Recycled paper products as substitutes for gelling agent in screwworm (Diptera: Calliphoridae) larval diet

Absorbent recycled paper products were tested and compared with a sodium polyacrylate gelling agent, Aquatain, for solidifying larval diets of the screwworm, Cochliomyia hominivorax (Coquerel). A recycled newsprint product, Terra-Mulch, when mixed with water and dietary ingredients produced a diet medium of homogeneous texture that supported larval growth and development comparable to the standard diet prepared with the gelling agent. Biological yields as calculated from number of pupae per tray from both the diets were similar. Three other types of recycled paper products were tested. Although less homogeneous in texture, these diets were as good as the diet with Terra-Mulch. There were no significant differences in the mean larval and pupal weights, no. of pupae/tray, percent emergence, oviposition, percent hatch, and percent longevity. When tests were conducted through eight consecutive generations, no adverse effect was noticed in any of the life history parameters. Terra-Mulch is inexpensive and biodegradable compared with the gelling agent Aquatain that is expensive and not biodegradable. Replacing Aquatain with Terra-Mulch in the screwworm larval diet for mass rearing may result in an annual savings of more than half a million U.S. dollars in costs of material and labor.     

Gelled diet for screwworm (Diptera: Calliphoridae) mass production

A rearing system based on a diet gelled with Water-Lock G-400, a synthetic superabsorbent (poly(2-propenamide-co-2-propenoic acid, sodium salt)) (WL), was compared with the standard rearing system (liquid diet suspended in acetate fibers) for the mass production of screwworms, Cochliomyia hominivorax (Coquerel). The WL rearing system yielded 2% heavier pupae, 32% higher egg to pupa survival, and required 54% less diet and 88% less labor than the standard rearing system. Other advantages of the WL system include reduced susceptibility to suboptimal environmental conditions and labor practices and characteristics conducive to centralization and mechanization of rearing procedures.     

Gel extenders in larval diet of Cochliomyia hominivorax (Diptera: Calliphoridae).

Larvae of the screwworm, Cochliomyia hominivorax (Coquerel), were reared on meridic diets containing ground corncob material (grit) in test 1 and the cob grit or sawdust in test 2 to reduce or replace the amount of an expensive synthetic copolymer gelling agent. Biological parameters for the tests included pupal yield (total number), pupal weight (size), pupal malformation, adult emergence, adult malformation, egg weight, and egg hatch to assess for any significant differences. In the first test, larval propagation was successful for five consecutive generations on 24 of an original 37 formulations. Results indicate that a minimum concentration of 4% of the cob grit is needed to reduce the gel by 50%; at least 8% concentration is needed to reduce the gel by 80%; and at least 12% concentration is needed to replace the gel completely. In the second test, the larvae were successfully propagated for five consecutive generations when the copolymer gel was reduced by 50 or 75% in the diet by using either sawdust or the no. 60 cob grit size. The cost savings and applications of use of sawdust or the cob grit incorporated into the larval diet can be beneficial to a mass-production screwworm program.     

A new meatless diet for adult screwworm (Diptera: Calliphoridae)

Screwworm flies, Cochliomyia hominivorax (Coquerel), were fed on honey and spray dried egg product; honey, molasses, and spray dried egg product; honey and spray dried meat protein; as well as on a control diet of honey and horsemeat, which is the standard diet used for screwworm adult colony in the mass-rearing facility. In general, the weight of eggs laid by females fed on the diet of spray dried egg product was significantly higher than that laid by females fed on the standard horsemeat diet. Egg production declined when spray dried meat protein replaced the egg product. Partial replacement of honey with molasses in the egg diet did not decrease egg production, compared with the control diet. The use of spray dried egg diet has advantages over the horsemeat diet, such as storage, handling, preparation, feeding, and expense. A cost analysis suggests that replacing the horsemeat with spray dried egg product, and half of the honey with molasses, would reduce the cost of the diet by more than US $100,000 annually.     

A cellulose fiber-based diet for screwworm (Diptera: Calliphoridae) larvae

A highly absorbent cellulose fiber from recycled paper was tested and compared with a polyacrylate gelling agent, Aquatain, normally used for bulking and solidifying larval rearing medium of screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae). The absorbent fiber, when mixed with water and dietary ingredients, produced a diet medium of homogeneous texture that supported larval growth and development comparable with the standard gelled diet. Larval and pupal weights from two concentrations of cellulose fiber-based diet were significantly higher than those obtained using gelled diet. The number of pupae per tray, percentage of adult emergence, oviposition, percentage of egg hatch, and adult longevity obtained from the insects reared in the cellulose fiber-based diet were comparable or slightly better than the biological parameters recorded from flies reared in the gelled diet. Moreover, results indicate that a lesser amount of the cellulose fiber-based diet than the normal amount of gelled diet per tray would support normal larval growth. Physical properties and texture of the new diet seem to allow the larvae to move and feed more freely than they do on the semisolid gelled diet, resulting in less wasted diet. The cellulose fiber is biodegradable and inexpensive, whereas the polyacrylate gel polymer is not biodegradable and is relatively expensive. Replacing gel with cellulose fiber in the screwworm larval diet for mass rearing should result in substantial cost savings in material and labor as well as eliminating concern of environmental pollution due to diet waste disposal.     

Solidifying agent and processing of blood used for the larval diet affect screwworm (Diptera: Calliphoridae) life-history parameters

Spray-dried whole bovine blood and a sodium polyacrylate polymer gel as a bulking and solidifying agent are among the constituents of the current larval diet for mass rearing screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae). Locally available, inexpensive dietary materials could reduce rearing cost and address an uncertain commercial supply of spray-dried blood. We compared efficacy of diet prepared from fresh bovine blood after decoagulation with sodium citrate or ethylenediaminetetraacetic acid (EDTA) or after mechanical defibrination, with the diet containing spray-dried blood using either gel or cellulose fiber as the bulking and solidifying agent. Several life-history parameters were compared among insects reared on each of the blood and bulking agent diets combination. Diets containing citrated blood yielded the lightest larval and pupal weights and fewest pupae. EDTA-treated blood with the gel also caused reductions. EDTA-treated blood with fiber yielded screwworms that were heavier and more numerous than those from the diet with citrated blood but lighter than those from the control diet using spray-dried blood. A reduction in percentage of adults emerging from pupae occurred from diets with both bulking agents using citrated blood and the diet using EDTA mixed with the gel bulking agent. As a group, the cellulose-fiber diets performed better than the gel diets. Larval diet did not affect adult longevity, weight of the eggs deposited by the females that emerged or subsequent egg hatch. Parameter measurements of insects from both defibrinated blood diets were similar to those from the spray-dried blood diets, indicating that fresh, defibrinated bovine blood can successfully replace the dry blood in the screwworm rearing medium.     

Cytogenetic variation in natural populations of the Old World screwworm fly Chrysomya bezziana (Diptera: Calliphoridae).

The existence of sibling species in the Old World screwworm fly Chrysomya bezziana would raise serious problems in eradicating this pest if it entered Australia. Cytogenetic variation in C. bezziana was investigated by analyzing pupal trichogen polytene chromosomes. Natural populations of C. bezziana spanning its range from southern Africa to Papua New Guinea were examined as well as hybrids between a New Guinea laboratory strain and natural populations. No evidence of sibling species was found. All populations exhibited the same basic banding pattern as the standard sequence established from a Papua New Guinea strain. Extensive asynapsis of chromosome homologues was found in some hybrid crosses and was therefore measured in all populations and hybrids to detect systematic variation. Asynapsis levels in most hybrids could not be statistically distinguished from those present in the parent populations except for crosses between populations at the ends of the range. This result does not permit asynapsis levels to be used in establishing the origin of introduced flies by estimating their distance from known populations. One inversion polymorphism and six band polymorphisms spread over three chromosomes were analyzed. Populations in each sampled region had characteristic combinations of band polymorphisms. This may offer a diagnostic method for determining the origin of flies accidentally introduced to Australia.     

Alternative to sentinel animals for collecting egg masses from wild females of the screwworm (Diptera: Calliphoridae).

Egg masses from wild populations of the screwworm, Cochliomyia hominivorax (Coquerel), were obtained by collecting females from rotted liver, holding them for 3 d until egg maturation and then placing them on heated ground beef for oviposition. Nearly 50% of the females oviposited. Fertility of egg masses was 66 and 95% at the two collection sites. Decreased fertility was associated with collections of virgin females. Average weight +/- SD of the egg masses was 13.7 +/- 1.40 mg; the number of eggs per mass averaged 343. This method of obtaining egg masses from wild screwworm females is a cost-effective, sensitive alternative to the use of sentinel animals in the field. The use of rotted liver costs less, takes less time, and takes less equipment than the traditional sentinel animal technique.     

A simple larval diet for population studies on the blowfly Lucilia sericata (Diptera: Calliphoridae)

1. A simple artificial diet was devised for larvae of Lucilia sericata Meigen. 2. A basic diet of 20 g/l agar with 20% horse blood and added yeast sustained normal growth and development, as compared with a lamb meat control. 3. When yeast was added to the blood agar diet at 50 g/l, both peak and final larval weights were increased by 25-50% at higher larval densities (2-8 larvae/g diet) and the hatchling-adult development period was reduced by about 3 days. 4. No adult insects emerged on an agar-yeast diet without blood. Increasing the concentration of blood from 10% to 20% increased adult weight at emergence by 50%, though developmental period was not significantly affected. 5. Increasing larval density significantly reduced the weights of pupae and emergent adults (by up to 50%), both on the blood-agar-yeast diet and the lamb meat control. Percentage survival fell from 80-90% to below 10% on the blood-agar-yeast diet at the highest densities of 4 or 8 larvae/g diet, though survival on lamb was not significantly affected by density.     

Age determination in the adult screwworm (Diptera: Calliphoridae) by pteridine levels

Fluorescence spectrophotometry was used to assess the possible use of pteridines in the compound eyes to estimate the age of adult screwworms, Cochliomyia hominivorax (Coquerel). Factors affecting the quantities of pteridines include temperature and head size. No difference in pteridine levels was found among flies fed protein or carbohydrate. A regression model for estimating the age of female screwworms was constructed. The model uses head capsule size and relative pteridine quantities and assumes a constant body temperature of 30 degrees C. This regression formula has an r2 of 0.74. Our study extends the use of pteridine accumulation for age determination from obligate sanguinivorous Diptera to an autogenous species that feeds facultatively on nectar and wound exudates. The technique appears to provide a valid means to determine age of these flies.     

Oviposition by screwworm flies (Diptera: Calliphoridae) on contact with host fluids

Factor affecting oviposition by screwworm flies, Cochliomyia hominivorax (Coquerel), contacting different host fluids were examined in a laboratory bioassay. Fresh bovine blood, which does not release the attractive odors involved in host finding, nevertheless stimulated as many or more females to oviposit than did the other fluids tested. These other fluids included attractive fluid from screwworm-infected wounds (a favored oviposition site in nature) and cultures of Providencia rettgeri (a bacterium implicated in attractant production). Oviposition did not vary with batch of fresh blood or frozen storage, making blood a useful standard for comparing oviposition rates among studies. Oviposition did vary with the substrate to which the blood was applied, suggesting that an interaction between tactile and chemical stimuli is important for host recognition. Both insemination and darkness during bioassay increased oviposition rates, but the magnitude of these effects was small compared with that due to substrate. Age had no effect for at least 1 wk after females became gravid.     

Laboratory environment effects on the reproduction and mortality of adult screwworm (Diptera: Calliphoridae)

The New World screwworm, Cochliomyia hominivorax Coquerel, is mass reared for screwworm eradication initiatives that use the sterile insect technique. New methods for rearing have helped to reduce the cost of the eradication program. We examined the effect and interaction of three temperatures (24.5, 29.5 and 34.5 degrees C), two diets (2% spray-dried blood plus 0.05% vitamins and corn syrup carrageenan) and three population densities (300, 400, and 500 flies/cage) on egg production, egg hatch, number of observable fertilized eggs, mortality (male and female) and ovarian development. The three population densities did not affect any of the parameters monitored. Using the protein diet increased egg production at all temperatures. Diet did not affect egg hatch or female mortality. Male mortality was significantly greater when fed the protein diet and reared at 24.5 degrees C and 34.5 degrees C. Egg hatch was significantly less when the flies were reared at 34.5 degrees C. When exposed to high temperatures (37 degrees C and 40 degrees C) egg production, egg hatch, fertility and mortality were adversely affected. At the higher temperatures, yolk did not adequately form during oogenesis. When compared to the normal rearing photoperiod (12 L:12 D), short photoperiod (1 L:23 D) increased egg production, egg hatch and fertility but lowered mortality.     

Persistence of sterile screwworm (Diptera: Calliphoridae) flies at a release site.

Studies to determine the persistence of released, sterile screwworms, Cochliomyia hominivorax (Coquerel), were conducted in Belize, Central America, in 1987-1988. A total of nine releases were made, each consisting of 4,000 females marked with a fluorescent dust. Previous similar studies with baited fly traps indicated that flies died or dispersed from the release site within 3-4 d. For this study, flies were recaptured at sentinel animals. Recapture rates varied greatly between releases, a result that was not correlated with any weather parameter. The pattern of recapture indicated a sharp drop in the released population after 9-10 d, with a few flies persisting in the release area for up to 3 wk. The results suggest that survival of released sterile flies is comparable with estimated survival rate and life span of wild, native flies.     

Genetic variability in mitochondrial DNA of the screwworm,Cochliomyia hominivorax (Diptera: Calliphoridae), from Brazil

Restriction fragment length polymorphism (RFLP) analysis of mitochondrial DNA (mtDNA) was used to examine genetic variation and population structure of screwworm flies in four populations from São Paulo State, Brazil. The total DNA of 405 individuals was digested with 15 restriction endonucleases and probed with five clonedHindIII fragments representing the entire mitochondrial genome ofCochliomyia hominivorax. The survey revealed that four enzymes (HaeIII,HindIII,MspI, andPvuII) were suitable to detect mtDNA variation among all populations. Based on the fragment patterns obtained for these four enzymes, a total of 15 haplotypes in combination was detected. Heteroplasmic individuals for thePvuII pattern were obtained in one of the populations. The estimated average for nucleotide sequence divergence (δ) was 0.92%. The cladogram of the geographical distribution among the observed haplotypes suggests that the sampled screwworms probably belong to a single evolutionary lineage with populations interconnected by reduced gene flow.     

Monitoring adult populations of the screwworm (Diptera: Calliphoridae) with feeding stations baited with liver.

Populations of the screwworm, Cochliomyia hominivorax (Coquerel), were monitored by capturing adults with hand nets and rooted liver set on the ground. Adults were marked and released. During the 61-d study conducted in a tropical dry forest in the dry season (January to March 1989), 2,640 individual females and 460 individual males were recorded at the four liver-baited stations. The total number of visits by females was 5,769 and by males 510. The mean number of unmarked adults per day was 44.1 females and 7.6 males in an area of approximately 2.59 km2. The mean percentage of marked flies that were recovered was 64.3%. The mean number of feeding stations visited, times recaptured, and days in the study area by individual females was 2.1, 2.4, and 4.5, respectively. Daily visitational patterns by both sexes at the feeding stations were bimodal with peaks occurring between the hours of 0730 to 0859 and 1500 to 1629. Flies were most active when the mean temperature was 29.9 degrees C; the majority of the visits occurred when air temperatures were between 26 and 33 degrees C. Most females attracted to feeding stations were nulliparous (70.7%) and mated (69.7%). Our results suggest that observing adults at feeding stations is a reliable method of obtaining data on the behaviour and population dynamics of indigenous populations of screwworm adults in tropical habitats. Although this method was labor intensive, the amount of data gleaned from the manipulation of wild populations more than compensated for such costs.     

Field comparisons of attractants for the screwworm fly (Diptera: Calliphoridae) in a tropical dry forest in Costa Rica.

Field trials were conducted to determine the best attractant for adults of the screwworm, Cochliomyia hominivorax (Coquerel), in Costa Rica. The five attractants were wounded sentinel sheep, decayed fruit, rotted beef liver, rotted fish, and medium used to rear larvae of this blow fly. Significantly more adults came to rotted liver than any other attractant. No adults were captured in traps baited with rotted fruit. Significantly more males came to beef liver than were captured at other baits; males were rarely encountered at wounds. Ovarian development of females obtained with different attractants indicated that proportionally more gravid females visited wounded animals than were caught in traps with proteinaceous baits. Most females had mated by egg stage five and mating and and parity in the populations that visited different attractants were similar. Significantly more females (81%) marked initially at wounds returned to wounds than visited traps baited with liver or fish.