation or hygiene(i) Water-based nisms which occur inthe water a¢ a funtion of their fe cycle and ome into dtect contact with humans in wate. (iv) Water-related : microorganisms with life eyes associated it or breeding in wate. insects living “Transmission can eccut by bites or stings from the ‘Hudson eta. (1983) notes the importance of the contol of bacterial growth in water ‘nestment and detributon systems in order wo prevent the growth of chlvine-revistant “Table 2.1 fom Hudson eta, (1983) ss the coliforms of primary concer, organisms. TABLE 2.1: Gol concern In drinking water systems (after Hudson etal 1989) Species Presence wastes of warm blooded animals [Common rosrce Resistance rectors forer eharactevstes Escherichia oli |Womerous in| Fasces of warm |Sensieve co indicate sewage acces, llcoded animate |ebirine, non |potution, considered 3 mocoid, non |itestinal Faecal coliform lensapaulted parasite Klebsiella |Faeces, wine, [Freshwater, [Mucoid or [Potentially mevmonice sputum and fresh vegetables, encapsulated, fopportuistic orsanic-rich wastes of warm harboured in veh waters [blooded anima: [sine or sediments Enucrobacer |Faeces, fewer [Fresh vegetntion, [Forms a mucoid [survives longer closeae than Ecoli {joins and valves lor han E. colt and lor pumps snd [polysaccharide | may be more pineines coating, esisant co jencapssiates —|shlovination Gtvobacer [Widespread in Soil and water [May encapsulate, [May survive frewndt ature and to re grows in flonger than arying degrees sediments in |F.coli and may in itestn rains lve more resistant eset in humane and other animale [Bmerobocter [Faces of Fresh vegetation [May encaprulate [May survive Jacrogenes [humans and fand joins and longer thas other animals salves of pumps 04 pipetines le. col) and may lve more resistant to chlorinationaaa 2s Fagcal pollution of drinking water may introduce a variety of intestinal pathogens ‘which may be bacterial, viral or parasitic. ‘The presence of the pathogens is directs Felated tothe microbial diseases present inthe community. Known drinking water pathogens include strains of Salmonella, Shigella, entrotxigenic Escherichia cl, Vitvio cholera, Yersinia emerocolitca and Camplobeter fetus. These onganisms ‘may cause diseases ranging from mild gastroenteritis © severe and potetaly fatal dysentery, cholera or typhoid (World Health Organisation, vol 2, 1984). ‘The minimom infectious dose varies For diferent pathogens with relatively few organisms of Salmonella typhi causing disease and enteropathogenic E.coli doses of 10 resulting in tines, The sizeof infectious dose varies according to age, nutitionst satus and general alte atthe time of exposure (Word Health Organi 1984). “able 2.2 from Hints some bacterial diseases transmitted ty facaly contaminated water on, vat 2, "TABLE : 2.2 Diseases tranamited by faccally contaminated water (after Frobisher etal. 1974) Onganive Disease ‘Salmonella typhi Typhoid fewer ‘Spare ‘aratyphoid fever (ener Fever) S. rchounneler 'S. cholerar-rae Salmonella spticemias ‘Syphicnarium Salmonella gistroemerits Shigella dysenteriae ‘Shigeo (eilary dysentery) Water Anabssis “The recognition that microbial infections ct te water-borne has ed to the development of methods for routine examination of water, Although itis possible to detect the presence of many pathogens in water, ine methods of evumerstion are time consuming snd complex. It is therefore useful to select a species of bacteria 38 an indicator of the state of the water. It is now common practice to cetct and enumerate organisms normally present in the faeces of man and other mammmls as indicators of excremental pollution (World Health Organisation, vol 2, 198). Tae following requirements exist for bacterial indicators (The microorganiam must always be precet in large numbers (Carson, 1991).26 i) They must be easily detectable with routine methods (Carlson, 199) Their survival period should at last correspond to that of the pathogenic organisms in question (Carlson, 199) iv) Indicators should not multiply in water (Borrepy et 1980) CCarzon (1991) notes the difficulty of the third requirenent as it hasbeen found tht often pathogens are infectious for a longer period of -ime in the envionment thin the indicator bacteria. Asa result negative Findings for indicator bacteria are often insufficient proof ofthe absence of bacteria pathogem or orally transmitted viruses in water, A study by Camper et al, (1991) indicated hat growth of certain bacteria including E.coli, did occur in water in distribution stems. Ifthe proliferation of ‘certain microorganisms occurs in the water distibutionsystem after teatment, health risks can reach epidemic proportions. Am accepted indicator ofthe qulityof drinking water i Escherichia clio coliform bacteria Coliform organisms refer 1 sram-negaive, rod-shaped bacteria capable of growth i the presence of bile salts or ober surface-actie agents with similar growth-inibting Lbroperties and able to ferment latote at either 35 °C o 37 °C with the production of ‘eid, Ros and aldehyde within 24 to 48 hours. They are also oxidase~negative ad tnon-sporeforming (World Health Organisation, vol 2, 1984). Those with the same properties ata temperature of 44°C oF 48,5 °C are descibed as faeal(thetmotoerant) organisms. ‘There are varios methods of analysis of water for che detection of the indicator ‘microorganisms, the most accepted methods being the multiple-rube method and the rmembrane-filtration method, (The maltiple-tabe method is comprised of to separate tests. A series of tubes containing a suitable broth cultre medium isinoculated with est portions of a water sample. After incubation, each ube showing gas formation i ‘garded a8 prevumptive poste, asthe same gat fesction could nave been produced by other organisms or combination of organisms. A subsequent confirmatory testi requiced, For the second test, a moce selective eultre medium is inoculated with the material obsined from the positive toes. Tubes are egain examined for gus formation. The concentration of bacteria inthe sample can then be estimated from the umber of tubes inoculated, and the number of positive tubes obtained inthe coafiematry test. The most probable number (MPN) of bacteria present can then be extimated using specially devised statistical tbles and the results are recorded at 95 % confidence limits (World Health Organisation, vol 3, 1985), ‘The MPN procedure i applicable to ail water types, especially waters of which turbidity. The equipment is relatively inexpensive and positive results are« iy ww “ cexty to read, The revuts require 48 h, ierespective of waether a negative or 8 presumptive positive rele i obtained, This technique isan estimate and is therefore subject to errr. The membrane- filtration technique enabes the determination of direct count ‘of total coliforms and faecal coliforms present ina water sample, The method is based on the filtration of @ known volume of water through 0,45 sm pore Wert. Bacteria present in the water are retained on the ‘membranes and the membranes are incubated on a stlective. medium. (Characteristic acid or aldehyde producing colonies develop on the membrane and are counted as coliform or Faecal coliform organisms. Usually the sample is Filtered through two membranes and one incubated at 35 or 37°C and the ‘other ar 44 or 44,5 °C. The count it simplified as a direst estimate of the number of fusca coliforms present is possible at 44,5 °C. “The heterotrophic plate count (HPC) or the standard plate count (SPC) i @ ‘method used to determine the general bacterial count of weter or wastewater ‘often by means ofthe pour-pate procedure. It san estimate of the bacteria able to row at 35 °C. Viable cals ae detected on non-selective medium, Generally natural water bacteria grow between 20 1930 °C, Bacteria fom soil and faecal material grow at 37°C (Holden, 1920 in Geldeshuys, 1978). I many bacterin grow at 22°C this can indicate organic pollation. High ‘eounteat 37°C indicate faecal polltion (Cruikshank el, 1975 in Geldenbuss, 1978), Am important aspect of tht mathod is the changin counts at the Aitterent temperatures at different times atthe same place. This method is ‘most commonly used as a means of mositocing the effiiency of a water treatment proces (Grabow, 1983 ia Department of Water Affairs and Forestry, 1993), The Clarks presence-absence test wes single fermentation bottle containing the mediom and a sample volume of either SO or 100 me. The botles are incubated at 35 «0,5 °C for up to five days and examined daily for growth (turbidity), acid preduction and gas production (Pipes eta, 1986), The Colilert and Coligelk methods wee two sctive substrates, o-nitrophenyl-P-D-galactopyrancride (onrc) and ‘4 methylumbeliferyl-f-D- glucuronide (MUG), to simultaneously detect total coliforms and £. call, Total coliforms produce the enzyme P-galctosidase which hydrolyses ONPG and releases o-altrophenol which produces a yellow colour. £. coli produces the enzyme glucuronidase, which hydrolyses MUG to form a flvoreseent compound (Oise etal, 1991). This method determines the presence (by the change in colour) or absence of the bacteria in questionaa oa) ity oa ‘An immunological method using monocbnal antibodies 10 detect fenterobacteriaceae in drinking water was studied by Obst et al. (158). “Antigenic patterns of bacteria ae constant and do ot fluctuate asa result of environmental conditions, An enzyme-linked immucosorbent assay (ELISA) ‘ses a monoclonal antibody aginst he enterobctril common antigen (ECA) Which is part of any representative of the Erterobaceraceae, ECA it Tipiotysaccharse whichis inked tothe outer nembrane of the bacterial cl [A defined amount of antibody i mixed with the sample. The ECA and the ‘otibody bind, A defined amount of anti-ECA antibody is then added. The EECA ofthe samples trapped between antibodies andthe added enzyme linked antibodies. A fluorogenic substrate is then added. The antibody-Tinked ‘enzyme degrades the substrate and 2 dye is released. The intensity of, ‘orescence is corelated tothe amount of ECA-Linked enzyme. This s then related back to 8 standard such atthe Fluorescence from a known amount of Ecol The dicect viable count method, used by Adum etal (1990), involves the addition of yeast extract and naidinic acld © water samples followed by incubation (6), then Fixation in formali, sting with acridine orange and txamination by epifluorescence microscopy. In bacteria where DNA replication is inhibited, ool elongation can occur ut not cll division. This ‘means thatthe cells ar stil iving athouen they may aot be cultrable on selective media. ‘This method hasbeen proposedfor the purpose of determining viable cel, i, the cells which ae atleast thre times their normal size. This surmouets the problems of overestimation ofthe number of viable bacteria ‘when direct mieoncopie methods are used (Roszak and Colwell, 1987). The procedure can determine both viable clterasle and viable non-culturable ‘bacterial cells (Adams et al, 1991, pathogenic microorganisms (Berry and Norton, 1976, Scarpino, 1978; Borrego tnd Romero, 1985 in Borrego etal, 1990), it may be more reliable to use ‘hem as indicators of fecal pollution, In edition, Vaughan and Mtealf in ‘Borrego etal. (1990) claim chat quantitative plage assays ere cheap, easy and rapla Problems with Analyses Disadvantages of the analyses are ® Lenathy incubation times forthe MPN technique (Clark etal, 1991). This is problematic asthe health authorities need immediate nottiation should a waterbody be dangerously contaminated (Obstet al, 1989).(Gi) Potential interference by beterotrophic plate count bacteria (Clarke 1981) Interactions between heterotrophic pate count bacteria and coliform organisms can result in injury to the coliform population. This can reduce coliform ‘densities (LeChevaliee and MeFeters, 19850), A reason forthe reduction in coliform court could be competition for limiting organic carbon, (ii) Ditticulties in interpreting results (Clark et a, 1981). This cam be due © 1 From problematipltes nd conclusions ase¢ on indicator species (jv) Menttication of stressed pathogenic or faecal bacteria in Srinking water i Aiticule asthe physiological characteristics can be lost (Hubner eta, 199) (©) Viable butnon-cultrablebacteriaarea problem withallanahses. Populations undergo decreases in pate count but remain viable when analysed by the direct viable count procedure. acterin known to exist ic the viable but rnon-cuturtble sate are of the genera: Vibrio, Escherichia, Salmonela ‘Aeromonas, Legionella, Campylobacter tnd Shigella (Byrd e ah 1991). Byrd ‘tal, (1991) stae that present methods of monitoring public water supplies that are untreated or havea low free-chorine residual, may underestimate the presence of coliforms or total heterotrophs as some bacteria become injured and do not proliferate on selective agar. (i) Turbid waters cannot be analysed by membrane filtration asthe membrane filters become blocked (World Health Organisation, vol 3. 1985). (Wii) Toxic substances which may be present in the water may be absorbed by Iembrane filters and affect the growth of the coliforms (World Health Organisation, vol 3, 1985). (ill) Analyses often do not allow sufficient time or nutrients of the recovery of injured bacteria. Bacteria intl existing with adequate autrent to support frowth, appear to adapt to low mutrent enviconmens (By et aly 1991) (ix) The HPC does not give good indication ofthe genera land of aerobic and facultative anaerobic hetefotrphic bacteria in the water as the conditions selec for bacteria which may comprise only a small percentage ofthe bacterial population present (Reasoner and Geldreich, 1985) (%) _Tresetiance on an indicator may nt bean accurate indicaton of the presence of pathogen (Watkins and Cameron, 1991). | factor in the analysis of indicator bacteria for determination of the safety of drinking “water isthe problem of injured bacteria (Bisonnete eta, 1975; uckin eta. 1991; MoFeters etal, 1986; LeChevalier and MeFeters, 19853). Injury is the sub-lethal physiological consequence of exposure to stresses which enuse aks in the ability of ‘microorganisms to grow under normal conditions that are satisfactory for uninjured calls (LeChevallier and McFetes, 1985s), Bacterial injury is often defined as the inability of sub-lthally injured microorganisms to form colonies ona normaly suitable210 ‘medium, whilst retsining colony forming ability on another medium, such 38 2 non-selective medium (Hurst, 1977 ia Verville and Heron, 1989). Bacteria become stresed in the aquatic envionment asa result of the low nutrient conditions, Ta ‘water eatment processes and drinking water systems bacteria become stressed a 2 result of chemical ad physical factors (MeFeters eta, 1986). After exposure t0 ‘sresful factors, injured coliforms are susceptible to ingredients suchas desonycholte ‘and bile salts in culture media, The result of tigi tat certain bacteria are viable i, able to reproduce, but are non-culturable on selestve media, False-nepative resus are obtained which imply that fewer bacteria aw present in the water sample than the nomber that are actualy present, When injared bacteria enter regions of suitable conditions of nutrients nd temperature, eg-an animal gt, they may prliferate and can become infectious, These bacteria have been detected by the use of a new culture media, A stody by Buckin tal (1991) noted that injured bacteria failed to {row on m-Endo mediom but were ensmerated on m-T7 medium, MeFeters eta. (1986) compared two media (ai-Endo agar LES and s-T") and reported that m™-T7 ‘agar yielded eight t thirty elght fold more coliforms than did the m-Endo agar LES, “These stodiessresed that improved bacteriological surveillance be introduced inorder to achieve more accurate eatimatee of bacteria in wate, Jn addition to probleme associated with analysis of war is the problem of survival and growth of bacteria in distribution systems (Camper et al, 1991). Ina study by Camper etal. (1991), of the growth of environmental and clinical, i. laboratory produced coliform bacteria (Klebsiella preumoniae, Escherichia call, Enterobacter ferogenes, and Enterabacer cloacae) under the conditons typical of drinking water Gistibution systems, it was found thatthe Bacteria prolerated at carbon levels low ‘4.2 ma/t dissolved organi carbon, Tt was conclude that sufficient nutrients exist ‘in water distribution systems to support the growth o coliforms Watkins and Cameron (1991) note the problems sutoeited withthe ute of indicators in the verification of disinfected water. Fecal contamination of wat is traditionally eteced by the selective culture on media of indicator Sacera which demonstrate the fermentation of lactose, Such a culture requires the indicator tobe present in excess of pathogens and itis therefore by inference that the water is deemed tobe se for ‘human consumption in the absence ofthe indicator (Vatkins and Cameron, 1991). Microial pathogens consists of bacteria, viruses and intestinal parasites The bacteria cam be divided into two groups (Those which on entry tthe aquatic environment lose ther viability or remain viable but cannot be recovered by standard cultarltechaiques (eg. Salmonella 0p., Campylobacter spp. a04 E.coli. (ii) Those that are able to multiply inthe envieonnent if suitable conditions exist (e., Peeudomonads and Legionella spp.)“The latter group may grow to form biofilms in detritotion sprees, Biofilms re ‘own to accumulate i the presence of 0,8 mg/t of free chlorine and considerable sccumulations an enist at concentrations lees than 0,2 mg/t. — BiofiIms protect ‘microorganime From the effects of disinfectant: and may detach and produce turbid water, taste and odour problems, corrosion and can provide atronts for the growth of other microorganisms Watkins and Cameron (1991) note that changes in climate and rainfall will resut in changes in the microbiological quality of water, both aw and inthe distribution system. ‘They recommend that effort mutt be made (0 identify newly recognised organisms sand that methods for water analysis should be constandy changed and improved Mistablolosical Standards [Numerical Himits nave been applied wo the selected microbiological indicators. The Timits issued by the Eastern European Community (EEC), the South Afeican Bureau of Standards (SABS), United States, (US), Canada, the World Health Orgasisation (OWHO) and the Department of Water Affairs (DWA) ae shown in Table 2.3 where IS represents noa-specified standards according to the references consulted. [TABLE 2.3: Summary of standards of various counties and organlsations (Ian Baily, [Umgeni Water pers. comm, 1983) SABS FEC vus_[eanapal wo | pwa Guide | Max | Guide| Mex | Max [ max | Guide [ Guise vale value value | value fEnterovinwes | NS | NS | NS | ws | ws | NS | WS | 0 (oer 10 mo) nee 100) Faecal ws fos [ows | oo [os | oss [ns [m0 iver 100 me) Pate count | nS | ns | 10 | ws | Ns | NS | NS | AS pac iver me) late count | 00 | NS | 1 | NS | NS ns [NS sc oer mo) Frou colirorms| 0 ) 5 | ws | o | ns | ns | 0 | 0 (pcr 100 l22 2 According tothe Water Quality Guidelines (Department of Water Affirs and Forests, 1993) the microbiological water quality criteria include faecal coliforms or Ecoli at 1 target guideline range of 0 counts/100 mi; coiphages ata target guideline range of (ol counts/100 me andenterc viruses ata target guideline of less than] TCIDyy/10 © ‘The TCIDso teers to the dose of virus required to cause 50 % infection in tissue culture (Department of Water Affairs and Forestry, 1993) 1 must be noted that often different limits apply to different situations ia the water lreatment proces. Umgeni Water ia Natal apply different standards tothe water immediately a K leaves the water treatment works and the water that is stored in reservoirs and that leaves the taps (Table 2.4) (Batley, 1999 pers ) ‘Recommended He Total coliform bacteria per 100 me ° 0 Faecal catform bactaria per 100 me ° 0 Standard plate count per meat 22 100 000 incubated for 3 days [Standard pate count per mt at 37 0 100 incubated for t day DISINFECTION OF WATER Degermination isthe we of physical or chemical agents to reduce the concentration of microorganisms and viruses with the purpse of contro, disinfection or steiizaton. Sterilization refers tothe use of any agents to Kill (inactivate) all vabe microorganisms and viruses within a sample. isifection means the use of germicidal and viruscial gents to destroy the potential infectivity ofa material (his eed not imply elimination ofall viable microbes, eg in drinking water) (Schenck in Lorch, 1987). inking water teatment varies withthe mort common proces involving ls ‘eagulaton, flocculation, sedimentation, filtration and a second disiafecton ste, The process results in potable water which ir defined as water whichis free from pathogenic microorganisms and chemicals that are deleterious to human health (Frobisher ec al, 1978),28 Chemical disinfection is used to destoy or control microorganisms present in water ‘which would otherwise cause fouing, corrosion of equipment, or lead to disease from microbial activity (Heoper, in Lorch, 1987). Disinfection i not sterilisation, ie. not tll microcrganiams are destroyed. The us of chemical disinfection must be undertaken ‘ith care as various Limitations exist between different dsinfectan. ‘Two groups of antimicrobial chemicals exis the oxidising and the non-oxidizing agents. Oxidizing biocides include chlorine, ehloramines, chlorine Sixide, chore, ‘onone, bromide, bromines and certain organic bromine-contaiing compounds NNon-oxiizing biocides include formaldehyde, iso-thisalons, iko-cyanates, bromopropionamides, quaternary-ammonivm compounds, chlovinated phenol, oegano-sulphur and organo-tia compounds (Hooper, in Lorch, 198 The relative efficiency of the varios disinfecting compounds depends om the rate of dittusion of the active agent through the cell wall, Factors affecting disinfection cfficiency are noted by White (1972) (The nature of the disinfectant. (ii)__The concenteation of the disinfectant ii) Length of contact time with the disinfectant iv) Temperatare (9) Type and concentration of organisms (ee Theory of Disinfection Exporure of microorganisms to abnormal physical or chemical conditions will kill or crest growth. Death of the microorganism is defined asthe irreversible los ofthe bility to reproduce, When a mierobial population is expoted to lethal agent, the kinetics of death are usually exponential, This indicates that al members of the population ae of similar sensitivity, Probability determines the tine of death of any sven individual and mixing and contact of microorganism and agent are important. “The Chick-Watton model is the most widely-used tool used to determine microbial inactivation by disinfectants ws (3) = 10% aa t ratio of number of organisa at time = concentration of disinfectant (oPhich must be constant) 5 ‘= empirical constantau k = death rte Ny = initial number of organisms surviving number of organisms laration of sterlstion If the logarithm of the number of survivors is plated asa function of the time of exposure, a stright line should be oblained where the slope defines the death rae “The death ate lone only indicates the surviving Fraction of the inal population over the treatment perio. “The underlying hypotheses of disinfection ae (Langlais, eta. 1991) (The disinfectant concentration, Cis constant during the cesction time, Gi) The microors nisms must be a single strain of synchronous development (ii) The killing ation must be of a single-hit and single-site type ‘Chlorine CChorine i the major disinfectant used in drinking water treatment, Chlorine is vied cither at gateous chlorine of 35a hypochlorite salt (usually sodium). The production (of hypochlorous acid (HOC) is the key agent in disinfection Chlorination of water suppliss and polluted water serves primalily t0 destroy of Instivate dieeae-causing. microorganisms, secondary benefit is the overall improvement in water quality resulting ftom the rettin of chlorine with ammoat iron, manganese, sulfide and some organic substances (AHA, 198). “Total fre chlorine i define as the sum of the concentration of chone, hypochlonoes acid and the hypochlorite ion (de Leer, 1987). Free chlorine exis as hypoctorous acid and the hypochlorite ion in solution. (APHA, 1985), This form of chlorine can be distinguished analytically from free chlorine because of the lower disinfection efficacy (de Leer, 1987) Fige chlorine reacts readily with ammonia and certain nitrogenous compounds to form ‘combined chlorine. ‘The presence and concentration ofthese combined forms depend chiefly on pH, temperature, intial ehlorine-to-ntrogen concentration and reaction time (APHA, 1985). Both fee and combined chlerine can be present simultaneously. Residual cMorine i the chlovine present after disinfection has been achieved during specific contact tim. ‘The chlorine demand isthe difference between the chlorine concentration applied to water and the residual chlrine after certain contact time (de Leer, 1987). “The most important reaction of chlorine in water isthe Formation of hypochlorous sia (HOC)2s “This is cepresented in Figure 2.1 where the pKa of 7,548 shown “The HOC isa “weak” acid which undergoes pata dssciaton a follows to produce 1 hypochlorite jon and a hydrogen ion: hlorite action of total Hypoc FIGURE 2.1: Distribution diagram for chlorine in water (Wadley, 1992) The same reaction occurs when hypochlorite is wed instead of gaseous chlorine [At pH 1,3, the chlorine concentration present inthe chorinated water consists of fifty percent undissociated hypocMlorous acid and Fifty percent hypochlorite ion OCr The most important and complex chemistry of water chlorination i the reaction with various forms of nitrogen, If no nitigenous compounds are present in water then216 Reactions of chlorine with ammonia resus in the formation monochloremine dichloramine and nitrogen trichloride, The reactions occu ia stops invaving the chlorine substation ofeach ofthe hydrogen atoms in the ammonia molecule (White, 197, Its exeetial to understand the behaviour of chlorine solution as te HOCI and OCI have diferent germicidal efficiencies. 2.2.2.1 Effect on Racterla Chlorine acts reversibly with the enzymatic sytem of bacteria (Green and Stumof, 1946 in White, 1972), Wy (1962 im White, 1972) ascribes the effective mechanism | a the phenomenon of untalaneed growth. The destruction of pert of the enzyme system will alter the balance ofthe call and the cell dies. Corne is known to cause Dhysiclogicel damage to cell membranes asthe vee of respiration, lucose transport ‘nd energy (ATP) decreat in injured popslations (LeChevalir and McFeters, 1985s) ‘The form of chlorine preset is dependent on the conditions undet which the init chlorination is undertaken (Vervile and Hereen, 1989), AtpH 4, mast ofthe availible chlorine i present at HOCI, while at pH 10 gssocation to OC is complete therefore the disinfecting power of chlorine is inuenced by the pH at which tis applied. For this reason longer contact times are required For equivalent doses effective atthe pH where HOCL isnot readily formed (Hooper, in Lorch, 1987). Ina study on the effect of ee chlorine on Escherichia coli populations, Vevile and j Herson (1985) Found that a6: 1 ratio offre to combined chlorine caused a comple | population death within 60 s. Treatment with chlorine can injure bacterial cell According to Venkobachar eta, (1977), trestment with chlorine can induce leakage (of mscromolecules from cells thes indicating a change in the permeability ofthe cell membrane, Proteins and RNA were detected in the supernatent when cells were tweated with 1,5 mg/t of chlorine. Studies on the oxidative phosphorylation of the call free extract indicated the complete cesition of phosphate upake at a dose of (0.4 mg & eis concluded thatthe First step i the interaction of cHorine with E.coli is the reaction withthe cell membrane and thatthe changes occurring to the cell membrane after chlorine treatment affect the vital functions of the bacterium, ouame and Haas (1991) studied the inactivation of B.colt by the combined action of free chlorine and monochloramine, —Taey found that when free chlorine and ‘monochloramine exit ina continuous flow system, high inactivation isachieved. They ‘proposed thatthe combined action of free chlorise and monochloamine i synergistic Jetad previous been assumed tha the inactivation rate offre and combined chlorine as additive (Haas and Karra, 1984 in Kouame and Haas, 1991). It was proved by ‘Kouame and Hats (1991, thi the combined rate of inactivation was greater than the rates resting from the two individual forms of chlorine, They suggested that freeaaaa | Chlorine is capable of causing sub-tetal injury in coliform bacteria (Camper and MeFeters, 1979 in Kouame and Haas, 1991). If enhanced sensitivity monochloraine is manifested by sub-lethal injury (or visa versa) then synergism is occurring, Hypochlorous Acid (HOCI) isthe most gormicidlly effective form af chlorine, The structure of HOC! is similar to that of water. "Tho germicidal efficiency is due tothe relative ease with which HOCI can penetrate the cell wall. This eccurs because of itslow molecular weight and electrical aeurality which is comparablto that of water ‘The germicidal efficiency of free availble chlorine i also a function of the pH at the pH which establishes the amount of dissociation of HOCI to H* end the OCT- ins The Hypochlorite ton (OCI-) is a poor disinfectant dve 10 is iaailty o diffuse through the cell wall asa result ofits negative electrical charge. Furthermore, the inactivation energy for disinfection by HOC! is 7000 calories, whereas OCI has an inactivation energy of 15 000 calories. The disinfecting efficiency of this form of ‘chlorine decreaes with increasing pl By-product Formation CChorine reactions with naturally occuring humic and fulvis acids ia water produce toxic residuals during the chlorination process (Oliver and Cares, 1976). The reactions result i the formation of tialomethanes (THM). The term Total THM refers to the sum of the concentrations of chloroform, tromadichloromettane, Aibeomochloromethane and Womoform which are produced, The concentration of the chlorination by-products vary according to pH. ‘The relative concentrations of chloroform, riehloreacetie acid and dichloroaceie acid then vary with respect to one another (Drinking Water Health Erfects Task Force (DWTF, 1989). “Trihalomethanes are known to effect the liver and kidneys of vatious animals with ‘he severity ranging ftom mild serum changes 10 fatty infiltration, necrosis and irreversible damage. Decreased immune Fusction has also been revorted, The most foted effect resulting from exposure to THMs, chloroform in partcslar, carcinogenicity (Orme et al. in Jolley et a, 1980) hloine Dies CChorine dioxide is wied as bleaching agent inthe paper and pulp industry. Iisa ‘deep yellow gas which is toric under certain conditions and may seact explosively (APHA, 1985). In aqueous solution i is harmless, It is ely expalled from solution by aeration Chlorine dioxide vesct with water a follows Chlorine dioxide does not react with ammonia and therefore tihuomethanes are not formed during disinfection, The predominant resction of chlorine dioxide in water Js the reduction to chlorite at pH? and above28 Since most of the reactions involved in water Weatment take place within the natural water and wastewater pH range of 710 8, the toxic chlorite fon is a major C10, A, agtin trangretsing the co-existence line wile still not ‘vapourising. Such a liquid in te state A is now stid tobe superheated and is again ina metastable phase. Ifthe liquid is overstretched along the path Ay ~ A it reaches ‘he limiting tension it can withstand and cavitates, ie it vapourises, In sila way ‘Tiga taken along the path Az +A will eventually reach its superheating limit and will vapours. ‘Travena (1987) notes the ack of knowledge of the mechanism of the initiation of cavities. He claims thatthe action of tearing the liquid apart requires pressures of ‘thousands of bars (Temperey and Chambers 1946 in Trevena, 1987) which would be unlikely. He suggests that the mechanism relates tothe presence af nucle. These could be inthe form of tiny, free bubbles inthe liquid, The bubbles could disotve de to surface tension, A a revit, two models have been suggested to explain the persistence of nucle in aliquid (A tiguid may contin a large number of minute spherical ras bubbles which are stabilised aginst ditfusion by an immiscible skin f organi impurities. Gi) Gas may be tn ped in tiny crevices in the walls of the coxainer oF pipes “The first model was discused by Fox and Hersfild (1984 in Trovena, 1987), As the bobble begins 1 contact under the influence of surface tension, the internal pessore inereate and the gat will ditfare out through the surface, resuking inthe bubble ‘contracting sil further, tis then assumed thatthe butble has accumulated, by adsorption on its surface an organi film (such as fatty acids) vhich will form a complete skin when the bubble has become small enough and further shrinking is ‘mnbtes, The Dubble 1s then sud tbe stabilised. 1, ata Iter stage, tne tguld Is subjected to tension, the skin wll be torn apart and g88 will diffuse into the bubble as cavitation i initiate, “The second model was First suggested by Harvey eta, (1984 in Trevena, 1987). It is known that the application ofa large positive pressure, of the ader of 1 000 bar, 104 liquid for about half an hour wil eliminate cavitation nucle i the Fguid. When the positive pressures released, i is found that the liquid can then withstand a greater tension befor it cavitates than would otherwise be the ete. According tothe crevice theory, the explanation Lis in the fact that these are tiny crevices both in the walls ofthe contsnes nin the motes (smal solid impurities) present in the Liquid. A pocket of gas (at) trapped in rach a crevice can ext ina stable equilibrium in the ‘cravce ifthe various conditions are eight, Ifthe Figuid is wot pre-pressurise asthe pressure is reduce, rita tage willbe reached at which th presureof the trapped32 Bs ‘85 together with the negative pretsure in the lagi, wil overcome the inward stress ‘due to surface tension, At this stage, the interface whch is chen convex on the fiuid side moves out of the crevice and forms a cavity in the Hild (Figure 32). FIGURE 3.2: Bubble formation occurs upon subjecton of the pressre the liguld-gas interface (2) Is drawn out of the erence and (0) later ‘breaks off Sato the Hild to form a eavty (after Trevens, 1987) HYDRODYNAMIC CAVITATION ‘The liquid velocity varies locally in a Flowing liquid. At the points of highest velocity, low pressures and cavity formation occurs (Young, 1989), There are thre cases of flow cavitation (i) Traveling cavitation: This is when bubbles form inthe liquid and travel with the Fguid 2 they expand and subsequently claps. i) Fixed “This whens pocket or cavity attached othe rigid boundary of an immersed tody or a flow passage forms, and remains fied in position in an unsteady state (ii) Vortex cavitation : Occurs in the cores of vortices which form in regions of high shear. Often oscars om the Blade tips of shi? propellers. Hence the In cavitation nozzle, as in the cavitation vig, wavellng cavitation ovcurs. The ‘vation describing the formation of cavitation i given by Bernoulli where potential ‘energy i conserved du t0 negligible differences in height across the nozzle+6 Pt dev? = constant 32 where = static pressure of the Aid (Pa) = density of the fluid (g/m) Y= velocity of the Maid (75) In studies of hydrodynamic exvitation varius definitions of the cavitation number have been used. Yan and Thorpe (1990) deere a cavitation inception number(s) for an orifice which is calculated from equation 3.1. These authors observed bubble formation at the vera contracta and defined cavitation inception as the first flow regime transition Harriton and Pandit (19923, 19926) studied hydrodynamic cavitation ata means of Airing calle, They used equation 3.1 to ealeulate the cavitation mumbers and noted ‘that cavitation occurs below 1,0 to 2,5, the onset depending on the flow reduction, the sele of the apparatus andthe geometry of the constriction. ‘Tullis snd Govindarsjan (1973 in Ball tal, 1975) conducted expeiments on choked cavitation in ap orifice and they define the eavitation number as follows where °. = cavitation number Py downstream pressure (Pa) vapour pressure (Pa) upstream pressure (Pa) cavitation to the pressure differential or volsity head (Ball and Tullis, 1975) 1, in an expanding flow (Figre 3.3, the local velocity of the id reaches a critical ‘vale, the static pressue is lowered tothe saturated vapour pressure at the temperature of the fuid. At this point, vapour bubbles wil form and, if the flow is diffused, cavitation wil accu ‘The various stages of cavitation have been described by Ball eta (1975). Taetpent ‘aviation mark the transition from single-phase to two-phase bubbly low (Yan aad ‘Thorpe, 1990). ‘Thsrepresents aviation whore the noize consists of light, itermittont Popping sounds, Critical cavitation i characterited by light steady noise similar to frying bacon. Choking cavitation is where the mean pressure atthe constriction reaches the vapour presure of the fluid and there is no further increas in discharge, ata constant upstream pressure, with a decrease in pressure in the downstream conduit ‘Sepereaitation occurs once choking has been reached, if the pressure far downstreamMw is further decrated, the vapour pocket atthe valve increates in size (Tullis, 197). (Choking and supercavittion can both cur at the same discharge fora given value of upstream pressive, The difference isthe size ofthe vapour pocket, the location ‘of bubble collapse and the response of the system. For ehoked cavitation the maximum ‘damage potential exits atthe valve or constriction, Is super cavitation the vale is ‘eseatialy damage Free while the downstream pipe may be damaged (Tuli, 197). V = telacity Prats) Pate (a) Prote(ot Pato (out Vee to) Ve ou Flow Bubble formation collapse ‘Yan and Thorpe (1990) note that choked cavitation can be described by the relation between flow rate (Q) and the differential pressure across an orifice (P,~) [RES V+) area where Cy = discharge coeficent A= cross sectional aren of pie (n?) a ‘= cross sectional area of orifice (nt) P= density of water (g/m?)33 Mt “The intensity of cavitation i 8 function of the jet velocity, turbulence level andthe ambient liquid pressure. By increasing the woostyor decreasing th ambient pressure, the cavitation intensity can be increased. As the cavitation becomes more intense, Increased numbers of bubbles are created and they lend to coalesce into larger cavities (rats, 1971), ‘Supe cavitation i discussed by Yan and Thorpe (1990). They describe thre regions 7 Region A consists of a single cavity witha liquid jet in the middle of 1 ‘apour pocket i) Region B, where white clouds appear where the big cavity breaks into smaller cavities which colape. This repin ean be short (3-5 em), (Gi) Reon C, a clear liquid region where all the cavities have collapsed, Figure 3.4 shows the work by Tuli (1971) whe defined areas of cavitation, N- Nose = Incipient L- Light M~ Moderate He Henry VE Very Henry S- Super casliagon FIGURE 3.4: Curves showing areas of cavitation for a 2-Iach ball valve at Aitterent openings. Q represents flow rate and AP represent the pressure drop across the valve (Tallis 1971) ULTRASONIC CAVITATION “The history of ultrasound (aFreqvency beyond human hearing) dates back 100 years to the work of F. Galton who esblihed threshold levels of heating in animals and humans, He proded a whistle which generated soand of known Frequencies andFe found the norma limit of human hearing from 16 He to 18 KHz. The whistle isan example ofan ultrasonic transducer, a device which converts one form of energy (in ‘his ese gts motion) into another (ultrasouné) (Mason, 1991). “Two ranges of ultrasound have been identified as being of use in chemistry: dingostc ultrasound (for physical measurements) and power ultrasound (to iafuence chemical activity). ltrasound : The first commercial exploitation of ultrasound came aftr the fac diater of 1912 when a competition, organised to find methods of avoiding iccberes, found thatthe distance of an iceberg (rom a ship could be estimated from the time lapse between emitting & sound from a ship and receiving an echo From it. 1k was from thi technique that the first depth gauge was developed and use. Improvements of this led to SONAR (SOund Navigtion And Ranging) and fw detection in metals and other material More recent progres in electonic measurement techniques has resulted in the use of ulaiound in diagnostic medicine for foetal Power ultrasound: Power ultrasound produces chemical effets through ‘phenomenon of cavitation, Cavitation has been described asthe formation and coll ‘of microbubbles in aliquid when a large negative pressure i applied to it. This was first characterised atthe turh of the century by Sir John Thorajeroft and Sidney Barby who foond that due to an incorrect setting of the propeller blades on the serew-drven destroyer H.MS, Daring, the blades ofthe propeller became eroded due to cavitation ozcurring on the suction side ofthe blades. During the rapid metion ‘ofthe blades through the water, the tailing edge produced suicien: negative presure to pull the water molecules apart and create tiny microbubbles fevities), These bubbles subsequently collapsed withthe release of intense focal energy inthe form of high pressures and temperatres, Such extreme conditions produce bond fission for ‘vapours in the bubble or for molecules subjected tothe shock-wave of collapse. The of the blade as it causes erosion of the Blade (Mason, 1991). The mechanical and potential chemical effects of sonication on reactions can be demonstated with a simple ultrasonic cleaning tath (Mason, 1991), Ina non-flowing system the ambient pressure can be varied by sending sound waves through the Hquid. If the amplitude of the pressure variations is large enough to reduce the pressure lclly to, or Below the vapour pressure inthe negative parts of ‘the sound cycle, minute bubbles and cavities will grow (Young, 189). Ultrasound is generated by transducers, transducer isthe ame fora device capable ‘of converting one form of energy into another, ea loudspeaker which converts tlectrial energy into sound energy. Ultrazonic trandcer aro designed to convert ‘ther mechanical or electrical energy ito high frequency sound (Meson, 1991). There are three main types of transducers: gas-driven, liquid d en and eletromechanicaloy Gay Gas-deven transducers: These are simply whisles with «high frequency output, This transducer has not been found fo have any significant chemical toplication because itis not posible to achieve a suftiienly high intensity in airborne ultrasound, » Sound is attenuated rapidly on pasage dough ensities of a fraction of a wat per square centimetre ar obtained in ge, but the source mustbe very powerful, usally sonic hors. AC these intensities, it posible to use sltrtsound to break down foams, to clear Fine smoke and dusts by particle agglomeration and to acsserate drying processes Liquid-driven transducers: Thete ae effectively propeller blades operating nan inverse mode. Cavitation produced by a conventional propeller isthe ‘esultof the rapid movement of the blade through water. If a metal biade is held clamped and a liguid is forced rapidly across it then the blade is cased to vibrate with a frequency dependent on the flow rate. When the vibrations ae at ultrasonic frequencies the liquid caitates as it passes across the bade When 2 mixture of immiscible liquids is forced across the blade cavittionsl rixing produce extremely efficient homogenization. This principe has ben sed inthe food industry and more rvently chemists have sted the device to provide emulsions for polymerization or o eakance hydrolysis, lectromechanical_ transducers: Two main types of lectromechanical twansducers are based on either the piezoelectric or the magnetostrictive effet. Te most commonly wed are the piezoelectric wansducerstsed to power bat and probe type systems. 8) Magnetostrictive transducers: Magnetostricton refer toa change inthe dimension of a suitable ferromagnetic material, nickel 2 nickel alloy, by the application ofa magnetic Field, A magnetostrictive transducer is usally in the form of a rod acting asthe magnetic core within a solenoid. Applying 4 varying current to the coil produces a variation in the dimensions of the ‘The major drawbacks of these transducers are that they ae not efficient in terms of electrizal power consumption, and that their sof requency » ducers: These are ased for both the generation and the detection of ltrisund. The materials have two effets. The direct effect when pressre i applied across the large surface of the section, a charge is generated on each face equal ia size but oppose in sign. This polarity i reversed if tension is applied across the surfaces, The dnverse effec, if charge is applied to one face of the section and an equal but opposite charge 10 the other face, the whole section of crystal will ether ‘expand or contract depending on the polarity of the applied charges.(On applying rapidly reversing charges to a piezoelectric materi, fluctuations in ‘imensions wil be produced. This effect can be harnessed to transmit ultrasonic vibrations from the crystal section through whatever medium itis in contact wit. Early piezoelectric devices were based on quate transducers, but a quartz is fagile and difficult to machine, three substitutes are commonly used, barium titanate (BaTi0y), lead metaniotate(PONb,O,) and the mined crystal, lead zconate titanate ‘These materials cannot be obtained as large single crystal, so they are ground with binders and sintered under pressure at above 1 000°C to form ceramic. The rstlites ofthe ceramic are then aligned by cooling from the fereletrc transition temperature in a magnetic Fed ‘The ac. output from an clectonic oscillator and amplifier is coaverted 10 mechanical waves either by a magnetostrictive ora piezoelectric transducer of barium titanate or lead zirconate. The output fom the transducer is coupled tothe suspension undergsing teeatment bya half wave metal probe, which osvilates atthe cigcuit frequency (Hughes ‘etal. in Norris and Ribbons, 1971), ‘The amplitude of the wave generate is inversely related tothe diameter of the probe tip. When in use the probe & immersed a few millimetres beneath the surface of the fiqui, Its important to ol the suspension either by packing the sonication cup in ice or by circulating a soolant through @ jcketed cup, Parameters which affect ultrasonic cavitation have been desribed by Mason (1991) (Frequency : As the frequency of sound waves is increased the rarefaction hase shortens and this has two consequences. Firstly it s necessary to increase the amplitude (power) of sound iraation to maintain an equivalent amount of cavitation in the system. In other words more power i required ata higher Frequency ifthe same cavitation effects ae to be mainsined. Ten times ‘more power is required to make water cavitate at 400 kHe than at 10 KH Secondly, when the ultrasonic frequency is increased towards the megahertz region, the production of cavitation in liquids decrease. The simplest ‘explanation in qualitative terms is that at «very high frequency, where the rarefaction (and compression) cycles are extremely shor, the finite time required forthe rarefaction cyete is too short to permit the molecules to be pulled apart suicienty to generate a bubble. I should also be recognised that transducers which operate at these high frequencies are not mechanically capable of generating very high ultrasonic power. Gi) Solveat sco The Formation of voids oF vapour-filed microbubbles (cavities) im a Tigi requires thatthe negative pressoe in the rarefaction region overcome the natural cohesive forces acting within the liquid. Kt follows that cavitation should be more dificult to produce in Tiguids where such forces ate large. Since it fs aecesary for the rareacton pressure 10«iy oo wy wy ity aa overcome the cohesive force inthe liquid inorder to gener a bubble, any Increasing visosty or surface tension wil naturally Iead to a increase inthe mount of energy needed to separate the liquid (Lorimer in Mason, 1990) Solvent surface tenslon : Solvents with low surface tensions ka toa reduction inthe cavitation threshold Solvent vaposr pressare: It is more dificult to induce cavitation ina solvent ‘of low vapour pressure than in a more volatile solvent. Inadition, when @ ‘more volte solvent is used, mote vapour enters the cavitation bubble during formation and the bubble collpe is cushioned and less vient. Temperature: Increasing the ambient temperature will raise the vapoor pressure ofthe medium and so lead teaser cavitation but less violent colle. [At temperatutes approaching the solvent boiling point a large number of cavitation bubbles are generated concurrently. These will act as a barrie sound transmision and dampen the effective ultrasonic ene. ‘Babbted eas: Dissolved gas or small bubbles in a fluid can set as nuclei for cavitation. When sonication is iniited cavitation ours relatively easily but, seas is removed cavitation becomes more dificult. Increasing the fas content (of liga leads not only to more cavitation bet also to 2 reduction in the intensity of the shock wave released on collapse of the bubble. The removal of gas from a Higuid will reduce she available mucli and it wil become increasingly dificult to cavitate the liquid (Lorimer in Man, 1950). External (applied) pressure : Increasing the external pressure will require renter ultrasonic energy t indice cavitation, ie. greater rarefaction pressure i required, Raising the external pressure wil give rise toa Lager intensity of eaviationalcolapee and consequently an enhanced sonochemical effect, Inteosity: The intensity of sonication is directly related te the amplitude of vibration of the ultrasonic source. In general an increas in intensity will result in an increate in sonochemical effects, but ultrasonic energy input into the system cannot be increased indefinitely for thee reasons. Firstly, the transducer sed in the sonicator will eventually fail as the increasing mensional changes in the transducer fractore the materal. Secondly, at high vibrational amplitudes, the source of wlteasound cannct maintain conast ‘with the guid throughout the complete cycle. This is krown as decoupling ind results in a great lots inefficiency of transfer of power from the source to the mediom, Thirdly, when a large amount of ultrasoxic power enters a system a great number of cavitation bubbles are generated in the solution. ‘Many of these will coalesce forming larger, more stable bubbles. These may dampen the sound exergy through the lguid and also remove many of the smaller bubbles which would have collapsed to give sonocwemical effects34 aa (si) Attenuation of sown someot 0 Sound is attenuated as tprogreses though a median, is energy is dissipated inthe form of hat. The extentof atemuation is inverely related to frequency. Sound at I18 KHe is reduced to half ofits intensity after passing through 1 km of water, The distance required to achieve the same reduction of intensity for 20 KHz sound is much greater, 0 kn, ‘CAVITATION PHENOMENA ‘Noise, chemical ations ad luminscence are phenomers which sult from cavitation. Sond Cavitation is acompanied by in 8 nose, A greater increase in acoustic intensity {accompanied by a preter increase in noise level for lquids containing gas than for Aegassed guid. Since the wall of the cavitation bubble may atin supersonic velocity ‘uring collapse, cavitation should be accompanied by shock waves. maximum pressure atthe bubble wall upon colapse of 4 O00 atm has been quoted (Harrison, 1952 in Webster, 1963). The sound emitted by eavitetion can be measured using sound level meter ateached to an octave fiter set (Broadhurst prs comm, 1993) (Figure 33) FIGURE 3.5: A sound preeure level meter 1981) th an octave filter set (after Fader Sound presourelvels (SPL) are based on the ti of two power-Hke quantities using ‘sound level rater (Figure 3.5). SPL deierbes how much round is available from the sr atthe loation where the measurement ie made (Fade, 1981). A SPL of © €B. corresponds to a presi wave of 20 Pa root mea squared (en).| | Pa] where SPL the sound pressure level (6B) referenced to 20uPa P= anaverage pressure variation (Pa) Py = the reference pressure variation of Pa ‘A decibel (4B) isa number on a logarithmic sale used wo express, as level difference, the rato of two lke quantities proportional to power or energy. ‘The ratio i expresed in decibels by multiplying is common logaihen by ten, When measuring sound pressure levels (SPL), if the power doubles the SPL increases bby 3.4B. For example, in a zoom with two fans, with each fan producing 60 <8. ‘When both are switched on a SPL of 63 dB is obtained. If there were 100 fans, the power would increase 100 times, and the SPL would incre by 20 4B. Note that 235 sound power is proportional top Another method used to calculate differences in dB isthe Difference Table for Addl 4 (Table 3.1) “TABLE 3:1; Difference Table for Adding dB (aftr Fader, 1981) Difference between two lve ‘Add tothe higher level ey ae) 0 30 1 2s 2oe3 20 4 13 S10? 10 80r9 os Wor more 80sas sccording to the following eavation = where 13. SPL_ = sound pressure level difference (8) a, distance (m) 4 corrected ditance(m) ‘The sound emited from a point source in 4 room is reflected off the walls and the sound received by the sound level meter sa combination 2f direct sound and reverberant souad, Direct sound is travel directly to the sound level meter from the source, ‘whereas the reverberant sound constitutes a multtede of reflected sound waves. The n be predicted equation 3.9 8) SPL at any distance () from an omnidirectional sure ‘hich has beon plated in Figure. 36 (after Beranek, “une” Sains ow] Environment Free Field Environment Log r ( FIGURE 3.6: Effect of tance on sound measured i a free field environment room (after Beraneh, 1985)316 “The effect of distance on sound is represented by the Following eatation er. = 10 tg (ME) 6 10 ww Gta + oth a9 (er) aa oy SPL = sound pressure level in dB at distance (e) from source = density of ar (1,208 kg/m) = speed of sound (346 m/s) threshold of hearing (20s P2) sound power of source (W) Fm distance from source to point of measurement reverberation time (3) Y= volume of room (en) “The equation can then be simplified asthe fir erm isconstant fra particular source 1, ery a * TW) a sre = c+ 10 tog [ For the line in Figure 36 indicating the direct sound ina free field environment, the ste = e+ 10 we Gh ant For th line ofthe reverberant sound Forte Sine combining revebrant and diet sound ‘Where the reverberantsound lve is equal tothe diect sound, there isa 3 dB aiference ‘between the combined sound and the intersection of the two lines representing the direct and reverberant sound pressure level The sistance R, is called the radios of reverberation. Reveberation te (T) i the time(s) required for 2 nse level to reduce by 60 4. “The time required depends on the size and dampening ability of the room. Typical 08-125 :alecrte teat 15-188 :a-concert hall approx 2,5 5 sa large boratory with no wound absorbing surfaces ‘The radius of reverberation in specific room canbe calculated using the following eavation342 sa where Re © radius of reverberation (m) = volume of room (mi) T= reverberation time (8) [An octave band filter is wsed in conjunction with a sound level meter for the measurement of sound. It retricte the sound reaching the measurement section of the sound level meter so only thore Frequencies within a certain range of freqvencies ae measured. The cente frequency (0) canbe selected to be ether 31; 63; 125; 250, '00; 1000; 2 00; 4 000; $ 000; 16 000 and 31 $00 He. For example ifthe meter it seta frequency (7 of 1 000 Ha, the Bandwidth ranges fom 700 Hz (7/2) 401 400 Flr (BE). So, each octave Frequency band selects a range of frequencies, ‘Chemical Reactions ‘Webster (1963) notes that chemical reactions initiated by ultrasonic cavitation can be wed t0 (Accelerate reactions, eg. the hydrolysis of estes. (i) ikate reactions that would not otherwise occur Reactions that would not otherwise ascur would take pice pefominemtl in an aqueous ‘medium. It sumed by Lindstrom and Lam (1951 in Webster, 1963) that under the action of cavitation water decomposes into free rai “The hydcogen exits as an stom which ita free radical due to the pretence ofthe free “The products ofthese reaction are then cespanibie for secondary reaction involving solved substances. In the presence of dissolved oxygen the mie of Formation of hydrogen peroxide ie increased a illustrated in the following eavationsWeiser et al. (1950) studied the oxidation of potasium iodide solution exposed to tltrasonic waves (1 000 KHz). Richards and Loomis (1927 in Weiser et al, 1950) showed that iodine ie produced when wltrtsouad travel through a potassium iodide elution containing disolved sie. Tals was attributed to the formation of hydrogen Detonide which results from cavitation, Li and Wu (1934 in Weiser et al, 1950) determined that oxygen and cavitation are esental inthe rexction, and they Found that the addition of carbon tetrachloride increased the amount of iodine liberated ‘They believe thatthe carbon tetrachloride reacts with the activated oxygen to give free chlorine which then oxides the potassium iodide, Inthe absence of carton tetrachloride the oxiation of potassium iodide by ultrasonic waves is brought aboot by ultrazoniclly-preduced hydrogen peroxide Ho, + 2 KI 4 4, + 2 KOH Weisser etal. (1950) report the following contusions (The amount of iodine liberated by vltronic. waves from potasiom Jdide-carbon tetrachloride aqueous solutions depends on the dimensions and material of the rection vest (ii) Although dissolved oxygen has beet considered as essential, nitrogen and hetium serve almost aswell (ii) As the power input i increased, no iodine is produced until cavitation occurs ‘Then the yield is increases almost linearly if the volume of the solution i large enough, however na smal volume the yield increases and then decreases sharply (is) tntte absence of carbon tetrachloride the oxidation of the potassium iodide solution proceeds only about one-fiteenth as rapidly 3 in the presence of carbon tetrachloride. tah Sonoluminescence ie a weak emission of light cccurring in some sonially-cavitated liquids. Its most intense in the pressure antnodes of stationary wave systems. Tt manifests itself asa discrete flash of light onee every sound cycle. I is believed that the luminescence emanates from the bubbles during the final stages of collapse. The sonoluminescence is measuted with a photomultiplier (Webster, 1963). Luminescence has also been observed in non-acovstic cavitation (Schmid, 1939 in Webster, 1963) Jarman and Taylor (1964 1965) designed experiments fo analyse the emission of Tight {sith 2 photomoltipiee) from cavitation induced in ap water flowing through the constriction of venturi, Light was emitted as flashes of uration of tess than $18 with spectrum between 400 and 700 no with a maximum intensity at 470 nm, Shock waves were detected by miniature presture transducers implanted inthe wall of the tube. It war found that a shock wave wa associated with every light flash. The ‘emision of Light was two orders of magnitude less than the wltrasonically-induced35 38 9 semoluminescence from water, They sugges that the ight originates From within the collapsing cavity, Jarman (1960 in Jarman and Taylor, 1963) describes the emision ‘mechanism as thermal. The light can be described st iacandescenceresltng from the adiabatic compression ofthe contents of the eavity. Jarman and Taylor (1965) analyted the luminescence from hydrodymmically-indueed cavitation and found ito be two orders of magnitude weaker than aeusialy induced soncluminescence, They suggest that since the light emission isso weak iti key ‘thatthe formation of free radicals and their recombination products, such as hydrogen peroxide, is aso very slight. Henglein (1956 in Jarman and Tayo", 1968) note that there i 8 correlation between sonoluminesence intensity and the veld of hydrogen peroxide from sltrasonically induced cavitation BACTERICIDAL EFFECT OF CAVITATION easonic Cavitation ‘High intensity sound is used extensively in bology to destroy or ropare cells although the resie mechanism ofa the effects obeervedis nt yet Fully understood Prudhomme and Grabar, 1947 in Hughes and Nyborg 1962). AS a result, much work has been reported on wlrasonic disintegration of bacteria, viruses, Fungi and animal cel Ultrasonic cavitation can be discussed in terms of two general catogeres, i. transient ‘cavitation and stable cavitation, Transient cavitation connotes relatively violent ‘etvty (bubble collapse) in which hotspots of high temperature and pressure occur In very short (in the order of microseconds) bursts at highly Tocaisedfoeations in the sonicated medium. ‘These bursts may be accompanied by localised shock waves and/or the generation of highly reactive chemical species In contrast, the much less violent form of cavitation, stable cavitation i associated with vibrating gaseous bodies. The nature of thi form of cavitation consist of a gateus body that remains spatially tions ar entablised, the Hguidlike medium immediately adjacent to the gas bubble Flows or steams (tormed microstreaming). ‘Mictostreaming resulting from stable cavitation has been shown t produce sreses sufficient to disrupt cell membranes (Scherba et a.191). The mchanism proposed isthe ont of turbulence which creates vortices near which there exist shear rates higher thin the shear rates throughout the bulk of the Iiquid. Prichard et al (1966 in Hughes etal. in Norris and Ribbons, 1971) showed tht shear gradients exerted by ‘microstreaming around bubbles (created by vltrasound), at low sourd amplitudes were sufficient to degrade DNA, ‘The damage caused by shear sires is thought to depend ‘on erotion of the outer cell wall polymers, paticulely a weakened places such as division or budsing scare “The following mechanisms have been proposed for cell disruption resulting from transient (gaseous) cavitation+0 Forces due to sueface resonances ofthe cll wall ini mechanical Fatigue (Hughes and Nyborg, 1962), i) Shearing forces occur due to mictosreaming cf the cell uid (Hughes ané Nybore, 1962), (ii) Pressures and pressare gradients result from the collapse of gas bubbles 08 oF ‘ear the cll wall. Damage may resul from a siagle event or fatigue involving 4 threshold time (Hughes and Nyborg, 1962) (iv) Radiat resonance of bubbles create pressure ard pressure gradients (Hughes and Nyborg, 1962). (0) Pressure or relative velocity effects resulting from the direst sound beam which fe enerated from compression and rarefaction of sound waves (Hughes and Nybore, 1962), (i) Chemica attack in which a wide range of free radicals, expecially H and OM radicals ae formed in eavitating aqueous Tiqsis. Those may attack the cel wall and weaken i tothe point of rupture. Hawever, it has been shown that col rupture and free radeal formation occur independently of one another (vgnes and Rogers, 1999 in Neppias and Hughes, 1964) (it) Combined chemical and mechasica attack (Hughes and Nyborg, 1962; Scherbs eval, 19). “The mechanism by which glrasound inactivates bacria has not been conclusively established. The support of the various theries i presented ‘Shear streses: Shock waves resulting fom transient evens may give rise to localised shearing streses (Thacker, 1973). Ths view is supported by Hughes and Nyborg (1962) where its noted that coll breakage isnot depenenton the violent collapse of bubbies bot may bea result of shearing action associated with bubble-induced eddies and related mation. Thacker (1973) algo noted that bunble collapse leads to trbulent flow and consequently high velocity gradients in localied areas resulting in increased shea sesss. Cavitation cet: Thicker (1973) suggests that mechanical streses generated from avitaional phenomena constitute the destestive fore. Hughes and Nyborg (1962) scribe the disruptive effects of ultrasound to the lca effects of transient cavitation, The local effects of cavity collapse include local heting and electrical dicharges, sonoluminescence, chentoluminescence and free radical formation (Lindstrom, 1956, {nd Jarman, 1960 in Hughes and Nyborg, 1962). In aditon to such posible aston, tas bubbles are capable of resonance vibration, The reakdown of the cell wal may be due tothe production of radicals (Peteron and Zaliko, 1981), Free radicals may leo cause chemical changes in cell eomponen' released int solution after cell breakags (livgnes and Nyborg, 1962; Peterson and Zaliko, 1981),321 “The effet of free radicals is questioned by various authors. Thacker (1973) nates ‘that, although free radical formation occur with cavitation, the organic matter inthe sample can scavenge the free radical, Hughes and Rogers (196) in Hughes ané Nyborg, 1962) state that cel breahage is independent of Free radical formation. These studies contradict the previously mentioned suggestion of the breakdown of the cell wall by free radicals produced from cavitation (Peterson and Zaleio, 1981). ‘Streaming motions: Streaming can be induce bya vibrating needle and the production of sufficient shear to injure red ood cells and bacteria has been reported (Hughes ‘and Nyborg, 1962). The damage is small compared withthe eavtation effects of alrasound, despite the fact that treatment periods were long (up ‘0 1h) compared ‘with he time (3105 ein) required forthe complete disruption fF. cl with ultrasonic cavitation (Hughes and Nyborg, 1962). are change : The cll wall may be destroyed loally asa reel of the implosion fof s bubble immediately adjacent toa bacterium, AReraatively, low leal pressure tay exist causing the cell contents to vapourite end the col to burst (Peterson and Zale, 1981), esses Thacker (1973) proposes tht tensile stress experianced by the el He describes that larger, budding yeast cells willbe exposed to grater tensile stress ‘than smaller non-dividing cel Ultrmonic cavitation har been applied to achieve goals her than disruption of cells forthe extraction of intracellular constitents, These ieclude the use of ultrasound forthe cieperion of bacterial aggregates and analysis of the poteniat for the ue of sltrasound t inactivate bacteria in water and effluent. Shropshire (1947) used ultrasound (9 kHz) as a means of dispersing suspensions of bacteria, He believed that dispersion prior to enumeration would be valuable in ricrobiclogcal analysis techniques. Scherha ofa (1901) wadertaok a qvantittin acuemant of the srmicidal efficacy of ltsonic energy. Ultrasonic energy at a frequency of 26 KHz was used t expose aqueous suipensions of bacteria (E. col, S. aureus, B suis. P ceruginosa), fungi land viruses, This work by Scherba etal (1991) suggest that ultrasound in the low-kiloherte frequency range has some efficacy in inactivating some disease agents ‘The combination of ultrasonics (40 kHz) and ozonation was wsed to determine the inactivation of views and bacteria (Burleson et al, 1973). 1 was found that simultaneous treatments by ozonation and sonication reduced the contact time for ‘complete inactivation of microorganisms in secondary effluent. It was suggested that cavitation reduced the high surface tension eaused by the organic material resulting in an increased efficiency of ozonation,an In addition to the doubt regarding the mechani of destruction of bacteria by cavitation, the target of damage is sll uncertain. Gram-positive organisms usualy have a thicker and more tightly adherent layer of peptidoglycans than gram-negative bacteria. coll and P. ceruginoa). Sherbet al (1991) sought to determine whether the nature of the cell wall was adi feretiating factor inthe destruction ofthe bacteria They concluded that this morphologi feature was not a factor determining the sensitivity of the bacteria to utraound, Iwas suggested that the target of vltasonie damage may be the inner (cytoplasmic) membrane which consists of & lipoprot bilayer ‘Hughes etal. (in Norris and Ribbons, 1971) note the factors which influence cll ) Uteasound and ozone (Dak, 1976, Burleson eta, 1975) (9) Onone and hydrogen peroxide (Wolfe, 1989) (oi) Hydrosen parosie, UV, cavitation (Cav-ox) (Peterson and Zaleko, 1981) “This pect ofthe projet wat a preliminary investigation to determine whether hydrodynamic cavitation prior to ultraviolet irradiation would enhance the death rate of raw water bacteria by virtue of increasing the accessibility bythe disinfectnt/agent, ie. UY. The potential of both ultrasonic and hyrodynamie cavitation prior to treatment with UV irradiation in water eaten i described, ‘A igh pressure mercury lamp (HBO) with «254 nm fier was used asthe light source, The intensity of the light emitted by the HBO lamp was determined b water wat obtained from Wiggios Water Treatment Plant and emicalactinometry. Raw tated for various periods of time by either of the two methods of cavitation, After cavitation the samples were irradiated ‘with UV Fish prior 0 plating and enumeration of viable bacteria61 (CHEMICAL ACTINOMETRY Actinometry isthe proces by which the intensity of light absorbed by an irradiated solution is measured by determination of the quastam yields of photoproducts ‘Chemical actinometry involves the ute of s photochemical reaction for which the ‘qwantam yield of certain product at particular wavelength i known, The most commonly used method was developed by Hatchard and Parker (1956). This method is sensitive for wavelongths from 250 nm 10 $77.nm. Tt involves the Dhotodepradation of acidic solutions of potassium ferioxslate aecording othe following 2 Fe(C,09" > Fee 640) + 9 (09% + 2 0, Expowure of the atisometry solution to Tigh of wavelengths frm 250 nm to $77 nm ‘causes the simultaneous reduction of Fe%* to Fet* and the oxidation of the oxalate fon, The reaction product do not absorb an appreciable amount of the inidest ieradiaton. [After exposure to UV tight, complexation of the Fe ion to 1,10-phenanthotine i fan acetate buffered solution (pH 3,3) results in a red complex that absorbs at 510 aa. “The molar absorptivity at S10 nm ofthis complex is approximately 1,1 x 10-4/M/om (Calvert and Pitts, 1966). The concentration of the ferrous ions insolation can be ‘determined by absorbance measurements, The quantum yield for ferrovs ion formation in such solutions is 1,25 at 254 nm over the tomperature range to $t0 8 °C (Matchand and Parker, 1956). _As potassium ferroualte decomporesin the pretence of la I preparations involving ‘potusium ferrioxalte wore cari out in a dak room using red photographic light. Potassium ferrioraate crystals were prepared according to Parker (1938 in Ciemmer, 1992). A volume of 750 m¢ of 1,5 M potassium oxalate (AR grade) solution wis Aided 1 250 mi of 1,5 M foric chloride (AR grade) solution. The solution ws concentrated to a volume of 250 mi by heating and thus enabling the precipitation of ‘the green potassium oxalate crystals, The crystals were secrytalised thre times from water and air dried. The following solutions were prepared 1) Fertioxalate actnometer solution (0,02 M) : 24 g of potassium ferroxalste crptals were dissolved in 200 mé ultrapure water, After the addition of 25 me (of 0 M H,80q the solution was made upto 250 me in volumetric fas 1b) Acetate buffer: 360 me of 0,5 M H,SO, was aided t0 600 me of | M sodium Acetate and mide up 10 1 & ° 0.1 & of 1,10-phenanthrotine monohydrate was disolved in ultrapure water and made up to 100 mé612 3 Procedure ‘4.254 am fier was installed inthe HBO lamp. A volume of 3,5 mot the fersionalte ‘sctinometer solution was pipetted into a1 em quartz cuvete and placed the optical teain ofthe light. The solution was exposed to UY for 2 min. After removal frm the optical train, 0,5 me of the iradiated solution was transferred toa10 mé volumetric ‘ask, To this, 05 mtoF0,% 1,10-phenanthrolne solution and | ‘of acetate buffer were added and the resulting soltion made up to 10 mé with water. This solution ‘was Kept in the dark for one hour before measuring the absorbance at 510 am onan LKB Ultrspec UV/Vis spectrophotometer, The Blank forthe experiment conse ‘of 0,5 mf of the un-iradiaed actinometerroltion, 0,5 mé of 1,!0-phenanthrline solution and 1 me of buffer, ade up wo 2 volume of 10 me Results “Ten samples were iradiated for 2 min each and the absorbance "ead after 1 hour (Tabte 6., [FABLE 6.1 + Abworbance reading Sample ‘Abior [From abvorbance measurements the number of Fe ions (#Fe!*) ‘ormed during the itraiation was calculated (Appendix G), The intesity of ieradiaed Tight was ‘determined and the power output of the HBO lamp was calulaed tbe 1,07 mW (Appendix G). “The ate of the illuminated surface ofthe cuvette used for iradiaticn of the raw water sample was em! ‘Therefore the intensity of UV radiation t which the raw water was exposed is 0.214 mW on? In treatment of water or effluent with UV, dose is the wre used to describe the intensity of UV lnradiation andthe duration of irradiation. Since the intensity of the HBO withthe 254 nm fier was determined 10 be 0,2 mW/em?, the UV dose for 8 2 min radiation of a water sample is 25,2 mWs/em?62a ULTRAVIOLET IRRADIATION [A.254 nm Fter was installed inthe HBO apparatus in order to decrease the intensity ofthe iradiation whilst stil producing germicidaly active irradiation at 254 am for disinfection water. A volume of 17 mé of raw water war poured into a5 em quartz cuvette, plied in the holder of the HBO and ierdiaed. Samples of 0,1 mé and 1,0 me were withdrawn with steie pipettes at various intervals, died according the serial dilution technique, and plated to monitor the change it CFU/m¢ over & period of time Results Figure 6. shows the death curve for raw water bacteria iradated with light at 254 am tan intensity of 0,2 mW/em?. The inital count was 12 00 CFU/m Time (rn) [FIGURE 6.1: Death carve for raw water bacteria Irradiated with light a¢ 254 am and an Intensity of 0,2 aW/eo™ The duplicate samples exhibit an initial rapid death rate followed by a “ailing off” effect. After? min of iradiation 297.5% inactivation was achinved for a dose of 24 else?6s 63 ULTRASOUND AND UV RADIATION [Raw water (25 1 sample volumes) was sonicted inthe ultrasonic bath for varous durations, The water was then iradnted with UV at 254 am for various periods of time and sums were collected at selected intervals. 63.1 Reslts “The number of viable colonies per mile were counted, The dats was manipulated by dividing the final amber (N) by the orginal count (N) and taking the logarithm. Figures 62 to 6.5 show the death curves for sonicated raw water followed by UV irradiation for 10 min, The controle for each consisted of non-sonicated water exposed 1 UV irradiation, Time (ia) ed (10 min) raw water followed by Iradiation with UY at 254 nm and an intensity of 0,2 mW/em?66 Tie (rin) FIGURE 6.4: Som ted raw water (1 la) followed by UV ierdlation (6 in)Tire (in) FIGURE 6.5 : Sonicated raw water (1 mis) followed by UV iradiation (6 mia) Figure 62 shows a 99.8% inactivation after 2 min of ieradiaton for the sonicated sample and a 98,2 survival forthe non-sonicated sample. This apparent diference isnot significant due tothe variation inthe number of bacteria and the HPC technique, ‘The initial population for the experiment shown in Figure 63 was 2 000 CFU/m [After 2 min, 945 % of the bacteria were inactivated. This corresponds to 1,3 log reduction at a dose of 25,2mWs/em®. ‘The death rate is highest in Figure 62 and this. ‘ould be due tothe 10 min duration of sonication which. alone can inactivate bacteria (sttion 53). “The initial population For the experinment shown in Figure 6.3 was 2 400 CFU/mé With a dose of 25,2 mWs/omt, «92,1 % inactivation rested ia 1,2 log reduction, “The inital population fr the experiment shown in Figure 6.3 was AD CFU/me After 2 min alg reduction of | resulted for a dose of 25,2 mWs/emt. In Figures 63 t0 6.5 the non-sonicatd and sonicated death curves are very smi. From thi itcan be concluded that there is no significant enhancemen: in the inactivation of bacteria by UV in raw water which has been sonicated. This could be due tothe fact that sonication alone inactivates bacteria ia addition to dispersing clumps of6a ot HYDRODYNAMIC CAVITATION AND UV RADIATION ‘As hydrodynamic cavitation disperses aggregates and garticles in water (section 44) ‘twas decided to determine whether cavitted raw waterprior to UV iradition would result in enhance inactivation of the bacteria [Experiments were undertaken where the feed tank was Filled with raw water obtained from Wiggins Water Treatment Past, Hydrodynamic cavitation was initiated for 3'min. Heterotrphic plate counts were performed prior to cavitation and after the 53min period of treatment, A volume of 17 me of raw water was placed in aS om twvartz cuvette and irradiated for 6 min, Samples were withdrawn at various intervals and plate counts were performed o determine the chan in numberof viable bacteria over time, Log N/N, was determined based onthe initial CFU/me “Three experiments were performed atthe highest intonsty (C section 42.1) produced by the cavitation rig, 12. With a Puy of 472 KPR, An experiment was undertaken ata Tow intensity (Condition A), 8P mu of 22 KPa Results Figures 6.5, 6.7 and 68 show the results ofthe experinents where high intensity (C) cavitation was undertaken for 3 min prior to UV iraciation for 6 min. Figure 69 shows the results of low intensity cavitation (A) prior io ieraiaton, Tino (it) FIGURE 66 ed for 3 min fellowed by UV irradiation9 . ‘ “me ni FIGURE 6.7 : Raw water caitated for 3 min followed by UV eradation FIGURE 6. Time (nin) tated for 3 in followed by UY i6-0 Time (in) FIGURE 6.9 : Raw water cavtated for 3 min at x fow Intensity, folloned by UV readin ‘The initial population in the experiment shown in Figure 66 was 16 000 CFU/m4 “There ino significant difference between the non-caitated raw water andthe cavitated water, A lo reduction of 1,7 resulted after 2 min of irradiation. This corresponds toa 1.6% survival (ie, 8.984% inactivation) with a dose of 25.2 mWsjem? “The inital population in Figure 6.7 was 7 500 CFU/me Again there i no ditference resulted, corresponding to 4 991 % inactivation with 8 UV dose of 25,2 mWs/em?. From an initia! population in the experiment shown in Figure 68 of 7000 CFU/m, 4 log reduction of 1,7 resulted with an inactivation of 982% from a dose of 25,2 mWs/em From an inital population of 4 000 CFU/mé (Figure 69), 9 log reduction of 1.7 revolted, corresponding to 97 inactivation after a dose of 25,2 mWs/em? (2 min) The log seduction ie consistent with the reduoton elting From more intense cavitation and ther sno significant diference between the UV initivatin of raw water exposed {o hydeodynamis cavitation and non-caviated water.65 65a SUMMARY ‘UY Jeradiaton “The intensity ofthe radiation produced by the HBO lamp was determined and found tw be 0,2 W/om?, Literature reports inactivation in a variety of terms, the most common of which quotes the result in log reduction nits for speifie UV dotes or exposure times. The above work was done using doses ranging from 0 mWs/em? to 175 mWs/em? which coresponds to exposure times of 0 min to 6 mi. According t9 Conacher (1986), the minimum dost for the inactivation of bacteria is 30 mWs/em?, Qualls eta, (1985) found that the dose necessary 10 achieve 3,5 10g redaction units ranged from 9,6 10.52 mWs/cmt and average at 20 mWs/em for total coliforms “The UV system vied in thi work rendered range of log reduction units from 1.510 2.2 witha dove of 25,2 W/o? for aw water bacteria a determined by heterotrophic plate counts ‘The combination of treatment with ultrasound of an intensity of appreximately 1 W/cm? rior to UV iradiation didnot significantly enkanee the inactivationof bacteria. Both sonicated and non-sonicated raw water, exposed to UV, resulted in og reduction units ranging fom | 10 13 ‘Utiasonis cavitation alone inactivates aw water bacteria (as determined in section 5.3) thereore the sequential effect of toniation followed by UV irradiation will be ‘obscured by the inactivation ofthe bacteria by sonication, In other word, there will be Fewer bacteria to inactivate, and ae death is exponential, this results in a slower “The combination of hydrodynamic cavitation price to UV iraiaton did not enhance the inactivation of raw water bacteria withthe log redsction usits ranging from 1,7 02. Hydrodyaumic cavitation dispersed aggregates containing bacteria according 10 turbidity studies and the scanning electron micrographs. The degree of dispersion felates to the duration that the water remained stagnant. The Inger the water is stagnant, provided sufficient autrents are present, growth will occur in the water [As the water is stagnant, the limps wil ot be dispersed as they would atthe water treatment plant, "The potential forthe use of hydrodynamic cavitation as a means of Aispersion must be noted. Perhaps other “problem” water or effluent could be usedon LUV lnraiation is effective at inactivation of bacteria, and asthe turbidity of the aw ater was low (ranging between 0,6 NTU and 7 NTU), the protectin of bacteria by clumps and aggregates was minimal, The potent forthe combination of hydroymamic cavitation with other means of disinfection shouldbe sssested, “Thore isa difference between the death curves of the sonicated and control samples and the hydrodynamic cavitation samples of approximately | log uit indicating that the hydrodynamic cavitation rig was more effective than the ultrasonic bath. Ta the case of hydrodynamic cavitation, che water was transferred to the feed tank by means ‘of bucke's which resulted in mixing ofthe water andthe subsequent dispersion ofthe ‘lumps. In the case of the sonicated sample, the water was simply removed from the Storage veel by means of the submersion oft sample botle. No mixing or agitation occurred at this stage. Perhaps the action of dispensing the water into the feed tank Aispersed the agareentes and resulted in a higher efficiency of UY iaactvation.Chapter 7 DISCUSSION AND CONCLUSIONS “The objective of this project was to evaluate the potential of hydrodynamic cavitation for water teatment with the inactivation of raw water bacterin ae the primary aim, —Secondar/ and tertiary aims were to us ultrasonic cwvitation asa comparton and to determine whether cavitation pretreatment would enance inactivation with uiravielet radiation. Severl pois ‘emerged asa result of the experimental work undertaken i this projet. Fist, the choice of the wse of raw water for experimentation a8 oppoted to the conventional procedure of the Snoculaton of sterile water with a selected indicator microorganism willbe diseussed. Based on this, heterotrophic pate couats were selected as the method of analysis. Secondly, experimentation revealed that hydrodynamic cavitation alone is ineffective inthe inactivation of raw water bacteria although dispersal of aggregate aril ocurred, The results of ths Investigation are contrasted with thos of Harrison and Pant (1992,b), in which hydrodynamic cavitation was wed for cll disruption. Thirdly, the results obtained for ultrasonic cavitation willbe discussed. Fourthly, the intrinsic atTerences between the two forms of cavitation will be highlighted. Fifty, the potential forthe application of hyérodyamie cavitation ia combination with ultraviolet radiation was assested and will be dicusea 7 RAWWATER AND HETEROTROPHIC PLATE COUNT ANALYSIS Heterotopic plate count analysis was used to monitor the changes in umber of viable bacterin eaw water exposed to : hydrodynamic cavitation alone (Chapter 4, ultrasonic cavitation alone (Chaptee Sj; and cavitatea raw water followed by ultraviolet raulaticn (Chapter 6). The selection of this technique i valid a various authors (Wolfe eta 1989; and Turai etal, 1980) note the difference in susceptibility to disinfection of faturally occurring bacteria compared to that of laboratory cultured species. The bacteria occuring in vivers and streams often exist in a resistant, stationary state, and they can occur enmeshed in material, protected from harsh surroundings. AS a res, ‘ese bacteria require a lavger dose of disinfectant than bacteria cultured in the laboratory and used inthe stationary or logarithmic phase of development. Certain species of bacteria form spores when exposed o adverse conditions suchas low nutrient levels, unfavourable temperatures and the presence or absence of sufficient oxygen. Upon entering a favourable environment, these bacterin reproduce. The ute of the more resistant species of bacteria inthe determination ofthe efFiciency ofa disinfection process is therefore Favoured12 1 Routine water analysis takes the Fore of the analysis and enumeration of selected Indicator species of bacteria. The presence ofthe indicators such 8 E. coli leads 19 the assumption of faecal contamination of the wate. Upon disinfection, tke absence of F. coli leads to the conclusion that the water is safe for human consumption. Pathogens occurring in contaminated and pollsted water can exhibit resistance to Aisinfection applied at dots to eliminate indicator species. Watkins (1991) noted that often fate sense of security is obtained when indicator species are sed For the assesment ofthe efficiency of disinfection proces “The bacteria used in the water treatment work had been tansportd from the iver to the water treatment plants, pasing through pipes and pumps. By the time they reached the water treatment plant, the weaker, more susceptible species would have been inactivated. The experimental procedure wsed therefore sected the hardier species, which i representative of water treatment probles. ysis was selected as it constituted 4 est of the ficiency of hydrodynamic cavitation asa means of inactivation of bacteria. AS an ‘ficiency of process was being assed it was 20tthe absolute numbers of bacteria that was important, but the changes in the counts, No false indication of inactivation as obtained, which could have occurred had less resistant spaces of bacteria bee {sed for the experimentation, Had the colony forming unite reduced in number os A revultof hydrodynamic cavitation, this would have indicated that hoprocess warranted ‘anther investigation of the use of hydeodyeamie cavitation, Indicator species would have been selected, and the effects of the proces on different species would have bean established HYDRODYNAMIC CAVITATION (Chapter 4) Cantation bers (s9otion 42.2) are sed to describe the susceptibility of @ flow to avila The lower the cavitation aumber the more intense the ewvitation. The cavitation numbers produced by the cavitation rg ranged From OS 100,03. Harricon ‘nd Pandit (1992) used the same equation as sed inthis dissertation For cavitation number and report numbers ranging from 2,79 10 0,99. They noted that eavitation occurred at numbers less than 1,01 2,5. Figure 7.1 shows the operating areas of the current investigation relative to those of Chetty (1993) and Harssen (19920). 1 was neces in this work to quantify cavitation generated in the hydrodyramic cavitation rig, Cavitation (urasonie) has been characterised inverms of hydrogen peroxide production (Mazon, 1990) and sonaluminessence (Jarman and Taylor, 1964; 1965). Hydrogen peroxide analyst proved to be insufficiently sensitive for the hydrodynamic cavitation rig, and literature (Jerman and Taylor, 1965) indicated fonoluminescence from hydrofyaamic cavitation was orders of magaitude fess than the intensities measured for ultasonic caviation. The measurement of sonoluminescence was therefore not selected at a form of chartcteristion oF vantitiation inthis projec.Py ‘The cavitation number (0.) deseriber conditions For cavitation but doesnot indicate bubble size or apparent intensity, therefore one cavitation number can represent 3 variety of flow conditions. Itis difficult wo reproduce another researchers conditions based on the cavitation number; one wouid require information onthe temperature, ‘ow rate, nozzle and pipe diameter. ‘The cavitation sumber can show two cavitation flow regimes. Fistly the approach jon up tothe choked condition and scondly at choked flow where the velocity ible, The cavitation achieved inthe cavitation rig was intense because B 3 ed E le. = 4439928). Zo, mat Bor 8 rs on ° 0 200 300400 300 AP (kPa abs) FIGURE : 71 A modification of Figure 4.13 showing the postions ofthe cavitation sumbers uted in thie project relative fo the operating conditions used by Chetty (1993) and Hareisoa (19920) ‘Sound pressure level (stction 4.24 spectra generated for the hydrodynamic cavitation chartterised the cavitation and indicated a decreae in sound level with an decrease in the pressure drop across the nozzle. A maximum of 82 dB was recorded at @1 frequency band width of 4 O00 Hz atthe mort intense operating condition (condition C) compared to $0 dB for the same frequency at the low inte (Condition A). ty of cavitation Hydrogen peroxide (section 4.3.1) was not detected using the method by Baga eta (0988) in revere osmosis permeate which was cavitated at the mximum condition (o.of 0.03) for 18h, From this it an te concluded thatthe formation of free radicals and the subsequent reactions to form hydrogen perovde was below the detectabeliait of 0,06 mg/¢ as measured at the sample por. Dispersion of agsresates (tection 44.4) by hydrodynamic cavitation was shown by scanning electron microscopy. Inactivation of sufficient raw water bacteria to constitute 4 decree of at lest an order of magnitude was nt achieved by hyodynamic cavitation in this project econ 444). Huterotrophic plate counts were usd to monitor changes in the concentration of viable colony foring units in cavitatod and non-cavitated raw water. The technique ‘was proved succesful during monitoring of the reduction in colony forming unis in ‘aw water expoted to ultrasonic radiation in which log reduction units of up to 2 were etermined, Harrison and Pandit (1992.8) wed hydrodynamic cavitation to disrupt bacteria and ‘The following differences between the two applications of hydrodynamic cavitation must be noted Aim: The aim of Harrison and Pandit (199280) was to divupt bacterial and eat cll collect ntaceaar material, Conditions for vigorous cell growth ‘were provided and the cells were grown tothe stationary phase. The broth coltare was then hydrodynamicalycavitaed through avalv or constriction. ‘The concenteation of soluble poten release was determined by the Lowry Protein Assay. At no stage ws the viability of the population ofthe culture ‘of concern. In contrat, the inactivation of bacteria in raw water was the sim ofthis project. This involved the use of natural ram water which was bydrodsmamically cavitated, The intial count was then compared with the Final count. Inactivation of bacteria by cavitation would have resulted in & decrease in viability as would be indicated by 9 decline in colony form (Gi) Biomass tn the work on cll disruption high biomass concentrations (7,2 to 113 kg/m) of cultured bacteria and yeast were exposed to hydrodynamic ‘cavitation resulting in 004 0 0,06 kg soluble protein/kg ty biomass. These biomass vales ae relatively high compare to the biomass concentration of raw water (estimated to be 6 mg/?)