The decomposition process of human (and other mammalian) remains is influenced by numerous factors such as the environmental temperature, relative humidity, precipitation, geographical location of the remains, as well as the medical conditions of and any injuries sustained by the deceased person. The decomposition process generally follows a consistent pattern (fresh, bloat, active decomposition, advanced decomposition, and dry stage). Understanding the time-span of each stage of decomposition and the influence of the biotic and abiotic factors involved is imperative when trying to estimate the time elapsed since death (postmortem interval [PMI]). Over the course of decomposition, the tissues are gradually consumed by necrophagous insects and bacteria. The environmental temperature and its variations influence how insects colonize the remains, having a significant impact on their presence and developmental cycle. Additionally, the bacterial community colonizing decomposing tissues is also greatly affected by variations in environmental temperature. Because both the rate of decomposition of human remains and the relative abundance of certain insects and bacterial species are all temperature-dependent, insect and bacterial colonization data are useful as key points for the PMI estimation.This article reviews the current literature documenting the data collected on the occurrence and development cycles of predominant necrophagous Diptera (Calliphoridae, Muscidae, Sarcophagidae) and bacterial taxa involved in the decomposition of various carcasses at different temperatures. This review will impact the forensic community by providing an overview on the temperature, insect and bacterial records for the PMI estimation, seeking to aid forensic entomologists, microbiologists, pathologists, and the legal community.

We conducted the first pilot radio telemetry study of hematophagous arthropods by placing transmitters on wild-caught triatomine insects (‘kissing bugs’), vectors of the Chagas disease parasite. InTexas—a recognized hotspot for triatomine diversity and locally-acquired human and animal Chagas disease—we tagged five female and four male Triatoma gerstaeckeri (Stål) (Hemiptera: Reduviidae), as well as one female and one male Triatoma sanguisuga (Leconte) (Hemiptera: Reduviidae) in three counties from 2015 to 2017. In comparative trials, placement of the transmitter on the dorsal side of the abdomen underneath the hemelytra wings, with the transmitter antenna shortened to 3 cm, yielded the best results. We tracked the movements of the 11 tagged bugs over an average of 4.8 d (range of 1 to 12 d) and detected 18 movement events with an average distance of 3.8 m (range of 1 to 20 m). This pilot study demonstrates the potential utility for using telemetry as a tool for studying fine-scale non-flight movement of triatomines and the discovery of cryptic resting habitats. Future studies using this or similar technologies to study movement and behavior of triatomines could test for site-fidelity of resting habitats and provide novel insight into aspects of vector biology that could be targeted in disease risk reduction efforts.

The seasonal activity pattern of immature Ixodes scapularis Say (Acari: Ixodidae) varies geographically in the United States, which may affect the efficiency of transmission cycles of pathogens transmitted by this species. To study the factors that determine seasonality, a multiyear study at seven sites across the geographic range of I. scapularis systematically collected questing ticks by flagging/dragging, and feeding ticks by capture of their hosts. The observed phenology patterns were consistent with previous studies reporting geographic variation in seasonal tick activity. Predictions of seasonal activity for each site were obtained from an I. scapularis simulation model calibrated using contemporaneous weather data. A range of scenarios for life-cycle processes—including different regimes of temperature-independent behavioral and developmental diapause, variations in temperature–development rate relationships, and temperature-dependent tick activity—were used in model formulations. These formulations produced a range of simulations of seasonal activity for each site and were compared against the field observed tick data using negative binomial regression models. Best fit scenarios were chosen for each site on the basis of Akaike's information criterion and regression model parameters. This analysis suggests that temperature-independent diapause mechanisms explain some key observed variations in I. scapularis seasonality, and are responsible in part for geographic variations in I. scapularis seasonality in the United States. However, diapause appears to operate in idiosyncratic ways in different regions of the United States, so further studies on populations in different regions will be needed to enable predictive modeling of climatic and climate change effects on I. scapularis seasonal activity and pathogen transmission.

Aldrichina grahami (Aldrich) (Diptera: Calliphoridae) is a forensically important species mainly found in Asia and is one of the first species to colonize a corpse in early spring and late autumn when the ambient temperature is slightly lower. In this study, A. grahami was collected from the Yangtze River Delta region and reared at seven constant temperatures ranging from 16 to 34°C to investigate the indices of development duration, accumulated degree hours, and larval body length. Using these results, we generated an isomorphen diagram, thermal summation model, and isomegalen diagram for A. grahami. Equations of the variation in larval body length with time after hatching and variation in time after hatching with body length were also obtained. A. grahami can complete its life cycle in 16–28°C, while its development was incomplete at 31 and 34°C, where puparia failed to transform into adults and eggs failed to hatch, respectively. The mean (±SD) developmental durations of A. grahami from egg to adult at 16, 19, 22, 25, and 28°C were 685.5 ± 18.8, 540.3 ± 15.9, 454.3 ± 19.4, 388.8 ± 19.0, and 335.5 ± 8.7 h, respectively. The mean (±SE) developmental threshold temperature D₀ and the thermal summation constant K were determined as 4.33 ± 0.21°C and 7,985.9 ± 104.5 degree hours, respectively. The results of this study provide fundamental development data for the use of A. grahami in minimum postmortem interval estimations.

This study assesses the succession of insects on rabbit carcass.The study was conducted in sunny and shaded sites in the west of Iran during four seasons in 2016 and 2017. Based on the results of this study, various factors such as body size, carcass location, and carcass injury could affect the pattern of insect succession on carcass. In this study, we estimated the elapsed time since death (R² > 0.98, P = 0.00) based on carcass weight loss during the stages of decomposition and the cubic method. Jaccard analysis was performed to determine the similarity of insect taxa during decomposition of rabbit carrion in two different sites during a period of four seasons. Succession pattern analysis for necrophagous insects in both habitats showed a similarity between bloat and decay stages for each habitat. On the other hand, pairwise similarities in taxa were low at the fresh and dry stages, however increased at bloat and decay stages of decomposition. This study shows that succession has some limitations in determining the elapsed time of death. Therefore, the use of source of information such as a weight loss model seems to be essential.

Geographical regions have a major effect on the arrival times of different insect species on carrion. This means that data generated in one region should not be used to determine time of death in a different region. In the present study, we demonstrate the effect of geographical location on the diversity of carrion beetles in Saudi Arabia, whereas the mitochondrial cytochrome c oxidase subunit I (mtCOI) barcodes were used as a marker for molecular identification of the carrion beetles at a dry stage on sheep carrion. We analyzed 819 adult beetles belonging to nine species originating from Riyadh (609 beetles), Jazan (157 beetles), and Arar (53 beetles). In Riyadh, results showed the presence of six species belonging to three different families. On the other hand, in Jazan five species belonging to four families were collected. From Arar, five species belonging to three families were collected. By comparing between individuals of the same species from different regions, individuals of Necrobia rufipes DeGeer (Coleoptera: Cleridae) showed the highest intraspecific variations 0–20%, while individuals of Saprinus splendens Paykull and Saprinus semistriatus Scriba (Coleoptera: Histeridae) showed the lowest intraspecific variations 0–1%. Interspecific variability was also measured between collected and identified species, with differences revealed to be in the range of 3.8–29.8%.The results are important from an ecological point of view and for Medico-Legal Forensic Entomology.

The sand fly fauna in Hipólito Yrigoyen, Argentina, a locality where cutaneous leishmaniasis cases occur, was surveyed with zones of higher abundance of sand flies correlated to vegetation cover estimated through normalized difference vegetation index (NDVI). Sand flies were collected with 10 CDC traps during six nights, from December 2009 to January 2010. A map was built of expected sand flies abundance in which levels of NDVI were categorized. In total, 1,392 Phlebotominae (Diptera: Psychodidae) specimens were collected, comprised of the following species: Nyssomyia neivai (Pinto 1926), Migonemyia migonei (França 1920), species of the cortelezzii complex (Brèthes 1923), Evandromyia sallesi (Galvão & Coutinho 1940), and Psathyromyia shannoni (Dyar 1929). Positive correlations were found between the abundance of sand flies and the NDVI (P < 0.05) for buffer areas of <150 m radii from the trap location points, i.e., the sand fly abundance was greater where vegetation cover and density were greater. In this context, plant cover should be taken into account to prioritize surveillance and control areas within the program of sand flies control in northern Argentina.

DNA, the blue print of life, is densely wrapped around histone proteins to form chromatin. Chromatin remodeling ATPases unwind histone-DNA interactions to facilitate DNA transcription, modification, and repair. Four genes involved in chromatin remodeling, namely, imitation SWI (iswi), chromodomain-helicase-DNA-binding protein 1 (chd-1), DNA helicase INO80 (ino80) and mi-2 were silenced through the injection of dsRNA, and phenotypes were assessed in bed bugs. Bed bugs were injected with 0.2 μg dsRNA per insect between the last thoracic segment and first abdominal segment using a fine capillary tube fitted to a nanoinjector. We observed a significant reduction in reproductive potential with all the genes tested, suggesting the essential function of chromatin remodeling ATPases in many cellular processes including egg-laying and egg-hatching. Knockdown of mi-2 and iswi completely inhibited oviposition over time. Real-time quantitative polymerase chain reaction confirmed significant knockdown of targeted mRNAs for at least 30 d, which supports persistence of RNAi in bed bugs. In addition, we observed a significant depletion of targeted transcripts in eggs laid by bed bugs injected with dsRNAs specific to chromatin remodeling ATPases. This study demonstrates the importance of chromatin remodeling ATPase in bed bug reproduction.

The members from Culex pipiens complex are the primary vector of various arboviruses including West Nile virus and Eastern equine encephalitis in the United States. Within Cx. pipiens complex, there are three biotypes that differ largely in habitat, bloodmeal preference, mating behavior, and overwintering strategy. The three biotypes are Cx. pipiens form molestus (F.) (Diptera: Culicidae), Cx. pipiens f. pipiens (L.) (Diptera: Culicidae), and Cx. pipiens quinquefasciatus (Diptera: Culicidae) (also known as Cx. quinquefasciatus). It is clear that additional genetic markers, molecular markers based on single nucleotide polymorphisms (SNPs), were identified from the Cx. pipiens complex species. Genomic DNA was extracted from adult females of f. pipiens and f. molestus and was amplified by polymerase chain reactions. Thirty out of the 100 primer pairs showed amplification and were used for SNP identification. All thirty genes contained biotype-specific SNPs: 10 were located on the Cx. pipiens complex genetic map from previous genetic studies. We also tested a high-resolution DNA melting analysis as a biotype identification method by examining the SNPs in the two genes (CPIJ005487 and CPIJ002074). Our method provides a high confidence for biotype determination among the three Cx. pipiens complex mosquitoes.

Data deposition: NCBI SNP Database (ssSNP accession nos.: ss947848635–ss947848878)

Seven populations of Simulium parahiyangumTakaoka & Sigit (Diptera: Simuliidae), a geographically widespread nominal species of black fly in Southeast Asia, were morphologically and molecularly studied. Three morphoforms based on male and pupal morphological features, and two primary lineages based on the COI gene sequence analysis were recognized. Morphoform 1 (lineage 1) from Sarawak, Malaysia, is identified as S. parahiyangum sensu stricto and morphoform 2 (lineage 2) from Thailand and Vietnam, and morphoform 3 (lineage 1) from Peninsular Malaysia are each regarded as distinct species, although morphoform 3 is partially homosequential for the COI gene with morphoform 1. Morphoforms 2 and 3 are described as Simulium ngaoense sp. nov. and Simulium sazalyi sp. nov., respectively. Overall, S. parahiyangum is not a single geographic generalist but a composite of multiple species.

We investigated the quill mite fauna of the family Syringophilidae (Acariformes: Prostigmata) associated with Sunbirds (Passeriformes: Nectariniidae) in the Sub-Saharan region. Six hundred eleven host individuals belonging to 52 species were examined. Syringophilid parasitization prevalence (44 hosts of 15 species) ranged from 1.4 to 75% in particular host species. In the collected material, we have found seven quill mite species and among them four are new for science: 1) Aulobia afroanthreptes Skoracki & Zmudzinski sp. nov. from Anthreptes neglectus Neumann, 1922 and Anthreptes longuemarei (Lesson, 1831), both from Tanzania; 2) Syringophiloidus nectariniae Skoracki & Zmudzinski sp. nov. from Hedydipna collaris (Vieillot, 1819) fromTanzania; 3) Aulonastus nectariniphilus Skoracki & Zmudzinski sp. nov. from Anthreptes reichenowi Gunning, 1909 from Tanzania and Nectarinia tacazze (Stanley, 1814) from Ethiopia; and 4) Picobia hedydipna Skoracki & Zmudzinski sp. nov. from Hedydipna collaris from Kenya. Representatives of the genera AulonastusKethley, 1970 and SyringophiloidusKethley, 1970 are recorded on Sunbirds for the first time. The following new host species are given: Cinnyris mariquensis Smith, 1836, C. shelleyi Alexander, 1899 and C. regius Reichenov, 1893, all fromTanzania for Aulobia nectariniaeSkoracki & Glowska, 2008; A. neglectus and A. longuemarei, both fromTanzania for Neoaulonastus cinnyrisKlimovicova et al., 2014; Cyanomitra verreauxii (Smith, 1832) fromTanzania and Cinnyris chalybeus (Linnaeus, 1766) from South Africa for Picobia oritisSkoracki et al., 2009. Additionally, host-parasite relationships are discussed.

The species of the genus Mepraia Mazza, Gajardo Tobar and Jörg (Hemiptera, Reduviidae:Triatominae) exhibit polymorphism in their hemelytra. The females of all species are always micropterous, but the males can be micropterous, brachypterous, or macropterous. Mepraia gajardoi Frías, Henry and González (Hemiptera: Reduviidae) males are always brachypterous, Mepraia parapatrica Frías-Lasserre (Hemiptera: Reduviidae) males are macropterous or brachypterous and Mepraia spinolai (Porter) (Hemiptera: Reduviidae) males are macropterous, brachypterous, or micropterous. Here, we determined distinctive characteristics of the hemelytra of the males of these three species and found that the length of the hemelytra of the brachypterous males in these three species differs statistically, as does their shape and morphology. These differences are particularly apparent at the apex of the hemelytra, the anterior part of the corium, in the morphology and number of the ctenidia of coaptors, and presence or absence of the cross vein of the hemelytra. Based on this, we propose a key based on hemelytra characteristics to identify the males of the three Mepraia species.

Nyssorhynchus strodei (Root) is an understudied species of potential epidemiological importance, having been found naturally infected in Brazil with Plasmodium falciparum Welch, Plasmodium vivax Grassi & Feletti, and Plasmodium malariae Feletti & Grassi. It belongs to the strodei subgroup that includes Nyssorhynchus albertoi (Unti), Nyssorhynchus arthuri (Unti), Nyssorhynchus rondoni (Neiva & Pinto), Nyssorhynchus striatus (Sant’Ana & Sallum), and three unnamed species, Nyssorhynchus arthuri B, Nyssorhynchus arthuri C, and Nyssorhynchus arthuri D. As the accurate identification of vector species is of fundamental importance for public health entomology, the aim of the study was to provide additional information for the presence of seven species that had been previously misidentified as Ny. strodei. Bayesian phylogenetic analyses using DNA sequences of the genes COI mtDNA, white, CAD and CAT nuclear genes confirmed Ny. albertoi, Ny. arthuri, Ny. strodei, and Ny. striatus as distinct groups within the strodei subgroup and corroborated the presence of three undescribed species under the name Ny. arthuri. Results of the GMYC model analysis corroborated Ny. arthuri B, Ny. arthuri C, and Ny. arthuri D; however, they did not distinguish between Ny. strodei and Ny. albertoi. Predicted distribution of seven species based on maximum entropy in MaxEnt showed that each species has its specific ecological niche suitability.

Blacklegged ticks (Ixodes scapularis Say, Acari: Ixodidae) are the most commonly encountered and medically relevant tick species in New York State (NY) and have exhibited recent geographic range expansion. Forests and adjacent habitat are important determinants of I. scapularis density and may influence tick-borne pathogen prevalence. We examined how percent forest cover, dominant land cover type, and habitat type influenced I. scapularis nymph and adult density, and associated tick-borne pathogen prevalence, in an inland Lyme-emergent region of NY. I. scapularis nymphs and adults were collected from edge and wooded habitats using tick drags at 16 sites in Onondaga County, NY in 2015 and 2016. A subsample of ticks from each site was tested for the presence of Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti using a novel multiplex real-time polymerase chain reaction (PCR) assay, and deer tick virus using reverse transcription–PCR. Habitat type (wooded versus edge) was an important determinant of tick density; however, percent forest cover had little effect. B. burgdorferi was the most commonly detected pathogen and was present in ticks from all sites. Ba. microti and deer tick virus were not detected. Habitat type and dominant land cover type were not significantly related to B. burgdorferi presence or prevalence; however, ticks infected with A. phagocytophilum and B. miyamotoi were collected more often in urban environments. Similarity between B. burgdorferi prevalence in Onondaga County and hyperendemic areas of southeastern NY indicates a more rapid emergence than expected in a relatively naive region. Possible mechanistic processes underlying these observations are discussed.

Vector abundance plays a key role in transmission of mosquito-borne disease. In Hawaii, Aedes albopictus (Skuse) (Diptera: Culicidae), the Asian tiger mosquito, has been implicated in locally-transmitted dengue outbreaks, while Culex quinquefasciatus Say (Diptera: Culicidae), the southern house mosquito, is the primary vector of avian malaria, a wildlife disease that has contributed to declines and extinctions of native Hawaiian birds. Despite the importance of these introduced species to human and wildlife health, little is known about the local-scale drivers that shape mosquito abundance across lowland Hawaii, where forest, agricultural, and residential land uses are prevalent. We examined landscape, larval habitat, and climate drivers of Ae. albopictus and Cx. quinquefasciatus abundance in eight lowland wet forest fragments on the Big Island of Hawaii. We found that the abundance of both species increased with the proportion of surrounding developed land and the availability of larval habitat, which were themselves correlated. Our findings suggest that conversion of natural habitats to residential and agricultural land increases mosquito larval habitats, increasing the abundance of Ae. albopictus and Cx. quinquefasciatus and increasing disease risk to humans and wildlife in Hawaii. Our results further indicate that while source reduction of artificial larval habitats—particularly moderately-sized human-made habitats including abandoned cars and tires—could reduce mosquito abundance, eliminating larval habitat will be challenging because both species utilize both natural and human-made larval habitats in lowland Hawaii.

Tick-borne diseases are poised to devastate the North American cattle industry if infected ticks invade the country either by importation of an infested-animal or with natural host migration. Our research objectives were to identify sources for invasive-tick monitoring and use those sources to describe seasonal and regional impacts on infestation prevalence and burden of ticks on beef cattle.Throughout the state of Tennessee, we sampled 25% of the total herd size (or 10 animals) at three university-operated research and education centers (RECs) (sentinel sites), six livestock auctions (check-stations), and nine Extension agents at 21 producer locations (tick scouts) from 2015 to 2016. From 1,817 sampled cattle 740 ticks (Acari: Ixodidae) were collected including 573 Amblyomma americanum (L.) (77.4%), 125 A. maculatum Koch (16.9%), 35 Dermacentor variabilis (Say) (4.7%), and 3 Ixodes scapularis (Say) (0.4%). Western and middle Tennessee were significantly different in infestation prevalence and burden of A. maculatum. For A. maculatum and the species total, infestation prevalence and burden were greater in spring than fall. Auctions (check stations) and RECs (sentinels) had the greatest infestation prevalence of A. maculatum, and the greatest burden of A. maculatum and D. variabilis. High-risk locations clustered in western and middle Tennessee, with low-risk locations in middle and easternTennessee. Results from this study provide knowledge necessary to initiate control measures, including seasonal phenology and regional distribution of current tick threats. Use of RECs as sentinel sites and routine tick surveillance at livestock auctions serving as check stations should be used for mitigating invasive tick threats.

Flies of the family Calliphoridae play a variety of ecological roles. They carry various pathogens and cause myiasis in humans and livestock, but they are useful to forensic entomology and in larval therapy. Mesembrinellidae flies, formerly classified in the family Calliphoridae, are good bioindicators of human interference in natural environments. In this study, we carried out an inventory of the Calliphoridae and Mesembrinellidae at four collecting sites within the Jardim Botânico do Rio de Janeiro. Eight traps were set, four in the arboretum (sites A and B), where the public is allowed to visit, and four at an Atlantic Forest fragment (sites C and D), open only to researchers. From July 2014 to June 2015, a total of 35,890 calliphorid flies were captured in 10 species and 145 Mesembrinellidea in 3. The greatest number of flies was found at site A and diversity was higher at site C. Chrysomya megacephala (Wiedemann) (Diptera: Calliphoridae) was the most prevalent species, being present at all sites. Mesembrinella bellardiana (Aldrich) (Diptera: Mesembrinellidae), Laneella nigripes (Guimarães) (Diptera: Mesembrinellidae), and Huascaromusca purpurata (Aldrich) (Diptera: Mesembrinellidae) were present at sites C and D, which indicates a preservation of the area because they are asynanthropic species.There were significant differences between sites A and D and sites B and D.

Steam application is an effective and environmentally friendly method for controlling bed bugs, Cimex lectularius L. (Hemiptera: Cimicidae). While a few studies documented the bed bug control efficacy of steam treatment, the sublethal effect of steam treatment on bed bug behavior and female fecundity is unknown. In this study, we evaluated the effect of steam treatment on the movement, feeding, mating behavior, and fecundity of female bed bugs in the laboratory. Bed bug adults received a calibrated steam treatment that caused approximately 28% mortality.The surviving bed bugs were observed for their feeding and mating behavior at 1 d posttreatment, female fecundity during a 7-d observation period, and offspring hatching. Steam-treated bed bugs were less active based on the percentage of bed bugs with movement, moving distance during a 10-min observation period, and feeding rate. However, steam treatment had no significant effect on blood intake (amount of blood taken per meal) among the fed bed bugs. After blood feeding, the steam-treated bed bugs had similar mating events and egg production as the control bed bugs. Furthermore, parental steam exposure had no significant effect on the offspring hatching. In conclusion, steam treatment could temporarily decrease bed bug activity levels and feeding rate, but had no significant impact on bed bug mating behavior and female fecundity.

Old mosquitoes are the most likely to transmit pathogens due to the higher probability that they will be exposed to pathogens, and the time required before a mosquito becomes infectious, the extrinsic incubation period (EIP). However, old mosquitoes are rarely considered in the evaluation of control tools. This study evaluated the effect of mosquito aging on the repellent efficacy of N,N-diethyl-3-methylbenzamide (DEET) in two vector mosquitoes, Anopheles gambiae s.s. (Giles) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae), respective vectors of malaria parasites and arboviruses such as dengue, chikungunya, and Zika viruses. For both mosquito species, DEET-induced inhibition of blood-feeding was assessed in three age classes. Life-history traits related to mosquito fecundity and survival following DEET exposure were also measured. Results showed that, in both species, bloodmeal inhibition induced by DEET was significantly higher in old females (>18 d old) than in younger ones (<13 d old). Life history traits recording showed no combined effects of DEET and aging on mosquito blood engorgement, oviposition rate, number of eggs laid nor survival; however, age effects are reported for all these traits.These results highlight the need for taking into account mosquito age in the evaluation of integrated mosquito management tools.They also suggest that the control of vector-borne pathogens with a long EIP could be improved by targeting old mosquitoes and supports the efficacy of repellents in the fight against mosquito-borne diseases.

Ixodes scapularis (Acari: Ixodidae) is the principal vector of Borrelia burgdorferi (the etiologic agent of Lyme disease) in the eastern and midwestern United States. Recent efforts have documented the first established population of I. scapularis in South Dakota, representing a western expansion of the known species distribution. Our goal was to describe the current distribution of I. scapularis in eastern South Dakota and to survey for the presence of B. burgdorferi in questing I. scapularis. We surveyed for the presence of adult and nymphal I. scapularis in seven counties within South Dakota, including 13 locales from 2016 to 2017. We then tested all I. scapularis, including those collected in 2015 from a previous study, for the presence of B. burgdorferi sensu stricto, via quantitative and conventional polymerase chain reaction. Here, we document the presence of I. scapularis in four new counties in South Dakota, and report the first instance of B. burgdorferi in a questing tick in South Dakota. Coupled with data from the Centers for Disease Control and Prevention and Companion Animal Parasite Council our data show that the risk of contracting Lyme disease in South Dakota is low, but existent and should be an important consideration with regard to public health, pets, and wildlife.

The Pacific Coast tick (Dermacentor occidentalis Marx, 1892) is one of the most widely distributed and frequently encountered tick species in California. This tick is the primary vector of an unclassified spotted fever group rickettsial pathogen, designated currently as Rickettsia 364D, the etiologic agent of a recently recognized tick-borne rickettsiosis known as Pacific Coast tick fever. Despite intensified interest in this pathogen, important questions remain regarding its taxonomic status and possible variations in genotype among different strains that could influence its pathogenicity. Only the extensively passaged prototypical isolate (strain 364-D) is widely available to rickettsiologists and public health scientists worldwide. To achieve a larger, more geographically diverse, and contemporary collection of strains, 1,060 questing adult D. occidentalis ticks were collected from 18 sites across six counties in northern and southern California in 2016 and 2017. Fourteen ticks (1.3%) yielded DNA of Rickettsia 364D and from these, 10 unique isolates from Lake and Orange counties were obtained. Additionally, Rickettsia rhipicephali was detected in 108 (10.2%) ticks, from which eight isolates were obtained, and Rickettsia bellii in six (0.6%), from which three isolates were obtained. The panel of recently acquired, low-passage strains of Rickettsia 364D derived from this study could enhance opportunities for investigators to accurately determine the taxonomic standing of this agent and to develop specific diagnostic assays for detecting infections with Rickettsia 364D in ticks and humans.

In the United States, the lone star tick, Amblyomma americanum (L.) (Acari: Ixodidae), is an aggressive southeastern species whose range has reportedly been steadily expanding northward.The number of A. americanum specimens submitted to theTickTesting Laboratory (TTL) at the Connecticut Agricultural Experiment Station (CAES) increased by 58% from the period of 1996–2006 (n = 488) to 2007–2017 (n = 773), mainly from Fairfield County in the southwestern corner of the state. The greatest numbers of A. americanum submissions to the CAES-TTL were from the City of Norwalk and a few adjacent municipalities. We also report the discovery of a large infestation of adult and nymphal lone star ticks detected on a dead male white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), on Manresa Island, Norwalk, in June 2017, indicating a long established, undetected population along the southwestern coast. A sample of nymphal and adult host-seeking A. americanum collected July 2017 from Manresa Island were tested and a proportion were positive for Ehrlichia chaffeensis, Ehrlichia ewingii, and Anaplasma phagocytophilum. The A. americanum tick and its associated disease pathogens are expected to become an increasing public health concern in southern New England.

The blacklegged tick, Ixodes scapularis Say (Acari: Ixodidae), is an economically important tick that affects veterinary and public health, but it can be difficult to collect in Oklahoma. The primary goal of this research was to examine the diel activity of each species to help improve collection methods for future field research and test field-collected I. scapularis for endemic and nonendemic tick-borne bacterial genera in the southern Great Plains region. Questing behavior was observed using caged bioassays over 24-h periods throughout fall and spring, and field collections were conducted throughout the afternoon and evening in different locations across Oklahoma. Blacklegged ticks were found to be more active during late afternoon and evening hours, and more ticks were recovered in pastures in the evening. None of the pools of adult I. scapularis tested positive for Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) or Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae) DNA. Of the 46 pools of I. scapularis tested, 27 (58.7%) were positive for Rickettsia sp. with ticks collected from the same location infected with the same species of rickettsial endosymbionts. Results suggest that sampling times later in the day may benefit off-host recovery of I. scapularis in Oklahoma ecosystems.

Most species of mosquitoes consume sugar to survive and during sugar feeding can expectorate virus. Scientists have used this behavior to develop novel methods of mosquito control and arbovirus surveillance. In this study, we use sugar feeding and corresponding viral expectoration to develop an affordable method of monitoring individual mosquitoes for longitudinal data collection. Female Culex tarsalis Coquillett (Diptera: Culicidae) that consumed an infectious bloodmeal of West Nile virus were placed into separate containers and offered a sucrose-soaked cotton wick. Wicks were then collected daily and tested for virus with similar results to those from standard capillary tube method.This yielded a direct longitudinal estimate of the extrinsic incubation period, while using fewer mosquitoes. This approach could be used to further characterize variation in the amount and diversity of expectorated virus over the life span of individual mosquitoes.

Arthropod-borne virus disease cycles constitute interactions among three primary players—the host, the vector, and the virus—in which the virus needs to interact with the host and the vector to establish its survival. While in the host, some arboviruses replicate aggressively, resulting in host pathogenicity, and manifest as a disease condition. These viruses more often utilize the vectors as reservoirs before they are transmitted to the host and therefore do not amplify to as large titers as they do in the hosts. In spite of this, the vector undergoes stress and activates several of its defense systems, resulting in alterations in its physiology. The present study was undertaken to evaluate the physiological changes that the mosquito vector Aedes aegypti (L.) (Diptera: Culicidae) undergoes during the replication of an arbovirus, Chikungunya virus (CHIKV). After the mosquitoes were infected with CHIKV, dissemination of the virus into various tissues and physiological parameters such as fecundity, vector mortality, egg laying, survival rate, overall fitness were monitored throughout the lifespan of the mosquitoes. Our study reveals that there is a fitness cost to the mosquitoes due to the infection of CHIKV. This fitness cost is manifested as higher mortality and low survival rate of the CHIKV-infected mosquitoes. Further evaluation revealed that the egg-laying pathway was affected, resulting in lower number of eggs. Expression analysis of six transcripts in the egg-laying pathway revealed that these transcripts were downregulated during the gonotrophic cycles in CHIKV-infected mosquitoes as compared to normal blood-fed mosquitoes.

Zoonotic cutaneous leishmaniasis (ZCL) due to Leishmania major (Yakimoff and Schokhor, 1914) (Kinetoplastida: Trypanosomatidae) is known as a neglected tropical disease, transmitted by Phlebotomus papatasi (Scopoli, 1786) (Diptera: Psychodidae) in North Africa and the Middle East. The main reservoirs of ZCL are desert gerbils and the role of humans as a reservoir host of the disease is not clearly defined and is therefore investigated in this study. In order to achieve this objective, Ph. papatasi sand flies were allowed to take blood through indirect blood sources (human, Rhombomys opimus (Lichtenstein, 1823) (Rodentia: cricetidae) (great gerbil), and BALB/c mice) using artificial feeding or direct xenodiagnoses from ZCL patients. To detect Leishmania promastigotes inside digestive canal, blood-fed sand flies were dissected and examined under light microscope and then confirmed by nested-PCR. In indirect xenodiagnoses, promastigote forms of Leishmania parasite were observed in 7.1%, 12%, and 13.6% of sand flies which fed on Rh. opimus, BALB/c mice, or human blood sources, respectively. In direct xenodiagnoses, among 93 female lab-reared sand flies which fed on active L. major lesion(s), 2.1% Leishmania infection was subsequently observed in the sand flies. No infection was detected in those sand flies that fed on nonlesioned skin of CL patients. Humans can serve as a reservoir of ZCL since the data indicate that Ph. papatasi is able to acquire L. major parasites from an active lesion of ZCL patients and the parasites can complete metacyclogenesis in the sand fly.

Vector control is important to effectively prevent trypanosomiasis. Therefore, alighting and feeding responses of trypanosome-transmitting vectors were conducted in Nigeria. The aim of this study was to assess the landing preference, level of fly annoyance on cattle, and feeding behavior by direct observation of cows, bulls, and calves. The percentage of Glossina species, Tabanus species, and stomoxyine that fed on cattle blood after landing varied according to the age and sex of the cattle host. Although most flies left the cattle host before completing their meal because of host defensive behaviors, an average of 78, 19.6, and 10.2% of Glossina, Tabanus, and stomoxyines, respectively, were fully engorged. The longest mean feeding time was observed in Glossina. Over more than 90% of Glossina spp. landed on the belly and legs. Most Tabanus spp. (85%) also landed on the belly and legs while Tabanus par Walker (Diptera: Tabanidae) often landed on the back (45%). Half of Stomoxys species landed on the head and neck. Vector flies' feeding success varied according to cattle host age and was negatively correlated with defensive host behaviors and apparent fly density. Hence, fly defensive behaviors contribute to trypanosomiasis dynamics in a cattle herd, and expended energy from defensive behaviors could adversely affect productivity.

Shape variability among individuals is important to understand some ecological relationships, since it provides the nexus between the genotype and the environment. Geometric morphometrics based on generalized procrustes analysis was applied on 17 landmarks of the wings of Aedes albifasciatus (Macquart 1838) (Diptera: Culicidae) females collected from three ecoregions of Argentina (Delta and islands of the Paraná River, Pampa, and Patagonian steppe). This methodology was used to discriminate the shapes of individuals belonging to different regions. The population of the Patagonian steppe, which was the most geographically distant, showed the most dissimilar shape. Different local variations in wing shape could have been selected according to the environmental characteristics and maintained by geographic isolation. The individuals of the two ecoregions closest to each other (Delta and islands of the Paraná River and Pampa) showed differences in shape that can be explained by a lower gene flow due to the effect of geographic isolation (by the Paraná River) and the limited dispersive capacity of Ae. albifasciatus. The results allow concluding that both environmental diversity and geographic barriers could contribute to local variations in wing shape.

The presence of Aedes albopictus (Skuse) was first reported in the Florida Keys in 1993. Despite extensive surveillance, the Florida Keys Mosquito Control District (FKMCD) only collected specimens occasionally on a few islands, some years finding no evidence of the invasive species. In 2013–2017, FKMCD witnessed a sudden increase in population size and geographic extent of Ae. albopictus. Samples of Ae. albopictus have now been identified on 30 different islands in the Florida Keys. Three islands in particular (Key Largo, Big Pine Key, and Stock Island) have produced multiple positive samples during at least 4 of the last 5 yr, suggesting establishment of the invasive species. FKMCD continues to monitor Ae. albopictus throughout the Keys and make extensive efforts to reduce population abundance and geographic extent of this disease vector.

In June 2016, we continued surveillance for tick-borne viruses in eastern Kansas following upon a larger surveillance program initiated in 2015 in response to a fatal human case of Bourbon virus (BRBV) (Family Orthomyxoviridae: Genus Thogotovirus). In 4 d, we collected 14,193 ticks representing four species from four sites. Amblyomma americanum (L.) (Acari: Ixodidae) accounted for nearly all ticks collected (n = 14,116, 99.5%), and the only other species identified were Amblyomma maculatum Koch (Acari: Ixodidae), Dermacentor variabilis (Say) (Acari: Ixodidae) and Ixodes scapularis Say (Acari: Ixodidae). All ticks were tested for both BRBV and Heartland virus (Family Bunyaviridae: Genus Phlebovirus) in 964 pools. Five Heartland virus positive tick pools were detected and confirmed by real-time reverse transcription PCR (rRT-PCR), while all pools tested negative for BRBV. Each Heartland positive pool was composed of 25 A. americanum nymphs with positive pools collected at three different sites in Bourbon County. A. americanum is believed to be the primary vector of both Heartland and BRBVs to humans based upon multiple detections of virus in field-collected ticks, its abundance, and its aggressive feeding behavior on mammals including humans. However, it is possible that A. americanum encounters viremic vertebrate hosts of BRBV less frequently than viremic hosts of Heartland virus, or that BRBV is less efficiently passed among ticks by co-feeding, or less efficiently passed vertically from infected female ticks to their offspring resulting in lower field infection rates.

The primary aim of this study was to determine if the Asian tiger mosquito (Aedes albopictus (Skuse)) (Diptera: Culicidae), that vectors a number of arboviruses, is parasitized by water mites (Acari: Prostigmata: Parasitengonina: Hydrachnidiae). Larval water mite parasitism of adult mosquitoes has been shown to reduce mosquito fecundity and life span in the laboratory, and impact some populations in the field. Water mites have been considered for biocontrol purposes and used to age-grade mosquito populations. Consequently, there is interest in water mite–mosquito associations from public health and integrated pest management perspectives. In 2016, the Pennsylvania Department of Environmental Protection West Nile Virus Control Program examined 146,607 mosquitoes representing 31 species for parasitic mites. Ten mosquitoes representing four species rarely or not previously known to be parasitized by mites were saved from destructive West Nile virus and Zika virus testing. Twelve parasitic mites were identified to the lowest taxonomic level possible. An Asian tiger mosquito and an Asian bush mosquito (Aedes japonicus) (Theobald) were each parasitized by a Parathyas sp. Lundblad (Hydryphantoidea: Hydryphantidae: Euthyadinae) water mite, and are the first and second records of these associations in North America, respectively. Two Culex restuans Theobald (Diptera: Culicidae) mosquitoes parasitized by either a Euthyas sp. Piersig (Hydryphantoidea: Hydryphantidae: Euthyadinae) or a Parathyas sp. water mite are new associations, and a Culex pipiens Linnaeus mosquito parasitized by a Parathyas sp. water mite represents a new record for this association in North America.

In criminal cases involving the recovery of human remains, as well as in cases of myiasis and pest management, the expertise of a forensic entomologist has been requested more and more frequently in Sicily. Recently, research on the insect species of forensic interest in Sicily has been investigated.The aim of this research was to raise awareness of this discipline and to build a Sicilian entomofauna checklist during cool temperatures. The predominant species observed in this study was Calliphora vicina (Robineau-Desvoidy) (Diptera, Calliphoridae), the first to colonize the carcasses; Lucilia spp. (Diptera, Calliphoridae) were also present. We wanted to explore the potential insect activity on decaying matter both on lava fields, exposed to sunlight, and in dark subterraneous environments. No activity was observed in our dark environment for 20 d; then, C. vicina was observed on one of the carcasses, laying eggs in complete darkness at a temperature of 6.4°C. Larval development under these conditions was delayed and the mortality rate was high.This preliminary trial allowed us to improve our experimental design and helped map new sites where we can expand our research to collect new data on insect distribution and their activity in caves.

The aim of this study was to perform a molecular survey and determine the genetic diversity of Bartonella spp. (Rhizobiales: Bartonellaceae) and Rickettsia spp. (Rickettsiales: Rickettsiaceae) in cat fleas (Siphonaptera: Pulicidae) from Southern Chile. Fleas (n = 251) were collected from 150 cats in Valdivia city and identified using morphological keys. Fleas belonging to the same cat were pooled (two to seven fleas per pool). DNA was purified from individual (n = 92) and pooled (n = 58) fleas and submitted to conventional polymerase chain reaction assays targeting Bartonella spp. (gltA) and Rickettsia spp. (ompA). Selected positive samples were sequenced for Bartonella gltA (n = 19), Rickettsia ompA (n = 14), and Rickettsia gltA (n = 11) for speciation, phylogenetic, and diversity analyses. All fleas (n = 251) were identified as Ctenocephalides felis felis (Bouché) (Siphonaptera: Pulicidae). Bartonella and Rickettsia occurrences in C. felis felis were 39.3% (59/150) and 76.6% (115/150), respectively. From sequenced Bartonella spp., 47.3% (9/19) were identified as Bartonella clarridgeiae, 42.1% (8/19) as Bartonella henselae, 5.3% (1/19) as Bartonella koehlerae, and 5.3% (1/19) as Bartonella sp. Rickettsia felis was the only Rickettsiaceae species identified in both ompA (14/14) and gltA (11/11) products. B. henselae and B. clarridgeiae presented five genotypes. R. felis ompA sequences presented seven genotypes. On the other hand, R. felis gltA sequences showed only one genotype. Bartonella spp. and R. felis are described for the first time in C. felis felis fleas from Southern Chile, highlighting the importance of these vectors as a source of zoonotic agents.

Culex quinquefasciatus Say (Diptera: Culicidae) is a mosquito species that has attracted a lot of attention from a medical and veterinary point of view; however, little is known about the frequency of L1014F mutations that have been found in the sodium channel gene, with this being a target for DDT and pyrethroid insecticides. The distribution and frequency of the L1014F mutation in Cx. quinquefasciatus populations was determined in rural and urban areas of Yucatan, Mexico from January 2015 to March 2016. Nine hundred fifty adult females out of 17,727 immature states were collected and analyzed in all sites sampled (n = 10). Susceptible homozygotes were identified (L1014/L1014) in 12% (114/950), heterozygous individuals (F1014/L1014) in 34% (323/950), and mutated homozygotes (F1014/F1014) in 54% (513/950) during the dry and rainy seasons. In this work, study areas with a high frequency of L1014F mutation were identified. These findings may help guarantee a more effective and efficient use of the resources available for the control of this vector.

Aedes aegypti (L.) (Diptera: Culicidae) is a vector of viruses causing dengue, Zika, chikungunya, and yellow fever and subsequently pose a significant global threat to public health. While sampling live mosquitoes is useful for surveillance purposes, most traps targeting Aedes kill captured mosquitoes. The Biogents Sentinel (BGS) trap, the gold standard for capturing Ae. aegypti, is one such trap. In our study, we modified the BGS trap to increase the survival of captured Ae. aegypti by replacing the trap's catch bag with a catch pot that protects mosquitoes from desiccation by airflow. A sucrose-soaked sponge or nucleic acid preservative card can also be placed inside the pot to enhance mosquito survival and augment arbovirus detection. These modifications to the BGS significantly increased the longevity of mosquitoes captured with weekly survivals of 93% in a semifield structure and 86% in the field.These high survival rates resulted in 3.5 times more alive Ae. aegypti captured weekly in the modified BGS compared to the original BGS, despite 40% lower overall catch rates. These cheap and simple trap modifications facilitate easier specimen identification as well as experiments requiring live field-collected samples such as virus detection from mosquito saliva and excreta.

The Borrelia genus consists of spirochete bacteria known to cause Lyme disease (LD) and relapsing fever in humans. Borrelia pathogens are commonly transmitted via arthropod vectors such as ticks, mites, or lice. Here, we report the molecular screening of LD group Borrelia sp. from ticks (Acari: Ixodidae) collected from rodents trapped in recreational forests and a semiurban residential area in the Selangor state in Malaysia. Of 156 adult ticks surveyed, 72 ticks were determined as positive for Borrelia sp. by polymerase chain reaction (PCR). All Borrelia PCR-positive ticks belonged to the Ixodes granulatus Supino species. Borrelia sp. was not detected in other tick species examined, including Dermacentor sp. and Amblyomma sp. ticks. Thirteen Borrelia PCR-positive tick samples were selected for further sequence analyses. Phylogenetic analyses of partial flaB gene sequences revealed that the Borrelia sp. were closely related to the LD group borreliae, Borrelia yangtzensis; a novel Borrelia genospecies reported in East Asian countries including Japan, Taiwan, and China. To our knowledge, this is the first report of Borrelia sp. related to Borrelia yangtzensis detected in Malaysia and Southeast Asia. The zoonotic potential of the Borrelia sp. reported here merits further investigation, as it may explain the previously reported serological evidence for borrelial infections in Malaysia.

Bed bugs, nocturnal ectoparasites adapted to feed on humans, have demonstrated a global resurgence since 1990s, presumably due to increased international travel and insecticide resistance. Resistance to insecticides has restricted the ability to manage bed bug populations. We evaluated the susceptibility of Cimex lectularius L. (Hemiptera: Cimicidae) collected from five districts in Paris against bendiocarb, a carbamate insecticide. The susceptible strain of C. lectularius from London included in our experiments as a control. Mortality of adult bed bugs was assessed after exposure to a fixed concentration of bendiocarb (80%) at intervals ranging from 15 min to 72 h. Mortality in samples ranged from 14% in Bobigny to 0% in Paris 15th arrondissement and Drancy. Bioassays with bendiocarb insecticide on C. lectularius revealed high levels of resistance in Paris.

The bioefficacy of commercial mosquito coils containing four different active ingredients, namely metofluthrin, d-allethrin, d-trans allethrin, and prallethrin against Aedes albopictus (Skuse) (Diptera: Culicidae) from 10 states in Malaysia, was evaluated using the glass chamber method. In this study, Ae. albopictus exhibited various knockdown rates (50% knockdown time, KT₅₀), ranging from 2.50 to 5.00 min, 2.50 to 7.00 min, 3.00 to 8.00 min, and 5.00 to 17.00 min for metofluthrin, d-trans allethrin, d-allethrin, and prallethrin, respectively. Overall, all strains of Ae. albopictus were most susceptible to metofluthrin, with mortality rates >80%. On the other hand, mortality rates ranging from 5.0 to 100% were observed from all populations exposed to d-trans allethrin, d-allethrin, and prallethrin. In addition, significant correlations between KT₅₀ of metofluthrin and d-allethrin (r = 0.758, P = 0.011), metofluthrin and prallethrin (r = 0.676, P = 0.032), d-allethrin and d-trans allethrin (r = 0.832, P = 0.003), d-allethrin and prallethrin (r = 0.921, P = 0.000), and d-trans allethrin with prallethrin (r = 0.941, P = 0.000) were detected, suggesting some levels of cross-resistance within the pyrethroid insecticides. This study demonstrated that metofluthrin can induce high insecticidal activity in Ae. albopictus in Malaysia, followed by d-trans allethrin, d-allethrin, and prallethrin.

Biological control can assist in the management of disease vector mosquitoes. However, we urgently require the identification of novel and effective agents to aid population management strategies. Previously, predatory biocontrol of disease vector mosquito species has focused extensively on cyclopoid copepods, but prey size refuge effects have been identified as a hindrance to their predatory efficacy. Calanoid copepods have yet to be examined in the context of mosquito control, despite their high prevalence, diversity, and distribution. Here, we apply functional responses (FRs; resource use as a function of resource density) to examine predation efficiencies of a recently described ephemeral pond specialist species, the freshwater calanoid copepod Lovenula raynerae Suárez-Morales, Wasserman & Dalu 2015 (Calanoida: Diaptomidae), using different size classes of larvae of the disease vector complex Culex pipiens (Diptera: Culicidae) as prey. Lovenula raynerae effectively consumed Cx. pipiens larvae across their ontogeny. A potentially population destabilizing type II FR was exhibited toward both early and late instar mosquitoes, indicative of a lack of prey refuge across ontogenetic stages. Attack rates were greatest and handling times lowest for early instar larvae compared to late instar larvae. These traits contrast to other copepods commonly applied in biocontrol, which are only able to handle early instars, and in much smaller numbers. We thus advocate that calanoid copepods can exert particularly marked predatory impact on lower trophic groups, and that their use in disease vector mosquito control strategies should be further explored.

The presence of vector-borne pathogens of veterinary and public health interest have received little attention in Chile. In Easter Island, in particular, a Chilean territory in the southeastern Pacific Ocean, no information is available. To fill this gap, 153 rural dogs were inspected for ectoparasites during a sterilization campaign carried out in 2016. Fleas were observed in 46% of the dogs, and Ctenocephalides felis (Bouché, 1835) was the only species present. Morphological identification of fleas was genetically confirmed using conventional polymerase chain reaction targeting the cox2 gene. No tick was observed in any dog. The presence of DNA of Rickettsia sp. (gltA and ompA fragment genes), Anaplasmataceae (16S rRNA), and Bartonella sp. (16S-23S ribosomal RNA intergenic spacer) was investigated in blood samples of 70 of the dogs and in 126 fleas analyzed in 68 pools that included 1–5 fleas. Rickettsial DNA was detected in 97% (n = 66) of the flea pools. Of these, 57 showed between 99 and 100% identity for both genes with published sequences of Candidatus Rickettsia asemboensis (CRa), six with Rickettsia felis, and one with Candidatus Rickettsia senegalensis. For two pools, gltA amplicons were identical to CRa but ompB amplicions showed 99–100% identity with R. felis. Anaplasmataceae DNA was detected in 16% (n = 11) pools. Sequenced amplicons showed highest identity with the endosymbiont Wolbachia pipientis. Bartonella DNA, showing 99% identity to Bartonella clarridgeiae, was detected in one pool (1.4%). No positive reaction was observed for any dog. This is the first detection of members of the ‘R. felis-like’ group other than R. felis in Chile.