African animal trypanosomiasis (AAT) a parasitic disease of livestock in sub-Saharan Africa causing tremendous loses. Sub-Saharan continental estimation of mean prevalence in both large and small domestic animals, risk factors, tsetse and non-tsetse prevalence and drug resistance is lacking. A review and meta-analysis was done to better comprehend changes in AAT prevalence and drug resistance. Publish/Perish software was used to search and extract peer-reviewed articles in Google scholar, PubMed and CrossRef. In addition, ResearchGate and African Journals Online (AJOL) were used. Screening and selection of articles from 2000–2021 was performed according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Articles 304 were retrieved; on domestic animals 192, tsetse and non-tsetse vectors 44, risk factors 49 and trypanocidal drug resistance 30. Prevalence varied by, host animals in different countries, diagnostic methods and species of Trypanosoma. Cattle had the highest prevalence with Ethiopia and Nigeria leading, T. congolense (11.80–13.40%) and T. vivax (10.50–18.80%) being detected most. This was followed by camels and pigs. Common diagnostic method used was buffy coat microscopy. However; polymerase chain reaction (PCR), CATT and ELISA had higher detection rates. G. pallidipes caused most infections in Eastern regions while G. palpalis followed by G. mortisans in Western Africa. Eastern Africa reported more non-tsetse biting flies with Stomoxys leading. Common risk factors were, body conditions, breed type, age, sex and seasons. Ethiopia and Nigeria had the highest trypanocidal resistance 30.00–35.00% and highest AAT prevalence. Isometamidium and diminazene showed more resistance with T. congolense being most resistant species 11.00–83.00%.

Culex vishnui Theobald, 1901, a main vector of Japanese encephalitis virus (JEV), is widely distributed in the Oriental region where it often accounts for a great part of the culicid fauna. This species also has been found naturally infected with at least 13 other arboviruses of medical and veterinary importance. Females blood feed predominantly upon pigs and birds, but may readily bite cattle and humans. Because of its abundance, medical importance, and presence throughout ecological gradients among urban, peri-urban, and rural areas, Cx. vishnui potentially may serve as a bridge vector transmitting viruses from natural and wild hosts to humans. Being zoo- and anthropophagic, omnipresent in the Oriental region, and presenting strong resistance to many insecticide families, this overlooked mosquito species may pose a serious health risk in one of the most densely populated regions of the world.

Bartonella are vector-transmitted, intracellular bacteria that infect a wide variety of blood-feeding arthropods and their vertebrate hosts. In California, more than 13 species of Bartonella have been described from companion animals, livestock, and wildlife, of which four have been associated with human disease. Infections in humans cause a range of symptoms from relatively mild to severe, especially in immunocompromised individuals. Exposure to infected domestic animals and wildlife, and their ectoparasites, may increase the risk of cross-species transmission. The objective of this review was to compile and summarize published materials on human and animal Bartonella infections in California. Medical and veterinary case reports of bartonellosis were highlighted in an effort to increase the awareness of this poorly understood and potentially under-recognized disease among healthcare professionals and veterinarians.

The yellow fever mosquito, Aedes aegypti, is a growing global concern. An ecological parameter necessary for vector control is the dispersal, or flight range, of mosquito vectors. Information on mosquito dispersal supports identifying optimal sampling and control strategies to limit the invasion of adult mosquitoes. Attempts around the world to understand the movement and flight range of Ae. aegypti have used a variety of techniques, but there has been little agreement regarding flight distances of Ae. aegypti, specifically the average linear distance Ae. aegypti travel in their natural environment. To generate a more robust estimate of Ae. aegypti flight distance, we conducted a meta-data analysis with the aims 1) to provide a flight distance measurement and 2) investigate how mosquito flight range can be affected by study design and climatic factors. Published studies were retrieved from public databases and reviewed for mean distance traveled (MDT) or maximum distance traveled measurements of Ae. aegypti. Linear regression was used to assess potential relationships between Ae. aegypti flight distance and factors pertaining to climate, degrees of urbanization, and study design. MDT estimates were pooled from 27 experiments to calculate a weighted MDT of 105.69 m. This study addresses the average flight distance of Ae. aegypti with the intention of informing vector control programs in Ae. aegypti prevalent regions of the world.

Human head lice Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae) are ectoparasites that cause pediculosis, a global scale disease mainly found in school-age children. Previous works from our laboratory found nonanal, sulcatone, and geranylacetone as the main human scalp volatile components, and individually evaluated their attraction to head lice using an olfactometer. In this work, we compared how their blends at different concentrations attract head lice, and how their blended effect compares to the effect of isolated compounds. At the concentrations evaluated, individual components did not show attraction towards head lice, but a ternary mixture of them was attractive. Moreover, a solvent extract from the human head scalp was analyzed by GC-MS, finding that tetradecanoic acid, palmitic acid, oleic acid, palmitoleic acid, and squalene are the most abundant components. Attraction to these individual compounds at natural concentrations was tested by bioassays in a circular experimental arena. No attraction was observed when the components were tested individually, but when they were evaluated as a blend they attracted head lice. This work presents new information about how chemical signals are attractive at certain concentrations and proportions. This information could be used to better understand communication mechanisms in head lice and for the development of louse repellents.

Forensic Entomology uses arthropods to aid in legal investigations. This study checked the biological response of Chrysomya putoria pupae to submersion in fresh water for up to 6 d, evaluating the critical submersion time, survival rate, and development time of the flies. Adults were collected using fish baits in two typical traps. Seven hundred and twenty fourth-generation pupae with 2 d of development were used and separated into submergence intervals: 24, 48, 72, 96, 120, and 144 h. An additional 120 pupae were used as a control. Each treatment was done in triplicate, consisting of 40 pupae distributed in four tulle-sealed test tubes containing 10 pupae each. All tubes of each treatment were co-adhered in test tube racks and were submerged in mineral water in a container with constant oxygenation, except those of the control group, which were not submerged. The tubes were removed from the water according to their respective submersion interval, until 144 h was completed. The control group had a survival rate of 90%, while the 24-h treatment had 85% and the 48-h treatment had 35.8%. The critical submersion time for pupae was 72 h, with 100% mortality by 144 h. The average development time for the control group was 3.2 d, while the 24- and 48-h treatments developed in 4.3 and 6.3 d, respectively. The longer the individuals were submerged, the lower the survival rate was, while the development time increased. The data obtained in this study have potential in applications to estimate the interval of submersion of a cadaver.

The postburial interval (PBI) can be inferred by using necrophagous insects colonizing the buried corpse. In different seasons, the species composition and succession of necrophagous insects on swine carrion (0.5–0.75 kg) buried at the depths of 30 cm and 60 cm in a Populus alba var. pyramidalis (Bunge, 1854) (Salicales: Salicaceae) grove of Shenyang, China from 2017 to 2019 were investigated. A total of 21 species of necrophagous insects belonging to 5 orders, 17 families were collected. Among them, the species of Phoridae and Platystomatidae were dominant at burial depth of 30 cm and 60 cm in summer and autumn. The species composition and time of colonization of necrophagous insects on the buried baits varied with seasons. Platystoma mandschuricum (Enderlein, 1937) (Diptera: Platystomatidae) and Aleochara puberula (Klug, 1833) (Coleoptera: Staphylinidae), the first arriving insects in spring, occurred on the baits for the longest time, from early June to early December. This work could provide reference data for the PBI estimation in Shenyang and similar geographical areas.

Forensic entomology is an important field of forensic science that utilizes insect evidence in criminal investigations. Blow flies (Diptera: Calliphoridae) are among the first colonizers of remains and are frequently used in determining the minimum postmortem interval (mPMI). Blow fly development is influenced by a variety of factors including temperature and feeding substrate. Unfortunately, dietary fat content remains an understudied factor in the development process, which is problematic given the high rates of obesity in the United States. To study the effects of fat content on blow flies we investigated survivorship, adult weight, and development time (overall and by sex) of Lucilia sericata (Meigen) and Phormia regina (Meigen) on ground beef with 10%, 20%, or 27% fat. As fat content increased, survivorship decreased across both species with significant impact to P. regina. While P. regina adults were generally larger than L. sericata across all fat levels, only L. sericata demonstrated a significant (P < 0.05) difference in weight by sex. Overall development times did not vary by fat content, excluding 27% for P. regina. Additionally, development times did not vary by sex for P. regina but did differ (P < 0.05) for L. sericata with females taking longer to develop. These findings provide insight into the effect of fat content on blow fly development, a factor that should be considered when estimating an mPMI. By understanding how fat levels affect the survivorship and development of the species studied here, we can begin improving the practice of insect evidence analysis in casework.

Wing mites of the genus Periglischrus are ectoparasites exclusively associated with phyllostomid bats. These mites show high host specificity and have been studied to understand the evolutionary history of their bat hosts mainly by using a morphological variation. Through a phylogeographic approach, we analyzed the genetic diversity and population genetic structure of the ectoparasite Periglischrus paracaligus Herrin and Tipton which parasitizes Leptonycteris yerbabuenae Martínez and Villa (lesser long-nosed bat) in Mexico. By the implementation of a multilocus approach, we found that P. paracaligus populations were diverse for haplotype diversity, and had values ranging from 0.5 to 1. No genetic structuring in the P. paracaligus parasites was observed along with the distribution of the host, L. yerbabuenae, in Mexico, nor when populations or regions were compared, but our results revealed a process of historical demographic expansion in all the analyzed markers. We discuss possible scenarios that could explain the lack of population structure in the light of the data analyzed for the parasites and the biology of L. yerbabuenae, such as the interplay between parasite and host traits being responsible for the genetic make-up of parasite populations. We also inferred its phylogenetic position among wing mites parasitizing the two other species of Leptonycteris bats. Long-nosed bats' monophyly helps to explain the observed presence of distinctive clades in the wing mite's phylogeny in specific association with each long-nosed bat host species.

Bluetongue virus (BTV) causes a viral, non-contagious disease that mainly affects sheep, cattle, and wild and farmed ruminants causing damage to these animals and significant economic losses. Culicoides insignis Lutz, the major BTV vector in South America, is one of the most abundant species in Argentina and commonly associated with cattle farms. The morphological identification of Culicoides spp. is routinely carried out with the aid of morphological keys, which mainly refer to the wing patterns, sensillae of palpi, and antennal flagella. Molecular tools applied to taxonomy provide a rapid and efficient method of identification of vector species. An easy protocol for the extraction of total DNA from single midges is herein described, and a forward primer for rapid and reliably test detection by polymerase chain reaction of C. insignis is developed.

Tabanus variegatus F. 1805 has been called by the name Tabanus sulcifrons Macquart 1855 for over 80 yr; T. variegatus is one of the most common large horse flies attacking livestock in much of the southeastern U.S. Morphological, ecological, and molecular evidence indicates that T. variegatus is a distinct species, and we redescribe the female and describe the male. The Fabricius holotype, heavily damaged after over nearly 220 yr, is nevertheless taxonomically sound. Morphology (size, color, palp shape, and r₅ wing cell shape) can usually distinguish T. variegatus from T. sulcifrons, but some specimens remain difficult to separate, especially in and west of the Mississippi River Valley. Using geometric morphometric analyses of the wing vein arrangement and palp shape the two species are significantly different. The wings of T. variegatus females also have more microsetae and sometimes a “frosty” appearance. Where they are common and sympatric, as in eastern Tennessee, they are temporally separated such that T. variegatus flies later (August-October) than T. sulcifrons (June-August), minimizing opportunity for gene flow. Museum specimens allow the approximate range of T. variegatus to be compared with that of T. sulcifrons s.l.; T. variegatus is particularly abundant from the coast of the Carolinas and Georgia east to central Tennessee and south to about central Alabama. DNA evidence (COI gene) recovers T. variegatus and T. sulcifrons s.s. in separate clades. Further studies on the T. sulcifrons complex are needed to fully resolve the range of both species, assess the degree of genetic substructuring, and examine relationships with other members of the T. sulcifrons complex.

This paper deals with species A, C, D, and E of the Lindesayi Complex of Anopheles subgenus Anopheles (Diptera: Culicidae) that were recently recorded from Bhutan. Species B is not included due to insufficient data. Species A is Anopheles lindesayi sensu stricto, and species C, D, and E are new species that are formally described and named herein as Anopheles druki Somboon, Namgay & Harbach, Anopheles himalayensis Somboon, Namgay & Harbach and Anopheles thimphuensis Somboon, Namgay & Harbach, respectively. Morphological characteristics of the adults, larval and pupal stages of each species are provided and compared with An. lindesayi from Bhutan.

The Educator Group of the Melanoconion Section of Culex (Melanoconion) (Diptera, Culicidae) is reviewed. Currently, the group includes the following valid species: Culex (Melanoconion) aphyllus Talaga, 2020, Culex (Melanoconion) cristovaoi Duret, 1968, Culex (Melanoconion) educator Dyar & Knab, 1906, Culex (Melanoconion) eknomios Forattini & Sallum, 1992, Culex (Melanoconion) inadmirabilis Dyar, 1928, Culex (Melanoconion) rachoui Duret, 1968, Culex (Melanoconion) theobaldi (Lutz), 1904, Culex (Melanoconion) vaxus Dyar, 1920, Culex (Melanoconion) angularis Sá & Sallum n. sp., Culex (Melanoconion) spiniformis Sá & Hutchings n. sp., Culex (Melanoconion) longistriatus Sá & Hutchings n. sp., Culex (Melanoconion) aneles Dyar & Ludlow, 1922, Culex (Melanoconion) apeteticus Howard, Dyar & Ludlow, 1913, and Culex (Melanoconion) bibulus Dyar, 1920.The last three species were resurrected from synonymy of either Cx. educator or Cx. vaxus. Descriptions, differential diagnoses, bionomics, and geographical data are provided for each species when available.

We evaluated the morphometric variation of wing mite Periglischrus paracaligus Herrin and Tipton, along with the distribution of their host Leptonycteris yerbabuenae Martinez and Villa, in Mexico. A total of 115 female and 96 male specimens of P. paracaligus were used to conduct linear and geometric morphometric analyses. We assessed the influence of the geographic distribution of the migratory and nonmigratory populations of its bat host species on changes in size and shape on these parasites. Both analyses revealed high intraspecific variation in P. paracaligus, but subtle geographic differentiation. None of the approaches used identified a consistent pattern that separates unambiguously migratory from nonmigratory populations. Females presented more phenotypic variation than males and UPGMA analyses showed southern and northern colonies grouped in two distinct clades. Males on the other hand showed randomly grouped colonies with no geographic concordance. Interestingly, the most differentiated colony was the north Pacific colony of Jalisco. For both, males and females, isolation by distance (IBD) was not observed. We discuss these results as a possible scenario of contact between migratory populations located in northern Mexico with nonmigratory populations in other localities in central and southern Mexico conforming to a panmictic population along with their distribution range.

A new species of the subgenus Pseudoficalbia of the genus Uranotaenia is described from larvae found in a hole in a log at high altitude in montane forest in the Mongar District of Bhutan. Based on morphology, the larvae are sufficiently distinct from other species of the subgenus to be described and formally named here as Uranotaenia bhutanensis Somboon, Namgay & Harbach, n. sp. The species is a member of the Bimaculata Series. The chaetotaxy of the fourth-instar larva is provided.

Psorophora (Grabhamia) dimidiata Cerqueira (1943) was described based on the adult female and male. Later, descriptions of the male and female genitalia were published by Lane (in Neotropical Culicidae, vols. I & II. Editora da Universidade de São Paulo, S. P., Brazil, 1953) and Guedes et al. (in Catalogo ilustrado dos mosquitos da coleção do Instituto Nacional de Endemias Rurais. I. Gênero Psorophora Robineau-Desvoidy, 1827. Rev. Bras. Malariol. Doencas Trop. 12: 3–24; 1965), respectively. Here we describe the pupa and fourth-instar larva and redescribe the adult male and female genitalia and female cibarium. All stages (except the egg) are illustrated. Unambiguous recognition is provided. Distinctions from Ps. cingulata (Fabricius) and possible phylogenetic relationships are discussed.

Graphical Abstract

German cockroaches (Blattella germanica L.) harbor and disperse medically important pathogens and are a source of allergens that impact human health and wellbeing. Management of this pest requires an understanding of their distribution and dispersal. In this study, we collected German cockroaches from three apartment buildings in New Jersey, USA. We identified single-nucleotide polymorphisms (SNPs) from DNA extractions using next generation sequencing. We analyzed the SNPs and characterized cockroach population genetic structure using F_st, principal component, phylogenetic, and STRUCTURE analyses. We found significant differences in German cockroach population structure among the buildings. Within buildings, we found variable population structure that may be evidence for multiple colonization events. This study shows that SNPs derived from next generation sequencing provide a powerful tool for analyzing the genetic population structure of these medically important pests.

Tickborne disease cases account for over 75% of reported vector-borne disease cases in the United States each year. In addition to transmitting the agents of Lyme disease (Borrelia burgdorferi sensu strict [Spirochaetales: Spirochaetaceae] and Borrelia mayonii [Spirochaetales: Spirochaetaceae]), the blacklegged tick, Ixodes scapularis, and the western blacklegged tick, Ixodes pacificus collectively transmit five additional human pathogens. By mapping the distributions of tickborne pathogens in host-seeking ticks, we can understand where humans are at risk of contracting tickborne diseases and devise targeted strategies to prevent them. Using publicly available tickborne pathogen surveillance databases, internal CDC pathogen testing databases, and SCOPUS search records published since 2000, we mapped the county-level distribution of Borrelia miyamotoi (Spirochaetales: Spirochaetaceae), Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae), Ehrlichia muris eauclairensis (Rickettsiales: Ehrlichiaceae), Babesia microti (Piroplasmida: Babesiidae), and Powassan virus (Flaviviridae) reported in host-seeking I. scapularis or I. pacificus in the contiguous United States. We also updated recently published maps of the distributions of Borrelia burgdorferi sensu stricto and Borrelia mayonii. All seven pathogen distributions were more limited than the distributions of vector ticks, with at least one of the seven pathogens detected in 30 states out of 41 total states (73.2% of states) where vector ticks are considered to be established. Prevention and diagnosis of tickborne diseases rely on an accurate understanding by the public and health care providers of where people are at risk for exposure to infected ticks. Our county-level pathogen distribution maps expand on previous efforts showing the distribution of Lyme disease spirochetes and highlight counties where further investigation may be warranted.

Aedes-borne viruses (ABVs) such as dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV) contribute significantly to the global burden of infectious diseases, disproportionately affecting disadvantaged populations from tropical and subtropical urban areas. ABVs can be transmitted from female mosquitoes to their progeny by vertical transmission via transovarial and/or trans-egg vertical transmission and contribute to the maintenance of infected-mosquito populations year-round in endemic regions. This study describes the natural infection rate of DENV, CHIKV, and ZIKV in field-caught male Aedes (Sergentomyia) aegypti (Linnaeus) mosquitoes from Mérida, Yucatán, México, as a proxy for the occurrence of vertical virus transmission. We used indoor sequential sampling with Prokopack aspirators to collect all mosquitoes inside houses from ABV hotspots areas. Collections were performed in a DENV and CHIKV post-epidemic phase and during a period of active ZIKV transmission. We individually RT-qPCR tested all indoor collected Ae. aegypti males (1,278) followed by Sanger sequencing analysis for final confirmation. A total of 6.7% male mosquitoes were positive for ABV (CHIKV = 5.7%; DENV = 0.9%; ZIKV = 0.1%) and came from 21.0% (30/143) houses infested with males. Most ABV-positive male mosquitoes were positive for CHIKV (84.8%). The distribution of ABV-positive Ae. aegypti males was aggregated in a few households, with two houses having 11 ABV-positive males each. We found a positive association between ABV-positive males and females per house. These findings suggested the occurrence of vertical arbovirus transmission within the mosquito populations in an ABV-endemic area and, a mechanism contributing to viral maintenance and virus re-emergence among humans in post-epidemic periods.

Graphical Abstract

The aim of the present study was to explore resistance markers and possible biochemical resistance mechanisms in the Phlebotomine sand fly Phlebotomus papatasi in Esfahan Province, central Iran. Homogenous resistant strains of sand flies were obtained by exposing P. papatasi collected from Esfahan to a single diagnostic dose of DDT. The adults from the colony were tested with papers impregnated with four pyrethroid insecticides: Permethrin 0.75%, Deltamethrin 0.05%, Cyfluthrin 0.15%, and Lambdacyhalothrin 0.05% to determine levels of cross-resistance. To discover the presence of mutations, a 440 base pair fragment of the voltage gated sodium channel (VGSC) gene was amplified and sequenced in both directions for the susceptible and resistant colonies. We also assayed the amount of four enzymes that play a key role in insecticide detoxification in the resistant colonies. A resistance ratio (RR) of 2.52 folds was achieved during the selection of resistant strains. Sequence analysis revealed no knockdown resistance (kdr) mutations in the VGSC gene. Enzyme activity ratio of the resistant candidate and susceptible colonies were calculated for α-esterases (3.78), β-esterases (3.72), mixed function oxidases (MFO) (3.21), and glutathione-S-transferases (GST) (1.59). No cross-resistance to the four pyrethroids insecticides was observed in the DDT resistant colony. The absence of kdr mutations in the VGSC gene suggests that alterations in esterase and MFO enzymes are responsible for the resistant of P. papatasi to DDT in central Iran. This information could have significant predictive utility in managing insecticide resistant in this Leishmania vector.

Carboxypeptidase B (CPB) plays an important role in blood digestion in mosquitos, aiding the release of free amino acids. Anopheles CPB is a target to block malaria transmission because it facilitates Plasmodium invasion of the mosquito midgut. Our study aimed to discover inhibitors of Anopheles CPB to prevent Plasmodium development in the mosquito. The Anopheles gambiae cpb (Agcpb) gene without a signal sequence was cloned into the pET28b expression vector. The recombinant AgCPB protein was expressed in E. coli BL21(DE3) within inclusion bodies after induction with 0.5 mM isopropyl β-D-1-thiogalactopyranoside at 37°C for 4 h. The protein pellet was dissolved in 6 M urea, purified by affinity chromatography, and dialyzed in reaction buffer. The refolded recombinant AgCPB could digest the hippuryl-arginine substrate similarly to that of the commercial porcine pancreas CPB. The 20 top-scoring malaria box compounds from the virtual-screening results were then chosen for an in vitro inhibition assay against AgCPB. Four of the 20 malaria box compounds could inhibit AgCPB activity. The compound MMV007591 was the most potent inhibitor with an IC₅₀ at 0.066 µM. The results indicate that these candidate compounds may be utilized in drug development against mosquito CPB activity to curb malaria transmission.

Knock-down resistance (kdr) mutations in the voltage-gated sodium channel gene of Aedes species mosquitoes are biomarkers for resistance to pyrethroid insecticides. In the United States, few studies have reported kdr mutations among Aedes albopictus (Skuse) (Diptera: Culicidae) populations. In this study, we sought to compare the presence of kdr alleles among Ae. albopictus mosquitoes collected from Fort Bragg and Wake County, North Carolina. We collected 538 Ae. albopictus mosquitoes, including 156 from 4 sites at Fort Bragg, North Carolina and 382 from 15 sites in Wake County, North Carolina to compare the prevalence of kdr mutations. Of those successfully sequenced, we identified 12 (3.0%) mosquitoes with kdr mutations, all of which were attributed to variants at position 1534 within domain 3. All mutations were found in mosquitoes collected at Wake County sites; no mutations were identified in collections from Fort Bragg. There was a focus of mutations observed at the Wake County sites with approximately 92% (11 of 12) of the mosquitoes with the mutation coming from one site, where kdr mutations represented 24.4% (11 of 45) of all mosquitoes collected. We observed highly focal resistance in a suburban area of Raleigh, which may be attributable to peri-domestic mosquito control activities that involve area dispersal of pyrethroid insecticides. More robust surveillance is needed to monitor the emergence and spread of resistance.

Insecticides and repellents are routinely used in Brazil because of the high rates of arbovirus transmission and the nuisance caused by mosquitoes. However, few studies have assessed the effectiveness of repellents against mosquito populations that have been under exposure to xenobiotics, mainly insecticides and repellents. This study investigated the sensitivity of a field population of Aedes aegypti (Linnaeus, 1762) from a dengue-endemic area under high insecticide pressure to N,N-diethylmethylbenzamide (DEET), the active ingredient in common repellent products. The field (Laranjeiras, Sergipe, Brazil) and laboratory (Rockefeller) populations were characterized for the presence of the Val1016Ile kdr mutation, associated with pyrethroid resistance, and locomotor activity. Repellency bioassays were performed to assess the response of the mosquitoes to human odor by exposing them to 10% DEET applied to the skin in ethanol. Samples from the field population showed higher frequency of the kdr mutation, 21.9% homozygous and 21.9% heterozygous, greater locomotor activity and greater sensitivity to DEET than the laboratory population. These results suggest increased sensitivity to DEET in field populations and a possible interaction between insecticide exposure and sensitivity to DEET.

A larval medium for house fly (Musca domestica L.) suitable for house fly parasitoid production was developed from locally available and inexpensive food ingredients. Biological parameters and life table parameters were estimated for house flies treated with five different diets. It was found that percentage survival of 1st–3rd instar larvae of house fly, percentage of pupation, percentage of hatching adults, growth index, and life table parameters (net reproductive rate [R₀] the cohort generation time [T], intrinsic rate of increase [r], and finite rate of increase [λ]) were significantly highest on larvae produced on diet 5 (composed of rice barn and chicken feed). Diet composition and performance of house fly larvae in larval medium are discussed, based on the results. The results of Spalangia gemina Bouček (Hymenoptera, Pteromalidae) parasitization on house fly pupae revealed that the number of total parasitized pupa and number of parasitoids hatched from pupa reared with diet 5 (rice bran and chicken feed) were greater than with diet 1 (rice bran, powdered milk, dry yeast, fish meal, soybean meal, and chicken feed). Additionally, S. gemina offspring from diet 5 treatment had a higher proportion of females. Our results indicated that pupal size resulting from larval diet was an important factor for parasitization.

Host feeding patterns and the prevalence of infection with Rickettsia parkeri were determined for the primary vector, Amblyomma maculatum Koch as well as sympatric tick species A. americanum (Linnaeus) and Dermacentor variabilis (Say) collected from a reconstructed prairie in the Piedmont region of North Carolina during 2011 and 2012. The occurrence of R. parkeri among A. maculatum adults and nymphs was 36.9% (45/122) and 33.3% (2/6), respectively. Rickettsia parkeri was detected in a single male A. americanum 2.3% (1/43). A PCR-reverse line blot hybridization assay of a 12S rDNA fragment amplified from remnant larval and nymphal bloodmeals of host-seeking ticks was used to identify bloodmeal hosts. Of the tick samples tested, bloodmeal host identification was successful for 29.3% (12/41) of adult A. americanum and 39.2% (20/51) of adult D. variabilis. For A. maculatum, bloodmeal host identification was successful for 50% (61/122) of adults collected from vegetation and 100% (4/4) of nymphs removed from cotton rats (Sigmodon hispidus Say and Ord). The cotton rat was the most common bloodmeal host with 59.0% (36/61) identified for adult A. maculatum. No statistically significant association was observed, however, between bloodmeal host and pathogen prevalence for any tick species. While the cotton rat was an important bloodmeal host for A. maculatum nymphs, this vertebrate did not appear to be the primary source of R. parkeri infection for A. maculatum.

Very little data exist on the biology of an afrotropical rainforest mosquito Eretmapodites (Er.) in a world undergoing dramatic changes due to deforestation. The aim was to assess the efficacy of different trapping methods in the collection of Er. mosquito in forested area.

This was a longitudinal study involving collection of mosquitoes for over two years. Multiple collection methods (grouped into two categories), were used; i) net baited and un-baited traps to collect adults, ii) techniques that target immature stages subsequently reared to adults. All males were identified by genitalia dissection. Five thousand seven hundred and four mosquitoes representing 11 genera among which 2,334 Er. were identified. Mosquito abundance was highest in the net traps (n = 1276 (56.4%)) and sweep nets (n = 393(17.4%)) respectively. The abundance was highest in green colored net traps (435(34.09%)) with significant value of χ²= 40.000, P < 0.001 and in pigeons baited traps (473 (37.06%)) with significant value of χ²= 42.000, P = 0.003. The diversity ranges from H' = 2.65; DS = 0.84; SR = 24; ACE = 24.77 in sweep net to H' = 0; DS = 0; SR = 1; ACE = 1 in rock pool among males mosquitoes. While for females, H = 1.14; DS = 0.71; SR = 5; ACE = 5.16, in sweep net to H = 0; DS = 0; SR = 1; ACE = 1 in rock pool, tarpaulin, resting cage.

Net traps, bamboo pot, and sweep netting are efficient in collecting high abundance of forest mosquitoes in the Talanagaye rainforest.

The western black-legged tick (Ixodes pacificus) is the most frequently identified human-biting tick species in the western United States and the principal vector of at least three recognized bacterial pathogens of humans. A potentially pathogenic Rickettsia species, first described in 1978 and recently characterized as a novel transitional group agent designated as Rickettsia tillamookensis, also exists among populations of I. pacificus, although the distribution and frequency of this agent are poorly known. We evaluated DNA extracts from 348 host-seeking I. pacificus nymphs collected from 9 locations in five California counties, and from 916 I. pacificus adults collected from 24 locations in 13 counties, by using a real-time PCR designed specifically to detect DNA of R. tillamookensis. DNA of R. tillamookensis was detected in 10 (2.9%) nymphs (95% CI: 1.6–5.2%) and 17 (1.9%) adults (95% CI: 1.2–3.0%) from 11 counties of northern California. Although site-specific infection rates varied greatly, frequencies of infection remained consistently low when aggregated by stage, sex, habitat type, or geographical region. Four novel isolates of R. tillamookensis were cultivated in Vero E6 cells from individual adult ticks collected from Alameda, Nevada, and Yolo counties. Four historical isolates, serotyped previously as ‘Tillamook-like’ strains over 40 yr ago, were revived from long-term storage in liquid nitrogen and confirmed subsequently by molecular methods as isolates of R. tillamookensis. The potential public health impact of R. tillamookensis requires further investigation.

Higher temperatures expected in a future warmer climate could adversely affect the growth and development of mosquitoes. This study investigated the effects of elevated temperatures on longevity, gonotrophic cycle length, biting rate, fecundity, and body size of Anopheles gambiae (s.l.) (Diptera: Culicidae) mosquitoes. Anopheles gambiae (s.l.) eggs obtained from laboratory established colonies were reared under eight temperature regimes (25, 28, 30, 32, 34, 36, 38, and 40°C), and 80 ± 10% RH. All adults were allowed to feed on a 10% sugar solution soaked in cotton wool; however, some mosquitoes were provided blood meal using guinea pig. Longevity was estimated for both blood-fed and non-blood-fed mosquitoes and analyzed using the Kaplan–Meier survival analysis. One-way ANOVA was used to test the effect of temperature on gonotrophic cycle length, biting rate, and fecundity. Adult measurement data were log-transformed and analyzed using ordinary least square regression with robust standard errors. Increasing temperature significantly decreased the longevity of both blood-fed (Log-rank test; X²(4) = 904.15, P < 0.001) and non-blood-fed (Log-rank test; X²(4) = 1163.60, P < 0.001) mosquitoes. In addition, the fecundity of mosquitoes decreased significantly (ANOVA; F(2,57) = 3.46, P = 0.038) with an increase in temperature. Body size (β = 0.14, 95% CI, 0.16, 0.12, P < 0.001) and proboscis length (β = 0.13, 95% CI, 0.17, 0.09, P < 0.001) significantly decreased with increasing temperature from 25 to 34°C. Increased temperatures expected in a future warmer climate could cause some unexpected effects on mosquitoes by directly influencing population dynamics and malaria transmission.

Chagas disease is endemic in ∼70% of Ecuador. Rhodnius ecuadoriensis and Triatoma carrioni (Hemiptera: Reduviidae) are the primary vectors of Chagas disease in Southern Ecuador. This study tested the effectiveness of selective deltamethrin application of Domiciliary Units (DUs) infested with triatomines, coupled with community education activities and a community-based surveillance system. Ten communities were selected in Loja Province, 466 DUs were examined, of these, 5.6% were infested with R. ecuadoriensis (Density [D] = 4 triatomines/DUs searched, Crowding [CR] = 71 triatomines/infested house, Colonization Index [CI] = 77% infested DUs with nymphs) and 8% with T. carrioni (D = 0.6, CR = 7, CI = 64%). Infested DUs were sprayed with deltamethrin. Subsequent visits were conducted at 6 and 12 mo after spraying. At each time point, new entomological searches were carried out in all DUs. All entomological indexes dropped significantly for the primary vector species one year after the initial intervention (R. ecuadoriensis: I = 2%, D = 0.1, CR = 7, CI = 100%; T. carrioni: I = 1.6%, D = 0.1, CR = 5.5, CI = 50%). Fifteen min educational talks were conducted in every DUs and workshops for schoolchildren were organized. Community-based surveillance system was established. However, there is a high risk of DUs reinfestation, possibly from sylvatic habitats (especially of R. ecuadoriensis) and reinforcing educational and surveillance activities are necessary.

We report the multi-year collection of the Gulf Coast tick, Amblyomma maculatum Koch (Acaridae: Ixodida: Ixodidae) in Staten Island, NewYork City (NYC) as well as their detection in Brooklyn, NYC, and in Atlantic and Cumberland counties in southern New Jersey, USA. The first and most common detections were of adults, however in Freshkills Park on Staten Island larvae were also collected in a following year. The presence of larvae indicates that adults are successfully finding hosts in Staten Island. While it is still unknown how A. americanum reached Staten Island, immatures of this species often parasitize migratory birds, which are now often seen in Freshkills Park. We describe the landscape features of the area in Staten Island where populations were highest and larvae were detected, which could have facilitated the establishment of A. maculatum. Notably, we also report the presence of human pathogens Rickettsia parkeri in 5/10 (50%) of adults tested and R. felis in 1/24 (4.17%) of larvae tested. In addition to established populations in Staten Island we found evidence of A. maculatum in NJ and other NYC boroughs, suggesting current or future establishment is possible. The failure thus far to detect established populations in these areas may be due to inherent difficulties in detecting low density, spatially heterogeneous incipient populations, which could require targeted surveillance efforts for this species. We discuss the consequences to public health of the establishment of A. maculatum and detection of two additional rickettsial pathogens in the densely populated northeastern United States.

Over the past decade, Haemaphysalis longicornis, the Asian longhorned tick, has undergone a geographic range expansion in the United States, from its historical range in east Asia. This tick has been characterized by its frequent parasitism of livestock, an ability to reproduce through parthenogenesis, and its ability to transmit a variety of vector-borne pathogens to livestock, wildlife, and human hosts in its native geographic range. Thus far in the United States, 17 states have reported H. longicornis populations, including 38 counties in Virginia. These numbers come from presence-absence reports provided to the U.S. Department of Agriculture, but little has been reported about this ticks' seasonality inVirginia or its habitat preferences. Our current study detected H. longicornis populations in seven of the nine surveyed counties in Virginia. Haemaphysalis longicornis were observed in multiple habitat types including mixed hardwood forests and pastures, with abundant H. longicornis populations detected at one particular pasture site in Wythe County. This study also attempted to investigate environmental conditions that may be of importance in predicting tick presence likelihood. While sample size limited the scope of these efforts, habitat type and climatic metrics were found to be important indicators of H. longicornis collection success and abundance for both the nymphal and larval life stages. This current study reports useful surveillance data for monitoring these tick populations as they become established in the western half ofVirginia and provides insight into their current distribution and maintenance over a large study region.

Tick-borne diseases are a growing public health problem in the United States, and the US northeast has reported consistently high case rates for decades. Monmouth County, New Jersey, was one of the earliest jurisdictions to report Lyme disease cases in 1979 and reports several hundred cases per year nearly 40 yr later. In the time since, however, tick-borne health risks have expanded far beyond Lyme disease to include a variety of other bacterial pathogens and viruses, and additional vectors, necessitating a continually evolving approach to tick surveillance. In 2017, Monmouth County initiated an active surveillance program targeting sites across three ecological regions for collection of Ixodes scapularis Say (Acari: Ixodidae) and Amblyomma americanum L. (Acari: Ixodidae) as well as testing via qPCR for associated bacterial pathogens. During the first five years of this program (2017–2021), we report high levels of spatiotemporal variability in nymphal density and infection prevalence in both species, limiting the granularity with which human risk can be predicted from acarological data. Nonetheless, broader patterns emerged, including an ongoing trend of A. americanum dominance, risks posed by Borrelia miyamotoi, and the frequency of coinfected ticks. We present some of the first county-level, systematic surveillance of nymphal A. americanum density and infection prevalence in the northeastern US. We also documented a temporary decline in Borrelia burgdorferi that could relate to unmeasured trends in reservoir host populations. We discuss the implications of our findings for tick-borne disease ecology, public health communication, and tick surveillance strategies in endemic areas.

Some species of Culicoides Latreille (Diptera: Ceratopogonidae) can be pests as well as pathogen vectors, but data on their distribution in Ontario, Canada, are sparse. Collecting this baseline data is important given ongoing, accelerated alterations in global climate patterns that may favor the establishment of some species in northern latitudes. Culicoides spp. were surveyed using UV light traps over two seasons in 2017 and 2018 at livestock farms in southern Ontario, Canada. Two Culicoides spp. not previously recorded in Canada were identified, C. bergi and C. baueri, representing new country and provincial records. Unlike some congenerics, these two species are not currently recognized as vectors of pathogens that pose a health risk to humans, livestock or wildlife in North America. However, the possibility that these Culicoides species may have recently expanded their geographic range, potentially in association with climate and/or landscape changes, warrants ongoing attention and research. Furthermore, our results provoke the question of the potential undocumented diversity of Culicoides spp. in Ontario and other parts of Canada, and whether other Culicoides spp. may be undergoing range expansion. The current and future distributions of Culicoides spp., and other potential vectors of human, agricultural, and wildlife health significance, are important to identify for proper disease risk assessment, mitigation, and management.

The tick genus Amblyomma Koch, 1844 (Acari: Ixodidae) has received little attention in Malaysia; therefore, its associated hosts and distribution records are poorly known. In this study, we collected six Amblyomma sp. individuals (two larvae and four adults) that infested a common treeshrew, Tupaia glis (Diard, 1820) (Scandentia: Tupaiidae) caught in a recreational area in Sungai Lembing, Pahang (West Malaysia). The adult female ticks were morphologically identified according to taxonomic keys prior to molecular identification using cytochrome oxidase subunit I (COI) and 16S rDNA genes. The ticks were genetically verified as Amblyomma geoemydae (Cantor, 1847) with 98%–99% similarity to the available GenBank sequences. Neighbor-joining (NJ) trees indicated that A. geoemydae was clearly distinguished from other Amblyomma ticks and this was supported with high bootstrap values. This paper is the first to report A. geoemydae ticks infesting T. glis and provides a new tick-host record from West Malaysia. This information is significant for further investigation, specifically on this tick species as potential vector of tick-borne disease (TBD) agents.

Flies and other arthropods mechanically transmit multiple pathogens and a recent experimental study demonstrated house flies, Musca domestica L. (Diptera: Muscidae), can mechanically transmit SARS-CoV-2. The purpose of this study was to explore the possibility of mechanical transmission of SARS-CoV-2 by domestic insects and their potential as a xenosurveillance tool for detection of the virus. Flies were trapped in homes where at least one confirmed human COVID-19 case(s) resided using sticky and liquid-baited fly traps placed inside and outside the home in the Texas counties of Brazos, Bell, and Montgomery, from June to September 2020. Flies from sticky traps were identified, pooled by taxa, homogenized, and tested for the presence of SARS-CoV-2 RNA using quantitative reverse transcription PCR (RT-qPCR). Liquid traps were drained, and the collected fluid similarly tested after RNA concentration. We processed the contents of 133 insect traps from 40 homes, which contained over 1,345 individual insects of 11 different Diptera families and Blattodea. These individuals were grouped into 243 pools, and all tested negative for SARS-CoV-2 RNA. Fourteen traps in seven homes were deployed on the day that cat or dog samples tested positive for SARS-CoV-2 RNA by nasal, oral, body, or rectal samples. This study presents evidence that biting and nonbiting flies and cockroaches (Blattodea) are not likely to contribute to mechanical transmission of SARS-CoV-2 or be useful in xenosurveillance for SARS-CoV-2.

Haemaphysalis longicornis Neumann, a vector of various pathogens with medical and veterinary importance, is a recent invasive species in the United States. Like many tick species, discerning H. longicornis from congeners can be a challenge. To overcome the difficulty of morphological identification, a Taqman quantitative real-time PCR based on the internal transcribed spacer gene (ITS2) was developed for quick and accurate identification of H. longicornis with a detection limit of as low as 19.8 copies. We also applied the assay to 76,004 archived ticks and found 37 ticks were H. longicornis. One H. longicornis was submitted from Warren, Somerset County, New Jersey in June 2015, 2 yr earlier than the initial report from the United States. None of these 37 H. longicornis was positive for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, B. miyamotoi, B. mayonii, Babesia microti, or Ehrlichia muris–like agent.