Original Article. E Oz 1, *H Cetin 1, J E Cilek 2, O Deveci 3, A Yanikoglu 1

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1 Irnin J Pul Helth, Vol. 39, No.3, 2010, Irnin pp J Pul Helth, Vol. 39, No.3, 2010, pp Originl Article Effects of Two Temperture Storge Regimes on the Efficcy of 3 Commercil Gel Bits ginst the Germn Cockroch, Blttell germnic L. (Dictyopter: Blttellide) E Oz 1, *H Cetin 1, J E Cilek 2, O Deveci 3, A Ynikoglu 1 1 Dept. of Biology, Fculty of Art nd Sciences, Akdeniz University, Antly, Turkey 2 John A. Mulrennn, Sr. Pulic Helth Entomology Reserch nd Eduction Center, College of Engineering Sciences, Technology nd Agriculture, Florid A & M University, Florid, USA 3 Dept. of Biology, Fculty of Science, Ege University, Izmir (Received 26 Mr 2010; ccepted 11 Aug 2010) Astrct Bckground: To compre the effectiveness of 3 commercil gel it formultions contining fipronil (Golith ã Cockroch Gel 0.05% AI), chlorpyrifos-a (Clen Bit ã Gel, 2% AI), nd chloropyrifos-b (Serp ã Gel 2% AI) ginst Germn cockroches (Blttell germnic) when stored t 23 C nd 30 C fter tretment. Methods: Lortory iossys consisted of plcing groups of fifteen cockroches ( rndom comintion of dult, mixed sex nd lrge nymphs-stge 6) into 5 L cylindricl plstic continer with one drop of product (vg 0.10 g ± 0.01 g) pplied to 76 mm x 26 mm glss microscope slide ffixed to the ottom of ech continer (one product tested per continer). Cumultive mortlity ws ssessed t 6 h, 1, 2, 3, 4, nd 5 d fter ppliction. To determine the further effects of storge temperture fter tretment on residul ctivity of the gels, drop of ech product ws pplied to seprte glss microscope slides nd stored t either 23º C (wrm) or 30º C (hot) under drk conditions for 0, 1, 7, 14, 30, 45, 60 nd 90 d fter ppliction. Results: Freshly pplied its (dy 0) contining fipronil provided complete cockroch mortlity (100%) within 5 d wheres chloryrifos-a nd chlorpyrifos-b provided»72% nd 88% mortlity, respectively. Generlly, cockroch mortlity ws greter when gels were stored t 30 C compred with 23º C. Conclusion: The fipronil gel formultion proved to e s efficcious s the chlorpyrifos gels nd in some instnces surpsed the ltter formultions depending on storge time nd temperture y providing»90% mortlity t 90 d post tretment. Key words: Blttell germnic, Chlorpyrifos, cockroch control, Fipronil, temperture Introduction The Germn cockroch, Blttell germnic L. (Dictyopter: Blttellide) is one of the most prominent urn pests in the world. This species is known to trnsmit vst rry of pthogenic orgnisms e.g. cteri, fungus, worms, nd viruses (1, 2). Moreover, Tsi nd Chen (3) reported tht 40% of ronchil sthm ptients in Chin were llergic to this cockroch species. Germn cockroches my lso contminte humn food nd leve persistent, ojectionle odor in infested res. Often times these pests re found in kitchens nd throoms ecuse they fvor wrm (27-28 C), humid res especilly when they re in close proximity to food nd wter. Snittion is key to reducing infesttions s it limits the pest s ccess to food resources. Primrily, insecticide spry pplictions re often used for control of B. germnic ut sometimes the presence of grese or custic floor cleners, especilly in commercil food estlishments, cn dversely ffect their effectiveness. Contmintion of food hndling res is lso mjor concern with these types of insecticide formultions. Moreover, continul ppliction of residul insecticides hs resulted in cockroches ecoming resistnt to severl of the commonly used insecticide clsses such s orgnochlorines, crmtes nd synthetic pyrethroids (4-7). 102 *Corresponding Author: E-mil: hcetin@kdeniz.edu.tr

2 E Oz et l: Effects of Two Temperture Storge Regimes on The development of it formultions hs llowed more precise site-specific plcement of toxicnts into cockroch hrorges where spry pplictions hve hd limited usefulness. A recent option for cockroch control is gel it formultions nd hs een reported to perform well ginst field popultions of the Germn cockroch (8). Gel its generlly contin vriety of ttrctive ingredients s crriers with high moisture content. These it formultions hve the dded dvntge over dry its in tht they cn e pplied in smll mounts in vriety of loctions tht cn improve it efficiency (8, 9). Moreover, they re re esily pplied, sfer, nd exhiit longer residul ctivity (10-13). Moreover, its cn e removed when control is ccomplished. However, if it ppliction occurs in res tht routinely hve high tempertures, mny gel its my run nd/or rpidly lose efficcy fter period of time. For exmple, the temperture in kitchens or food preprtion res (such s resturnts) is often greter (30 C) thn the mient temperture of the dining room (23 C). Therefore, fter initil ppliction, we wnted to determine the residul efficcy of severl different commercil gel it formultions ginst B. germnic popultions with prolonged exposure (i.e. storge) to these two temperture regimes in lortory iossys. Mterils nd Methods Insects An insecticide susceptile strin of B. germnic, originlly otined from Dr. Oner Kock (Hcettepe University, Pesticide Test Lortory, Ankr, Turkey) nd mintined in the lortory for 20 yr, ws used in these experiments. The colony ws mintined t 26±2 o C, 60±10% reltive humidity nd photoperiod of 12L12D. Gel formultions Three commercil gel its were used: Golith ã Cockroch Gel (0.05% AI, fipronil, BASF Turk Kimy Sn. Tic. Ltd. Sti. Istnul, Turkey), Clen Bit ã Gel (2% AI, chlorpyrifos-a; Kuko Science Co. Ltd., South Kore), nd Serp ã Gel (2% AI, chloropyrifos-b; Mylv S. A. C./Verdi Brcelon, Spin). Experimentl set-up Before testing, 15 cockroches ( rndom comintion of dult, mixed sex nd lrge nymphs, stge 6) were plced into 5 L cylindricl cler plstic continer 48 h efore testing for cclimtion purposes. (Durier nd Rivult (14) previously reported tht cockroch ge generlly does not ffect efficcy performnce in gel it ssys.) The verticl wlls of ech continer hd een previously coted with tlc (Utlk Pudr ã, Kurtsn İlclrı A.S., Merter/Istnul, Turkey) to prevent cockroch escpe. During the cclimtion period (nd during testing) hlf of sugr cookie, cotton ll sturted with wter nd crdord tue (dimeter 4.5 cm x height 10 cm) ws plced in ech continer tht provided food nd hrorge. After cclimtion, one drop (or ed) of product (vg 0.10 g±0.01 g) ws pplied to 76 mm x 26 mm glss microscope slide nd plced on the ottom of continers t 0900h. Only one gel formultion ws tested per continer. Tests were conducted t 26±2 o C, 60±10% reltive humidity, nd photoperiod of 12L12D in lortory. Becuse fipronil is often slow cting nd my possess secondry kill properties due to coprophgy, necrophgy, or cnnilism (15, 16), mortlity ws ssessed t 6 h, 1, 2, 3, 4, nd 5 d fter ppliction. Cockroches were considered ded when they were unle to right themselves. To evlute efficcy under opertionl conditions, ded cockroches were left in continers until the end of test (5 d). In order to determine the effects of storge temperture on residul ctivity of the gels fter plcement, one drop of ech product ws pplied to seprte glss microscope slides nd stored t either 23 o C (wrm) or 30 o C (hot) under drk conditions for 1, 7, 14, 30, 45, 60 nd 90 d fter ppliction. A sufficient numer of slides were treted so tht none were tested more thn once. Tests were repeted on four different dys. 103

3 Irnin J Pul Helth, Vol. 39, No.3, 2010, pp Sttisticl nlyses Men percent mortlity dt were sujected to one-wy nlysis of vrince (ANOVA) nd Student-Newmn-Keuls (SNK) to determine differences within nd etween products, storge time nd temperture (17). Differences were considered significnt t P< Cumultive 5 d men dt re presented in tles. Results Comprison of storge time nd temperture on individul product efficcy Generlly, cockroch verge mortlity to chlorpyrifos-a its stored t 30 o C for 30, 45, nd 90 d ws significntly greter thn t 23 o C (Tle 1). Conversely, t 23 o C mortlity ws significntly greter to chlorpyrifos-a its stored for 14 d nd lest to its ged 90 d compred with the other storge time periods. At 30 o C mortlity ws gretest to this product when stored for 1 nd 30 d. At 14 d mortlity significntly declined to 66%, the reson for this sudden decrese is unknown. Cockroch verge mortlity to fipronil it ged 1 d ws not significntly greter thn it ged through 60 d t either storge temperture (Tle 1). During this period verge percent mortlity rnged from 95 to 98.3%. However, when the it hd een stored for 90 d t 23 o C, mortlity hd significntly decresed to <60%. Wheres, fipronil it stored for the sme mount of time t 30 o C continued to produce 90% mortlity. Averge mortlity to chlorpyrifos-b it stored t vrious time periods t either temperture ws highly vrile (Tle 1). Mortlity levels generlly followed similr ptterns of decline seen in chlorpyrifos-a s storge time incresed. However, cockroch mortlity to chlorpyrifos-b it stored t 23 o C for 60 d suddenly incresed significntly to»87% compred with 56% t 30 o C. In this instnce, evportion of gel moisture my hve resulted in concentrtion of the ctive ingredient. Cockroch mortlity to chlorpyrifos-b it stored for 90 d continued to decline t oth tempertures to >47% nd were not significntly different from one nother. Chlorpyrifos-B its stored t 23 o C for 1 nd 14 d exhiited the gretest cockroch mortlity, ut were not significntly different from one nother, when compred with the other storge periods t this temperture (Tle 1). However t 90 d, efficcy ws significntly reduced compred with the other periods. At 30 o C, chlorpyrifos-b its exhiited the gretest mortlity t dy 14 compred with the other storge periods. At 60 nd 90 d post ppliction, mortlity ws significntly less thn the rest of the storge periods ut not significntly different from one nother. Product efficcy comprison Cockroch mortlity to fipronil or chlorpyrifos-b its stored t 23 o C for 1 dy ws significntly greter thn chlorpyrifos-a (Fig. 1). However, no significnt difference in toxicity etween ll 3 products ws oserved when its were stored t this temperture for 14 d. Fipronil gel stored for 30 nd 45 d t 23 o C continued to provide significntly greter mortlity (³ 95%) thn the chlorpyrifos gels. At 60 d cockroch mortlity to chlorpyrifos-b nd fipronil ws not different from ech other ut were greter thn chlorpyrifos-a. At 90 d mortlity to the fipronil gel hd declined to»60% ut ws still significntly greter thn the chlorpyrifos gels. At 30 o C, fipronil mortlity ws firly consistent nd rnged from 100 to 90% through 90 d (Fig. 2). Cockroch mortlity from chlorpyrifos-a gels stored t this temperture were not significntly different thn fipronil t dys 1 nd 30 while chlorpyrifos-b gels only performed s well s fipronil on dy 14. At dys 45 nd 60, fipronil hd performed significntly etter thn the chlorpyrifos formultions ut t 90 d there ws no difference etween chlorpyrifos-a nd fipronil when stored t 30 o C. 104

4 E Oz et l: Effects of Two Temperture Storge Regimes on Tle 1: Evlution of 3 commercil gel it formultions ginst Bltell. germnic stored t 23 o C nd 30 o C under lortory conditions (5 dy cumultive men percent mortlity±se). Clen Bit Gel (Chlorpyrifos-A) Golith Gel (Fipronil) Serp Gel (Chlorpyrifos-B) Stored dys 23 o C 30 o C 23 o C 30 o C 23 o C 30 o C ± 6.3 x B y 71.5 ± 6.3 A ± 0.0 B ± 0.0 B 88.3 ± 3.0 C 88.3 ± 3.0 B ± 3.3 C 98.3 ± 1.8 C 98.3 ± 1.8 B ± 0.0 B 98.3 ± 1.8 D 81.5 ± 5.0 B ± 1.8 D 66.3 ± 2.7 A 94.8 ± 1.8 B ± 0.0 B 96.5 ± 2.0 D ± 0.0 C ± 4.1 B ± 0.0 C 95.0 ± 3.1 B 96.0 ± 2.0 B 67.8 ± 1.8 B 71.5 ± 6.4 B ± 3.1 A 83.3 ± 4.3 B 96.5 ± 2.0 B ± 0.0 B 69.5 ± 2.2 B 74.8 ± 4.4 B ± 2.7 B 81.8 ± 3.4 B 96.8 ± 3.3 B 96.5 ± 2.0 B 86.5 ± 4.7 C 56.5 ± 4.1 A ± 4.3 A 81.5 ± 4.2 B 59.8 ± 5.9 A 90.0 ± 1.7 A 34.8 ± 1.8 A 46.8 ± 4.7 A x Mens followed y the sme lower cse letter on the sme store dy etween storge tempertures for ech product (row) re not significntly different (P> 0.05). y Mens followed y the sme cpitl letter within n ech product nd temperture etween storge dy (columns) re not significntly different (P> 0.05). Men mortlity c Stored dys Chlorp 1 Chlorp 2 Fipronil Mens followed y the sme letter on the sme dy re not significntly different (P> 0.05). Fig. 1: Lortory evlution of gel it formultions ginst Blttell germnic stored t 23º C (5-dy cumultive men numer ded±se) 105

5 Irnin J Pul Helth, Vol. 39, No.3, 2010, pp Men mortlity c Stored dys Chlorp 1 Chlorp 2 Fipronil Mens followed y the sme letter on the sme dy re not significntly different (P>0.05). Fig. 2: Lortory evlution of gel it formultions ginst Blttell germnic stored t 30º C (5-dy cumultive men mortlity±se) Discussion Our results showed tht lthough ll gel its were highly effective they were not eqully efficcious ginst B. germnic. Freshly pplied its contining fipronil provided complete cockroch mortlity (100%) within 5 d followed y chlorpyrifos-b nd chlorpyrifos A (Tle 1). These dt re in greement with tht reported y Kkeh et l. (18) who reported tht fipronil ws more toxic tht chlorpyrifos. Indeed, complete mortlity to fipronil ws chieved t 2 d wheres the chlorpyrifos its never chieved this level even t 5 d post ppliction evlution. Ghlhoff et l. (15) reported the LT 50 of fipronil gel its ws 1.8 d nd for chlorpyrifos (MxForce FC) 8.5 d. It ppered tht mortlity in our study ws greter thn tht reported y those uthors. However, their study lso found tht secondry kill of mle Germn cockroches vi cnnilism of nymphs tht previously fed on fipronil or chloryrifos its cused significntly greter mortlity thn when they fed upon untreted nymphs. Recent work y Buczkowski nd Schl (16) on horizontl trnsfer of fipronil hs confirmed this oservtion. Generlly, we found tht cockroch mortlity ws greter when the gels were stored t 30º C compred with 23º C. It is possile tht the ctive ingredients in these 3 formultions my hve een concentrted s result of incresed evportion of the gel moisture t the higher temperture. In ny event, fipronil gel proved to e s efficcious s the chlorpyrifos gels nd in some instnces surpssed the ltter formultions depending on storge time nd temperture. Indeed, field trils y Ree et l. (19) found tht fipronil gel its pplied in food hndling estlishments where tempertures cn rech or exceed 30º C reduced B. germnic popultions y 90.9% in Koren resturnts, 96.4% in Chinese resturnts, nd 89.4% in eer-hll kitchens for 4 wk. Moreover, Miller nd Peters (8) reported tht t 12 wk Golith cockroch gel pplied t the rte of 1 drop (0.03 g) per m 2 chieved 94.8% reduction of field popultions of Austrlin nd Germn cockroches. While Srinivsn et l. (20) found tht this sme fipronil formultion (0.03% AI gel) pplied t rtes from to g/m 2 on wood, cement, mud nd thtch surfces gve etween % percent mortlity of dult Germn cockroches fter 1 h exposure in lortory iossys. 106

6 E Oz et l: Effects of Two Temperture Storge Regimes on Ethicl considertions Ethicl issues (Including plgirism, Informed Consent, misconduct, dt friction nd/or flsifiction, doule puliction nd/or sumission, redundncy, etc) hve een completely oserved y the uthors. Acknowledgments The uthors re thnkful to Akdeniz University, The Scientific Reserch Projects Coordintion Unit, Antly, for finncil support. The uthors declre tht they hve no conflicts of interest. References 1. Fotedr R, Shriniws UB, Verm A (1991). Cockroches (Blttell germnic) s crriers of microorgnisms of medicl importnce in hospitls. Epidemiol Infect, 107: Lopes RB, Alves SB (2005). Effect of Gregrin sp. prsitism on the susceptiility of Blttell germnic to some control gents. J Inverter Pthol, 88: Tsi JJ, Chen WC (1999). Different ge of sthmtic ptients ffected y different erollergens. J Microiol Immunol, 32: Ad-elghfr SF, Appel AG, Mck TP (1990). Toxicity of severl insecticide formultions ginst dult Germn cockroches (Dictyopter: Blttellide) control. J Econ Entomol, 83: Cochrn DG (1989). Monitoring for insecticide resistnce in field collected strins of the Germn cockroch (Dictyopter: Blttellide). J Econ Entomol, 82: Ldonni H (2000). Permethrin resistnce rtios compred y two methods of testing nymphs of the Germn cockroch, Blttell germnic. Med Vet Entomol, 14 (2): Rust MK, Reierson DA (1991). A survey of chlorpyrifos resistnce in Germn cockroches from resturnts. J Econ Entomol, 84: Miller PF, Peters BA (1999). Performnce of Golith cockroch gel ginst Germn cockroch (Blttode: Blttellide) nd mixed popultion of Americn cockroch nd Austrlin cockroch (Blttode: Blttide) in the field. In Proceedings of the 3rd Interntionl Conference on Urn Pests. (W. H. Roinson, F. Rettich, nd G.W. Rmo, eds) Prgue: Grfické Závody Hronov, Czech Repulic. pp: Durier V, Rivult C (2003). Improvement of Germn cockroch (Dictyopter: Blttellide) popultion control y frgmented distriution of gel its. J Econ Entomol, 96: Reierson DA, Rust MK, VnDyke A, Apel AG (1983). Control of Germn cockroches with midinohydrzone it. Insectic Acr Tests, 8: Bennett GW, Runstrom ES, Bertholf J (1984). Exmining the where, why nd how of cockroch control. Pest Cont, 48: 19-22, Milio JF, Koehler PG, Ptterson RS (1986). Lortory nd field evlutions of hydrmethylnon it formultions for the control of Americn nd Germn cockroches (Orthopter: Blttellide). J Econ Entomol, 79: Lee CY (1998). Control of insecticide-resistnt Germn cockroches, Blttell germnic (L.) (Dictyopter: Blttellide) in food-outlets with hydrmethylnon sed it sttions. Trop Biomed, 15: Durier V, Rivult C (2008). Comprisons of toxic its for controlling the cockroch, Blttell germnic: ttrctiveness nd feeding stimultion. Med Vet Entomol, 14: Ghlhoff Jr JE, Miller DM, Koehler PG (1999). Secondry kill of dult mle Germn cockroches (Dictyopter: Blttellide) vi cnnilism of nymphs fed toxic its. J Econ Entomol, 92: Buczkowski G, Schl C (2001). Method of insecticide delivery ffects horizontl trnsfer of fipronil in the Germn cockroch (Dic- 107

7 Irnin J Pul Helth, Vol. 39, No.3, 2010, pp tyopter: Blttellide). J Econ Entomol, 94: SPSS Inc (1999). SPSS Bse 10.0 for Windows User's Guide. SPSS Inc., Chicgo IL. 18. Kkeh W, Reid BL, Bennett GW (1997). Toxicity of fipronil to Germn nd Americn cockroches. Entomol Exp Appl, 84: Ree HI, Lee IY, Jeon SH, Yong TS (2006). Field tril on the control effect of fipronil it ginst Germn cockroches. Koren J Prsitol, 44: Srinivsn R, Jmulingm P, Surmnin S, Klynsundrm M (2005). Lortory evlution of fipronil ginst Periplnet mericn nd Blttell germnic. Indin J Med Res, 122: