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The seasonal activity of adults of eight forensically important blow fly species [Calliphora croceipalpis (Diptera: Calliphoridae), Jaennicke, Lucilia sericata (Meigen) (Diptera: Calliphoridae), L. cuprina (Wiedemann) (Diptera: Calliphoridae), Chrysomya chloropyga (Wiedemann) (Diptera: Calliphoridae), Ch. albiceps (Wiedemann) (Diptera: Calliphoridae), Ch. marginalis (Wiedemann) (Diptera: Calliphoridae), Ch. putoria (Wiedemann) (Diptera: Calliphoridae), Ch. megacephala (Fabricius) (Diptera: Calliphoridae)] was quantified from collections in Makhanda, South Africa. Flies were collected in traps baited with chicken liver and placed in the field at eight locations for four consecutive days each fortnight for 14 mo. The seasonal distribution of each species compared well to published seasonal distributions of these blow flies elsewhere in South Africa, with evidence of year-to-year variation within seasons that might be explained by weather. This information is important for determining when and where certain species are likely to occur and will be of use in forensic investigations and myiasis management plans., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The superfamily Oestroidea, comprising ∼15,000 species, is a large and ecologically diverse clade within the order Diptera. Among its six commonly recognized families, Calliphoridae seems to be crucial for understanding evolutionary relationships in the group, as it is recognized as a controversial paraphyletic grouping. To further investigate this matter, the ITS2, 28S, COI and 16S regions were used to infer phylogenetic relationships in Oestroidea with maximum-parsimony (MP), maximum-likelihood (ML) and Bayesian inference (BI) methods. For the BI analyses, a deep evaluation of different data partitioning strategies was conducted, including consideration of structural conformation (ITS2 and 16S) and codon position (COI) information. Results suggest the existence of two main clades in Oestroidea: (Tachinidae+Mesembrinellinae) and (Rhiniinae, (Sarcophagidae+Calliphoridae sensu stricto)). Oestridae was recovered as sister group of the remaining Oestroidea in the MP trees while it was placed closer to the (Rhiniinae+Sarcophagidae+Calliphoridae sensu stricto) group in the ML and BI trees. A paraphyletic Calliphoridae was recovered, confirming the exclusion of Rhiniinae, a clade recently promoted to family status and therefore already excluded. Mesembrinellinae could also be considered a distinct group apart from Calliphoridae, although further studies are required. Consideration of structural and codon position information led to a significant increase in the log-likelihoods of the analyses, which were accompanied by small changes in the inferred topologies, branch lengths and posterior probability support values. However, as model complexity increases, so does uncertainty across the estimated parameters, including tree topologies, and phylogenies inferred under very parameter-rich models may be less reliable even when possessing higher log-likelihoods., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Reinvestigation of mitochondrial haplotypes previously reported to be shared between the Afrotropical blowflies Chrysomya putoria Weidemann and Chrysomya chloropyga Weidemann (Diptera: Calliphoridae) revealed an error resulting from the misidentification of specimens. Preliminary amplified fragment length polymorphism (AFLP) analysis of the original and additional individuals again failed to find reciprocal monophyly, leading to a re-examination of the specimens for diagnostic male genitalic characters that were first described following the earlier study. Four of the original study specimens were found to have been misidentified, and definitive analysis of both mtDNA and AFLP genotypes using phylogenetic analysis and genetic assignment showed that each species was indeed reciprocally monophyletic. In addition to correcting the earlier error, this study illustrates how AFLP analysis can be used for efficient and effective specimen identification through both phylogenetic analysis and genetic assignment, and suggests that the latter method has special advantages for identification when no conspecific specimens are represented in the reference database., (© 2011 The Authors. Medical and Veterinary Entomology © 2011 The Royal Entomological Society.)

To highlight some issues regarding data quality that are significant in estimating post-mortem intervals (PMI) from maggots, the developmental constants of thermal summation models for development of Chrysomya megacephala Fabricius (Diptera: Calliphoridae) were calculated from incidental data gathered from 12 published studies, and from data generated specifically for the purpose in a single experiment. The focused experiment involved measuring the timing of five developmental landmarks at nine constant temperatures with a sampling resolution of 6-12 h, which is characteristic of other published studies. Combining data from different studies produced inconsistent results because of statistical noise introduced by (at least) disparities in temporal precision, descriptive statistics, geographical location and rearing diets. A robust experimental design to estimate a developmental model should involve at least six constant temperatures, starting at about 7 degrees C above the relevant developmental zero (D0) and going almost to the upper critical temperature, and a temporal sampling interval with a relative precision of about 10%, which requires sampling about every 2 h until hatching, about every 3 h until first ecdysis and about every 6 h until second ecdysis.

Developmental curves for the sister species Chrysomya chloropyga (Wiedemann, 1818) and Chrysomya putoria (Wiedemann, 1830) (Diptera: Calliphoridae) were established at eight and 10 different constant temperatures, respectively, using developmental landmarks and body length as measures of age. The thermal summation constants (K) and developmental threshold (D(0)) were calculated for five developmental landmarks using a previously described method. Isomorphen and isomegalen diagrams were also constructed for the purpose of estimating postmortem intervals (PMIs). Chrysomya chloropyga had an average developmental threshold value (D(0)) of 10.91 degrees C (standard error [SE] = 0.94 degrees C, n = 5), significantly lower than that of C. putoria (13.42 degrees C, SE = 0.45 degrees C, n = 5) (paired t-test: t = - 4.63, d.f. = 8, P < 0.00). Similarly, K values for C. chloropyga were larger than those for C. putoria for all developmental events except onset of the wandering phase. These are the first data that can be used to calculate minimum PMIs and predict population growth of C. chloropyga and C. putoria in Africa; the data indicate that developmental data for one of these species cannot be used as surrogate data for the sister species.

Complementary nuclear (28S rRNA) and mitochondrial (COI) genes were sequenced from blowflies that phenotypically resembled Lucilia cuprina (W.), Lucilia sericata (Meigen) or exhibited characters of both species. The aim was to test a long-held hypothesis that these species hybridize under natural conditions in South Africa (Ullyett, 1945). Blowflies were obtained predominantly from the Cape Town metropolitan area, but reference samples were acquired for L. sericata from Pretoria. Several L. cuprina-like flies were shown to possess a conflicting combination of nuclear and mitochondrial genes that has also been seen in Hawaiian specimens. Homoplasy, sampling of pseudogenes, hybridization and incomplete lineage sorting are discussed as possible hypotheses for the pattern and the latter is concluded to represent the most likely explanation.

A proliferation of molecular studies of the forensically significant Calliphoridae in the last decade has seen molecule-based identification of immature and damaged specimens become a routine complement to traditional morphological identification as a preliminary to the accurate estimation of post-mortem intervals (PMI), which depends on the use of species-specific developmental data. Published molecular studies have tended to focus on generating data for geographically localised communities of species of importance, which has limited the consideration of intraspecific variation in species of global distribution. This study used phylogenetic analysis to assess the species status of 27 forensically important calliphorid species based on 1167 base pairs of the COI gene of 119 specimens from 22 countries, and confirmed the utility of the COI gene in identifying most species. The species Lucilia cuprina, Chrysomya megacephala, Ch. saffranea, Ch. albifrontalis and Calliphora stygia were unable to be monophyletically resolved based on these data. Identification of phylogenetically young species will require a faster-evolving molecular marker, but most species could be unambiguously characterised by sampling relatively few conspecific individuals if they were from distant localities. Intraspecific geographical variation was observed within Ch. rufifacies and L. cuprina, and is discussed with reference to unrecognised species.

Chrysomya chloropyga (Wiedemann) and C. putoria (Wiedemann) (Diptera: Calliphoridae) are closely related Afrotropical blowflies that breed in carrion and latrines, reaching high density in association with humans and spreading to other continents. In some cases of human death, Chyrsomya specimens provide forensic clues. Because the immature stages of such flies are often difficult to identify taxonomically, it is useful to develop DNA-based tests for specimen identification. Therefore we attempted to distinguish between C. chloropyga and C. putoria using mitochondrial DNA (mtDNA) sequence data from a 593-bp region of the gene for cytochrome oxidase subunit one (COI). Twelve specimens from each species yielded a total of five haplotypes, none being unique to C. putoria. Therefore it was not possible to distinguish between the two species using this locus. Maximum parsimony analysis indicated paraphyletic C. chloropyga mtDNA with C. putoria nested therein. Based on these and previously published data, we infer that C. putoria diverged very recently from C. chloropyga.

One major aspect of research in forensic entomology is the investigation of molecular techniques for the accurate identification of insects. Studies to date have addressed the corpse fauna of many geographical regions, but generally neglected the southern African calliphorid species. In this study, forensically significant calliphorids from South Africa, Swaziland, Botswana and Zimbabwe and Australia were sequenced over an 1167 base pair region of the COI gene. Phylogenetic analysis was performed to examine the ability of the region to resolve species identities and taxonomic relationships between species. Analyses by neighbour-joining, maximum parsimony and maximum likelihood methods all showed the potential of this region to provide the necessary species-level identifications for application to post-mortem interval (PMI) estimation; however, higher level taxonomic relationships did vary according to method of analysis. Intraspecific variation was also considered in relation to determining suitable maximum levels of variation to be expected during analysis. Individuals of some species in the study represented populations from both South Africa and the east coast of Australia, yet maximum intraspecific variation over this gene region was calculated at 0.8%, with minimum interspecific variation at 3%, indicating distinct ranges of variation to be expected at intra- and interspecific levels. This region therefore appears to provide southern African forensic entomologists with a new technique for providing accurate identification for application to estimation of PMI.

Larvae of Sarcophaga (Curranea) tibialis (S. tibialis) were reared at constant temperature on chicken liver treated with a steroid or a barbiturate at concentrations that would be lethal, half-lethal and twice-lethal doses for humans. Trends to greater mortality at higher drug concentrations were not statistically significant. Larvae exposed to either drug took significantly longer to reach pupation compared to those in the control, while larvae exposed to sodium methohexital passed through pupation significantly faster than those in the control. No systematic relationship was found between drug concentration and development time of larvae or pupae. The total developmental period from hatching to eclosion did not differ between treatments, implying that estimates of post mortem intervals- (PMI) based on the emergence of adult flies will not be affected by the involvement of these drugs in a case. On the other hand, anomalous pupation spans may indicate the presence of barbiturates. These findings are compared with patterns found in another fly fed other contaminants.

An upsurge in African horse sickness (AHS) in the Eastern Cape, South Africa, from 2006 led to an epidemiological reassessment of the disease there. Light trapping surveys carried out near horses, donkeys and zebras in 2014–2016 collected 39 species of Culicoides midge (Diptera: Ceratopogonidae) that are potential vectors of AHS. To establish if these midges fed on equids, DNA sequences were obtained from the gut contents of 52 female midges (35 freshly blood‐fed, 13 gravid and four parous), representing 11 species collected across 11 sites. Culicoides leucostictus fed on all three equids. Culicoides bolitinos, Culicoides imicola and Culicoides magnus fed on both horses and donkeys. Culicoides onderstepoortensis fed on donkeys, and Culicoides similis and Culicoides pycnostictus fed on zebras. Bloodmeals from cows, pigs, warthogs, impalas and a domestic dog were also identified in various species, but none of the midges tested had fed on birds. These results contribute to knowledge of the vectorial capacity of several species of Culicoides with regard to AHS in the Eastern Cape and point to potential reservoir hosts, of which donkeys, zebras and domestic dogs have previously been found to harbour AHS. Blood‐fed midges were also obtained throughout winter, indicating the potential for endemic AHS in the province. [ABSTRACT FROM AUTHOR]

We evaluated the bacteria occurring externally on Musca domestica and Chrysomya megacephala, the two most common synanthropic flies which may be found at many refuse sites throughout the world. Bacteria cultured from 10 specimens of each species were isolated, Gram-stained and examined microscopically, and divided into morphologically distinct 'pseudospecies', to avoid excessive duplication of genetic identification. About 350 bp of the 16S ribosomalRNAgene was amplified from genomicDNAextracted from each 'pseudospecies', sequenced, and bacteria identified using BLASTn. Nineteen different types of colony were identified from M. domestica, with Pseudomonas sp. and Swine Manure Bacterium SP14 being most abundant. Chrysomya megacephala yielded 15 distinct pseudospecies with total colony counts approximating to 10 000 from 10 plates, where 80% of colonies were non-pathogenic Bacillus pumilus. A total of 18 species were identified genetically: three shared by the fly species; four unique to C. megacephala, and 13 unique to M. domestica. Half of these 18 species were pathogenic, two or three others were food spoilers and the rest were environmental or commensal bacteria from soil or plant matter. This study added three new pathogenic strains of bacteria and one new environmental strain to the list of bacteria reported to be vectored by these flies. [ABSTRACT FROM AUTHOR]

A variety of temperature thresholds for larvae, pupae, and adults of seven African species of carrion-feeding blowflies (Diptera: Calliphoridae) was measured and compared to understand their basic thermal biology and the influence of temperature on their behaviour. Calliphora croceipalpis (Jaennicke) had consistently lower temperature thresholds than all other species tested for all larval (42.9 °C), pupal (16.6 °C), and adult (45.6 °C) stages. Larvae (50.1 °C) and adults (53.4 °C) of Chrysomya marginalis (Robineau-Desvoidy) had higher upper lethal temperature thresholds than all other species and weighed more than all other species. Pupae and adults of both Chrysomya albiceps (Wiedemann) and Lucilia sericata (Meigen) had similar temperature thresholds, whereas Chrysomya putoria (Wiedemann), Chrysomya chloropyga (Wiedemann), and Chrysomya megacephala (Fabricius) had inconsistent rank temperature thresholds between the larval, pupal, and adult stages. With a few minor exceptions, the nervous activity, muscle activity, and death thresholds in female adult flies responded at higher temperatures than conspecific male flies for all species tested. Similarly, female adult flies weighed consistently more than conspecific male flies for all species tested, except Ca. croceipalpis. These data suggest that there is a phylogenetic component to the thermal biology of blowflies, because Ca. croceipalpis belongs to a primarily Holarctic genus and shows adaptation to that climate even though it inhabits Africa. Comparisons between these temperature thresholds and the distributions of blowfly species present on three rhinoceros carcasses suggest that blowfly larvae with high upper lethal temperature thresholds (particularly C. marginalis) dominate in interspecific competition on the carcass by raising the temperature of the amassed maggots above the thresholds of other carrion-feeding blowflies, through metabolically generated heat. [ABSTRACT FROM AUTHOR]

Chrysomya chloropyga(Wiedemann) andC. putoria(Wiedemann) (Diptera: Calliphoridae) are closely related Afrotropical blowflies that breed in carrion and latrines, reaching high density in association with humans and spreading to other continents. In some cases of human death,Chyrsomyaspecimens provide forensic clues. Because the immature stages of such flies are often difficult to identify taxonomically, it is useful to develop DNA-based tests for specimen identification. Therefore we attempted to distinguish betweenC. chloropygaandC. putoriausing mitochondrial DNA (mtDNA) sequence data from a 593-bp region of the gene for cytochrome oxidase subunit one (COI). Twelve specimens from each species yielded a total of five haplotypes, none being unique toC. putoria. Therefore it was not possible to distinguish between the two species using this locus. Maximum parsimony analysis indicated paraphyleticC. chloropygamtDNA withC. putorianested therein. Based on these and previously published data, we infer thatC. putoriadiverged very recently fromC. chloropyga. [ABSTRACT FROM AUTHOR]