A geometric morphometric analysis was performed on the right wing of adult Calliphora vicina (Robineau-Desvoidy) collected across 4 altitudinal levels in Sicily. The objective of this study was to assess differences in shape and centroid size (CS) between females and males and across elevations. The wings analyzed in this study were removed from adults of C. vicina collected with baited traps at 20, 700, 1,153, and 1,552; for this study, 19 landmarks were identified in each wing. The coordinates of the landmarks were aligned and superimposed to prevent variations due to position, orientation, and scale; they were then scaled to the same CS and recentered. CS and Procrustes differences were, respectively, used to assess variations in size and shape. Significant differences were observed in wing shape between males and females but not between all altitudinal levels. Female wings were found to be significantly larger than males (P < 0.01). Wings of flies collected at the highest altitudinal level resulted in significantly larger wings than those collected at lower altitudes (P < 0.001), with CS values ranging from 12.1 to 14.1. Variation in wing shape can impact thermal regulation, and therefore, oxygen content, temperature, atmospheric pressure, and solar radiation can have an effect on an insect\'s body and activity levels. At high elevations and lower temperatures, larger wings could mean less energy expenditure when flying to increase body temperature.

Tail docking is a husbandry practice widely incorporated in sheep farms around the world. It is an irreversible mutilation that impairs animal welfare, both immediately and in the longer term. The defence of tail docking as a practice is centred around the perception that doing so contributes to the promotion of local hygiene, allowing the use of the wool, facilitating reproductive management and reducing the chances of myiasis, a disease caused by the invasion of blowfly larvae in the tissues of warm-blooded animals. However, current understanding of farm animal welfare questions the need to maintain practices such as tail docking. Thus, the aim of this study was to evaluate the effect of tail docking on the incidence of Cochliomyia hominivorax myiasis in sheep in an experimental flock in Brazil during a six-year retrospective cohort study. Relative risk, odds ratio and incidence rate ratio were the association measures adopted. A total of 4,318 data-points were collected and supplied the analytical model. Tail docking did not decrease the risk and, on the contrary, was found to increase the chances of sheep being affected by myiasis. The results support the hypothesis that tail docking is not a protective factor against the occurrence of myiasis and further fuel calls for a rethink of tail docking being deployed as a blanket measure in the prevention of myiasis in sheep.

Ivermectin is one of the most widely used drugs for parasite control. Previous studies have shown a reduction in the abundance and diversity of \"non-target\" coprophilous organisms due to the presence of ivermectin (IVM) in bovine faecal matter (FM). Due to its breadth of behavioural habits, Calliphora vicina is a suitable dipteran species to evaluate the effects of IVM in FM. The aim of this work was to evaluate the effect of five concentrations of IVM in FM (3000, 300, 100, 30, and 3 ng/g) on the development of C. vicina. The following endpoints were evaluated: survival (between the first larval stage and emergence of new adults), larval development times to pupation and pupation times to adult, and adult emergence (% sex) and LC50. Sampling was performed from larval hatching at 60 and 120 min and at 3, 4, 5, and 12 h, and every 24 h specimens were weighed until pupae were observed. Data were analysed by ANOVA using a non-parametric Kruskal-Wallis test and as a function of elapsed development time and accumulated degree hours (ADH). Mortality at 3000 and 300 ng/g was 100% and 97%, respectively. There were statistically significant delays in adult emergence time (p = 0.0216) and in the ADH (p = 0.0431) between the control group (C) and 100 ng/g. The LC50 was determined at 5.6 ng/g. These results demonstrate the lethal and sub-lethal effects of IVM on C. vicina, while highlighting the usefulness of this species as a bioindicator for ecotoxicological studies.

We explored the potential influence of temperature and precipitation on the abundance of two nest-dwelling ectoparasites (blowflies and mites) of Eurasian blue tits (Cyanistes caeruleus) during a period of 21 years and compared the results with those of a shorter period. The abundance of blowflies was negatively related to precipitation, which could prevent flies from locating their host, and laying date. In addition, blowflies were positively related to brood size (more food implies more parasites) and the interaction between precipitation and temperature. The highest abundances of blowfly pupae were attained in conditions of increasing precipitation and decreasing temperature, which should be more common at the beginning of the bird breeding season. Mites were significantly and positively related to laying date and the interaction between average precipitation and temperature but only for the larger dataset. Higher abundances of mites were related to intermediate values of temperature and precipitations, conditions that are found at the end of the breeding season. These results imply that optimal conditions for both parasites differ, with blowflies preferring earlier breeders and colder and more humid conditions than mites. Thus, the effects of the climatic conditions studied on parasite abundances are non-monotonic and can vary with years and parasite species. Finally, the fact that average temperature and precipitation decreases across the years of study is probably due to the advancement in Eurasian blue tit laying date because we calculated those variables for the period of birds\' reproduction. This earlier nesting does not affect parasite abundance.

The blowfly, Aldrichina grahami (Diptera: Calliphoridae), is a well-known forensically important insect. Basic data related to A. grahami have been well documented; but despite the pivotal role of gut microbes in various facets of insect biology, little is known about its gut microbiome. To investigate the gut bacterial community of A. grahami and explore its stability, diet and developmental stage were selected as the two variables. Larvae were reared on bovine liver, swine manure, and chicken manure, and high-throughput sequencing of the 1st, 2nd, and 3rd instar larvae and the newly emerged adults was performed. According to our results, the alpha diversity of the gut bacterial community did not significantly vary among different diets and developmental stages. Principal coordinate analysis revealed that the gut microbiome of A. grahami clustered together among different diets and developmental stages. The main phyla in the gut microbial community of A. grahami were Firmicutes and Proteobacteria, and the dominant genera were Vagococcus, Providencia, Lactobacillus, and Morganella. These findings characterized the gut microbiome of A. grahami and demonstrated that the gut bacterial community is fairly stable. The dominant genera Vagococcus, Providencia, Lactobacillus, and Morganella have the potential to serve as core microbiomes within the gut microbial community of A. grahami.

Male-killing bacteria are found in a broad range of arthropods. Arsenophonus nasoniae is a male-killing bacterium, causing a 80% reduction of the male progeny in infected Nasonia vitripennis wasps. Although the discovery of A. nasoniae dates from the early 80\'s, knowledge about the biology and ecology of this endosymbiont is still scarce. One of these poorly studied features is the ecological factors underlying A. nasoniae incidence on its Nasonia spp. hosts in different geographical locations. Here, we studied the prevalence of A. nasoniae in Iberian wild populations of its host N. vitripennis. This wasp species is a common parasitoid of the blowfly Protocalliphora azurea pupae, which in turn is a parasite of hole-nesting birds, such as the blue tit (Cyanistes caeruleus). We also examined the effects of bird rearing conditions on the prevalence of A. nasoniae through a brood size manipulation experiment (creating enlarged, control and reduced broods). Both the wasp and bacterium presence were tested through PCR assays in blowfly pupae. We found A. nasoniae in almost half (47%) of nests containing blowflies parasitized by N. vitripennis. The prevalence of A. nasoniae was similar in the two geographical areas examined (central Portugal and southeastern Spain) and the probability of infection by A. nasoniae was independent of the number of blowfly pupae in the nest. Experimental manipulation of brood size did not affect the prevalence of A. nasoniae nor the prevalence of its host, N. vitripennis. These results suggest that the incidence of A. nasoniae in natural populations of N. vitripennis is high in the Iberian Peninsula, and the infestation frequency of nests by N. vitripennis carrying A. nasoniae is spatially stable in this geographical region independently of bird rearing conditions.

UNASSIGNED: Lucilia sericata (Diptera: Calliphoridae) is used in larval therapy for wound healing. Netrin-A is an enzyme secreted from the salivary glands of these larvae, and has a central role in neural regeneration and angiogenesis. This study aimed to produce the recombinant Netrin-A protein from Lucilia sericata larvae by the baculovirus expression vector system in the Sf9 insect cell line.
UNASSIGNED: The coding sequence of Netrin-A was cloned, amplified in the pTG19 vector, and then cloned in the pFastBac HTA vector. It was then transformed into DH10Bac, and the recombinant Bacmid was subsequently transfected into Sf9 cells. The recombinant Netrin-A was purified by Ni-NTA agarose. The evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay.
UNASSIGNED: The molecular weight of this protein was 52 kDa with 404 amino acids. The signal peptide was located between amino acids 24 and 25. The concentration of Netrin-A was calculated to be 48.8 μg/ml. It reaffirmed the characterized gene codes of Lucilia sericata Netrin-A in a previous study.
UNASSIGNED: The generation of recombinant Netrin-A could be used in larval therapy, and as a biomarker in certain diseases. The netrin-A of Lucilia sericata was unprecedentedly cloned and expressed in a eukaryotic cell line. Given that this larva is FDA-approved, and non-pathogenic, it conduces to research on the development of maggot therapy in future.

We present a genome assembly from an individual male Calliphora vomitoria (a bluebottle; Arthropoda; Insecta; Diptera; Calliphoridae). The genome sequence is 708 megabases in span. Most of the assembly is scaffolded into six chromosomal pseudomolecules, including the assembled X sex chromosome. The mitochondrial genome has also been assembled and is 16.2 kilobases in length. Gene annotation of this assembly on Ensembl identified 12,917 protein coding genes.

Blow flies (Calliphoridae) play essential ecological roles in nutrient recycling by consuming decaying organic matter. They serve as valuable bioindicators in ecosystem management and forensic entomology, with their unique feeding behavior leading to the accumulation of environmental pollutants in their cuticular hydrocarbons (CHCs), making them potential indicators of exposure history. This study focuses on CHC degradation dynamics in empty puparia of Lucilia sericata under different environmental conditions for up to 90 days. The three distinct conditions were considered: outdoor-buried, outdoor-above-ground, and indoor environments. Five predominant CHCs, n-Pentacosane (n-C25), n-Hexacosane (n-C26), n-Heptacosane (n-C27), n-Octacosane (n-C28), and n-Nonacosane (n-C29), were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS). The findings revealed variations in CHC concentrations over time, influenced by environmental factors, with significant differences at different time points. Correlation heatmap analysis indicated negative correlations between weathering time and certain CHCs, suggesting decreasing concentrations over time. Machine learning techniques Support Vector Machine (SVM), Multilayer Perceptron (MLP), and eXtreme Gradient Boosting (XGBoost) models explored the potential of CHCs as age indicators. SVM achieved an R-squared value of 0.991, demonstrating high accuracy in age estimation based on CHC concentrations. MLP also exhibited satisfactory performance in outdoor conditions, while SVM and MLP yielded unsatisfactory results indoors due to the lack of significant CHC variations. After comprehensive model selection and performance evaluations, it was found that the XGBoost model excelled in capturing the patterns in all three datasets. This study bridges the gap between baseline and ecological/forensic use of empty puparia, offering valuable insights into the potential of CHCs in environmental monitoring and investigations. Understanding CHCs\' stability and degradation enhances blow flies\' utility as bioindicators for pollutants and exposure history, benefiting environmental monitoring and forensic entomology.

Performing quantitative sampling and determining faunistic analyses of dipterans is of fundamental importance in the analysis of ecological behavior, such as population dynamics and diversity, among other factors, for exotic and native species of necrophagous dipterans, so it is important to observe the type of bait used in traps to capture these dipteran species. This work aims to study structural parameters and faunistic indices of the diversity of Calliphoridae and Mesembrinellidae species as well as the abundance and diversity of species attracted to liver in two stages of decomposition: fresh liver and liver at 48 h of putrefaction. A total of 2826 dipterans were collected during the period from May 2021 to February 2022. We observed that liver in decomposition for 48 h was more attractive in the forest and rural environments, while fresh liver showed greater attractiveness in the urban environment; however, no statistical difference was evidenced between the attractiveness in the different environments. The Mesembrinellidae family and the species Lucilia eximia (Wiedemann, 1819) were collected mostly from deteriorated liver, while Cochliomyia macellaria (Fabricius, 1775) showed no preference for any liver decomposition stage. The Wilcoxon test indicated that there is a significant difference between the preferences for putrefied bait in Mesembrinellidae, while in Calliphoridae, there was no preference for type of bait. The faunistic analysis showed that richness in the forest area was always higher when compared to the urban and rural areas. Laneela nigripes (Guimarães, 1977) and Mesembrinella bellardiana (Aldrich, 1922) were abundant and exclusive in the preserved environment, showing themselves to be good environmental bioindicators.