Laboratory Evaluation of Novaluron As a Development Site Treatment for Controlling Larval Horn Flies, House Flies, and Stable Flies (Diptera: Muscidae)

University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln

U.S. Department of Agriculture: Agricultural Publications from USDA-ARS / UNL Faculty Research Service, Lincoln, Nebraska

2012

Laboratory Evaluation of Novaluron as a Development Site Treatment for Controlling Larval Horn Flies, House Flies, and Stable Flies (Diptera: Muscidae)

K. H. Lohmeyer USDA-ARS, [email protected]

J.M. Pound USDA-ARS

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Lohmeyer, K. H. and Pound, J.M., "Laboratory Evaluation of Novaluron as a Development Site Treatment for Controlling Larval Horn Flies, House Flies, and Stable Flies (Diptera: Muscidae)" (2012). Publications from USDA-ARS / UNL Faculty. 1049. https://digitalcommons.unl.edu/usdaarsfacpub/1049

This Article is brought to you for free and open access by the U.S. Department of Agriculture: Agricultural Research Service, Lincoln, Nebraska at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Publications from USDA-ARS / UNL Faculty by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. VECTOR CONTROL,PEST MANAGEMENT,RESISTANCE,REPELLENTS Laboratory Evaluation of Novaluron as a Development Site Treatment for Controlling Larval Horn Flies, House Flies, and Stable Flies (Diptera: Muscidae)

1 K. H. LOHMEYER AND J. M. POUND

USDAÐARS, Knipling-Bushland U. S. Livestock Insects Research Laboratory, 2700 Fredericksburg Road, Kerrville, TX 78028

J. Med. Entomol. 49(3): 647Ð651 (2012); DOI: http://dx.doi.org/10.1603/ME11220 ABSTRACT A granular formulation of novaluron (Novaluron 0.2G, 0.2% [AI]), a newer benzoylphenyl urea insecticide, was evaluated for its efÞcacy in controlling the larval stage of horn ßies, Haematobia irritans (L.); house ßies, Musca domestica L.; and stable ßies, Stomoxys calcitrans (L.), in cow manure. Various rates and insecticide placement locations (top, middle, and bottom of manure) were evaluated in this study and all combinations of these variables reduced adult emergence of all three species when compared with the untreated controls. The presence of deformed pupae indicated that novaluron had an insect growth regulator effect on the developing ßy larvae. Top, middle, or bottom application rates of 0.125, 0.195, 0.25, and 0.375 g novaluron onto manure samples, reduced adult horn ßy emergence by Ͼ90%. Middle and bottom application rates of 0.195, 0.25, and 0.375 g novaluron reduced adult house ßy emergence Ͼ93%. All rates and placement combinations resulted in Ͼ98% reduction of adult stable ßy emergence. The level of control efÞcacy observed against these three ßy species along with the ease of use of a granular formulation, make this product an ideal candidate for use in an integrated livestock pest management program.

KEY WORDS Haematobia irritans, Stomoxys calcitrans, Musca domestica, benzoylphenyl urea

Horn ßies, Haematobia irritans (L.) (Diptera: Mus- control because of the limited time they spend on cidae), and stable ßies, Stomoxys calcitrans (L.) hosts as well as the ability to disperse long distances (Diptera: Muscidae), are two of the most econom- (Taylor et al. 2010, Pitzer et al. 2011). Traditionally, ically damaging ectoparasites of pastured cattle. An- stable ßies were considered pests of conÞned cattle nual losses in cattle production because of reduced operations such as dairies and feedlots, but over the weight gain and poor feeding efÞciency are esti- past few decades the use of round hay bales has mated at over two billion dollars each year (Kunz et complicated stable ßy control in rangeland and pas- al. 1991, Taylor et al. 2012). House ßies, Musca do- tured cattle operations. Round bale technology mestica L. (Diptera: Muscidae), are pestiferous to makes it more convenient for producers to feed humans and cattle and can mechanically transmit, or pastured and rangeland cattle during winter serve as reservoirs for helminth eggs, bacteria, and months, but also increases amounts of wasted hay. viruses that cause enteric infections, eye infections, This wasted hay mixes with cattle manure deposited and skin infections (Greenberg 1973, Emerson et al. at feeding sites and creates an ideal development 1999, Sasaki et al. 2000). medium for larval stable ßies (Broce et al. 2005, Historically, Þlth ßy management has been based Talley et al. 2009, Taylor and Berkebile 2011). on the use of insecticides applied as dusts, sprays, Reliance upon insecticides for ßy control has led to pour-ons, feed additives, ear tags, structure sprays, an increased incidence of resistance that is the even- and boluses (Foil and Hogsette 1994, Butler and tual cause of product failure. As the incidence of Okine 1999). Stable ßies are particularly difÞcult to resistance increases and traditional chemical insecticides fail, the need is becoming crucial for compounds with novel modes of action that improve integrated This paper reports the results of research only. Mention of a com- pest management (IPM) programs. mercial or proprietary product in this paper does not constitute an endorsement by the U.S. Department of Agriculture. In conducting Novaluron (1-[3-chloro-4-(1,1,2-trißuoro-2-trißuo- the research described in this report, the investigators adhered to romethoxy-ethoxy) phenyl]-3-(2,6-dißuorobenzoyl) protocol approved by the USDAÐARS Animal Welfare Committee. urea) is a newer insect growth regulator (IGR) in the The protocol is on Þle at the USDAÐARS, Knipling-Bushland U.S. benzoylphenyl urea group of insecticides and has very Livestock Insects Laboratory, Tick Research Unit, Kerrville, TX. 78028. low toxicity to mammals, birds, and earthworms 1 Corresponding author, e-mail: [email protected]. (Barazani 2001, Ishaaya et al. 2007). Novaluron is a 648 JOURNAL OF MEDICAL ENTOMOLOGY Vol. 49, no. 3 chitin synthesis inhibitor that is active through in- under Ϸ5 cm of manure. These treatment placement gestion and contact, in particular targeting larval locations would also allow determination of how evenly insects that actively synthesize chitin. Previous novaluron would need to be distributed within manure work has shown that novaluron is larvicidal against for controlling larval horn ßies developing in manure sand ßies and several species of mosquitoes (Mulla pats, should this product be formulated as a feed-thru et al. 2003, Mascari et al. 2007, Arredondo-Jime´ nez product. and Valdez-Delgado 2006). A study conducted by Each rate/location was applied to 100 g of manure Cetin et al. (2006) showed that a 1% emulsiÞable contained in 532 ml plastic Solo brand cups (Solo Cup concentrate of novaluron caused Ͼ80% mortality Company, Lake Forest, IL). For top treatment 100 g when fed to a Þeld strain of house ßy larvae, but no of manure were placed in the cup and the appropriate other work has been done to evaluate efÞcacy of amount of granular novaluron was sprinkled on the novaluron against larval dipteran pests of cattle. The top of the manure. For middle treatment 50 g of ma- objective of the current study was to evaluate efÞ- nure were placed in the cup, the novaluron was sprin- cacy of a granular formulation of novaluron for kled on top, and the remaining 50 g of manure were controlling immature stages of horn ßies, house ßies, added to the top. For bottom treatment: the appro- and stable ßies in cow manure. priate amount of novaluron was sprinkled onto the bottom of the cup and 100 g of manure were added to the cup. A similar quantity of untreated manure was Materials and Methods used as a control. Three replicates of each rate/loca- Fly Rearing. Flies used in this study were obtained tion combination and untreated controls were per- from colonies of insecticide-susceptible horn ßies, formed. Bioassays for horn ßies, house ßies, and stable house ßies, and stable ßies reared at the U.S. Depart- ßies were initiated on separate days (14 September ment of AgricultureÐAgriculture Research Services 2010, 26 October 2010, and 11 March 2011, respec- (USDAÐARS), Knipling-Bushland U.S. Livestock In- tively). After manure and product had been added to sects Research Laboratory (KBUSLIRL), Kerrville, each cup, 100 ßy eggs were applied to the top of the TX. Laboratory colonies were maintained at 24Ð28ЊC, manure. Cups were covered with facial tissue held in 60% RH, and a photoperiod of 12:12 (L:D) h. Colony place with a rubber band, maintained in a rearing horn ßy larvae were reared on a cow manure and room held at 28ЊC, 60% RH, and a photoperiod of 12:12 peanut hull diet (Lohmeyer and Kammlah 2006). Lar- (L:D) h for 7Ð10 d, and observed for pupation. Pupae val stable ßies and larval house ßies were reared on a were then collected by ßotation and numbers of nor- diet of Purina Fly Chow (Purina Mills LLC, Gray mal and deformed pupae recorded. Pupae were held Summit, MO). Colony adult horn ßies and stable ßies at 28ЊC, 60% RH, and a photoperiod of 12:12 (L:D) h were fed citrated bovine blood, and adult house ßies until adult emergence. The number of emerged adults were fed a sugar/milk solution (50 g of powdered milk was recorded for each sample. and 20 g of table sugar per liter of water). Statistical Analysis. Before analysis, manure bioas- Cattle Manure. Manure used for ßy bioassays was say data were corrected using AbbottÕs formula (Ab- collected from untreated steers maintained in stanchion bott 1925). Analysis of the corrected AbbottÕs formula at KBUSLIRL. Steers used for manure production variable was performed using the GLIMMIX proce- weighed Ϸ136 kg and were fed 5.4 kg of 5.1 cm square dure in SAS (SAS Institute 2009). The response alfalfa cubes per day and water ad libitum. Fresh manure variable (number of emerged adult ßies) was non- was collected and placed in plastic bags and was frozen normally distributed in each bioassay, and was deter- until needed for bioassays. Animal care procedures ad- mined to follow the structure of the Beta probability hered to the Guide for the Care and Use of Laboratory distribution, which uses the logit link function as its Animals, as promulgated by the Institutional Animal Use linearization approach. To have complete estimability and Care Committee of KBUSLIRL. of the interaction and main effects, the AbbottÕs values Larval Bioassays. Granular novaluron (MANA No- were assessed in a 0Ð1 scale where values equal to valuron 0.2G: a purple granule (0.4Ð1.68 mm) formu- 100% (i.e., complete control) were replaced with 9 ϫ lated as 0.2% [AI] in wood Þber/granite; Makhteshim 10Ϫ9. Rate, location, and their interaction were con- Agan of North America, Inc, Raleigh, NC) was applied sidered Þxed effects in the statistical model. Estimated at Þve rates (0.0625, 0.125, 0.195, 0.25, or 0.375 g prod- means, SEs, and differences of mean were calculated uct per cup. In the Þeld, a granular product for house using the LSMEANS option. Multiple comparison P and stable ßy control would be scattered over areas values were controlled for type I error using the Tuke- where larval ßies are suspected to be developing. In yÐKramer adjustment with P Ͻ 0.05. most cases the material may not remain on top, but may be worked down into the soil/manure to varying Results levels by animal movement or deposition of manure as cattle are standing around a food source. Therefore, Horn Flies. All application rates and treatment loca- three treatment placement locations were selected at tions of novaluron reduced adult ßy emergence com- each application rate to simulate Þeld conditions: top pared with the untreated controls (Fig. 1A). Statistical of manure, product remains where it was originally comparison of the treatments revealed a signiÞcant ef- placed; middle of manure, product ends up under Ϸ2.5 fect for the rate of novaluron applied (F ϭ 32.54; df ϭ 4, cm of manure; and bottom of manure, product ends up 29; P Ͻ 0.0001), the placement location of the novaluron May 2012 LOHMEYER AND POUND:CONTROL OF DIPTERA IN MANURE USING NOVALURON 649

Fig. 1. Effects of various rates and treatment placements of granular novaluron on adult emergence (A, B, C), number of pupae recovered (D, E, F), and pupal deformity (G, H, I) of horn ßies, house ßies, and stable ßies.

(F ϭ 16.18; df ϭ 2, 29; P Ͻ 0.0001), and the interaction horn ßies, house ßies, and stable ßies in the laboratory. of rate and placement location (F ϭ 5.86; df ϭ 8, 29; P ϭ All of the rates and insecticide location placement 0.0002) on adult ßy emergence. Deformed pupae were combinations evaluated in this study reduced adult observed at all application rates and treatment locations horn ßy, house ßy, and stable ßy emergence. The (Fig. 1G). presence of deformed pupae indicated that novaluron House Flies. All application rates and treatment has an IGR effect on the development of the three ßy locations of novaluron reduced adult ßy emergence species tested, resulting in larval mortality before they compared with the untreated controls (Fig. 1B). Sta- reached pupation or deformed pupae from which tistical comparison of the treatments showed a signif- adults never emerged. icant effect for the rate of novaluron applied (F ϭ It was evident that novaluron middle and bottom 46.28; df ϭ 4, 29; P Ͻ 0.0001), the placement location application at Ͼ0.195 g affected both horn ßy and of the novaluron (F ϭ 111.04; df ϭ 2, 29; P Ͻ 0.0001), house ßy development, as this combination reduced and the interaction of rate and placement location adult emergence of these ßies by Ͼ90%. Interest- (F ϭ 10.48; df ϭ 8, 29; P Ͻ 0.0001) on adult ßy emer- ingly, a few of the more effective rate/placement gence. As with horn ßies, deformed pupae were ob- location combinations for controlling adult horn ßy served at all application rates and treatment locations emergence were not as effective against adult house (Fig. 1H). ßy emergence (i.e., middle application of 0.125 g Stable Flies. All application rates and treatment novaluron and top application of 0.195, 0.25, and locations caused Ն98% reduction in adult ßy emer- 0.375 g). This was not unexpected because these gence (Fig. 1C). Deformed pupae were not observed ßies do not develop in similar environments in na- for the bottom application rates of 0.0625, 0.125, 0.195, ture. Further, the efÞcacy we observed against 0.25, and 0.375 g; however, it appears that mortality house ßies is similar to the efÞcacy observed by may have been caused in larval ßies before they Cetin et al. (2006) for higher rates of a 1% emulsion reached the point of pupation (Fig. 1I). concentrate formulation of novaluron mixed into larval house ßy diet. The granular formulation of novaluron evaluated in Discussion this study was particularly efÞcacious against stable Novaluron, formulated as a granule (0.2% [AI]) and ßies. A dose affect was not apparent as all rates and applied to cow manure, is efÞcacious against larval placement combinations in this study exhibited Ͼ98% 650 JOURNAL OF MEDICAL ENTOMOLOGY Vol. 49, no. 3 reduction in adult stable ßy emergence. This is an the knowledge of Diptera. Association Publishers, interesting phenomenon as the rate of insecticide Gainesville, FL. needed to kill stable ßies is usually much higher than Cetin, H., F. Erler, and A. Yanikoglu. 2006. Larvicidal the rate needed to kill horn ßies (Frazar and Schmidt activity of novaluron, a chitin synthesis inhibitor, 1979, Schmidt and Kunz 1980, Miller et al. 1986). The against the houseßy, Musca domestica. J. Insect Sci. 6: IGR effect of novaluron on larval stable ßies appears 50Ð54. to be robust as few to no pupae were recovered from Emerson, P. M., S. W. Lindsay, G.E.L. Walraven, H. Faal, C. Bogh, K. Lowe, and R. L. Bailey. 1999. Effect of ßy most rate and treatment placement combinations, and control on trachoma and diarrhea. Lancet 353: 1401Ð a very small number of stable ßies survived to emer- 1403. gence. Foil, L. D., and J. A. Hogsette. 1994. Biology and control of In all likelihood, a granular product placed in the tabanids, stable ßies, and horn ßies. Rev. Sci. Tech Pff. Int. Þeld would not remain on the surface of the treatment Epiz. 13: 1125Ð1158. site, but would be worked down into the substrate by Frazar, E. D., and C. D. Schmidt. 1979. Susceptibility of animal movement or deposition of manure, as is the laboratory-reared horn ßies and stable ßies to selected case with wasted hay and other debris around cattle insecticides. J. Econ. Entomol. 72: 884Ð886. feeding sites. Though some variation in efÞcacy was Greenberg, B. 1973. Flies and disease, vol. II. Biology and observed for the different placement locations, it ap- disease transmission. Princeton University Press, Prince- pears that the granular formulation of novaluron ton, NJ. would work well regardless of where it is spread pro- Ishaaya, I., A. Barazani, S. Kontsedalov, and A. R. Horowitz. 2007. Insecticides with novel modes of action: Mecha- vided it is applied at a rate that is efÞcacious for nism, selectivity and cross-resistance. Entomol. Res. 37: controlling that particular ßy species. 148Ð152. In conclusion, the granular formulation of novalu- Kunz, S. E., K. D. Murrell, G. Lambert, L. F. James, and ron used in this study appears to deliver adequate C. E. Terrill. 1991. Estimated losses of livestock to levels of active ingredient for effective control of horn pests, vol. 1, pp. 69Ð98. In D. Pimentel (ed.), CRC ßies, house ßies, and stable ßies in treated cow ma- handbook of pest management in agriculture. CRC, nure. The efÞcacy observed against stable ßies is par- Boca Raton, FL. ticularly promising, making this formulation of no- Lohmeyer, K. H., and D. M. Kammlah. 2006. Improved mass valuron a viable candidate for use by cattle operations rearing techniques for the horn ßy, Haematobia irritans that use round bales. Future studies are being planned (L.) (Diptera: Muscidae). Southwest. Entomol. 31: 83Ð to evaluate granular novaluron in the Þeld for use at 85. round bale feeding sites and to evaluate longevity of Mascari, T. M., M. A. Mitchell, E. D. Rowtan, and L. D. Foil. 2007. Evaluation of novaluron as a feed-through insec- efÞcacy. Studies are also needed to evaluate the po- ticide for control of immature sand ßies (Diptera: Psy- tential of the product for controlling house ßies de- chodidae). J. Med. Entomol. 44: 714Ð717. veloping in dairies and to determine treatment appli- Miller, J. A., D. D. Oehler, A. J. Siebenaler, and S. E. Kunz. cation techniques for using the product for horn ßy 1986. Effect of ivermectin on survival and fecundity of control in the Þeld. horn ßies and stable ßies (Diptera: Muscidae). J. Econ. 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