INT'R.

ODUCTION·

1

H .. Denis Burges and Keith A. Jones

Some pest~ddal and other OOnefic.ial org,anisms have been very effective ~n the laboratory only to faUat some stage in the f~e]dteven after development ,of a p[oduct for marketing, Common causes ,afthis demise are poor stability of the product during storage prior to appUcanon.~oo ~itt1e active material actuaUy reaehtng the ne1d ta[get~. and rapid d.egrada~· t~on. of the active material on the'ta.rg~t. Fon:n~J.danon plays a vital role in helpIng to solve tnese problems andm making an. organism effective 1111. p'I'actke. However, th~s m.ust be achieved ina eose-effective manner if the final product is to survive co:mm:erciaUy ..

What is formulatIon? Deflned collectively, Iormuletion comprises aids to preserving organisms, ~o delivering ~hE!m.to thelr targets and - ,once there ~. to improving theiractivlUes. A~echntcal correentrate of an org.ani:iim tha~ ha~s been formulated is termed. a formolation, or a. product, w.hkh maJ¥ be stored and put on sale commercially, A product often does net fully serve aU the" requiremen ts ,of use on all crops. More additlve.s may be needed to aebieve opUmumappllca,tion on some ,crops. These are normally added just before application and the final Formulation applied is termed a tank mix.

This book deals spedficaUy with formulat~on of beneficial microorganisms and nema tooes. These are divided into six groups; (I) microbial insecticides; (2) microbes that destroy, i:nact~vat~ or compete wHh plant

pathogens; (3} mlcrobi,a~ herbicides; (4), bene-ficial ·mgan.i:s.m:$ tha.~ improve plant uut:ri~ion; (5) microbes applied ~o seeds; and (6) entomo'phiUc nematodes. Form ulation hastendedto be considelled and research has tended to proceed indepena,ently in these sb:. groups. However, they have common str'e'ngths and technologieal weaknesses, as well as unique features, so' WQu1d. benefit from cross-fertibzaDon and! assimilation ofideas and data. The varices gt1DU ps rOf microorganisms are summar~,zedand compared ~n Table 1.1.

General prin.dples ·of form./I,datruonare esta b.. lished in Pad 1 of th~s book. Formu.lation is d€l~ermifil~d not o:n~y by sdenUfic requirements •. e.g,. uniform. spray cav,er on. fo.U:ager but also by commercial requitemen~s. e .. g. user friendliness. Products must be easy to store, easy .to Mse and eompatlble with users' equipment.

Critical formula tion requi rements are determined by features of the organisms themselves and of thei'f environm.en is. An overddill'lg f:eatur·e ,is mode of action: it dictates the' formulaeor's ultima.te objecti.ves and, therefore, ~s used as the basis for thre€' sepi.'l~· ra Ie pa.m in the book. Some organisms, sudil as insect :pathog~n~c bacteria and viruses, ad through the gut and must be eaten to take effect; these are dealt with in Part 2. Part 3- covers others - saeh as insect pathogenic fungi. bacteria and fungi. that control plant pathogens and weeds, Rhl:zIJbiun'r moculants

.Formul.alion of Mkrobi'tll BlopesUc:ides; Bemificial mit'roorganism±i, w)'matod~';\; tln,d ~li'IJ:d lUll'.tmm/s. Edited by H.. D. aur~es. Pt!bI~Slhtld i:n. 1998 by Kiu'W·e:r Ac,ademk P\~hl:ishe:rs, Doro~ht [SBN () 412 6:25~O 2~

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2: l.ufroduction

Spo~fonni.ng baclterilal in:sedidde

Prot02.oa!n lMecUdde Jnsect viruses Miyrnimec:tkrd,C'

Cr}'Sb~ tOiXlJ1!. dllrr:abJe s.par:e

DuraMe SPOIJ1P

Du!raMe in:clllls~on bodiy Relatil\fely del!icatc or d.uta.t)lcspore Relatively de~icat.e spore

[)e:]kate baderial cen~ Wl;lgh spore

Ouriabh~ or deMcail.e spore. rnt'Cileriaio!l' adi:nomY(pte ~n Bacterium. delicate

Mycoherbidde

FMgi,. bacteria combating phm!t pathQogen5 Bacteri.a~1 fUf1lgal~ Syllli!lbion~ Efilito~ophUi(' nematodes

Infoctivestage (arid asseciated bact.eda) deJk,atc

Sro~ach poi~ irllfection

l:n~ects vtag:uf Infeds Villli gJut ~n(cclts on contact

Plants,urfares~. wateT, SQi~ Pilant sudaces Plant SI!lI.tif,aces

s.o~.I. plant surfaces, w.M~If. insect rut~dc P~ant surfaccs~ soil

Intects on contact

Inf,ectso:r mhilbib QD (lOntad

Sail

and iI\I'indenl sua.ins ofpl,a:f1It pathogens - whkh ad, .~n:fecl Of CQlonilJe aft~:.r contact with ,external surfaces of the pest 0.[ plant. Part 4- de.aJs with e:ntomophmcnematodes which have a power of se.arch before infecting ('[able 1.1). Thus the IDm1ul,aior must rnncent~ rateon ,enoour:aging the pest to eat the organisms .. or on faciUtating conbd between the OFg~nism. and the extemal surfaces of pesf a.ndplant~.or on. preserving search mobility" as wen as provid~nga food. base for proUferation of the organisms in the~r new surrounds. Contrasting prob~ems and ctlaUenges are often presented to tne fomu.da'tOF by U'liese [eq'lilirements~ together with the needs of the environmentsto which the organisms are applied and the p.urposesfor which they 31re used. The organisms areappl:iedto target insects" or plants or the~r suibstrales, which. :iindude fo~iage,soil, "vater and commodities :in ft:J<'.W..1 stores.

Various Uk stages andloxins of the organisms are applied and. these v.ary great~y .in their robustness. Their fomrldat~Oll requirements are more strin~ntthanthose for dlif~m:i.cal. products. The activeingrrooien.t is

ofltern.a living organism. which must be kepI alive arndin g'ood heal.lh in order to ach~@ve the desired effect. It should not,thefe~~o:r!e# be subjoctedlioharSh chemka.1 or physicalkeat~ ment during the fomnu]llatianprocess.There may a.lso, be criti)ca~ .nub'.~t~oillal orphyskaI requiJremen~:s fo[' org~nism ma.mte.nanoe. Organisms ar,epartirula:~e~ wh~ch further complicates fomudation. Since beneficial organisms are regl1li'dedasenvironmentaUy fr~endly ~ .it is desirable that anyforrnulati!on addifiv,€ also should be ,enviro:n:mentaUy benigrn.morde.r to [Ietailn th.is advallta.Ee.

On the posUiive side~many organ~sms multiply and spread from the site of app't1:ca~ lion. AJso~ thr.ougb g:€li1eHc ,engineering, impwvements can be made in activity~ stability or distril!:ndi,o:n o.f an. orgamsm. I(o.r its active iogrediemt}.Any of these features can red ace the~omi'lu.lati.on .requirements for effectiv!e acti\r~ty. The insertion of geneHc factors .~nto the plant genometo prOOJJ.II.oe a transg,enic plant is regarded. .forthe purposes of this bCKlkaslbeu]timafefonnulation,anal& gnus f,o a systemk iln:sectkide but superior as it lasts the life ·ala plamt and is b'arnsmlift'ed. in

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the seed .. This method made a b~g impact in 1'996.~ itslfitst year of wid.e ,commerdaluse~ and should encourage the use of other spedfic 'COl1ltrO~ measures, induding microbial insecticides, in jntegrated p€'St management (IPM) systef[1is.Howev,er:~together wiihr,ecmt new chemical insecticides, itVJiU cause severe competi tion wi th.B.adllus thurin:giensis,lhe le'adir'1goonvenHonal mierobtel, Just when the biopesticide manufaetusers areleaming how to compete withchem.icaI pesticides, these itworun_llov.a6ons Jernd ill note ofursency to further Impreve the effectiveness ,of conv'ennonal microaialproduets bybetter formulatiion.

Despite the central role that. effective formu[aNon plays fn the practical and eeoncmic success of eonventiona 1 mlerobtalprcd ucts, the formulation of organlsms is a neglected areawhen comparedto fundamental aspects ofreseareh .. Al though. th.s may be due part~y loa. lack of published mlormationr,esultin:g from commercial secr,€'Cy (usua.Uythe most effective wa.y or "protecting' formulattcn infonnaHon), it is du.e also to a. lack of acknow ledgement of the Importance of the subje<:t.Thls. is compounded by some reported field trials which ,apparent~y mdicated that the formulation did not drastically .improve fie[dactivity {Bull, 1978; Couch and Ignof60, 1981; Payne. 19,86). However, such trials are subject to the normal extensive variations of field plots whlch often mask the desired effects, a handicap to field experiments that. is ,ClQunter,edm the foUQwing chap~eTS by-assembling and assessing as much data as pessible,

Reeview Uterature on formulationrequirements 0.1 organisms has not keptpace with the development of the subject. Formulation of ml.cmbia~.pestidde:shas been reviewed by Mos~ and Quillian (1986) and [}evisetty (1988,). and to·r a wider range of orgaJ1ll:sms by Rbodes(1'993), amongst others. These reviews, however, have concentrated on. generelprinciples and have notpresented data ebout formulation pfooesses, addlttves or

hlt.mduclio.n3

effects, Some earlierreviews ghl"iiilg Ilmlted data have been. ,confined~o products aimed at specific narrow target ranges, and 'even these lack practkal information. on. specific fermulatirm ingredients and methods (e.g. Angus-and Lu thy I' 19?'1; IgnoffoandFal(Jon, 1978; Couch and l.gnoffo~ 1981; Soper and Ward, 198]L~ Yoangand Yearian, 1986; Connick et a1.#t99fJl; Diagle and. Connickr 1990; Mdntyf:l~ and Press, 1991)..

The present volume endeavours to remedy these shortcomings in depth, btl t ~n a readerfriendlyfashion, Here are some suggestions about how best W use iLPad 1 describes types of applicatlon machinery OGmmon. for m.any groups 'of organisms and for chemicals toe, ,exp~aLning the types of formulation needed for efficient use of products. In su:bsequent Parts. the arrangement of organisms ingroups around mode of action facititatespenetrati ve assessment of the bask requirements of formulanon in fieiation~(!I targetorgarusm.afild environment, by ,eX:p'la.in~ng in. an analytical way why each gr-oup needs somewhat different formulatien, Thebookbrings~ogether information ln a. compareti ve and. critica 1 manoer so thateach group of organisms can benefit from ideas., technolugy and. progr;ess in. the other groups.,withiTI the framework of a maier reference work. Much effort has been made to .insert cross-references between chapters to speed comparisons: the text has been. designed to facUitate navigati!Il[1l and locating figures and tables quoted incross-refereaces, particwady by givmg~e section in which each occurs. R.ead,erswith 'laded Interests are serviced,ranging 1111lm those probing theory and science Ideveloped in the text oif chapters), to those desk~ng practical detail on individualadditives. Detail ts partitioned into tables and appendices. Atilention ts direetedto a series ,of recipe-type tables which ,exp~aim how to rnakespecimen formu larions.WitihiiJ'il. tbe chapters comparab]'e data are assembled, assessed andcompared as far as possible. In. (loriltrast" in. Appendix I additives .~ which are the baste tools offQrmulation-afe ~isted

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alphabeticaUy fore.asy n?feren(£. Since there is a bewiMerin.garrayo,fmost sorts of additiv{'S, only those .actuallytriedwUh. organisms are induded~ not only tbose that haive been successful, but also those that have failed# together with Information. on harmful ,effec!ts af unexpected! beneficial effects. More data on each additiv,€ can De sougb~ byreference to theIndex, which haspu.rposely been .made comprehensiee, mevitably, mrmulation terminology relating to :rnkrobials has become somewhat specialized. The glossary in Appendix m defines relevant additives and terms:oplusa v,nriell' of additional useful words. Obviously a global system is desirsble for termrnol!ogy offo.nnILlLat~oI1l1pr:od.u(t types for microb~als and it seems reasonable to (O]h)i"!il' the intematlenal system recommended forchemkalpesti:cidest so these terms are Hsted in. the glossary. Research needs and the future are assessedat the end of each chapter, then reviewed"oompared and discussed in Part 5; Ollsis possIbly fhemoot important aspect of the book.

R.EfEItENICES

Angus, T ... A. and. Luthy I P. (1971) F(Jrmulation of mkrobi.al fru;cctiddes~ in Microbial CO'utr;o1fJ/ llfseds a.r.ra M.r.trs (eds H. D. Burges a,l'!Id N. W. H!tIiSSeYl, Aca:dernicPress., wmdoo, PI"· 61J-.:38.

BulL D.. L nm) F~nno~ati()n ,of Olicroblal~nsectkiides; mi.croeru::apsulation and .adjuvants. Misc .. Publ. Ent. Soc. Ihn. lO~ 11-20.

Connicik. W. J • .h~ Lewis, J. A. and Quimby. P. C. Jr .. (1990) Fonnu.latian. of bhx(i!l1Ibol ,agent$: lOfUse in p~anif patho~ogy. iI1I Neal nir:octio,,$ .iu Bial!I1gfcill OmtroJ (eds It R. B.lker 3!nd P.E. DunlO). Alan Liss. New "(:ork.,pp .. 345-72.

Couch, T. L.and Ignon·o:. C. M. (1981.)Formu~ati011l ,of insect pathQg~ns, in Microbial GI'Jltroi oj Pes's {;1M Ptau/ Disea~ (ed .. H. D. Borges)# Ac,ademk Press, London .. pp.621~35.

Dev:~t:ty, B. N. (l9SS) Microbial ~()rmOlatiOOiS - op'Por~lJnilies and chaUenges,in Pestidde Fl'rr:mu~ lafip'IS (IUd A:pplimUt;lu Sy$t.fms~ Vol. S (005 n A. Hovde and C. E. &estman). Arru'!dca!n Society fur 'fe5ling Mllterjails (ASfM STI? 980). Philadel~ phia, pp. %-64.

Diagte, D. J. and Connkk. W. J.):(. (I'm) Fonnuta~ ti.ornand. applkation technology for microbial weed (OOtr,o!). in Mk~ aud Micmbitll Products a,s Hl!rbldd'eS (ed!.R. E. Hoagland), ACS Symposium Serles No. 09 .• A1l1lelican ChemkalSooiety # W.ashington DC., pp.2.86-304.

fgnofto~ C. M. and Fakoo.,l. A. (Ied;s) (978) For!inl,daitiOJl and appUcat~o!il. of microbia; insect:. lcides, Misc. Pubi., rut. Soc .. Aun"ncn 10, 1-80.

Mdntyrn, J. L and Press, L. S. (] 9\H) Fonm.datiiOriI. del~v.ery sy.sternsand matkeUng of biooon'lrol agenls and plamt gl'Pwth promoting rhiz,oha(:~ tenia {rGPR) •. in Tbe RJdzosplrere (1ud .PJ'Qrd Grm~rlh t{eds D. L ~lsterand P. R Gregan.). Kh,lwer, The Nether1a;nds. pp. 289-~.5 ..

Most" B. H. and Qui:lillan. R. J. (1986) Fonillulatton of b~o~og~cal pestkides, in. filrraarJ~errh'd alJd AppUed Aspects of lu~'i'rltbffltel?ntllo/~ (OOs R.. A. SamSOfil, J. M. Vlak and D. Peters). in Prlooeedings of fhelV Inlemal:ilOOOl CaUoquh .. un (llii! b1l\i~rtebfaw PalhOI.ogy, Wag~iflgen.,. Society for blrvertebr.ate

Pathology, pp. :624,-:7. .

Payne. C C. 1(1'986)1 Insect p'athoc~rikvirosesa:nd pest control ag.ents, ttl ffiarogical Plau' n.ml HmUl, ProJ,rc.tfcJU - Bwl10giml Con:troIo/ P,laut PlSis ,auil of Viffl.ats of Hunm~r al~d .Anima.lDisease,hogress in ZooJogy V,ot 32 (ed. J. M .. Frainz)I, Gustav Fisher, Stuttgart. Pl'. 1 ~200.

RluxJ,cs:. D, J(1'993} Formulation of b~oh'l;gkal rolil~ trn~agentsl in E:tploi'alirnl of Mi~.rQO":8'QtJ.i$n.l$ (ed. D. G. Jo.li'I.eS)l, Chapman ,at Ha~l. l..cmdo[1l,pp. 4U-:39.

Soper:. R S. and. Wani,M. G. (981) Production. .f~rmulaUo.lil and. arpilkation of fungi ((lit i~ ~ontml,in Bi~IClgiml Can1rol in Crop Pr('Jf,ediou, BARe Symposh.l!rn. No.5 [ed, G .. c..Papavizas), AUalil_held, OHawa~ pp. 161-80.

Young. S. Y. and Yearian" W. C. (1986) FOflT1iu~ation and application of barukwiifl.l~"S .• in Tlw Biolngy of Bncrll('lvirustS .• Vo!. 2 (cds It R. Granados alld B. A.Fed~dd).t CRCPress, BocaRat(m~ pp. 15~-79.

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PART ONE

-P>R'·-I'N· ···.C·· ··I·~P'·-LE··'S·') 'O.·;F·· ·F···',.O·c ·RM· •• ' .··IU'L! A··T:·I'O· .·N:'

'" '_', _._ ._' . __ ::~,_I ~'I_~~_.' ,_ ..... _ , __ "';_', ,1_," __ ' , , " ,=-,1,.'. _' . ...:_ __ ';. I, ._

Ponnulated organis.ms are suspended ~n a suitable carrier, which is supplemented by additives to maxirruze survival in store, optimize appHcaUon to the target a.nrl protect the organisms afterapplication, Incontrastto chemical active Ingredients, they are 'particulate and live or proteirteus in nature, making them flf;latlvely sensiUve to storagecondihons and the environment.

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2

TECHNOLOGY O'E FORMULATION AND APPLICATION

Keith A. Jones and H .. Denis Burges

OONtENTS

2.1 Introductiol'il

22 Functh:ms of funnulil~jQn 2 .. 2..1 StabUization

2 .. 2.2 Handling and appHcation 1fi

Z.2..3Environmental persistence 19

2 .. 2..4 Improvement of action 2:l

2.3 Types offonnulation :2:1

2..3 .. 1 Dry products 22

2.3.2 Liquid suspensi!ons 24

2.4 Condusions '11

References 2'l

2.1 INTRODUCTION target irtl~he mostapprcopda~e- manner and

Fonnul,ation le(hnoiogy m.ust be ~onsidered at all stages from production otenorgaetsm to its eventual action. ofil.lne ta,rgeL The method of production allen dictales subsequent formulation adivities.~ which in tum may lead to alteradons being made in the production procesa.The .range of organisms is given 'in Table l.t, section L'l,

There are four bask functions ,of ~orm ulation, These are:

to stabilize the organism durtng prod uctton .. distribution and storage;

to aid handling and. application ,of the prod uctso that lit iseasi ~y de~ive:red. to the

form;.

to protect the agent {(,Om hannft_d environmental factors aft!h@ target site .• thereby ~ncreasing persistefl(!€:;

to enhance activi ty .of the organism attl1le target site by incr,eas~ng Hsactivity,repc,O'" ductioT\, c(mtad andinte1:'actiGn with the target pest or d lsease organ] sm.

A wiJde vadety ofapproachesare available to the formelator to achieve these bask functioM1rangmg from productton .of Uquid suspensions ~o sol idbdquettes,and even incorporation of the agent in a Uving organism .. The final produd devetoped d(!pends on.

.fWMlltjll1fWJ1fJ! MrcmlJi.d B:,!pf;;h'c,Mw B~~,~pdl,llrmj;~,,&a~'i~'n~. rIClI1I!'1I!1d~·.~II~,d~d f.~h'J{!'.n.l·s"Edi'!~ bv H- [)_ DlI~es" P:ub~ish.ed. i[ll 1'998 rbv K~lJwer .A.cademk Pubhs.hersr Dordrociblt [SBN .0 412 6:2520 2.

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S Tedmology Qfformrdationaud appUcali(m TaMe 2:.1 Prob~fl'mlsfac(-d1 by a fOffilul.iI:tor

neduction of mate'nal bulk..

Divisionin,to partidesabte to pass spray no!Ules

Pr,e\1~"l growtb ·ofagetlltand ,contaminant microorga:nisms. Pl:lev£lIIt pmteaSl?S denaluring acHve agent

Avoid powders caking d.ueto m:l;!lsWl\e uptake ..

Centrol v]S{;'osi:ty ,of liquids to keep p1.trtk~~es in suspenslon without aggregation so that they flow.

Retahll viabHity

EnsUifie good performance ~nappnc:arors tor dusls/ powders/greneles, M.a.intain. appr~priale vi;scosily .of~iql.!lids to form sp.ra)' drop~et:s. EI'ISUfie goodpedormanCE' o~ spiTay,eirS without makmg roam

Ensul'l\' good rove.r:age of ta.rget and good:proouct l'IetentiOfi. R.oouoophyska~ '~05Sbyra~n or otbermeans.

Protect agenls knm ma'iIi"atii~ faCfOts, ,e .. g .. sun.

Ensure deposU~$ pil,latabJe and preferably ,attractive to tatgetpest

S\~rage

several factors, such astype and .iocat:ion of target~a\'anabUity of formulation materials and app~icatj,on.equipment" as well as user preference, A fortTIlI.labClr Is, therefore,fa!c@d with several pr,oblems and ,chaUeoges., summarized .. ~n T'abl,e L.I.The aim of a. formulation process .15 to address these problems.

'Qptimization of formulation toaddress one problem may result in an adverse effect on another aspect oftheprOOuct's.f!Uncfion. Formulatlon therefere nO:rn1laUy represents a compromise between these opposing effects. Due totlile d.iverse' nature of d imatic sJituaUons.~ targetsand_ user prefereaces, a. single organism wiUonlen. be formulatedin several diffefient forms, each aimed at a. particular market,Broadly formulations oforg~n~sms can be splittntotwogroups, dry solid and liquid .e. AU~ormulatiof1liS .• however, must still

k.-. ........-k .,~ . md . •. .. .'. n. ·'bi f-

.L)'I:: pm..,; ~lc;aL an.' eeonomica _ y VUlt Ie. .. or

example, it is not practical to develop a. prod ~ octthatindudesapmtecnvE' dye that also stains the skin of th.e operator. I~.is not €COnomica Uy ".iable to add expensive sunscreens to a product that wiU be applied at a lidgh volume per unlit area unless they are formuI!ated!o stay in. dose juxtaposition. to' the organlsmto minim. me lhequantity required (Appendix U. fig .. II ,6). Further ,challenges are

presented by a prod uct that m.uSf be eaoen by a target pest . .II must be di:slribu~ed ev,enly in. the area wn.ereapest feeds, or - to attract pest to, active orgafilism - the product must be p't',e~efilentiaUypa~atabfetothe feedmg s,lage ,of a pest. In. contrast .. an. Qrganisma:cting by oontad can attack aUstage.sofa pest and. may be pickedup as the target pest movesaoout, so d ~stributjon is less critical and palatability is nota eoneemas long as the product is not repellent,

T.his chap~er eleboratesthe four bas.icru.TIIl:tiions of a formulatlon and how theyaffect (ormwat~on approach; it also reviews the main forrnulaf~on types suitable for use with orgllnisms!. Both function and. type are in~errelated..eac.h influencing the other. However:, other consideratiens must be taken into account, General priindplres and approaches are d~scussed., and. descriptions of p,artictdar prod Il.tcl:s used for dUiierent organisms are covered .in. ensumg Chap~ers. Formulation types for appUcation to soilare reviewed in section '7.5 .. Technology for seed treatment is descrjbed in Chapter 8. Appendix Ill lists and. defmes types of fo.rmulation and additive .. Appendix I describes individual addinves known to have been tried with microbia ls,

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1.2 fUNCfIONS OF FORMULATION

2.2.1 Sf AlintlZA 1l0N

The lo~g.est. period of time m. the life of a product elapses during storage, ie.lheti:me between manufacture and eventualuse jn the Held. This period can range from several weeks to Y'l?a rs. Organlsms a re requtred to remain viable during s~orag,e# with minimum loss of potency!adivity a:nd without loss or breakdown of the desired formulation properties.

Couc.h and .Ignoffo (1981), state tba.t a shel."ute of 1.8 months is the miNmumpracticai fo'r mkrobiid pesticides. However, most currently available commercial chemical pesmcides require a minimum shelf-life ,of :2 years, and Rhodes (1993) indkales that up to 4 yeats is desirable, This should be acbievable at temperatures at whichthe product may be stored. In the tropics it is not unusual for temperawres in somepesticide stores to reach 40 °e' o'r more tor extended pErioos. FOIr chemicals .. thls tar;get cannot beachi,eved"..Tith many unstable a,dive ingredients withoutapplOpriate additivesto prevent breakdown, e.g, antioxidants. Biopes.tiddes and beneficial microcrgasusms (T able 1.1 in section 1.1) are usually live, albeit aftenin a dormant stage, :SO are generally less stable than chemicals and cannot easily be alter,ed ,chem:k,aUyloimpr,OvlE' st:abiUty (secti'on 10~2). Jmpreved stability can. however, be achieved by tn:-afm!ent before fermulation,for examph! by appropriate growth conditions dUfLOg production (e.g. Whipps and McQui.t~e.n, 1'99'3; sections 4.7.3, '9.2) .. by appropriate storage prior 'h) ~onm.datio.n (e.,g. Georgisand Hag:ue, 1'99 t) or byappropaate pr,ooess~ng after pr,od UCtiOIil.. such as drying (e.g, secticns ,S .. 2..1~ 3.2.2~ 4~7.3; Jo:rlu~,s, 1'994). lnadditWn~ 'or i.ltemati v,ely ~' additives a re irscluded tolimpfov,e stability,.

Depending on the organisms Jrwolved, different problems mu.st be addressed 1'0 improve stabUity. Some organisms, such as nematodes .• may require air or free oxygen

2.2.1 5.ta,biiizali0I19

(section 9.3.1) to remain viabi.e. as weU as [,equiring appropriate 'energy stores. Depletion of energy stores in this case may be limited through restricting movement during storage (section '9.3.2:). Others" such. as fun.gi. may be stored as a resting stal,ge. ,and additives that prevent premature growth may be needed or those wi th a nutritive value avoided (sections 4.6,.4" 4 .. 6.6:); also moisture may be a crit:ieal factor (section 4.6.3)., Som.e additives that .inhibit germination of 'the organisms after application. must be avoided (sections 4.3.7~, 65; Roberts and Yendol, 1971)"

Willi dry products, deterioration aceeler,atesif the-prod uct m,oisture ,content is a lloWied to increase prowessiv,ely abeve 5% i(,e.g. Couch and J~offQ,19tn). When exposed freely to ambient air. the moisture content of a materialalways approaches an equilibrium with theair humidity. Equilibrium moisture conteat increases as the air humidity increases and may be exeesslve for hygroscopk materia.ls. Even ~hose not nonrudlyre,garded as hygroscopic may moisten enough. to cause particles to adhere together. sometimes inducingcaking and .sometimes deflerioration, pa rticularly at high temperatures. Similarly. moisture above 5% can impairprod urn wi til oil as canter, as has been shown with insect pathogenic fungi (5&00.111 4.!6.5). It is therefore importan.t to store products im w',ater' vapourproof containers, sametimeswith ahygroscop~c additive such as silica gel (sections 3 . .3.6.4.6.3). Removmg water from, or dry'ing, oil carriers by heating prior to incorporation of the microergarusm into the formuJation may be necessary.

The pH .of a product must be stabilized within certain ranges, Very higJR and low pH ,conditions wUI nonnaHytl:ilzu::tivate ag.ents (e,.g. Grim~hs~ 19,82; Salama and MOlTris,.1993; sections 3.3.6; '4.6.6'b) . .A buffer may th.erefo're ber,equired, and certain additives with extreme pH values must be avoided. Maintenanee of opttmal pH may improve sheEH.ife (Date, 1970).

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Products based on water, such as some baeulovlms formulations, may support growth of Ciontaminant micr,oorgani:s:ms~ mos(wi.th.alkaline or near neutral optima. Contaminants may indudeantagonists of the formnlated arganitsm; they may cause pH changes due to accumulation of waste products; ord1ey may produce enzymes ha,rm,ful tothe orgarldsm. Gases from .microhial. activity can. cause explosive release of the product prior to~ or on, Qpening. Initial levels of c.Q;ntam~na.nts and the range of species present may be I,imited by appropria~e condltions orlre,atment during prodlilCnon. andtim:imlg,as wen. as handling; of harvest (McKinley et al.~ 19189; Lisansky et al.~ 199'31; the problem may be: minim.ized by ensuring fllat inocu lants or substrates used in. theproou,ctjon are ... as far as possible, free from contaminants, e .. g. use of food-grade ingredients (Shennan. 19'85). A useful method of minimizing ,growth 'of ·contamiflaots in a product is to maintain the pH of tbe suspension at a va,lueoutsidethe ,optima for growth of contaminants~ but notinlllbUory of infectiofl,gwwtb. or replicafion of the biocontrol organism af~er applkation (section 3.3.6; Jones and Burges. 1997). In addition" or a~tematively! it may also be necessary '~O iindude an additiv€fhat restricts the growth. of ~ontaminants. 'Caution is reql!J!ired. however, in tne use of medkaUy important anUbiofics which by widEl-sca~e applicaUon in the ,environment might cause res:lslance to develop im mkroorg,anisms pathogenilc ~o huma;nsand other vertebrates, Growth o.f c-antaminan.ts can be avoided by drying the prodact,

Many eommercial products rnft1enHy avaUabl.e avoid part ·of the problem by recommending immedilateuse or storageililooI1l~ trolled condirions, e.g.re.frigeraltio.n (Rhod.es, 1'99.3i; jones and Burgesof (997). Howeve.r,this may not bepractical orpossib~@ in many situations~ sl!l!ch as in developmg countries. Moreover .• i.t increases costs, thereby reducing competitiveness with chemical altematives ..

Rhodes (1993) points to the commercia.) success of seme o.rganisms desp.irelh.e need for restricted storag.e' l'equ.irements! but their tot.al market va.hi:l!e, now and in the {ublre~is limited .. Special storage condi.ti.OrIS shou~d Dot be regarded as an aUemaHv,e 'toappropria,te fomu:dati:on.

Stabm.zingagenm may also be needed to pro~ocldil.e agent from the ha.rsll physi.ca.lor chemical treatment sometimes required in. the formulation pl"('lC@Ss~(or example to reduce shear effectsm grmdimg or mixing, or 10 proteet the organism &om hea.tinacfivation dUF~ ing sptay drying (section 4,;6.2).

The producr UseU may require stabilization.

For examp!e~granules and bdquenes may need blnderstamaintam their integrity over a periodof time. Uquid e.muls.i:ons require Sllabilizers ,to maintain the emutsi.on.

2.:2-:2 HANDUNG AND APPUCA1ION Add.iti.ves ensurethat the product is easy to handle and ap'ply. For example~ in suspen+ siornsthickeners or suspenders hel.pmaintaim even distribution of the organism in the carrier, prevent dumping nEttle organism and ensure itsready resuspensionafter p'mlonged storag.e.Dusts and w,ettaJble pOlwdeuoontaill additives 'fro prevent dumpilllg and caking (sections 2 .. 3. Ia, 2.3J.c). Additiv!e! types are lisredand defined m AppendIx HI and additives tried with microbiads are d.escdbed. in. Appendix L

E1ffeclive and econemic use .of a product r.eqldresthe active ingfOOh:mt torearn the target: no mlaUe.r bow good the p:rOOUd, if it does not reach the target it win not perform: the ooquired function. With some chemical insecticides ttids problem was partly solved thJ\ough the development o.facfi.ve ingredients or formulationseble to move within. the planl- tr,ans.1amina,r or system ie action, Nematodes apad"m.ovement of microbial agentsa.fter 3pplicatimll lis not possible using conventionalbOCMology, ,emphasizing nile importance of ef£ectiv,e appl ica.tinn. Genetic

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E'rnlgirneermg has developed transgenic plants and mjcroo~gamsms a'b~e lo express microbial products such. as the ~oxms ,of ,Bacillus: thurina gitn5is. These can be considered as highly spedanced formulations,. equivalent to systemk chemical insecticid.es. Hnwever I beneficial orgaulsms have Ito beapplied insome way, u:suaUy- but not always- requiring the use of app~kation 'equ~pm,enl.

JEffecnv,eapp~icat:ionis infilnenced bytwo main parameters: appropriate equipment and fcrmulatton. These two pa:rame~ersare in~err~lated; 'equipmen,t wmnot perform at an. optimum if the characteristics of a product are not .sui.1able,and 'vice 'versa (e.g .. Appendix Tables IIA, II.?; Figs 1I.41 n.S). Thefo'lnlulalOr aims to prov~de the proouct in the most suit,abl:efonnfofopfimalperiormance ,of the appltcation ,equipment.

By far the most research andinroifmaULlJrl! on appUcationted:mologycon.cemsrnemi.cal pestid..::les,.Ther>e are comprehenslv'c:l' gener:al r~views (e.g. SoldhoomtJe,. 1985; Matthews~ 1992)~ reviews on specific appHca.non techrl.i+ ques I~e.g. MaaSI 1971; ReardoIil.. 11.99'1)" and. others on appJi1cation of specific groups of pesdcides (e.g. Rossand Lembi, 1985). Some reviewsoonrentrate orrformulation principles and tbephyslcs ofpesHcide appltcation '(HarUeyand Grnham-Bryce, 1'980; Badow .• 1985) .. However, many ,of theprtnclples (an be extrapolated to microorganisms. Some criticism has been. made that app~lcation of mkrobialpe;lticides hasrelied too hea.vi.ly ,em technologyd~eloped :ilor chemiealpesticides, and more research should be devoted to, novel methods forth.e biologicals. Even. so,m most situations a farmer {parlku]arlyon a smaU scale) isunlikif]yto buy special equipment ~Ilr microbials .. w~ereasequ~pment used. for chemicals1most1yhydrauUc nozzle systems used at low '0 high volumes (Tabrue 2.3 below )~Is already nkE'~yto be on site, and c{)mpati'bilUy of mkrobes with. such sy:s~ems is more likely to Jead~otheiruse. AppU,cafion of microorganisms has been discussed by Smith and Bouse (1981); Diagle and Connick

2.2.2 HandliHgaud .applicatioll 11 (1m); Reardon (199]); Jones (l'99,3);Bateman (1994).

A pesticide is brcadcest im. the region wile-lie the target is located: somewill impact on. (If near the target,; much wm misscompletely, Forexample, Himel let al .. (1990) estimate that as Iittleas 5% of the tlotal active lngredl,ent applledreacbes ·lhetarget si~e,and Graha.mBryce (197:7) estlmates that >97% of applied pesticides are .1001 tothegenera~environment. Much can. be I,ostwaugh being Mown or drifting offta:rget (Wod.ageneh and Matthews, 1981; Appendix Table 11.3)1 and a signUi:C'ant amcunt .ofproduct csnbeunce or be reflected. hum atar8~t. W.ith micreorganlsms there is also some suggestion that particles are 'lost" between the spray tankandta£get"as estima tes for the volume of suspension reeovered and nu.mber of mkr(~orgarusms do nol'taUy (Smith and. Bou5e,l9Bt; Richards" 1984).

Applica.tion equipment is designed bJ maxim~~ the effectiveness o.f a product through accurateand safe delivery within. the cons~:raints of practicaii:t:y and efficiency. Equipmentfor so~idand. llq u.idfo.rmula.tions ranges (rom small hand-heldappa:ratus~olarge maclhinery mounted ontraetorssndaircraft, Types of equipment suitable fOif use wim microorganisms are sum;mar.izedin Table :2,:2. Types of formulation are discussed. in. section 2.3.

2 .. 2.2a A,ppl irn.tionoj so.lid!or,nulatiolls These consist .of dustsl grnnules and bdqu.ettes; the.ir properties are descrilbedi. in. section 2~3 .. 1. WaIk.er (11976) summanzed the requirements ofa good. granule applicatores being ableto deUver accura~[ycahbrated amounts of produc! and. to spread them evenly ~without damagingpreductsby grinding or impacocm. The machine sh:ouhi be' robust and easy to handle"calibr,ale ana. repair, as wen as inexpensive, These criteria can be appHednot anlyto gr.anu~e applicators, but also ito ,aU types of apphcator ingeneral,

Dry pred li1ctsl parfieularl y granules and bri.quettes, have the advantage thatlhey C3.riI.Ve

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easi~y distributed by hand. Speda,~ized equipment r,a,nges from simple hand-held devices - such as a 'pepper-pot shaker' r which is partkulady suited to small-plot, ~ow-input agr~cu.~t!ilreand gene'raUyaUows the product ~o be placed a{1cura~e~y an the target - to' large traC'to:r~bo:r.ne ,equipment and airc:raft su.ited to application over large areas (Table 2.,2). Available machinery is described in mere deta.H by M,atttiews (1992}.

·2..2.2b AppJica,tion (if liquid lamm/,a,liorlS In these the organism is camed in EI. ~iquid~ normally oil O.F water. Addltion of sur:factant (section 2 .. 3 . .2) or oil and emulsUier to w.a.~e.rr or use ofpun~ on, forms drops of mO.re eV€,.F1I size (Appendix 111 Fig. HJ) thaa those .of water alone, with eonsequently betser controlled spray + OHls pre~ferred~Qr ultra low~vo~ume (UL V} spray:s, water is normally used as the difuent at tdgher voh u mes, Talble 2.3 classifies volume appHcaHon r.a.~es. low volumes are preferred, in areas where water issc,a,rc~I' because they redllc,e the vohsmeend weight to be transported and the Hme needed for appUc:a.UOfi't. Higher vclemes are aimed a.t pro-v~ding romple't.~ wetling of a target surf.a!cf# a ~tho!:lgh this also causes h~gh run ~off ·of spray from the-target. The general trend has there.f..:ne been toward reduction of v()~ume.With

chemicals. control has been best' with low volumes (MaUhews, 1~2) .. WIth. microh~a~s, re.s~.~ts have varied (e.g. 'SmUh and Bouser 1981; Topper d al., 19'84~ Jones, 1994~ Jones et al.t 1997), but this may be dille to a ~acl of a tterl.tion~o efficient appHcat1on.

Reduction of volume increases the need to optimiae spray droplet' size to max~mize eoverage of the target Coverage is influenced by drop~€t' v~scos~tyl' ~mpaction andretenHOIl, and depends an several factor.s. Impaction is ~nfl uenced by comp~.El;l( in~€.racdonsbetwEle.n. dmple-t size and velocity, as well as obstacles ~n its path {Johnstone ,et tIL; 1977; :R!~a.rdlJnlf 199'l;: Matthews, 1992). CoUectio.ri1 efficiency of a ta.rget in am airstreamis defined as ~~e ratio. of droplers stdking the object ~o tlli1le number that would strike it if the air was not deflected by the obj:&t. Collection increases wit~. drop~@~ size and velocitytand. decreases 'WIth increase' fntarget size, Himel cl ai. 0'990). dI vided drop sizes ~nto two main dasses: smail drops {<1rnJ.",.150 p,m d~ameter) and large drops (>l50J1m). The small drops are pdmarUy transportedon turbulent eddies and therefore' penetra~:e through foUagecanop[esl whereas the large drops are pr:i,marUyaffeded by gra:vity and hnpi~g~ onth.efirst substrate inth.eir path. A spray cloud [!IQrmany oonta:iJnsa range of droplet' sizes (e.g. Appendix HI Hg" n.u The droplet spectrum :is measured in terms of volume med~an diameter (VMD} or number medien diameter (NMD). VMD represents the d,ropiet diameter al which half the liqUid vetume is cQnta,ined ~n smaner drops .

NMD is the d.iameteI' below whidil.balf the numlber of: drops is smaUei. VMD is more oommonly 'Used. Spray uniformity (evenness 0·£ drop size) can be measured asthe .Fat~o VMD /NMD.fa. ratio of ~2 represents controUed~dmp~et .a.pp]ication spray. The

High v'Oh.U'fU~ ,(MV) Med~um v(}Jum~ (MV} Lowvdume (l,v),

Very low volume (VlV) Ultra ~ow volume .rUt V)

>600 200-600 ~200 5-50 <5

>1000 &IO-UIOO 2!~iQ!) 5&-200 <50

3=5 'l15"'"3

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statistics for relating measured droplet spectra a re discussed In more detall by Bateman (199.3). .

. A droplet !impacting 0[11 a surface can be retained, or lost through bouncing or reflec- 600 Ifdiscus:sed in detail by spmman~ 1984h Retention is greatly influenced by the abiUty of the droplet to wet fbe surface, largely determined by formulation (Appendix II. Fig. 11.6),,, For example.jhe spread of a droplet on a leaf is increased 8--16 times; and bounce off the leaf on impacticn is prevented or red ueed wUh formulation in oil. or as an oH emulsion (E. M. Chadd" Imperial" College, Sllwood Park, AS(1ot~ personal cemmunication),

It is critical. 1'0 ensure that tlhe correct amount of active ingredient reaches the target; this fs mainly influenced by the number of drops available for Impaction (Appendix Tables II.S, 11.61 uz. Figs U.21 [1 .. 7;. GrahamBryce" 197'7). Low volumes require the generation of smaller d rop~ets" limi too by the increasing importance of effects such as evaporaHo,n of drop~e~ and reduced collection efficiency such that an optimum is reached Oohnstone.. 19.8.5).. Table 2.4 summarizes optimum droplet size ranges for selected targets. With micr(libial pesticides the particu.,ate na ture of the active ingredient roay infl U!ence this optimum. Primarily aneppropdate number of active pa riides must be contained within a ,dr:oplet (Appendix 'rabies It5~ n.6). for 'example,a~ow- i.e, suJbletha~ _, number of B. tl1uri"gietlsls spor,es/toxin crystals ina small droplet may stop insect feeding before lethal poisoning. and the insect may recover and survive, It may be physkaUy impos'sible to obtain enough active p,amdes in very small droplets. lncontrast, with a v1ery activ!€ product, used ,at I,ow conceotranon, it is likely that a signi ficant number of smaller droplets would contain no particles at all (Appendix Table U.6). In th~s case the distribution of particles between d roplets should follow a Polsson distribution (Amesel1um let 0,1." 199,0).. The relationship between amount of active ingredient and droplet size is discussed further;

2.2.2 Hmldlin,g alld applicatio.n 1'7 TaMe 2.4 Droplet size ranges for optimumapplication to selectedtargets"

F~y1ing i!l1lSel;'rl:S Insects on fo~iage Fo~~ag,e

Soil

10-.80 so-so 40-100

250...!500

-After Matlh~ws (992); Bal-eman (1991.~.

wifh examples, in Appendix n and section 3.3.3a"

EffechvEn,e,S$ of add iti yes in a form tdation

may also be inRuem:led by d rop~et size. .For' example, the degree of H1trati.onby a sunscreen win depend both on ltsconcentradon and on droplet size.,whkh determine the amount of screen covering the organisms (Appendix II" Fig, U.16). Similarly the influence of a feeding attractant may vary accordlng to coneentration and number of point sources (drops),

The production of optimum sized droplets from a sprayer is greatly infmuen~ed by formu ~ lation, Drop~et. formation and size are influenced by viscosity (Appendix Table 11.41, Fig. 11.4). volatility and toa lesser extent surface tension of a liquid suspension (Sundaram, 1'988) .. The influence of 'organisms on vtscosity depends on their ccncentranon, Normal concentratiens of M,etarn.iz.iuUl /I,avovi,ride conidia had negH,gibl'e 'eHec~, 'but high concenrretion ('5@gspof,es/l) mcreased viscosity of a ULV spray in etl from 5-8cenUpo~ses ,(c..p.) (Bateman, in press], Been use ()f the pseudoplastte behaviour ot suspensionconcentrates, simple cup viscometers may give a more rehable predktion.of flow in spinn.~ng disc sprayers than more sophistlcated vlscometers (Bateman!. 1996~.

Partic~.dar sprayer types mayn€ed spray UquidwithphyskalproperUes conftned ~"itruncertain ltmlts. For example. a. ULV sprayer requi res a solu lion /suspension wUhwn a predefined viscosity ra.nge (Appendix II, Figs n.3, 0..4, 115) .. This is achieved by formulation in an ,appropdate carrier.

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18 Tochuo.I'ogy of Ji'lrmu.latiou a.uQapplication normally a mixture of v,egetab~e 8indmineral olls (Appendix Table n.of)., aUnougn wa.ter~ based. lJLV sprays (very low-volume .• VLV), are also possible. fdal1lY other sprayer .types are available (rab!e 2.l), fequ.iringform.ulations with. different ph.ysical. properties. Several types, :indudingknapsa.c'k a.ndh'actormounsed sy.stems~ ,eommonly .ba.ve hydraulk nozzles o:f various type:s# an re~ying om pressure to force liquid through .ana.rrow orm~e to fonn droplets. Some microbIal products may cootatm large partides, e.g, insect debris in viral insecticides. La.rge pa:rtides calli block nozzlescansing in£'!om.pieteor poor appheation~ avoidable by grinding dry produots.W€I' mill_gor filtration. lo. other sprayers droplets are formed bra spinning cag{l inside which Eormulatwnmust prevent particles ~omjng 'caked'. Sprayer .types are described in 'Iab1e 2;2, and. in more: detail. by Matthews 0.99.2); Reardon. 1(1991) ,gives a. comprehensivereview of aerial application, in.cludin,glhe application of fl. thuring.iensis.

The finalsi:ze ofa droplet l1eaching the larg~t depends also on theamollnt of evaporation. The gen.era.i ,cbaracteristi.cs of eva.pora.lion from water~ba,sed droplets have been fairly wen described by a theoretic,al model; evaporadcn rates from B.flurringiensis fOrmulations· wereessmtiaUy the same as those from wale.r droplets until the droplets crysta.llized (Luoet aL, 1994). .oil evapo.rates much less than water as a result of different vapoorpft!SSUle and v iseosi Iy. Also" evapora~ lion rate increases as droplet size decreases due fo increas:itn,g surface to volamemnos. For these reasons ail. cameraare used most often tor ULV sprays, If water is used .. especially with smaU droplets, an anti-evapo:rmt or h.Wl1I~tlnt is added. Anli~vaporanfs may f:orm a film at the droplet surface to reduce evaporation: oftemthese are water-sohJ.ble polym.ers (Appendjjx J). Humectants, such as gly:crerol, glyc:ols or molasses, increase hygrCl-" scopi.dtyto rOO.uce evapora non.

Suspemded. particles, such as microbes, also alterphysiealaspects of dr,OJ'S' Theirinflll-

eno€,particulady on spr:ay dlsil1ltegratlon,is reduced if theyarewe«ed. (Smilli and Bouse, ]981). Mittoor,ganjsms can be hyd.rophobico.r byd.rophilic, which win affect both cheiee of carrier and wetter (section .2 .• 3.2a; Appendix 1abtes 1.1-1.3, I.S.).

fogs or aerosols are the equiv,alml o.fthe lowes:l volume ULV spray with.lfI.e ;nnest droph~ts. Therearetwe types .offogging machine, cold and thermal, earners such. escottonseed oil or water can be used with. coldfoggers. Thermal .fqg_ge[S generaUy use specialized chemical carriers; it must be detemdn.edthat .n@;ithe'f carrier nor jhe process ,of fogging inactiva.test.heorg~nj:sm.One camer (VK2) conta.ining methanol and. ettmx.yethanoi} added. to water is Iimited in volume per acre by phytotoxicUy.

Fogs a~e particulady suited for use in greenhouses because gentle wind does not carry the~ogawa:y(rom the target area .. and highin-hm:.u;e .humidity delays evaporanon o.f w.a~er from drop~els.However"fOgs shm:dd not be applied in windy w,eather because stromg w.indl b:lows fog through spaces between sheets of glass in modem metalframed houses, reducing depos,it of drop~ets on foliage towindward .. This does not eccur in plastic-sheeted .houses.

P,edormal1lC€ of the produ.ctfoU.o,wing appUcatio.n is Improved by several different types of additive; these .iru::bJlde sunscreens and stickers (soction 2.3; Appendix Tab~e 1,,6),. However, additi:v,esabo mfl.uen.ce~tures sueh as viscosity and the wetting ability of the suspension. [de,aUy#th.erefore;.. choice of tlliickeners# humectaotsand wetting agents should be made inthe context of the formulano:n.asa whole ..

Additiivesmay be mcorporated into furmulated.products or added later to spray tank mixes. For ease 0'£ use.and to ensure that aU necessary ingrediel1l.ts are present in theoorreet proportiOfl!S,t!he muneris prefl!rabl~ designed for the most amenable pestl plantoombirnation, but with. the :intention that further addiUons should. be made to

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~anl(.-.mix torm!!.datiar;ls :for mor,eintrl.lctab~@ com:b~natiQns.

Dyes are addedexperimentally in trials to V1SU.aUze' deposits on folia,ge' or target cards sited hl target areas (Append.ix Table E. 9). Although nor a normal part of a. fiormulatioo, their presence may affect the pedo.rma:nCf! ,of a produdl.e.g. by additional pro,tect]on from U\l light. This should be taken in.to account when assessing the I1esults oftrl,als using these dyes. Wa~er~ or on~ensi.tive pa,pe:r is an a.ltemative,

1.2,2c AppliiclJU011 to' water .In:itialtya pmduct must F,eadUy per:tetra~e through tllile water surface. This h;ls~end,ed to lead to the use of larger drop~et sizes wi~h sprays,. Further req.uil1f[!m@nts depend on jhe type o,f water body andihe habits of tbe targ~t wIfhin, For example; target~arvae ,af mosq1:lUoos and. simumd v1,addUes. feed. try fiUerin:gparticulate food from. the water (lalble 3.19 in section 3.6. ).. Btackfty Iarvae attach themselves by silken. threads~o a substr,a.~e in rapidly flo'w'~ng water and tra.ppa.rticles .moving past. In contrast, mosquito Iarvaea:re m,ooilel .Hving mainly In static or slow flowing waters. These wange from. large dversto srnan s,trea:ms and from swamps to small bodies Jikie water buttst car tyres and leaf a:l(Us, Lo,gistil.cs range from. applicanon across large areas with or without dense :6oHage" to searehmg for a h.os,t 0.£ small and varied oontainers..The sa.me target species may occupy different habita.ts.50me speci.esare surface feeders; others bottem feeders (Table 3.19 in section 3J6).. Appropriatl'!' formulations mustth.erefore be designed to delive'rtne correct dose oft:he orgarusm~o the zone harhourIng target speeies, ,o:ften avera penod of time. Thuss~.ow release formulaHons$ sach as bdquettes, may be deslgned!~o disiinteg,rate into some parth:Jes that I',ema~n at"o.f just l~bw ~ the wate'F surfaee and others that :slow~y sinkeo the bonom. Slow release farmula.tionsare notgen~raUy suiltoo.to fast flowing streams.

2 . .2.3 Env'Jronmcnial persi:sftm(;,1! 19 2.2.3 ENVIRONMENTAL f'ERSJSl'.ENCE

Normally an organtsmmust rematn active for a. time after applica.tion, .ide'(lUy throughout the period that a. pest is lik:ely~o attack. the erupT or fn so.U thrO!lgh.otlt the crop ,cycle, Mh::.r.obesare inactivated by several environmental factors, including sun, high temperature, humidity r ~ea.f suda:ceexudat.es and com.petitors (sections 3 .. 4:.1. b) 3.4.4<:,4.3.3, 4.3.4" 704) .. Also they may 00 lost physkaHy' from the target I,ocation by the action of w~nd# ratn or le,a,ching '0 ones and McKinleyJ 198'7; sections 3".;1.1, 3.41.2, 4..3.3 to 4 .. 3.6,404..2).

. The re~ati.velmpo.rtan.C'e of each .of these factors deeends 0'- whv '-,,-d' whc.-.c .- ·.00· .. iet

... '~""~"'-·-.r,",-· .. ·n .. .r an _. __ ere ... p1:1 ... uc.

is used .. Forexampje, inacUv,ation by sunHght: is the mostJmportant facter :r'rouc~ngpers:iJstenee all microbes appUed to Eo.Hage, whereas fleld temperan.ues and humidIty have relatively Utde effect ex.cept on fung:~ andnematedes (J aque51 198:5; Jones and McKinley, 1987). In contrast, su:nUghtis not important when. mkrobialins:ect.~ddes a.re app1ied to pests in stores or for eentrol ,of .soH d welUng. pests; here~E'mJ>eratur,e, humidity and soU biota are more Uke~ytoaffe<:t· persisteace, FonnulaUon:s ncrmally contain .addit~ves to protect agents from rava.ges of the environmens. The ma.~n e(fectsare discussed below.

2.2.3a InalcUvaf.to'nby suuUghf The most hannfl.d wav,el.engths wachl:ng. the Earth's surface aile between :280 and 3.20nm. (UVB); 320.400nm (U-VA) are' less damag,lng, hut x8

- -·te· •..• 11· , la, if'W H.· ,w c·v·r th c.- -,; - b c.

grea .. f _I"t qunu~,:1" .. _0 ... e e~lue.re may .. e

sensiti.viUes of some o:rganlsmslowave~ lengths outside this raZ1lg~ '~e .. g. Harm, 11980). The powe.r of Ugbt to penetrate into. solids increases with increasing wavelmgth, UVC{ <280 nm.)·~ UVB·~ UVA ~ vis]b.leHght·~ infra red rays (Shaath, 199Qar b). Greenhouse glass transmits n.o uva 3Indsoll'l.E' UVA rang~n_g from neady zero at 320nm to 90% at 400 nm (pilkmg.tonBrQs Ltd, St Helens, UKlc personal com.municaUan). SimUa"rly some .p~astks do no,ttransmn UVB (K. JOllies,

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20 1i!(:JmQ.log$l ,Q/ form ul,atlcm and appH,~~tiou unpublished rcesults)I.. Penetration of UVB~n water ]8 also limned.

To COlllnt:er harmful effects, sunscreens are often added to a formtdaHon (sections 3 . .4!.4e, 4.3.3; reviewed by Sha,a.th, 1990",.. b). Sunscreens ad by physically reflecting and scatle.ringJ' or by se~ecUvely ,absorbing radiaHon, ,eaniverting. short w ave~englhr; in harmless longer O!J1!es. Reflt;'Ctors include zinc oxlde, m,ani.l.lm oxide, sHka.te and talcum, Aboor:bentscemprlse most of the ma terlals tested w:ilthm.icroorgan.~sm~s (Ta.bie :3.1.6 ~n section 3.4.4; Tilb~e 4.4 in seetlon 4.3) ~ndUJding: specialized dyes and chemicals, such as Cong~ Red and sUI:bene derivatives, which abserb specific wavehmg:ths g,iven in Appendix Tables I.9-t 12r as w,eH as cheap and readiiy ava.na:b.~.e add.ilives tnata.bsorb over a wider ra:nge .. e.g. mclasses ..

2.2.3b Effeclof t'empemlu rt'1'emperahlre' i:s important for the shel£-Hf:e oJ mIcmbia.~ products (section 4.6.1) andIt can a:ffect th.ei:r actlvtty enee appUed. Temperatuf'e" optima and limits vary with nil.e microorganism. rf'abl:e 4.~ in. :s-oct.~Ofil. 4.6) .. For examp~e, insect pathogenic viruses are unaffected by 1. ~lO GC and <:O"'C has Uttle eff!'ect (Tanada, 1953; David and Gardinerl 196'l'i David er ei., 1971).. At the ot~.er end of the scale, e%PQsoJ'e to 6O"C .far morethan 10 min i:nacUvates these viruses (Entw~sUe and EvansJ 1985).lnfed~on proceeds at f1eldtempe'ratures in the plant grow~ng season, but slows at lower ~emperatures. Insect pathogenic f'u~gI can be frozen; surv~val 1:5 improved by add.~t~ves (sed~orll 4.6.1). Although iuactliv,ated byhi.gh temperaeures (TaMe 4.5 In section 4.6)fungi~. if protected by addi HVe5, can survive spray d.rying (section4,!6,2). StrainSllsed in pest eorurol nonnaUy willno,tgruw at 3'7°C, a featureof tneir' safety to mam:[nals,.R:h:~zobja iaeculanes naJv,e well deUned optima for growth, il'odare sensitive to high temperatures, whkh may Hmlt· th@.i!r use in the tropics (oomasega.f'an et a.I., t 984), althoagn this m.ay also be an ef~ect of deslccation ilthigh temperatures.

Gen~mlly; most prelonged ~~mpe'ra~ulre ,effects cannot be counteeed through formulation, A more promisiegappeoach i8 selection

f . f'· . .

c stralns or nucroorgasusms ,a!cbve over a

wider :r,ange of temperatures.

2.2.3c Effr,'(., of hmuidityAvn'tl?rapaift~'bNfty As w€'H as theeH'ect: 0.£ moisture (10n~ent on storage stabiHtY(Se{'tions4.6.31 4.65h some olgi;llli~smsrn.ay ~dso have c@rta.~n moisture needs fGractl vi~y.. ThusTFilemaltndes often need free water to survtve on a.~arget surface and 10 move andIocate thE':.~r hosts. AntagpnisUc becteria used agpInst plant patbogens. need the p~aliJJl: surface tiO be wet in order ~o estsblish them ",oilv' . .,,, ~·" ... ction 5 .. 3·1C:) F-·j",n·g·: ""1

_-_,u-l'l.:!',~-l ,I, h._ I ",O""~'.''!i..,~ ,\,r.71I...!I;,,;;IIIILV .. 'I'.",J,,~ ,!!irlII",.:U,.

spores RormaHy need h~gh humidUy ~o g~rmiFilate. (section 4 . .3.4). These needs may be overcome hy approprlate forml.daUon in oil (lit oil em~lsion. ~)r by the addition of a humectant ~o the product (sections 4.3.4, 4.6.5). lngene.raI there is Uttle direct effect of relative humldlty on the acHvi:ty of V~:r!JJ.se5 and sporeforn1ll:ng bade.r1al (l\lmada. 1973; Jaques, 1977); probably, however, .hu.midity d<)es influencethe rate or extent to w hich these organismsa.re inactl v ated by (lither factors, such as heat, leaf-susfaee c.hemictds (next sect~aFil) and sun (JOlil€S, 1'988).

2.2.3d Effe,rt~f cbenl.icals at tfu?'argd substrate Chemicals on ()iI in the target substrate may 10Huence organ~sm:s in. various ways (secHon 3.4.3).. These ~ndude chemicals present in the son, as weU as plant secretions and chemicals on the phyno,p~ane. For example, on cotton and many other xerophytic plants, htgh levels of magnesium, calcium .and. mang,ane.se as carbonates and bicarbonat~·s are exuded onto the leaf surface. Th~s can raise the pH of the leaf surface to. vahres as high as 10 or 11 (Harr e.l al., 1980; Ioaes and McK:~nle'Y; 1981; Jones# 1988). The presence of these ions orthe assoelated pH value, when w,eftecl by dew or water spray $ can inhibi t or inactivat~ same organisms ,( Andrews and Sikorowski, 1973; Ell@man and En~wisHef 1980.) .. Thua a product

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~ay be formulatedin microcapsules to provide physical protection from. ~hechemjcal effect.

_ Plant extracts c-an also jiarm erganisms, Some extracts depress the growth of B. tlu~fringien8is (Morris and Moore .. 1975).. However, since B. thu.ringiensis acts more -by poisoning larvae w'ith.the 'crystal toxinthan by~infectioF'l. the bactericidal e.ffect of leavesprebabl y has a relaHvely minor effect on pest- control. Plant extracts have also directly or indirectly inacti:vated baculovtsuses, ora t least affected activity (Uchida et ai., ]'984; Richter et ,al., 198!; SanHago-Ah/,erez and Ortez-Garda, 1992). S~mHady, chemlcals om. ~he insect cuticle may suppress the' growth .of fungal spores (St Leger, 1993). Thus addit:b"e8 deslg;ned to overcome antimicrobial acti'Jl~ty mal}' be needed to improve the activityo.f proo ucts on these target substrates (see ~tjons3.4.3, 3_4.51 3.4-.6)-

2..2.3£ .Physical loss~.iljcroorga,nisms can be lost from target areas through acUon of wind and ,F,ai_o~ physkalabrasion or flowing water, toss varies with type and properties of the formulation. The ·effeds of drop size and. .Iiqu~d properties on spray retention are discussed in section 2.2.2. SimHar[y i particle size in dry SQlid formulations alsoinfluerilces retention (sections 2.3.1a, 2.3Jbbelow) .. Pine dmsts readily. adheretoleaf surfaees, Lafger and heavy solid formulations will no~ easily be washed awa,y in meving water.

Stickers (Appendix Table L6) ~mprove adherence of erganisms to foHag,e and persistence dlu.ing wind andrain, EffectiveneSis varies .. from the delaying action of water-soluble materials such as molasses, to the fastness of materials such as resins; which dry tobecome insoluble. Stickers (App~ndix Table 1.6) may double up as thickeners (Appendix Table ,fA,) f l.e.addiHves such 3:S gums - and molasses to I~:c_rease sp.ra.yv:~scosUy and reduce evapQra~ non from drops, or theym,a.y double up as phagostimu~a,nts such as. molasses (Appendix

Table 1.7). -

2.2.4 IU1provemeut of ac/l0H 21 2,2.4 IMPROVEMENT OF ACTION

W1.t'n. organIsms that ,a.ct after ingesUon,phagosUml.dants encourage pests ~o ea t more of ~he. organlsm be{ore~t detertcrates on foH~ge er b~forea. microbial tmdn arr~st:steed~ng (section 3.4_3)._~nsed larvae are encouraged ~o eat more rapid~y and may be attracted to the formulated products .... partku~ar~y baUs: neonate larvae. may flnd .n softl n~tri,tjous prcduceeastsr tobfiOilj'vse on than the hard s:~rface of aIeef. Ba.it is g.enerally a more efficient method of d()Sag~tTansfer t~afl. blanket application and F'andomtransfe~r (Ca udwell, 1993 ). although it Is ~nferiar~o d ust on ants (section 4.04 .. 3). PhagClstimulan:ts can be added to~a :spray_tank mix or incorporated into a solid bait. Plant extracts and liquid or ground plant products are often used _ However, a sthnulant must be selected with care as its effect may. not be .th~ same with an target pests Oones, 1990). Common solid baits Indudebran and calcium alg i.fiIa te I wheat mixtures for g[asshoppEl~rcontrol (section 4.4.;3)..

Some chemiealssynergize the action of an organism, 'e.g. socii urn tetraborat.e increases the potency ,of some ililsec~ viruses (section 3.4.6).. SimiladYI certain mixtures of organ'~ isms or mixtares wi th chemical pes:tk.~des cause pol:ential'iolil. However,caution is ~eeded as some pesticides antagOFllI2!€ organtsms, and the ·eHed 'Cali! vary with pes,Uc~de dnse (sections 3,4.6~ 6 .a 5), Morooverl the formu]a.t~r is adding toxic chemicals ~o a product~ whi,cn may negate the envirenmental benefits of using safe mkroorganisms,

2.3 TYPES OF FORMULA. TION

l'hel'e.an~" a wide vari@~y of formulation types, both h,gmdand solid ~GIFAPr 1989,; Appendix ~n). The main types currently used for -organ~sm5 have been classified by Rhodes (1993) into dry products (dusts, gramnes and brtqnettes) and suspensions. (oil- or water-besed a.ndemu~sions). A w.ide.rr'ange of fonmslation

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n Tec1~.tlQJogy oj /ormuJtllian ami appli(i1ticm type5,~ogetlhlerwlfh additive types, are listed and defined in Appendix In using nomenelaWIt! recommendedfcradoptien worldwide'. AddiUonal speciaUzed methods may be used, such aasponges for nematodes, Chap~er7 describes .formWlatiQn types from thev.iew~ point of appUcationto soil.

2.3.1 DRY PRODUCTS

These comprise dusts, gr.anules and briquertes, a ·clas5lification based 00 partide or aweg~tesi2e (Appendj,)( III) .. Alsomduded in this group are wettable powders. which are formulated as dry powder designed to be .addedtoa liquid carrier .. liIonnaHy water, just before application.

2.3 .. 1 a DuslsBasedl on mert diluents or carriers, norm.aUy of low absorbent capacity, these have particle sizes ranging from 5-20 mmc Particles of <:10 mm are abrasive' and jnsectieidal, also an lfllhalation hazard, but the smafler p.am.c~esadh.erebest.

Minerals such as days are often the firsl choice 'of di[uent~but silica minerals are also used (Append~x Table t2h varyjngthe proportions~o obta.in the desired bulk density (Matthews, 1992). Diluents wi.th high surface addityor aU<annitya,re usuaHy ,avoided ,as they tend t:ofonn an UJilstable product (Polon., 197.3) .. Inert filletS (Appendix Tab~e [.2.) used. with il:ilsectpathogensan~: listed by Angus and Luthy (1971), also Couch and Igno:Uo (1981J; ~oJilge:r Iists of ava ~~able .oHers are given byPa.lon (1973) and Becher (1973).

Dusts typical.~y eenta ~n <110% of an 0rgani:sm by weigM.They are normallyprepared by tieedmg theorganism into an airstream for mixmgwith the-mineral. diluent in a ribbon blender Dr Muson mixer (e.g. Table .3..Sin section 3~3.1).Part:klJ(l' s:ize,bulk density and Dowabilityar,€ exroemely important-The proport.ion of ,components is variedjo form a. free-f1ow~ng, .Huffy powderwluch does not stick to machinery (section 4.4.,3) or aUow sepa ration of the organjsm_s from the dUuent

duringtranspon" storage andappUcation. Separationoccur:s if dilueml particles are nQt wi~hin 10 mm of the size of the organism paetiides.Bul.k: density should be 3~ kg 1m3 I(Polon, 1973).

At applicat~on" dust partidl€'sare carried on air ,current:sto penetra~epianf cenopies, Smaller particles collect o.nlargd surfaees .. lar~r ones fall li1lrough. Usually ,only 10% ·Qf the particles adhere to the surface, SHekelS can be .mc.l.uded, sueh as mil.k powder (which is hygr'll)S("(ilpic :50 the air must be dry during mtxing)1 or 8. d:esiccant such as :sodiium sulphate (added to pf>event caking).

2. .• 11 b Granules,petiets, capsules and briqnelt,es Granules are discrete masses 5-l0mm3~n s.i:ze..peHets are >10 mm3 ,and briquettes are large blocks uptoseveral cubk centimetres; the sizes are defined in. Appendixm.Uke dusts, these products contain ,an inert carrier hoJding theorgan~s,m5. Carriers indude day minerals, sta rch polym.e1"8" dry femliz'cr.sand ground p~antre.s~dues (Ross and Lembi, 11.'985; section. i.6Jl}.OlQlce of carrier depends on sorption (more Important for formulating slarries ·of ,organisms) r hardness r bulk density and. product disinteg;r.aficm rate in water (;Po~on" 1973). Soft carriers, ,e.g. bentonite. d isintegtatc qukkly tor,e~easethe org~nl5m. 'The pn:duct: can. be coated with various materialsfQ slow arildrontml the rate ofrelease~whk~ also depends en unit size,

TypicaHytbe concentration. of ,()~ganisws is 5-20%, usually <15%. The"ear,et:lwee types ofgrantt!les! (1) the o:rg:arusmsare attachedto,the outer surface of a.granular ,ca:n'ie:r marota~ing drum bya sneker (fable 3.20 in section 3.6.3); (2)lhe organisms are sprayed onto a rotating granular ~amer\Vithout a sHekel' !(T,abh:~ .a.21in section 3.6.4); (3) the organismsa:re incorporaledintoa ,carrier paste or powder whkh sets as ametrix (fable 6.8~n section 6.8; Tabl.e 7.6 in section 7 .. 5 .. 3)~ size being controlled by passing the product through a SIIE'Ve •. Type 3 is the most common

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with nUrifying microorganisms. When ~he carder forms a protective ceataround a core aggrega:~e of organisms, the unit is termed a cap.sule. A code .of nemenclature f,Of gra,nulest matrices and capsules is described in section 3.3.11..

The,s;e products present no inhala Han hazan:ir do not ,r:eadH y drUt in the windand can be measured eas.iJlyor weighed out. in oamrast to dusts. Althougb the smaller products can be-applied to surfaces .. such as :foliag~ and ~eafaxHs, much material falls to the ground, The larger productsareappH@d ma.~n.lyto water bmHes1having the adva,ntag~ o:f penetrating a ,~oHage- canopy, Briquettes, used fot mosquUocontrol, can be p~aced indi viduaH yinwater CD-n tainers ..

2.3.1 c Wettabie powders These predominated among eady commercial products and comprise tedmkal powders (e.g., Table ,3..2 in seeticn .3.1.1; Table 3,,3 in.sectiofl .3.,,2 . .2) blended with additives to make them stable during Sitorag@ on the shelf and readily niscib ]' - with w ater M which tih .. p. - wder l,!:;,~.

m.L",. 1 ,.e ~ _ , . ac_ ~ K_ _ _ __ ,.... . I) . , __

added shortly before spray.ing (Table 3 .. 1 lin section 3.3.41).. As with other forrnulatlens uSLng water ascarrierJ chlcnnated water must be aVQlided (OF the water allowed to stand toevaparate~he chlorine) for feer of da,ma,ging the organisms.

.Most wettable powders contain 50--80% ~ocM,ka.~ powder', 154,5% filler, l-lO% dJsper:santa:l'ild 3--1-5% su.rfactant: by we~ght (w@trer; section 2.;3.2a,; Table 3.11 in. section 3.,a, 4).. The fHI.er should be ~ne:rt and hydroph inc to mix wen wUn water. Normally a minaral such as silica .is added to prevent d ump~ngan.d .fus.ing during grinding (gdnd. ,irng may not be possible wi th some organ.~sms) anda,ids flow abili~ by m~n~miz]rlg caking: du.ring storage. Ca.kin,greducesw,ettabdny ~fid d~)gs spray r:!O:lz~es. 'However, me silica mapfloouct .s.h.ou.~d be kept to' a min~muma;s sihca is abrasive and wears io:rm,u.lation equipment, as well a:s spray .f1IOZ' zles, Other materials such as days or Jildose

2.3.1 Dn) p.roducis 23 {Appendix I able 1.2) may also be used as fiHer.s, w~th or wIthout silica (e.g. Mc:Kinl~y et et; 1989) ..

The dispe~rsant (A ppendix T,abieL4) neutralizes a~tractiv'e interactions of .Iike parUdes and ensures that particles .of the technical product do notattract each other, but suspend uniforml y 1111 a water column wi~hout settHng. ThoeWorld Health Organ:isa,t;iIClI,n (WHO) recommends that a product suspended In ,a g:radl..hlt~d cylinder for 30 min should ha veil cencentra non of at le,!3:st50% of the orig~nal cencentration at a point halfway up the cyUnder. Polan fI973) lists dispersants suitable for use w.ithchemlcal pestiddes, Excessive amounts of soaporsalts (sod~um,calcium) must be avoided with Iignosulpha te dispersants to avoid killing or flocculating spores (Darvan, 19178)..

A dry powder, put: Into a liquid" must penetrate the snrfaceand overcome surface tension at the liqj~~d~oHd interface, The surfaceant helps to red uce this tension and ellowsllquld to displace atraroimd pa rUd.es.. Too much s~rfactantl however, can eaese excessive foamIng, preventa ble to some degree by sillconeandfeams and by using low-foam surfactants, As a gerllera l ru~e no, more tha.f1 1.0 ml of foam should remain In a 100 ml cylinder .5 min after mixing. Foam should be avoided not: onl y because it is ill nuisance, but also because some organisms, such as spores.separa te d U~ ferenUaUy Into foam. W.he.riI rain re-wets sprays dd.oo on fo.Ha.ge, surfactants also ad as detergents, Inc.reasing the vuln-erabHity of organisms to wash-·aff. Ofterrone add.~tlve acts as both dlspersantand surfadant.

Usually, wettable powders lend ~o mix slowly into wa~er and separa~€ mixers may be needed before .fIHlng spraiy tanks, since tankagaaJors are often not~O[1cefulefiloug~. Powders form unweHed baHs,tI. few of which may pe:rs~st even a.her protracteel mix~ng.. A mter, p:re.fera.My of ny~on mesh" is essentialin the sp~ay[ine to prevent nozzle h~ockim1lg.

Mixingp'rob~ems can large! y be solved by dry b~ending a powder wift.a binder and!

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24 'redmology of fo.nnu,l~tionQnd appl.ical.i(l'n .fornllng the mixmto water~di:spersiblle granules .. These break surfaeetension more easUythan.powders. Theyallow high concentratiOI1lS of ,orgamsms~Oowmgfreelr wIth ltttie dust'and. can. be accurately measured by v'olume Uke a Uquid.. However, production costs are high, more' agUanon is needed for dispersion in cold watJerin the spray tank, ami smaUparru:le s~~es may be diffiadtlo achieve. These gr'anwes are gaining popularity: a .newrnngeoIIAho:x" wetting and dispersing agents has been. develeped fortheiruse (Appendix Table 1.5).

When a we.ttable powder or water-dtspersiible granule is mixed Into wat,er to' form a spray ( solutes are dissolved, These jnclude enzymes, bac~eriial nutrientsaJnd additives .. such as surfactantsand sugars, which may stiimu1ate germination ·of sememicrobes, The spray must not beallewed to stand, as deterJoratien may be sl,gn.ific,a_filt in 1..:2 days and (.10U ld beaacel.erated by some surfactants harmful to organis.ms.

ParticlesIn these pFioduc:ts settle rapidly in spray tanks.'lso tanks fitted with agita:torsmay beessennal, Settling can be reduced by millimizingpadide size and adding thickeners to .increase v.iscosity of the sp~y tank mlx,

2.3.2 LIQUID SUSPENSIONS

This range ·of fermulations usesa liqUid as carrier, usuaUy warer or ,oU .. but, solvents are alsopossible, The commonest are suspension concentrates end 'emulsii:ons'l al though there are aloospedalized types such as micrccapsules (Appendix UO.

2.3.2a Suspensio.n (flOWllbld conceu.tmles These are essenttally suspensions of particulate a'rgarusms in. liqukis. Particles account fOT 11040%., suspender m;grlooietlt 1~3%" dispersant 1--:S%,5uriadant3r8% and. carrier Uquld (oi~or\\!,a~er)3~% by weight (e.g. Tables 3~9 MIld 3.10 in. section 3.3 .. 3). Viscosnyshould roughly equal the seWing rate ·ofthe pa rtjdes. This is achieved b:ytl1ie use of ()oUQ~dal days,

po~ysacd1addegums, cellulose or synthetic polymers (Appendix Table l4;BaHi:sta, 1'915; Theng, 1979). Apeodect s.hou.ldresist a smaU forcesuch aspartlcie settfmg during storage, but should f1owfree~yunder a greater force, Dispersants reduce the rate of sedimentation of solids, which is increasedthmughreverslble dumping ,(tlocadaHom). Sudacta[il.ts aiet as ,emulsifiers as well as wetiers OM spreaders; fhey are described by Anderson (1'983) and Smjth. (199.3) among other addU:ivesl with lists of available pr,oducts.The oldest .types of surfactantaresoaps,nonnaHy sodium or potassium salts of weak faUy adds, e.g. sodium: s~eara~, whichhaviI? a polar hydrophilic (waler-:soluble) and a non-polarHpophilc( oiH;oh:lble) end ofllbemo,locule .. Although soaps are ,effective anionic surfa.ctants, fhey precipitate-as an msoluble scuen in. hardwater or actdic seluttons and surfactant properties are lost

:Synthetic surfactanes are spllttnto anionie .• cationic and non-lome types (Appendix Table 1 . .5). They are .sj>mi~aF to soap in possessing a non-poler, Hpophificgnrup Ooog12-18 hydroc,arbon dlain)'~ and apolar hydrophilic group. The hydrophilic ,group differs - {rom that in. soap... Anionic surfectants are y,ery goed wenera and derergents; they indude alkylarylsutphonates" fatty a.loohQl sulphates and alkylsulphllna~e5. Cationic sudaC'tu~ts are derfved from ammonia and. are generally phytoto.xic:and effective bacterldd!es;they precipitate in hard water and arepoor detergIffllts,generaUy unsuitable for 'use wiiliD1loSl, if not aU, microorganisms. A third group, amphoteric su.riactants"act ,as,ei.ther anicnic or cationic depending 00. the acid Ry.

Nen-ionle stl!Ifadants do not :ionize in sclutlo.u and so there is no detrimental effect !Qf hard wa~erand highly add solutions" They are good dispersing ,agents and. detergents and can form staM.e emulsions. wetting can. be improvedb1addi't~o:n of aruonic surfactants, Non-ionic aurfactaotsare mainJy dedvatives or po~yoxyetheylene' andpolyoxf~ propylene, They have a lipophi.tichydliocar-

Copytnlghted material

bon chain, e.g, fa tty' acid, and corresponding alcohcl aUached~oa second, hydrophiUc, oxyethylene cham, The length of h.ydroph.ilic chain and the nature or length of hydrophobic chain determme the surfactant properties, the relationship betw'eenth,e lengths cf fhese chains is the hydrophile-UpophUe balance or HLB scale. The HLB scale is arbitrary r rang:ing from 1 (lipophilic) to 20 (hydrophHk) (Becher, 11.973). Table 25 summarizes properties of surfaetan ts on the scale. Nen-ienic surfaetants are the most useful type' for use In formulation of microorganisms.

S!ilsperu;~on (>f particles in the carrier requires a. surfactant (Appendix. Table 1.5). Depending on the :r,€',actions of the org~nism and carrier used, the sur.ta,ctant needs to be hydrophilic, lipophilic or mid~way between these, BG.r example, UpophiUc :fungal spores can be suspended in oil using a UpophiUc surfactant; highly purified baculovimses e.asUy suspend In water with hydrophHic wet:. ters, but for suspension Q1f unpunfted insect viruses the :surfactant needs both hydrophiUc and. lipophilic: properties because they contain much fatty rnsec:t material. Similarly, Iormafinn of a stable emulsion '(see be~.o,w' requires a. surfactant which is padly hydrophHic and part~.y lipophilic Crable 2.5).

As well. as the~r problems 'of .foaming and wash-off (section 2.3.1c), some surfactants harm or inhibU microorganisms. Althcugh

Table 2.5 Hydroph:i.lk-UpophHk balance (liIH) range and p'J\opertie:s of aon-tonic sur.folctants (ranges shown in bold a~nl not s:uitable for formula.t:i,on of bene.fldal mkroorg~:ni&ms)l·

HLB range Property

H Inv~.r-t {wa~e~·ij'H)n emulsien)

'7-9' WcUin,gag.enl

8-15 NO'lmaJ (oll~in~water) emllltsion. Good fo, wetting Qrgan!$ms with hydrophilh:: ,ilnd lipophiUc properties

13-15 Detergent

1.5-18 Solubilizei!'

.. From Anderson (]9B3}.

2,3,2 Liquid Sllspensioln~!J: 25 there is litUe experimental work designed tn ,(dUcally test sudac~an~s" repeated good hioassayand/or field l1esuUs WIth Individual wettars accumulate to aU ow significant (00- elusions to be drawn, However; labo.ra.tory results should be extrapo],a~ed to the HeM with caution because field condltions dHter wi th, for example, v arlable exposure times, b~.odegradation~ concennatlon of surfactants bydryilllg of drops and their periodk rerno~stening to make active solutions. Non-ionic surfactantsere less likely Ul!an ionic surfactants ~o damage microorganisms. Theis selection is discussed, In sections 3.4.1; 4.,3.6 and 4.6.6, and by Ward (1984). Surfactant manufacture.rs can provide HLB values as well as advice GIl suitable uses and com patibHities of their products. Surla.ctantsadded 10.:1" storage stability should be minimized because, due to long exposure, theya.n1~ more likely to harm organisms than those put in the spra y tank, w hi.cncan be varied according tothe weUabi1~ lUes of different leaf surfaces.

F - J'J1JJlati I of a-c f1'"':"""rrate by· . . '-Us ~ - ndi n

o~ on _ . __ co_ .. ,",~,_ r ' s __ pe g

fine I y ground dry technical powder in on mClIeases sedimentation compared wah suspending in water .. because of relatively larger partkle size and ~o'W densHy of the .oil carrier. Of o~~s used w.~th micrebial pesHcid.es (Appendrux Tab~e l.l; Cou.c.h and Ig110ffOr 1981), vegetable oilsare preferable to mineral oils to mhumize phyt~to:xidty ifra:ncidity and soUdifying dudng storage areavcided. Liquid fwet') mining ala. product in oH results in smal.ler parUdes that stay in suspension longer and are easily resuspended. Weinbe.f'gff and Greenha1 gh (l'984)g~ve contentanalyses of f~US from the eeolegiral viewpoint,

2.3.2b Emu/sliMs Em ulsifica tion red uces sedimentatien of parHdes during storage and in the spray tank, because b~oyan,cy of ~tle oH coenreracts high rela ti,VI€ densities of th.e particles.

Approprla te surfactants (emulsiflers) a:l1€ required for mhe formation ()f a. stable ernulsion. Their molecules contain stnH:turally dis-

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26 Tcch,wlqgyof /om.lulatioll and applicat.ion similar groups - with opposing tendencies >that enable them, in dilute solution, to adsorb a t the in terface of the solution and a phase in contact with it (section 2.3.2a). They modify properties ,of the interfaces to allow formation of emulsions and to enable the solvens.e.g. water I more effective!y to wet both particle and target surfaces, as wen as to spread on those sorfaces.

Once formed an emulsion needs to be maintained. Separation and creaming can occur a t temperature extremes, and emulsions are also a ffeeted by the ha rdness of water and pH.

With normal emulsions the OilW5 the dispersed phase and the Iiq ldd in which it is suspended; water. the continuous phase; vice t't'rsa for invert emulsions. Invert emulsions have high viscosity. so are less Uke~y to separate. They produce larger drops with most sprayers and. because theexternal phase is 011, ev apora eion from d rap-sis minirnal, both factors combining to present less drift hazard. Invert emulsions can be made by mixing two phases at the spray nozzle; or are pre-mixed with the addition of :fatty amine salts, The stability and possible phytotmddty of these formulations need to be carefully assessed,

2.3.2c Em::dps.lth1h·d formulafions Microcapsules contain the organism and are compared w! th granules in section 3.3.1 .. Sher (19'77) lists 22 commonly used encapselattng materials, including gelatin. starch,ceHulose and several types of polymers. Capsules give good protection from. environmental factors, such as sunlight and leaf-surface chemicals, Dyes can be incorpora ted in to capsule walls jo increase UVprote<: tion, a ]::;0 stickers and wetters can be adsorbed to their surfaces to im prove retention on the-target. Mlcrocapsules are usually formed by three methods:

Pha sesepar a tion, A suspension of the organism (core phase) is emu lslfled in a sol utlon of the wall materia] in an immiscible continuous phase. A change; such as pH

or temperature, makes the wall material separate and deposit around theorganisms in [he emulsifiedcore phase. Spray drying! f] uidized bed spray c-oating can also be used w here a film-.forming polymer, dissolved in a liquid phase; is sprayed on or with the organisms, The pelymer coats the organisms and dries to form a solid m icrocapsule.

Interfacial reactions. Reaction between chemicals in an organic phase and those in an aqueous phase forms a wall at the interface between the two phases. Wall forrnanon can be inl tia ted by h.eat.~ng or by adding a ca ta.~yst. fartkle size, wall thickness a-nd permeabUity can be aIte'red by changing veriables such as the amount of an organIsm or the amount of wall-formmg agent.

Ph yska] methods.Muln-orifice centrifugation and electrostaticencapsulation, A solution of the capsule wall js fed through specialized nozzle orific@s with a suspension of the organism. Fluid i rods' are formed sheathed in a fl uid shell, w hich breaks to form microcapsules. Size oftne microcapsules is determined by flowra res of sol utionand suspension. For electrostatic encapsulatlon, two aerosolsare produced separately with opposite charges on the core and coating particles. These coalesce and solidify on COOling. A problem with these-physical method s is that some of the o-rganisms may be on the outside of the mierocapsule,

'Encapsulation' .of the .Bacillus thur:ingiensis toxin has also been achieved by genetically engineering and fixing other bacteria (section 3..3.5)

Depending on the rna terial used for encapsulation, the wall is broken jo release the organisms by crushing, pr-essure within the capsule. dissol u tion tQ'f hydrolysis. Unlike chemicals, diffusion cannot be 'used to release the organlsrns.

Once formed, microcapsules .need to be formulated as liquid suspensions, e.g. flowable concentrates, or as powders, granules etc.

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:U CONCLUSIONS

This chapter iUustrates the wide range of approaches to fonn ulan.on. and the manytechnlques a:va.Uab.te. Therea.fe many additivesr some with. dual o.rtri,ple roles. n.is often dIfficillt to. r-ecngniz.e or measure effects of individ ual additi ves, so the performence of complexes shouM be tested In terms of~he end product under conditions ,of intended usaand compared wlth min:imal mixtures. Wherepossibl.e, tests shouldalso be carried out overa range of ambient envfronmental cnndidon:s., 111 . e ()ve'.rridjng pdncIp Ie for any successful fonnu1at~Ofl" however,~s that .H mast be ,effective,ecQf1!omical arid practical,

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