Using COI barcodes to identify forensically and medically important blowflies.

The utility of cytochrome oxidase I (COI) DNA barcodes for the identification of nine species of forensically important blowflies of the genus Chrysomya (Diptera: Calliphoridae), from Australia, was tested. A 658-bp fragment of the COI gene was sequenced from 56 specimens, representing all nine Chrysomya species and three calliphorid outgroups. Nucleotide sequence divergences were calculated using the Kimura-two-parameter distance model and a neighbour-joining (NJ) analysis was performed to provide a graphic display of the patterns of divergence among the species. All species were resolved as reciprocally monophyletic on the NJ tree. Mean intraspecific and interspecific sequence divergences were 0.097% (range 0-0.612%, standard error [SE] = 0.119%) and 6.499% (range 0.458-9.254%, SE = 1.864%), respectively. In one case, a specimen that was identified morphologically was recovered with its sister species on the NJ tree. The hybrid status of this specimen was established by sequence analysis of the second ribosomal internal transcribed spacer (ITS2). In another instance, this nuclear region was used to verify four cases of specimen misidentification that had been highlighted by the COI analysis. The COI barcode sequence was found to be suitable for the identification of Chrysomya species from the east coast of Australia.

Food consumption and individual lifespan of adults of the blowfly, Calliphora stygia: a test of the 'rate of living' theory of aging.

Following eclosion, adult Calliphora stygia were individually housed (at 22 degrees C and 73% RH) in 125 ml plastic vials and provided with ad libitum access to either 0.125 or 0.2M sucrose as a food source and daily food consumption measured throughout their adult life. All blowflies were weighed daily and food consumed is determined by weighing individual food dishes. Blowflies provided with 0.125 M sucrose (N=59) consumed daily a significantly greater amount of the sucrose solution than those provided with 0.2M sucrose (n=55) such that the average rates of sucrose consumption were, respectively, 1.72 and 1.96 mg sucrose day(-1). There was no significant difference in the survival curves of the two populations with respective average (+/-SEM) lifespans being 25.4 (+/-1.2) and 26.5 (+/-1.2) days. The respective ranges of individual lifespans were 4-53 and 5-50 days. There was no statistically significant relationship between mass-specific rate of sucrose consumption and lifespan in either population but there were highly significant (P<0.0001) correlations between lifetime sucrose consumption and lifespan in both groups of blowflies. These findings contradict the predictions of the 'rate of living' theory of aging. In both populations of blowflies, body mass and the rate of food consumption were relatively constant through the adult life of blowflies, except that a few days before death both sucrose consumption and body mass showed a dramatic decline.

The utility of mitochondrial DNA sequences for the identification of forensically important blowflies (Diptera: Calliphoridae) in southeastern Australia.

The applicability of mitochondrial DNA (mtDNA) sequencing was investigated for the identification of the following forensically important species of blowflies from southeastern Australia: Calliphora albifrontalis, C. augur, C. dubia, C. hilli hilli, C. maritima, C. stygia, C. vicina, Chrysomya rufifacies, Ch. varipes and Onesia tibialis. All breed in carrion except O. tibialis, which is an earthworm parasitoid. Emphasis was placed on Calliphora species because they predominate among the carrion-breeding blowfly fauna of southern Australia and their immatures are difficult to identify morphologically. A partial sequence of the mitochondrial COII gene was determined for all species and for COI for C. albifrontalis, C. augur, C. dubia and C. stygia only. Five other species of blowflies, Chrysomya albiceps, Ch. rufifacies, Protophormia terraenovae, Lucilia illustris and L. sericata, for which sequence data were already available, were also included. Analysis of the COI and COII sequences revealed abundant phylogenetically informative nucleotide substitutions that could identify blowfly species to species group. In contrast, because of the low level of sequence divergence of sister species, the data could not distinguish among taxa from the same species group, i.e. the species within the C. augur and C. stygia groups. The molecular data support the existing species group separation of the taxa within Calliphora. Because of the speed and accuracy of current nucleotide sequencing technology and the abundant apomorphic substitutions available from mtDNA sequences, this approach, with the analysis of additional taxa and genes, is likely to enable the reliable identification of carrion-breeding blowflies in Australia.

The forensic application of allozyme electrophoresis to the identification of blowfly larvae (Diptera: Calliphoridae) in southern Australia.

Most known carrion-breeding species of blowflies in southern Australia are of the genus Calliphora. The morphological similarity of the immatures of these species means that correctly identifying them poses a challenge for forensic entomologists. This study investigates the potential of allozyme analysis to assist with this task. Molecular profiles of third-instar larvae and adults representing four of these carrion-breeding species, Calliphora stygia, C. dubia, C. hilli hilli, and C. vicina, were compared at 42 allozyme loci. The two life stages were found to display almost identical allozyme profiles in each species (93% of loci were expressed in both life history stages), enabling the reliable identification of larvae in these four species. Integration of these results with data from a previous study indicates that allozyme analysis would also be suitable for rapid, species-level identification of the larvae of six other carrion-breeding Calliphora species occurring in southern Australia. This is the first report of the application of allozyme data to the identification of forensically important blowflies.

A key to the adults of species of blowflies in southern Australia known or suspected to breed in carrion.

The reliable morphological identification of carrion-breeding blowflies is important ecologically, as well as for medical, veterinary and forensic reasons. To date, no comprehensive key has been available to make this possible for workers in southern Australia. An illustrated key is presented to the adults of all species of blowflies (Diptera: Calliphoridae) south of 30 degrees S known or suspected to breed in carrion (species exclusive to Queensland and/or the Northern Territory are excluded).