1-Dr Sloayman RTL Iranian J Arthropod-Borne Dis, (2008), 2(1): 1-6 S Sulaiman et al: Effect of Triflumuron and … 1 Original Article Effect of Triflumuron and Pyriproxyfen on Musca domestica L Larval Stages in the Laboratory S Sulaiman 1 , Sh Hidayatul 2 , Mustakim Syed Mustaffa 1 , J Jeffery 2 1 Department of Biomedical Science, Faculty of Allied Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul aziz 50300 Kuala Lumpur, Malaysia 2 Department of Parasitology and Medical Entomology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz 50300 Kuala Lumpur, Malaysia (Received 15 Apr 2007; accepted 28 Jul 2008) Abstract Background: The residual effect of triflumuron and pyriproxyfen on Musca domestica L larval stages was studied in the laboratory. Methods: Both IGRs at varying concentrations ranging between 0.5 to 2.5 mg/L were placed inside beakers with mice chow and vitamin B complex and water as food for the 1 st , 2 nd and 3 rd instars M.domestica larvae. Results: Both IGRs inhibit M. domestica adult emergence of 98-98.5% when applied at the lowest concentration of 0.5 mg/L on the 1 st instar, 93-97% of adult emergence inhibition on the 2 nd instar,and 91-97% of adult emergence inhibition on the 3 rd instar larvae respectively. There was no significant difference between triflumuron and pyriproxyfen on housefly adult emergence inhibition when fed to the 1 st , 2 nd and 3 rd instars M.domestica larvae (P> 0.05). However, there was a significant difference between the IGRs and the control (P< 0.05). Conclusion: Both triflumuron and pyriproxyfen are effective in inhibiting adult emergence of housefly M domestica and therefore should be recommended for fly control particularly in chicken farms and dumping grounds in Malaysia for housefly control activities. Keywords: Triflumuron, Pyriproxyfen, Adult emergence inhibition, Musca domestica L. Introduction In Malaysia, Sulaiman et al. (1988a, b, 1989, 2000) isolated parasites and enteropa- thogenic bacteria from the external body sur- face and gut lumen of Musca domestica L., Musca sorbens Wiedemann and Chrysomya megacephala (Fabricius). As they also visit clean food, they are important mechanical transmitters of pathogens causing human dis- eases (Reid 1953, Greenberg 1973). In fly management programme chemi- cals are extensively used. Combined use of in- sect growth regulator (IGR) and parasitoids would yield a better reduction in fly density (Srinivasan and Amalraj 2003). Axtell and Edwards (1983) utilized cyromazine as feed additive to caged layering hens under field conditions in high rise, wide span and nar- row poultry houses, the chemical effectively controlled house flies (M domestica) and soldier flies (Hermatia illucens). William and Berry (1980) evaluated the IGR cyromazine as topical spray and feed additive for con- trolling the housefly M. domestica breeding in chicken manure and provided a 70% re- duction of native house flies and 100% inhi- bition of laboratory reared house flies. The objective of this study was to evalu- ate the residual effect of IGRs triflumuron and pyriproxyfen on the larval stages of M domestica L.in the laboratory. *Corresponding author: Prof Dr S. Sulaiman, Tel: +039 2897416, E-mail: salsul@medic.ukm.my Iranian J Arthropod-Borne Dis, (2008), 2(1): 1-6 S Sulaiman et al: Effect of Triflumuron and … 2 Materials and Methods The M domestica L. used was bred in the insectarium of Department of Biomedical Science, Faculty of Allied Health Sciences, Universiti Kebangsaan Malaysia. The two IGRs evaluated were triflumuron (Starycide SC480) provided by Bayer Malaysia Sdn Bhd and pyriproxyfen (Sumilarv 0.5% G) sup- plied by Sumitomo Chemical Company Ltd Tokyo Japan. In each of the glass beaker of 250 ml capacity was placed 5g mice chow and vitamin B complex, 5 ml distilled water and 5 ml of triflumuron or pyriproxyfen of varying concentrations viz 0.5, 1.0, 1.5, 2.0 and 2.5 mg/L. 25 1 st instar M. domestica L. 1 st instar larvae were placed inside each beaker using the forceps. The beakers were then covered with nettings. For the control, only mice chow and vitamin B complex with distilled water were placed in the beaker. After pupation, the pupae were collected and transferred to a small bottle and covered with cotton wool for adult emergence. The mortality of larvae and the number of formed pupae and adults emerging were recorded. Pupae were allowed to emerge into adults for 10 days, if unemerged they were assumed as dead. The efficacy of the IGRs triflumuron and pyriproxyfen were determined by counting the numbers of dead larvae, pupae and adults daily and removing them until mortality and adult emergence were completed. There were 8 replicates per treatment. The above experiment was repeated using 2 nd and 3 rd instar larvae of M domestica .Statistical analysis with ANOVA followed by Pos Hoc Tests using LSD Multiple Comparison. Results The effect of triflumuron on 1 st instar M. domestica larvae indicated adult emergence inhibition of 98.0-100% at concentrations of 0.5-2.5 mg/L and 23.5% adult emergence inhibition for the control, respectively (Table 1). Effect of pyriproxyfen on 1 st instar M. domestica larvae indicated adult emergence inhibition of 98.5-100% at concentrations between 0.5 to 2.5 mg/L and 18.0% adult emergence inhibition for the control respectively (Table 2). There was no significant difference be- tween triflumuron and pyriproxyfen on the housefly adult emergence inhibition when fed to the 1 st instar larvae. However, there was a significant difference between the IGRs and the control (P< 0.05). The effect of triflu- muron on 2 nd instar M. domestica larvae in- dicated adult emergence inhibition between 93.0 to 100% at concentrations of 0.5-2.5 mg/L and 19.0% adult emergence inhibition for the control (Table 3). Effect of pyriproxyfen on the 2 nd instar M. domestica lar- vae indicated adult emergence inhibition of 97.0-100% at concentrations varying between 0.5 to 2.5 mg/L and 13.0% adult emergence inhibition for the control respectively (Table 4). There was no significant difference between triflumuron and pyriproxyfen on the housefly adult emergence inhibition when fed to the 2 nd instar larvae (P> 0.05). However, there was a significant difference between the IGRs and the control (P< 0.05). The effect of triflu- muron on 3 rd instar M. domestica larvae indi- cated adult emergence inhibition of 91.0-99.5% at concentrations between 0.5 to 2.5 mg/L and 6.5% adult emergence inhibition for the con- trol respectively (Table 5). Effect of pyriproxy- fen on 3 rd instar M. domestica larvae indi- cated adult emergence inhibition of 97.0- 100% at concentrations varying between 0.5 to 2.5 mg/L and 8.5% adult emergence inhibition for the control respectively (Table 6). There was no significant difference between triflumuron and pyriproxyfen on the housefly adult emergence inhibition when fed to the 3 rd instar larvae. However, there was a signifi- cant difference between the IGRs and the control (P<0.05). Iranian J Arthropod-Borne Dis, (2008), 2(1): 1-6 S Sulaiman et al: Effect of Triflumuron and … 3 Table 1. Residual effect of triflumuron on 1 st instar M domestica L.larvae and emerging pupae and adults in the laboratory Concentrations (mg/L) No. of L1 larvae Mortality of L1 larvae (% in parentheses) No. of pupae formed (% in parentheses) Adult emer- gence (% in parentheses) Adult emergence inhibition (%) 0.5 200 98 (49.0) 102 (51.0) 4(2.0) 98.0 1.0 200 112 (56.0) 88 (44.0) 2(1.0) 99.0 1.5 200 156 (78.0) 44 (22.0) 0 (0) 100 2.0 200 140 (70.0) 60 (30.0) 1(0.5) 99.5 2.5 200 163 (81.5) 37 (18.5) 0 (0) 100 Control 200 32 (16.0) 168 (84.0) 153 (76.5) 23.5 Table 2. Residual effect of pyriproxyfen on 1st instar M domestica L.larvae and emerging pupae and adults in the laboratory Concentrations (mg/L) No. of L1 larvae Mortality of L1 larvae (% in parentheses) No. of pupae formed (% in parentheses) Adult emer- gence (% in parentheses) Adult emer- gence inhibi- tion (%) 0.5 200 23 (11.5) 177 (88.5) 3 (1.5) 98.5 1.0 200 26 (13.0) 174 (87.0) 1(0.5) 99.5 1.5 200 28 (14.0) 172 (86.0) 0 (0) 100 2.0 200 44 (22.0) 156 (78.0) 1 (0.5) 99.5 2.5 200 61(30.5) 139 (69.5) 0 (0) 100 Control 200 22 (11.0) 178 (89.0) 164 (82.0) 18 Table 3. Residual effect of triflumuron on 2 nd instar M domestica L.larvae and emerging pupae and adults in the laboratory Concentrations (mg/L) No. of L1 larvae Mortality of L1 larvae (% in pa- rentheses) No. of pupae formed (% in parentheses) Adult emergence (% in parenthe- ses) Adult emer- gence inhibition (%) 0.5 200 56 (28) 144 (72.0) 14 (7.0) 93.0 1.0 200 65 (32.5) 135 (67.5) 7 (3.5) 96.5 1.5 200 80 (40.0) 120 (60.0) 2 (1.0) 99 2.0 200 93 (46.5) 107 (53.5) 3 (1.5) 98.5 2.5 200 102 (51.0) 98 (49.0) 0 (0) 100 Control 200 23 (11.5) 177 (88.5) 162 (81.0) 19.0 Iranian J Arthropod-Borne Dis, (2008), 2(1): 1-6 S Sulaiman et al: Effect of Triflumuron and … 4 Table 4. Residual effects of pyriproxyfen on 2 nd instar M domestica L.larvae and emerging pupae and adults in the laboratory Concentrations (mg/L) No. of L1 larvae Mortality of L1 lar- vae (% in parenthe- ses) No. of pupae formed (% in parentheses) Adult emergence (% in parentheses) Adult emer- gence inhibition (%) 0.5 200 18 (9.0) 183 (91.5) 6 (3.0) 97.0 1.0 200 19 (9.5) 181(90.5) 5 (2.5) 97.5 1.5 200 24 (12.0) 176 (88.0) 1(0.5) 99.5 2.0 200 26 (13.0) 174 (87.0) 1(0.5) 99.5 2.5 200 26 (13.0) 174 (87.0) 0 (0) 100 Control 200 14 (7) 186 (93.0) 174 (87.0) 13.0 Table 5. Residual effect of triflumuron on 3 rd instar M domestica L.larvae and emerging pupae and adults in the laboratory. Concentrations (mg/L) No. of L1 larvae Mortality of L1 larvae (% in pa- rentheses) No. of pupae formed (% in parentheses) Adult emergence (% in parenthe- ses) Adult emergence inhibition (%) 0.5 200 16 (8.0) 184 (92.0) 18 (9.0) 91.0 1.0 200 25 (12.5) 175 (87.5) 8 (4.0) 96.0 1.5 200 40 (20.0) 160 (80.0) 4 (2.0) 98.0 2.0 200 62 (31.0) 138 (69.0) 4 (2.0) 98.0 2.5 200 89 (44.5) 111(55.5) 1 (0.5) 99.5 Control 200 6 (3.0) 194 (97.0) 187(93.5) 6.5 Table 6. Residual effect of pyriproxyfen on 3 rd instar M domestica L.larvae and emerging pupae and adults in the laboratory Concentrations (mg/L) No. of L1 larvae Mortality of L1 larvae (% in parentheses) No. of pupae formed (% in parentheses) Adult emer- gence (% in parentheses) Adult emergence inhibition (%) 0.5 200 11(5.5) 189(94.5) 6(3.0) 97.0 1.0 200 11(5.5) 189(94.5) 5(2.5) 97.5 1.5 200 16(8.0) 184(92.0) 1(0.5) 99.5 2.0 200 14(7.0) 186(93.0) 2(1.0) 99.0 2.5 200 15(7.5) 185(92.5) 0(0) 100 Control 200 10(5.0) 190(95.0) 183(91.5) 8.5 Iranian J Arthropod-Borne Dis, (2008), 2(1): 1-6 S Sulaiman et al: Effect of Triflumuron and … 5 Discussion Kelly et al. (1987) evaluated cyromaz- ine against insecticide-resistant field strains of M domestica. No tolerance to cyromazine was found, neither was there a direct correlation between larval responses to cy- romazine and adult responses to other in- secticides. It was concluded that cyromazine has a good potential for the control of house- flies that exhibited high levels of resistance to other insecticides. Vazirianzadeh et al. (2007) studied the effects of oral application of cyromazine and triflumuron on housefly larvae. Both IGRs had sufficient effect on larval mortality compared with their controls. It was concluded that cyromazine should be used in a larvicidal programme to control housefly rather than triflumuron. In the present study both triflumuron and pyriproxyfen had residual effects on all the larval stages of M. domestica. Both IGRs inhibit adult emergence of 98 to 98.5% when applied at the lowest concentration of 0.5 mg/L on the 1 st instar, 93 to 97% adult emer- gence inhibition on the 2 nd instar and 91 to 97% adult emergence inhibition on the 3 rd instar larvae. According to Hatakoshi et al. (1987) S-31183 (now called pyriproxyfen) was more effective than methoprene and diflubenzuron against 4-day old larvae of M. domestica in artificial medium and more ac- tive than methoprene against eggs and 4- day-old larvae of M. domestica in the chicken manure medium. Kostina (1999) studied the influence of pyriproxyfen on preimaginal stages of M. domestica and Aedes aegypti. The presence of 10, 20, and 40 grams preparation per square meter of surface the number of fly puparia was 62.2%, 50% and 38.7% respec- tively. The emergence of adults was 0.5% in the case of 10 gram per square meter, and it was completely absent in the latter cases. According to Zhang and Shono (1997), the toxicity of pyriproxyfen gradually increased from the early stage of 3 rd instar larva of houseflies to the white pupal stage. Further studies indicated that the white pupa was the most susceptible stage to the chemical and the mortality of pupae decreased sharply after the white pupal stage. In conclusion, both triflumuron and pyriproxyfen are effective in inhibiting adult emergence of housefly M domestica and therefore should be recommended for fly control particularly in chicken farms and dumping grounds in Malaysia for housefly control activities. Acknowledgements We wish to thank Faculty of Allied Health Sciences, Universiti Kebangsaan Malay- sia for providing research facilities. We ap- preciate Bayer Malaysia Sdn Bhd for provid- ing triflumuron and Sumitomo Chemical Com- pany Tokyo, Japan for providing pyriproxyfen. References Axtell RC, Edwards TD (1983) Efficacy and nontarget effects of Larvadex as a feed additive for controlling house flies in a caged-layer poultry manure. Poult Sci. 62(12):2371-2377. Greenberg B (1973) Flies and Disease, Vol II. Biology and Disease Transmission. Princeton University Press, New Jersey. Hatakoshi M, Kawada H, Nishida S, Kisida H, Nakayama I (1987) Laboratory evaluation of 2-[1-methyl-2-(4-phenoxy- phenoxy)-ethoxy] pyridine against larvae of mosquitoes and housefly. Jpn J Sanit Zool. 38(4):271-274. Kelly JA, Stubbs MR, Pinniger DB (1987) Laboratory evaluation of cyromazine against insecticide-resistant field strains of Musca domestica. Med Vet Ento- mol. 1(1):65-69. Kostina MN (1999) Influence of pyriproxy- fen on preimaginal stages of Musca do- Iranian J Arthropod-Borne Dis, (2008), 2(1): 1-6 S Sulaiman et al: Effect of Triflumuron and … 6 mestica and Aedes aegypti. Proceedings of the 3 rd International Conference on Urban Pests, 1999 19 - 22 July, Prague, Czech Republic . p. 626 Reid JA (1953) Notes on house-flies and blowflies in Malaya. Bull Inst Med Res. Fed of Malaya. 7:1-26. Srinivasan R, Amalraj DD (2003) Efficacy of insect parasitoid Dirhinus himalayanus (Hymenoptera: Chalcidae) and insect growth regulator, triflumuron against house fly, Musca domestica (Diptera: Muscidae). Indian J Med Res.118:158- 66. Sulaiman S, Sohadi AR, Yunus H, Iberahim R (1988a) The role of some cyclorrhaphan flies as carriers of human helminths in Malaysia. Med Vet Entomol. 2:1-6. Sulaiman S, Aziz AH, Yunus H, Sohadi AR (1988b) Isolations of enteropathogenic bacteria from some cyclorrhaphan flies in Malaysia. Malays Appl Biol. 17(2): 129-133. Sulaiman S, Sohadi AR, Jeffery J (1989) Human helminth parasite burdens on cyclorrhaphan flies (Diptera) trapped at an aboriginal settlement in Malaysia. Bull Ent Res.79:625-629. Sulaiman S, Othman MZ, Aziz AH (2000) Isolations of enteric pathogens from synanthropic flies trapped in down- town Kuala Lumpur. J Vector Ecol. 25(1): 90-93. Vazirianzadeh B, Jervis MA, Kidd NAC (2007) The effects of oral application of cyromazine and triflumuron on house- fly larvae. Iranian J Arthropod-Borne Dis. 1(2): 7- 13. William RE, Berry TG (1980) Evaluation of CGA 72662 as a topical spray and feed additive for controlling house flies breed- ing in chicken manure. Poult Sci. 59(10): 2207-2212. Zhang Li, Shona T (1997) Toxicities of pyrip- roxyfen to susceptible and resistant strains of houseflies. Jap Soc Appl Entomol Zool. 32(2): 373-378.