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Identification of Oviposition Attractants of the Secondary Screwworm, Cochliomyia Macellaria (F.) Released from Rotten Chicken Liver

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Identification of Oviposition Attractants of the Secondary Screwworm, Cochliomyia Macellaria (F.) Released from Rotten Chicken Liver Identification of Oviposition Attractants of the Secondary Screwworm, Cochliomyia macellaria (F.) Released from Rotten Chicken Liver Junwei J. Zhu, Muhammad F. Chaudhury, Khanobporn Tangtrakulwanich & Steven R. Skoda Journal of Chemical Ecology ISSN 0098-0331 Volume 39 Combined 11-12 J Chem Ecol (2013) 39:1407-1414 DOI 10.1007/s10886-013-0359-z 1 23 Your article is protected by copyright and all rights are held exclusively by Springer Science+Business Media New York (outside the USA). This e-offprint is for personal use only and shall not be self-archived in electronic repositories. If you wish to self-archive your article, please use the accepted manuscript version for posting on your own website. You may further deposit the accepted manuscript version in any repository, provided it is only made publicly available 12 months after official publication or later and provided acknowledgement is given to the original source of publication and a link is inserted to the published article on Springer's website. The link must be accompanied by the following text: "The final publication is available at link.springer.com”. 1 23 Author's personal copy J Chem Ecol (2013) 39:1407–1414 DOI 10.1007/s10886-013-0359-z Identification of Oviposition Attractants of the Secondary Screwworm, Cochliomyia macellaria (F.) Released from Rotten Chicken Liver Junwei J. Zhu & Muhammad F. Chaudhury & Khanobporn Tangtrakulwanich & Steven R. Skoda Received: 5 September 2013 /Revised: 25 September 2013 /Accepted: 13 October 2013 /Published online: 31 October 2013 # Springer Science+Business Media New York (outside the USA) 2013 Abstract The secondary screwworm, Cochliomyia macellaria elicited significant EAG responses from antennae of gravid (F.), is an important blowfly species affecting both livestock females. Oviposition assays showed that the 7-component and humans. It can transmit pathogenic disease agents blend stimulated gravid females to lay significantly more eggs mechanically and is an agent of facultative myiasis, which than the other combinations tested. This 7-component blend leads to economic losses. The adult flies are attracted to may have potential for use in monitoring and sampling decomposing carcasses, carrion, or rotten meat in order to populations of secondary screwworm and their associated deposit their eggs, and the hatched larvae develop on these disease epidemiology. decaying organic materials. This research was aimed to identify volatiles emitted from rotten chicken livers that were Keywords Oviposition attractant . Blow flies . Forensic reported previously to attract gravid females. In laboratory veterinary pest . Solid phase microextraction . Livestock oviposition assays, gravid females laid significantly more disease . Economic entomology eggs on rotten livers than on fresh livers, and rotten chicken liver was more attractive than rotten beef liver. Volatiles from the two livers were collected using solid phase Introduction microextraction. Significantly different volatile profiles were detected from the rotten livers of beef and chicken. The secondary screwworm (SSW), Cochliomyia macellaria Electroantennography (EAG) was performed to determine (F.), belongs to the family Calliphoridae. The species can be antennal responses to chemicals released from the most found in almost every known terrestrial habitat throughout attractive chicken liver that are candidate oviposition the Western Hemisphere. In the United States, secondary attractants. Seven compounds from rotten chicken liver screwworms are of particular importance to both humans and the livestock industry because of large economic losses resulting from pathogen transmission and myiasis (Anderson J. J. Zhu (*) and Huitson 2004). Adults of secondary screwworm appeared USDA-ARS, Agroecosystem Management Research Unit, to be morphologically indistinguishable from the primary 305B Entomology Hall, UNL-EC, Lincoln, NE 68583, USA screwworm (PSW), Cochliomyia hominivorax (Coquerel) in e-mail: [email protected] earlier studies until 1933, when Laake and others discovered M. F. Chaudhury characteristic differences between these two species (Cushing USDA-ARS, Screwworm Research Unit, 105B Entomology Hall, and Patton, 1933; Laake et al. 1936). Their larval habitats are UNL-EC, Lincoln, NE 68583, USA distinct in that PSW feeds and develops on living tissues of animals and humans (obligatory or primary myiasis) whereas K. Tangtrakulwanich Department of Entomology, University of Nebraska, Lincoln, SSW breeds in carrion or necrotic flesh (Laake et al. 1936). NE 68583, USA Although the larvae of SSW typically develop in decomposing carcasses, carrion, or rotten meat; they can also cause S. R. Skoda facultative myiasis by infesting wounds of animals and USDA-ARS, Screwworm Research Unit, Knipling-Bushland U. S. Livestock Insects Research Laboratory, 2700 Fredericksburg Road, humans, often following invasion by primary myiasis- Kerrville, TX 78028, USA causing flies (Broce 1985; Mullen and Durden 2002). Recently Author's personal copy 1408 J Chem Ecol (2013) 39:1407–1414 SSW has gained more recognition in the field of forensic refrigerated until used in the experiments. Livers were entomology as the principal species used to determine removed from the refrigerator and kept at room temperature postmortem interval estimations (Boatright and Tomberlin (25°C)andhumidity(55%R.H.)forabout1hr,and 2010). Furthermore, SSW has been considered as an invasive afterward were used as fresh liver; rotten livers were prepared pest that was intercepted from cargoes entering Asia from by leaving the liver pieces exposed at room temperature Mexico, Peru, and Panama (Deng et al. 2009). (25 °C) and humidity (55 % R.H.) for 3 days after purchase. In nature, adults of PSWand SSW occur together, but SSW greatly outnumbers PSW in liver, and in the traps baited with Landing Response to Oviposition Substrate Two sets of attractant that lures contain blends of compounds identified experiments were conducted using methods similar to those from infected animal wounds, “swormlures”, which are used described for the primary screwworm by Chaudhury et al. to monitor PSW adults (Coppedge et al. 1977; Jones et al. (2002) in wire mesh cages (30×30×30 cm) with solid metal 1976;Lindquist1938;Mackley1986). In the southern United base. In the first set, 25 eight-day-old male and female flies States, decaying animal remains (e.g., rotten beef and chicken were introduced in the cage using a 20 cm long and 1 cm livers) have been reported to be attractive to gravid females of diameter glass tube aspirator through the front opening that SSW, with egg-laying also observed (Byrd 1998). This was secured with a stretched cotton/polyester sleeve. Flies indicates that olfactory cues associated with these livers play were allowed to acclimatize in the cage for 10 min. a role in female SSW oviposition site selection. Meanwhile two clear plastic containers (11 cm top diam× The primary goal of this study was to isolate and identify 9 cm bottom diam×8 cm high) were prepared by lining the volatiles emitted from rotten beef and chicken livers that are bottom of each with a 9 cm filter paper on which 10 g of fresh attractive to gravid SSW females and stimulate oviposition. chicken liver were placed, and this served as the oviposition Results can contribute to the development of synthetic bait substrate. A piece of dental wick (1 cm) was placed on top of systems for monitoring this economically-important pest the liver in each container. The dental wick of one of the species for quarantine purposes and in control strategies to containers was treated with 1 ml of the 7-chemical blend reduce their abundance in urban recreational settings. (test) and that of the other was treated with an equal volume of ethyl alcohol (control). The prepared test and control containers then were introduced into the cage and placed on Methods and Materials the base of the cage diagonally and about 10 cm apart. Fly behavior was observed for 15 min when the flies were Insects Secondary screwworms used in this study originated observed to land in the containers. Flies that landed in each from pupae obtained from a laboratory colony in the USDA- container were then captured by placing the container lid on ARS laboratory in Fargo, North Dakota, USA; progeny were top of the containers. Flies were immobilized by cooling in a maintained in a laboratory colony at the USDA-ARS, Lincoln, freezer, and the number trapped was recorded according to Nebraska,. Larvae were reared under laboratory conditions sexes for each container. (30 °C, 60 % RH and 16 L: 8 D) using an artificial diet The second set of experiments was conducted as in the first consisting of 60 g of spray-dried whole egg (California Spray series except 25 four-day-old females and 8-day-old females Dry Co., Stockton, CA, USA), 160 g of powdered milk were used. For identification purpose, flies were previously (Carnation®, Nestle, USA), 40 g of sodium polyacrylate gel marked with acrylic paint on the thorax with white for 4-day- (Waterlock G-400, Grain Processing Corp, Muscatine, IA, old and red for 8-day-old. Flies also were dissected after USA), and 140 ml of citrated bovine blood (collected from a experiments to determine the ovarian stage. Each experiment local abattoir). These ingredients were mixed with 600 ml of was replicated six times. The positions of the treated and water with 1 ml of 37 % formaldehyde solution (Sigma- control containers were randomized each time. Aldrich, St. Louis, MO, USA) as a fungistat to prepare 1 l of diet. Eggs were collected from 8-day-old gravid females over Oviposition Assays To determine oviposition preference, tests rotten liver, and were placed directly on the diet in a 20×15× were conducted in 30×30×30 cm wire mesh cages as 12 cm rearing tub (~200 mg eggs per liter diet). Pupae were described in the above section. Fifty 8-day-old gravid females collected after 5 days in a 26×20×6 cm tray of vermiculite were transferred from the colony cage to each test cage placed under the rearing tub.
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