Medical and Veterinary Entomology最新文献

The aim of this work was to analyse the spatiotemporal heterogeneity of Rhipicephalus microplus (Canestrini, 1888) (Acari: Ixodidae) resistance to chemical acaricides at intra-farm level under different environmental (favourable and unfavourable areas for tick development) and management (different schemes of acaricides applications) conditions using ivermectin as a model. The in vitro larval immersion test (LIT) was used to determine quantitatively the levels of resistance to ivermectin in the different populations and subpopulations of R. microplus analysed. In the first case study, differences in resistance levels among tick samples within the same paddock across time and among tick samples from different paddocks were recorded. These results stress the importance of taking successive samples so that they represent the spatial and temporal variabilities in the levels of resistance that can occur within the same farm. In a second case study, the evolution of resistance in R. microplus subpopulations subjected to strategic and threshold control methods based on the application of three annual treatments with alternation of chemical groups was compared. No changes in resistance/susceptibility status were observed in both tick subpopulations. These results show that the application of a particular drug once a year within a scheme of alternation with other chemical groups could be an appropriate strategy to delay the development of resistance. Data of the third case study showed that environmental constraints is not only a key modulator of R. microplus abundance but could also affect the evolution of resistance in the tick populations. The decreasing trend of LC₅₀ values in the tick subpopulation not exposed to chemical treatments but also in that subpopulation exposed to three annual chemical treatments under unfavourable environmental conditions allow us to hypothesise that environmental constraints can modify the levels of resistance in a tick population because it can constitute a stronger selection factor than the treatments themselves.

Vector-borne diseases represent various sicknesses that are increasingly significant in human and veterinary health. Among the zoonotic agents transmitted by ticks, infections caused by the intracellular pathogens from the Anaplasmataceae family and piroplasmids (Babesia/Theileria spp.) are particularly notable due to the substantial economic losses they cause in the livestock sector. A study was carried out to assess the prevalence of these pathogens in ticks from ruminants in the province of Valencia (Eastern Iberian Peninsula). Between 2019 and 2022, 1674 ticks were collected from 163 domestic and wild ruminants. The tick species were identified using molecular and morphological features and included Rhipicephalus spp. (Rhipicephalus bursa and Rhipicephalus sanguineus) (88.7%), Haemaphysalis sulcata (5.9%), Ixodes ricinus (3.6%) and Dermacentor marginatum (1.2%). The Spanish ibex (Capra pyrenaica) was found to be at greater risk of infestation by Haemaphysalis compared to other ruminants. Polimerase Chain Reaction (PCR) analysis was performed on 108 individual ticks from different hosts, with 16.7% testing positive for members of the Anaplasmataceae family and 39.8% for Theileria spp. Sequencing of 10 positive samples from each pathogen group revealed the presence of Anaplasma ovis, Anaplasma platys, Ehrlichia spp., Theileria ovis and Theileria capreoli. No Anaplasma phagocytophilum or Babesia spp. were detected. Further research is essential for improved management of ticks and the diseases they transmit in the study area.

Aedes albopictus (Diptera: Culicidae), commonly known as the Asian tiger mosquito, is an important vector transmitting dangerous arboviruses to humans. This study investigated the phenotypic and genetic variation of this species in Thailand through wing geometric morphometric (GM) and mitochondrial cytochrome c oxidase subunit I (COI) gene sequence analyses. A total of 236 Ae. albopictus specimens from 12 populations in Thailand and 89 specimens from invasive populations in Florida, Hawaii and Brazil underwent wing GM analysis. The centroid size (CS) of Ae. albopictus populations in Thailand ranged from 2.00 mm in Bangkok to 2.36 mm in Chanthaburi, while in invasive populations, CS varied from 2.25 mm in Brazil to 2.47 mm in Florida. Pairwise comparisons of wing shape revealed significant differences for most population pairs, with distances ranging from 1.63 to 10.02. The clustering tree indicated distant relationships in wing shape between native and invasive populations. Additionally, partial COI gene sequences were amplified from 108 specimens, revealing a mean haplotype diversity of 0.842 ± 0.025 and a mean nucleotide diversity of 0.002 ± 0.001. The results from neutral Tajima's D and Fu's Fs tests indicated negative and statistically significant values (-2.159 and -33.846, respectively), suggesting population expansion. Further examination of haplotype relationships between Thailand and other countries identified two distinct groups: a Southeast Asia group, with Thai haplotypes clustered exclusively within it, and a non-Southeast Asia group. These findings highlight the phenotypic and genetic variation of Ae. albopictus in Thailand, providing essential insights for disease control strategies and tracing the mosquito's origins across regions.

The 158 bug species that make up the subfamily Triatominae are the potential vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. Despite recent progress in developing a picture-based automated system for identification of triatomines, an extensive and diverse image database is required for a broadly useful automated application for identifying these vectors. We evaluated performance of a deep-learning network (AlexNet) for identifying triatomine species from a database of dorsal images of adult insects. We used a sample of photos of 6397 triatomines belonging to seven genera and 65 species from 27 countries. AlexNet had an accuracy of ~0.93 (95% confidence interval [CI], 0.91-0.94) for identifying triatomine species from pictures of varying resolutions. Highest specific accuracy was observed for 21 species in the genera Rhodnius and Panstrongylus. AlexNet performance improved to ~0.95 (95% CI, 0.93-0.96) when only the species with highest vectorial capacity were considered. These results show that AlexNet, when trained with a large, diverse, and well-structured picture set, exhibits excellent performance for identifying triatomine species. This study contributed to the development of an automated Chagas disease vector identification system.

The Canary Islands are an excellent natural laboratory for understanding ecological and evolutionary processes such as biogeographical colonisation. The morphology of the larva, puparium and adult of the endemic Canarian copper fly, Calliphora splendens, is described, illustrated and contrasted with those of the other species of Calliphora that occur in Africa, the Iberian Peninsula and Macaronesia. Partial cytochrome oxidase I sequences show a connection between C. splendens, Calliphora vicina, Calliphora loewi and Calliphora croceipalpis, but more distant relationship with Calliphora vomitoria. Calliphora splendens produced unisexual offspring in captivity. This work confirms the relict character of the Canarian copper fly associated with the endemic laurel forest habitat.

Macaques are widely distributed in Thailand with remarkable adaptation to living close to humans in residential, religious sites, markets and tourist areas. They play an essential role in the persistence of pathogens in the environment. As reservoir hosts, they are exposed to hematophagous vectors that secrete saliva, a cocktail of bioactive molecules including antigenic components stimulating host antibody production. Subsequent to the detection of mosquito-borne pathogens in macaques living in national parks, we aimed to determine the seroprevalence of antibodies to crude salivary gland extracts (SGEs) from culicine mosquitoes (Aedes aegypti [Linnaeus, 1762], Ae. albopictus [Skuse, 1895] and Culex quinquefasciatus [Say, 1823]) and compare individual titres between macaque species/national parks (33, Macaca arctoides [I. Geoffroy Saint-Hilaire, 1831] [Primates: Cercopithecidae] from Kaeng Krachan, 23 M. leonina leonina [Blyth, 1863] [Primates: Cercopithecidae] from Khao Yai and four M. fascicularis [Raffles, 1821] [Primates: Cercopithecidae] from Mu Ko Ranong). The anti-mosquito SGE antibodies found in 60 macaques from three national parks indicate varying levels of host-vector exposure. Macaque antibody titres were high against culicine mosquitoes. However, the significant difference among national parks (or macaque species) was only observed against Cx. quinquefasciatus. Correlation analysis of titres between Aedes SGE and arboviruses revealed a significantly more intense immune response against Ae. albopictus in DENV3-positive M. arctoides. Current findings support the concept of salivary biomarkers using accessible SGE, available from mosquito colonies of interest. However, we observed cross-reactivity between Aedes species because of crude SGE containing species-shared proteins. Nevertheless, a potential risk of pathogen transmission is emphasised between national park visitors and macaques via mosquitoes as bridge vectors. This information contributes to preventive measures against mosquito bites, including those implemented in tourist areas.

The control of zoonotic and vector-borne pathogens is challenging due to the limited availability of intervention tools. West Nile virus (WNV) is an example of a globally distributed zoonotic arbovirus that circulates between Culex species (Diptera: Culicidae) mosquitoes and avian hosts, with spillover transmission to humans, resulting in disease cases. Interventions delivering systemic insecticides to vertebrate hosts used by vector species, known as xenointoxication, are potential tools for managing vector populations by creating toxic bloodmeals. In this study, we evaluated the impact of two systemic pesticides (ivermectin; Ivomec® Pour-On and fluralaner; Bravecto®), and one anthelmintic (fenbendazole; Safe-Guard® Aquasol) on the mortality of Cx. quinquefasciatus Say (Diptera: Culicidae). We found no significant difference in the feeding rates of mosquitoes that fed on treated chickens compared with those fed on untreated chickens, suggesting that the treatment did not repel mosquitoes. The mortality of Cx. quinquefasciatus mosquitoes feeding on fluralaner-treated chickens was significantly higher (p < 0.01) than those fed on control chickens at 3 and 7 days post-treatment, but this effect was not observed in mosquitoes fed on chickens treated with fenbendazole or ivermectin. No differences in mortality were observed among the groups at 14, 26 or 56 days post-treatment. These data support fluralaner as a xenointoxication tool to control Cx. quinquefasciatus populations and decrease the risk of human exposure to their associated pathogens.

Ground water tanks are known to be preferred Aedes aegypti oviposition places providing opportunities for adult and larvae control. Therefore, a dual-effect insecticidal coating (IC) (alphacypermethrin/ pyriproxyfen) with a slow-release mechanism and safe for users could be applied within Aedes spp. breeding sites, representing a promising option. Bioassays were designed to determine the mortality and sterilizing effects on gravid mosquitoes exposed to IC. The effect of inhibition of emergence was evaluated in eggs, larvae and pupae exposed in different containers. For the water safety assessment concentrations of active ingredients were determined by reverse phase high performance liquid chromatography (RP-HPLC) and the health risk was calculated. The IC applied to the interior walls of water-holding containers showed efficacy against Ae. aegypti in terms of high gravid-female mortality (81% at 24 h, p < 0.01), sterilizing effect (inhibition of oviposition by 63%, p < 0.01) and emergence inhibition (100% in eggs, L3 and L4; 97% in pupae). The offspring rate was reduced [only 0.15 (38/250) new adults emerged per exposed gravid females as against 11.90 per unexposed female (2976/250) at baseline]. Emergence inhibition was recorded up to 12 months and adult mortality >80% up to 6 months. The use of water stored in treated containers, either for washing or drinking, is not expected to pose a health risk to users. IC applied to domestic water containers has dual and complementary action that reduces Ae. aegypti densities (immature and adult stages). This represents baseline information for a cluster randomized efficacy trial in Colombia.

Culex species, such as Culex quinquefasciatus and Culex nigripalpus display a range of feeding habits and act as vectors for pathogens that can cause diseases in both humans and animals. Understanding their feeding habits is pivotal for enhancing disease prevention strategies. The present study introduces the application of two multiplex real-time PCR (qPCR) followed by high-resolution melting (HRM) as a cost-effective and time-efficient alternative. This investigation involved the development of two multiplex qPCR-HRM: assay 1 detects human, dog and chicken, while assay 2 detects cat, cattle and horse in Culex sp. engorged female mosquitoes. The qPCR-HRM reactions showed a detection limit of one copy of genomic DNA when performed as single and multiplex qPCR-HRM. The reaction efficiencies were 97.96% for human, 100.60% for dog, 99.03% for chicken, 99.92% for feline, 99.18% for cattle and 97.68% for horse. The qPCR-HRM method, employing multiplex 1 and 2, was applied to field-collected mosquitoes and demonstrated the ability to detect DNA from multiple blood sources within a single sample. By analysing both multiplexes, it was possible to identify up to five distinct blood sources in Cx. quinquefasciatus and Cx. nigripalpus, and up to two sources in Culex coronatus. Sequencing corroborated the qPCR-HRM results, confirming the presence of DNA from one to four different blood sources with 100% accuracy. The development of these molecular methods may contribute for identification of blood-feeding patterns in mosquitoes. It contributes to studies on the dissemination and transmission of pathogens among various animals and humans, thereby bridging the gap between epidemiology and vector monitoring.

The diversity and abundance of vectors are essential parameters in the transmission dynamics of West Nile virus (WNV) between its avian reservoirs and clinically susceptible mammalian species. Knowing the determinants of vector abundance could be thus useful in preventing West Nile fever (WNF) cases and associated socio-economic impact. We designed a survey at the wildlife-livestock interface to test the hypothesis that variations in environmental favourability between anthropized and wild scenarios modulate WNV vector abundance and transmission risk. In a continental Mediterranean region where WNF has recently emerged, we selected nine sampling sites and allocated three areas to every site with a decreasing gradient of wildlife-livestock interaction: A₁-a horse farm where interaction is maximal; A₂-a zone of intermediate interaction 500-1000 m from the farm; and A₃-an entirely wild zone of low interaction 1-5 km from the farm. At a fortnightly frequency, we estimated mosquito abundance at each of the 27 study sites in May-December 2018 and April-July 2019. We estimated bird and mammal abundance, collected meteorological information and characterised mosquito habitat at the site scale. Thereafter, we studied the determinants of Culex spp., Culex pipiens sensu lato (s.l.) Linnaeus, 1758 (Diptera: Culicidae) and Culex theileri  Theobald, 1903 abundance by constructing negative binomial generalised linear mixed models. We identified 20 mosquito species, with a notable predominance of Culex spp. and, particularly, of Cx. pipiens s.l. We found differences in the spatiotemporal distribution of Culex spp. abundance and confirmed our hypothesis by finding important effects of local environmental variations in abundance. The accumulated rainfall in fortnights 4-14 and the mean temperature of the two fortnights before sampling were positively and statistically significantly associated with the abundance of Cx. pipiens s.l. (Z = 13.09, p < 0.001, and Z = 9.91, p < 0. 001, respectively) and Culex spp. (Z = 13.35, p < 0.001, and Z = 6.99, p < 0.001, respectively), while the mean temperature of the two previous fortnights was a positive statistically significant predictor (Z = 14.69, p < 0.001) of the abundance of Cx. theileri. The farm environment was the most conducive predictor to hosting Culex spp. compared with wild settings. Our results indicate that continental Mediterranean environments are favourable for WNV circulation and maintenance, especially the environment of anthropized rural settings such as farms. These results will have an impact on the spatiotemporal risk prediction of WNF emergence in continental Mediterranean environments.