Medical and Veterinary Entomology最新文献

Culicoides biting midge species (Diptera: Ceratopogonidae) of the Obsoletus Group and the Pulicaris Complex are considered the major vectors of bluetongue and Schmallenberg viruses in Europe. Overwintering strategies of these arboviruses are controversially discussed, with the ongoing activity of vector species and a non-disrupted transmission cycle during winter being a plausible explanation. Although data on Culicoides winter activity are relatively scant, a seasonal vector-free period (SVFP), during which adult Culicoides are not or hardly active, is questionable. To determine winter activity and define SVFPs according to the EU Commission Regulation No 1266/2007, adult Culicoides were trapped weekly with UV-light traps from October to April 2019/2020 and 2020/2021 inside and outside stables on 16 farms throughout Germany. Temperature measurements were taken regularly at each trapping site since the temperature is a known driver of biting midge activity. In 960 indoor and outdoor catches, 32,377 Culicoides were trapped, with 90.9% of them belonging to the Obsoletus Group, 6.1% to the Pulicaris Complex and 3.0% to ‘other Culicoides’ according to morphological identification. The majority (61.3%) of Culicoides were trapped indoors, with substantial numbers of specimens collected from October to December, in March and in April, and only a few or no specimens in January and February. Obsoletus Group biting midges were active indoors for almost the entire winter. Outdoors, Culicoides numbers decreased from October to December, few or no specimens were caught from January to March, and high numbers were captured in April. Of the collected Culicoides, 2028 were blood-fed, of which 94.6% were trapped in the stables. The indoor SVFP, although calculated for blood-fed instead of parous females, lasted for almost 4 months (late November until mid-March) in winter 2019/2020 and 2 months (January and February) in winter 2020/2021. The outdoor SVFPs covered almost the entire study period in both winters, with slight differences between the onsets and the ends. The Culicoides activity significantly depended on temperature. Specimens of the Obsoletus Group were caught at an average temperature of 7.4°C (minimum 0.3°C) and of the Pulicaris Complex at an average temperature of 10.3°C (minimum 1.2°C). These temperatures were reached inside the stables over more extended periods than outside. The average indoor temperatures were 1.2 K higher than the average outdoor temperatures, although absolute temperature differences of up to 9.0 K were recorded. Based on Culicoides activity, the results of the present study indicate an almost continuous potential for virus transmission in winter within livestock houses.

Research into various bacterial pathogens that can be transmitted between different animals and may have zoonotic potential has led to the discovery of different strains of Bartonella sp. in bats and their associated ectoparasites. Despite their enormous species diversity, only a few studies have focussed on the detection of bacterial pathogens in insectivorous bats of boreal forests and their associated Macronyssidae and Spinturnicidae mites. We collected and molecularly analysed mite samples from forest-dwelling bat species distributed all along the boreal belt of the Palearctic, from Central Europe to Far East. Ectoparasitic mites were pooled for DNA extraction and DNA amplification polymerase chain reaction (PCRs) were conducted to detect the presence of various bacterial (Anaplasmataceae, Bartonella sp., Rickettsia sp., Mycoplasma sp.) and protozoal (Hepatozoon sp.) pathogens. Bartonella sp. DNA was detected in four different mite species (Macronyssidae: Steatonyssus periblepharus and Spinturnicidae: Spinturnix acuminata, Sp. myoti and Sp. mystacinus), with different prevalences of the targeted gene (gltA, 16-23S ribosomal RNA intergenic spacer and ftsZ). Larger pools (>5 samples pooled) were more likely to harbour Bartonella sp. DNA, than smaller ones. In addition, cave-dwelling bat hosts and host generalist mite species are more associated with Bartonella spp. presence. Spinturnicidae mites may transmit several distinct Bartonella strains, which cluster phylogenetically close to Bartonella species known to cause diseases in humans and livestock. Mites with ubiquitous presence may facilitate the long-term maintenance (and even local recurrence) of Bartonella-infestations inside local bat populations, thus acting as continuous reservoirs for Bartonella spp in bats.

Chagas disease is considered one of the most important human parasitosis in the United States. This disease is mainly transmitted by insects of the subfamily Triatominae. The chemical vector control is the main tool for reducing the incidence of the disease. However, the presence of triatomines after pyrethroids spraying has been reported in some regions, as in the case of Triatoma infestans in Argentina and Bolivia. The presence of insects can be explained by the colonization from neighbouring areas, the reduction of insecticide dose to sublethal levels due to environmental factors, and/or by the evolution of insecticide resistance. In the last two scenarios, a proportion of the insects is not killed by insecticide and gives rise to residual populations. This article focuses on the toxicological processes associated with these scenarios in triatomines. Sublethal doses may have different effects on insect biology, that is, sublethal effects, which may contribute to the control. In addition, for insect disease vectors, sublethal doses could have negative effects on disease transmission. The study of sublethal effects in triatomines has focused primarily on the sequence of symptoms associated with nervous intoxication. However, the effects of sublethal doses on excretion, reproduction and morphology have also been studied. Rhodnius prolixus and T. infestans and pyrethroids insecticides were the triatomine species and insecticides, respectively, mainly studied. Insecticide resistance is an evolutionary phenomenon in which the insecticide acts as a selective force, concentrating on the insect population's pre-existing traits that confer resistance. This leads to a reduction in the susceptibility to the insecticide, which was previously effective in controlling this species. The evolution of resistance in triatomines received little attention before the 2000s, but after the detection of the first focus of resistance associated with chemical control failures in T. infestans from Argentina in 2002, the study of resistance increased remarkably. A significant number of works have studied the geographical distribution, the resistance mechanisms, the biological modifications associated with resistance, the environmental influences and the genetic of T. infestans resistant to pyrethroid insecticides. Currently, studies of insecticide resistance are gradually being extended to other areas and other species. The aim of this article was to review the knowledge on both phenomena (sublethal effects and insecticide resistance) in triatomines. For a better understanding of this article, some concepts and processes related to insect-insecticide interactions, individual and population toxicology and evolutionary biology are briefly reviewed. Finally, possible future lines of research in triatomine toxicology are discussed.

Culicoides oxystoma Kieffer (Diptera: Ceratopogonidae) has been vectoring several arboviruses, protozoa and nematodes, leading to mortality and morbidity of livestock and wild ruminants in the tropics and subtropics. Insight into the bacterial communities associated with the vector species must be worked out. This work tries to inventorize the bacterial communities associated with this important vector species. Acquisition of gut microbiota may be the parental origin, while some are obtained through feeding during larval stages. Culicoides oxystoma possesses semi-aquatic life cycle strategies for egg-laying and larval survival. The bacteria associated with C. oxystoma were compared throughout (i) life stages: egg, larval instars, pupa, adult: male and female obtained from laboratory colony; (ii) field-collected adult: male and age-graded females; and (iii) natural breeding substrate and artificial rearing substrate. The culture-dependent bacteria were identified by Sanger sequencing of 16S rRNA, and haemolytic bacteria were screened on blood agar. Results show that Firmicutes and Proteobacteria are the predominant Phyla, of which Bacillus spp. was the most abundant across the life stages. Across the life history, Bacillus cereus, Bacillus pumilus, Bacillus tropicus, Lysinibacillus sp. and Paenibacillus sp. were retrieved routinely. Bacillus cereus and Alcaligenes faecalis were detected in the lab-reared specimens and shared between the natural breeding site and rearing medium. From the adults trapped across two locations, B. cereus, Bacillus flexus, A. faecalis, Enterococcus faecium and Pseudomonas sp. were isolated. The bacterial species associated with this vector may influence various physiological traits, such as vectorial capacity, digestion and larval development, which need further investigation.

Most blow flies (Diptera: Calliphoridae) species are sarcosaprophagous during the larval stage, primarily feeding on the soft tissues of carcasses during the early stages of decomposition, making them valuable forensic indicators for minimum post-mortem interval (minPMI) estimations. Like other insects, their developmental rates are strongly influenced by the environmental temperature. Although several studies have examined the influence of temperature on the development of different blow fly species, the impact of cold temperatures remains largely unstudied, despite its potential forensic implications. The present study investigates the effect of three cold temperatures (0, -2.5 and -5°C) on the survival of Calliphora vicina embryos of five different ages (0%, 20%, 40%, 60% and 80% of the total embryonic development) and two exposure times (6 and 24 h). Our results revealed significant differences in egg survival at the earliest embryonic stages (0% and 20% of the total embryonic development), resulting in high mortality rates. While at 20% of the total embryonic development high mortality was only observed under -5°C, at 0% of the total embryonic development high mortality rates were observed at all the temperatures tested. Although C. vicina embryos demonstrate tolerance to cold temperatures once they have completed the first 20% of the total embryonic development, potentially mitigating the impact of cold weather events, the possibility of minPMI underestimations due to the death of the first egg batches should not be disregarded. Additionally, considering that the embryonic development stages may last for several days under low temperatures, caution should be taken in the analysis of entomological evidence if a cadaver is discovered following cold weather episodes.

Investigations of host feeding behaviour in haematophagous insects are critical to assess transmission routes of vector-borne diseases. Understanding if a certain species has ornithophilic or mammalophilic feeding behaviour can facilitate future studies focused on pathogens transmission to and from certain host species. Culicoides Latreille (Diptera: Ceratopogonidae) are vectors of several pathogens, which include arboviruses, bacteria and parasites to a considerable diversity of vertebrate hosts. However, most of the studies focused on feeding habits target Culicoides species that could transmit the Bluetongue virus, consequently with a mammalophilic feeding behaviour, leaving aside the Culicoides species that are involved in the transmission of vector-borne parasites to birds, such as Haemoproteus Kruse (Haemosporida: Haemoproteidae). This study aimed to investigate the source of blood meals of wild-caught Culicoides using molecular-based methods and to correlate our findings with the reports of Haemoproteus parasites in Culicoides species. Engorged Culicoides females were collected using ultraviolet (UV)-light traps at seven different localities in Lithuania in 2021–2023. Biting midges were dissected, and the abdomens of engorged females were used for molecular investigation of the blood meal source. A polymerase chain reaction (PCR) protocol that amplifies a fragment of the Cytochrome B gene of vertebrates was used. Obtained sequences were compared to available information in GenBank database to confirm the source of the blood meal. In total, 258 engorged Culicoides females, representing nine different species, were analysed. The source of blood meal was identified in 29.1% of them with most of the insects having fed on birds (74.7%) and the remaining on mammals (25.3%). Culicoides segnis Campbell, Pelham-Clinton was the only species to feed exclusively on birds; Culicoides from the Obsoletus group, C. pallidicornis Kieffer and C. punctatus Latreille were found to feed exclusively on mammals; C. festivipennis Kieffer, C. kibunensis Tokunaga and C. pictipennis Staeger had an opportunistic feeding behaviour, with the first two preferably feeding on birds. Due to their feeding behaviour and the presence of Haemoproteus parasites reported in the literature, C. festivipennis, C. kibunensis, C. pictipennis, and C. segnis play an important role in the transmission of those avian vector-borne parasite in the wild. These Culicoides species were already confirmed as being able to support the development of several Haemoproteus species and lineages. Future studies focused on understanding the epidemiology of avian pathogens transmitted by Culicoides should target these species.

Stomoxys calcitrans L. (Diptera: Muscidae), the stable fly, is a hematophagous insect of great veterinary importance, because it is a mechanical vector of diverse pathogens in livestock. The saliva of blood-feeding insects presents important pharmacologically active molecules that impair blood clotting, promote vasodilation and modulate the host immune system response, crucial processes for successful feeding. These properties also enable pathogens' transmission. In the present work, we describe an efficient protocol to dissect S. calcitrans salivary glands, their morphological characteristics and lipid profile. The mean length of the tubular gland is 3.23 mm with a bulbous posterior end and a narrow anterior end. Histological analysis revealed a monolayer of large polygonal epithelial cells with voluminous nuclei and high lipid content in their cytoplasm. Ultrastructural analysis showed that the epithelium is rich in mitochondria, free ribosomes, Golgi complex cisternae, presenting a great extension of rough endoplasmic reticulum that contains an electron-dense material. Lipid analysis by thin-layer chromatography showed that neutral fatty acids and phosphatidylcholine are predominant in the fly salivary glands. Lysophosphatidylcholine, an important signalling biomolecule involved in different metabolic processes, including host's immunomodulation and pathogens proliferation and differentiation, is also present.

Several species of the worldwide distributed genus Calliphora Robineau-Desvoidy (Insecta, Diptera, Calliphoridae) are medically important vectors and agents of myiasis. Furthermore, these flies are relevant in forensics because they are found in corpses. Information regarding the taxonomy, bionomics and distribution of Calliphora species endemic to South America, including Calliphora lopesi Mello, is scarce. To expand knowledge on C. lopesi, this study presents descriptions of eggs, larvae, puparia and developmental data at 14, 17, 20, 23 and 26 ± 1°C for the first time. Adult flies were collected from the field and kept in the laboratory to obtain samples for morphological and biological studies. Immatures were examined using light and scanning electron microscopy. To assess the growth rate, 10 specimens from each temperature group were randomly removed from the diet and weighed every 24 h from larval hatching until pupation. The minimum developmental threshold, thermal constant and linear development-rate equations were calculated for each stage. Considering weight gain records and survival rates, the optimum temperature for the development of C. lopesi ranges from 23 to 26°C. A key to third-instar larvae of known Neotropical species of Calliphora was also provided to assist in identification. The information provided in this study should be useful in expanding knowledge about Neotropical Calliphoridae species of forensic importance.

Estimating the age of immature blow flies is of great importance for forensic entomology. However, no gold-standard technique for an accurate determination of the intra-puparial age has yet been established. Fourier transform infrared (FTIR) spectroscopy is a method to (bio-)chemically characterise material based on the absorbance of electromagnetic energy by functional groups of molecules. In recent years, it also has become a powerful tool in forensic and life sciences, as it is a fast and cost-effective way to characterise all kinds of material and biological traces. This study is the first to collect developmental reference data on the changes in absorption spectra during the intra-puparial period of the forensically important blow fly Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae). Calliphora vicina was reared at constant 20°C and 25°C and specimens were killed every other day throughout their intra-puparial development. In order to investigate which part yields the highest detectable differences in absorption spectra throughout the intra-puparial development, each specimen was divided into two different subsamples: the pupal body and the former cuticle of the third instar, that is, the puparium. Absorption spectra were collected with a FTIR spectrometer coupled to an attenuated total reflection (ATR) unit. Classification accuracies of different wavenumber regions with two machine learning models, i.e., random forests (RF) and support vector machines (SVMs), were tested. The best age predictions for both temperature settings and machine learning models were obtained by using the full spectral range from 3700 to 600 cm^-1. While SVMs resulted in better accuracies for C. vicina reared at 20°C, RFs performed almost as good as SVMs for data obtained from 25°C. In terms of sample type, the pupal body gave smoother spectra and usually better classification accuracies than the puparia. This study shows that FTIR spectroscopy is a promising technique in forensic entomology to support the estimation of the minimum post-mortem interval (PMI_min), by estimating the age of a given insect specimen.

Triatomines of the species Triatoma sherlocki are considered sylvatic; however, household invasion appears imminent, potentially carrying Trypanosoma cruzi, the causative agent of Chagas disease. The aim of this study was to report the first occurrence of a colony of T. sherlocki infected by T. cruzi in a subsistence pig farm. Triatomines collected underwent polymerase chain reaction (PCR) technique for T. cruzi detection and determination of blood meal source. The 19 triatomines collected in the pig farm were of the species T. sherlocki, comprising 26.3% nymphs (5/19), 52.6% males (10/19) and 21.1% females (4/19). PCR showed that 15.8% (3/19) of triatomines were infected by T. cruzi. The only detected blood meal source in triatomines (n = 11) was the domestic mammal Sus scrofa, commonly known as domestic pig, indicating that T. sherlocki is an opportunist, feeding on available vertebrates in the environment, including domestic animals such as pigs. These results highlight the possibility of domiciliation of the species T. sherlocki and its potential role in bridging the transmission of T. cruzi between sylvatic and domestic environments.