Old World screwworm fly, Chrysomya bezziana, occurs as two geographical races

A morphological and molecular analysis was undertaken with the objective of identifying markers for geographical populations of Old World screwworm flies, Chrysomya bezziana Villeneuve (Diptera: Calliphoridae). The morphological analysis involved 192 adult flies from 14 countries, and the molecular analysis involved 45 larvae or adults from 14 populations in 11 countries. Principal components and cluster analysis of 10 morphological characters indicated that flies from Papua New Guinea (PNG) were a distinct group and most similar to flies from nearby Asian islands (Java, Sabah). There was poor resolution of other geographical regions, but some support for clustering of flies from Africa or India. Cladistic analysis of mitochondrial DNA sequences gave strong support for recognizing two races of Old World screwworm, one from sub-Saharan Africa and the other from the Gulf region and Asia. This latter race could be further divided into two lineages, i.e. one from mainland Asia (from Iraq to the Malay Peninsula) and the other from two islands of PNG.     

Phylogeography and recent emergence of the Old World screwworm fly, Chrysomya bezziana, based on mitochondrial and nuclear gene sequences

Abstract.  A previous study had identified an African and an Asian race of the Old World screwworm fly, Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), based on the 3' terminal 279 basepairs (bp) of the mitochondrial cytochrome b gene. The current study improved the phylogeographic resolution of cytochrome b for this species by characterizing more of the gene (the 3' terminal 715 bp) and by sampling more geographical populations, including Oman, Iran, Hong Kong and the Indonesian islands of Sulawesi and East Sumba. Strong support was found for recognizing an African race, but not for a monophyletic Asian race. The cladistic and genealogical relationships among the Asian populations were complex. There was sufficient genetic homogeneity throughout separate regions (mainland Asia and each Indonesian island) to suggest that there are no reproductive barriers within each region that might necessitate the production of more than one strain for control by the sterile insect technique (SIT). Primers were designed for the amplification by polymerase chain reaction of two nuclear loci, the highly conserved elongation factor-1α gene and the less conserved white gene, and the preliminary results indicated that these genes showed the same pattern of small-scale regional variation as cytochrome b . The cytochrome b haplotypes are useful markers for identifying the geographical origins of any emerging infestations of the species: the absence of Indonesian and African haplotypes in the Middle East demonstrates that the large-scale transport of livestock is not spreading Old World screwworm.     

Seasonality of Old World screwworm myiasis in the Mesopotamia valley in Iraq

Following the first recorded introduction of the Old World screwworm fly (OWS), Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), into the Mesopotamia valley in Iraq in September 1996, cases of livestock myiasis caused by OWS developed a distinctly seasonal pattern. The annual cycle of clinical OWS cases is explained here on the basis of environmental variables that affect the different life-cycle stages of C. bezziana. This analysis suggests that low temperatures restricted pupal development during the winter, whereas the dispersal of adult flies was constrained by hot/dry summer conditions. A restricted number of OWS foci persisted throughout the year. In these foci, pupal development was fastest during the autumn months. In autumn, rapid multiplication, lasting several OWS generations, allowed subsequent adult fly dispersal across the valley floor during the winter. Hence, the monthly incidence of clinical OWS cases in livestock peaked during December-January and was lowest during July-August. In addition to temperature and humidity, vegetation cover played a role in OWS distribution. Hence the majority of OWS cases were clustered in the medium density type of vegetation [normalized difference vegetation index (NDVI) values of 0.2-0.4] along the main watercourses in the marshy Mesopotamia valley. Although sheep were the host most commonly infested by C. bezziana, local sheep density was not found to be a major factor in disease spread. Satellite imagery and the application of Geographical Information System (GIS) tools were found to be valuable in understanding the distribution of OWS in relation to vegetation and watercourses. The presence of screwworm in Iraq, at the perimeter of the intercontinental OWS distribution, may give rise to major seasonal flare-ups.     

The potential geographical distribution of the Old World screw-worm fly, Chrysomya bezziana

The potential geographical distribution and relative abundance of the Old World screw-worm fly, Chrysomya bezziana Villeneuve (Diptera: Calliphoridae) as determined by climate, was assessed using CLIMEX, a computer program for matching climates. CLIMEX describes the relative growth and persistence of animal populations in relation to climate. The observed global distribution of C.bezziana was compared with the potential distribution predicted by CLIMEX. The differences in the two distributions indicate the areas at risk of colonization, with particular reference to Australia and the Americas. According to the model, the potential area of permanent colonization in Australia extends south to the mid-coast of New South Wales. Comparison of areas suitable for permanent establishment with the potential summer distribution indicates that large additional areas, carrying most of the continent's livestock, could be colonized in the summer months. Seasonal population growth indices are presented for three ports in Australia at which screw-worm fly specimens have been collected by quarantine authorities. They indicate the relative risk associated with introductions at different places in different seasons and so provide valuable planning information for quarantine authorities. The CLIMEX predictions for C.bezziana in North America are shown to be similar to the recorded distribution limits of the New World screw-worm fly, Cochliomyia hominivorax (Coquerel). The fly could also colonize South America, as far south as southern Brazil and midway through Argentina.     

Mean life expectancy of Old World screwworm fly,Chrysomya bezziana,inferred from the reproductive age-structure of native females caught on swormlure-baited sticky traps

Abstract. The reproductive status of native (wild) screwworm fly, Chrysomya bezziana, caught on swormlure-baited sticky traps in Papua New Guinea is described. A total of 1122 females and 25 males were trapped. Of these females 595 were scored for insemination status and stage of ovarian development (on a scale of 2–10) of which 20% were in suitable condition for assignment to first, second and third ovarian cycles. Of the nulliparous females, only 17% were inseminated at stage 3 of ovarian development, 70% at stage 4, 93-97% at stages 5 and 6, and all of stages 7–10 (gravids). All parous females were inseminated. More than half of the captured females were parous (58%) and only 7% of the total were gravid. Proportions of females in ovarian cycles 1, 2 and 3 were 41%, 50% and 9% respectively. Survival of female Ch. bezziana in the laboratory was adequately described by lognormal and Gompertz survival functions, for both of which the mortality rate is an increasing function of reproductive age. Analysis of the reproductive age distribution of native females estimated their mean life-expectancy at 9 days under the prevailing mean field temperature of 26.5^oC. This equates to completion of 1.7 ovarian cycles and an estimated mean lifetime fecundity of 146 female progeny. The survival models, which also allowed responsiveness of females to swormlure-baited traps (female trappability) to vary according to their stage of ovarian development, indicated significant age-dependent trapping bias. These findings are compared with similar data for the New World screwworm fly, Cochliomyia hominivorax.     

Specific detection of the Old World screwworm fly,Chrysomya bezziana,in bulk fly trap catches using real‐time PCR

The Old World screwworm fly (OWS), Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), is a myiasis‐causing blowfly of major concern for both animals and humans. Surveillance traps are used in several countries for early detection of incursions and to monitor control strategies. Examination of surveillance trap catches is time‐consuming and is complicated by the presence of morphologically similar flies that are difficult to differentiate from Ch. bezziana, especially when the condition of specimens is poor. A molecular‐based method to confirm or refute the presence of Ch. bezziana in trap catches would greatly simplify monitoring programmes. A species‐specific real‐time polymerase chain reaction (PCR) assay was designed to target the ribosomal DNA internal transcribed spacer 1 (rDNA ITS1) of Ch. bezziana. The assay uses both species‐specific primers and an OWS‐specific Taqman^® MGB probe. Specificity was confirmed against morphologically similar and related Chrysomya and Cochliomyia species. An optimal extraction protocol was developed to process trap catches of up to 1000 flies and the assay is sensitive enough to detect one Ch. bezziana in a sample of 1000 non‐target species. Blind testing of 29 trap catches from Australia and Malaysia detected Ch. bezziana with 100% accuracy. The probability of detecting OWS in a trap catch of 50 000 flies when the OWS population prevalence is low (one in 1000 flies) is 63.6% for one extraction. For three extractions (3000 flies), the probability of detection increases to 95.5%. The real‐time PCR assay, used in conjunction with morphology, will greatly increase screening capabilities in surveillance areas where OWS prevalence is low.