Diversity of Mosquitoes (Diptera: Culicidae) in the Bom Retiro Private Natural Heritage Reserve, Rio de Janeiro State, Brazil

This study registers the diversity of Culicidae in the Bom Retiro Private Natural Heritage Reserve (RPPNBR), Rio de Janeiro state, Brazil, based on the collection of the immature stages in natural and artificial larval habitats. Larvae and pupae were collected monthly at two sites of the RPPNBR from May 2014 to July 2015 using dippers and aquatic pipettes. The diversity of the mosquito community was described using the Shannon–Wiener Diversity Index (H′), as well as diversity, richness, and dominance of species found in different larval habitats (lake, bamboos, bromeliads, and artificial vessels). The Mann–Whitney test was used to calculate differences between the two natural and artificial habitats. Overall, 15,659 specimens belonging to 25 species, ten genera, and two subfamilies were collected. The most abundant species collected at sites that were reforested recently were Culex pleuristriatus Theobald, 1903, Limatus durhamii (Theobald, 1901), Aedes albopictus (Skuse, 1895), Culex neglectus (Lutz, 1904), and Culex retrosus (Lane & Whitman, 1951). In a forest preserved site, the most abundant species were Cx. neglectus, Culex iridescens (Lutz, 1905), Sabethes identicus (Dyar & Knab, 1907), Wyeomyia arthrostigma (Lutz, 1905), and Li. durhamii. With respect to larval habitats, 0.1% of the specimens were collected along the edge of a lake, 5.5% in bamboos, 35.9% in bromeliads, and 58.4% in artificial containers. Only 5.5% of the specimens were collected in the forest preserved site, with the remaining samples from the site with altered vegetation. A greater species richness and diversity were found in forest-altered sites compared to the forest preserved site. Several species were collected in the water accumulated in the nylon lids of plastic water tanks. Such vessels can promote an increase in mosquito population density in the environment surrounding the study area.

Effect of Urbanization on Presence, Abundance, and Coinfection of Bacteria and Protozoa in Ticks in the US Great Plains

Urbanization alters components of natural ecosystems which can affect tick abundance and tick-borne disease prevalence. Likely due to these changes, tick-borne pathogen prevalence has increased in many U.S. urban areas. Despite the growing public health importance of tick-borne diseases, little is known about how they are influenced by urbanization in North America, especially in the central U.S. where several pathogens occur at or near their highest levels of incidence nationally. To determine whether urban development influences tick infection with bacteria and protozoa, we collected ticks at 16 parks across a gradient of urbanization intensity in Oklahoma City, Oklahoma, USA and tested them using a variety of PCR assays. Adult ticks tested positive for Rickettsia parkeri, R. amblyommatis, R. rhiphicephali, ‘Candidatus R. andeanae’, Ehrlichia chaffeensis, E. ewingii, Panola Mountain Ehrlichia, ‘Borrelia lonestari’, Theileria cervi, Babesia spp. Coco, and Cytauxzoon felis. These results indicate the presence of a high diversity of tick-borne bacteria and protozoa across an expanding urban area in the U.S. Great Plains. Although there appeared to be some risk of encountering tick-borne microorganisms across the entire urbanization gradient, E. chaffeensis, E. ewingii, and T. cervi-infected ticks and microbe diversity decreased with increasing urbanization intensity. We identified a low rate of coinfection between different microorganisms, with coinfected ticks mainly collected from sites in the least-urbanized areas. This study suggests the need for awareness of tick-borne disease risk throughout urban areas in the central U.S., and highlights a need for studies of tick host habitat use and movement in cities.

Pyrethroid Susceptibility in Culex quinquefasciatus Say. (Diptera: Culicidae) Populations from Delta State, Niger-Delta Region, Nigeria

The development of insecticide resistance in different species of mosquitoes to Pyrethroids is a major challenge for vector-borne diseases transmitted by mosquitoes. Failure of Pyrethroids in control of mosquitoes would impact negatively on the gains recorded in control of mosquito-borne diseases in previous years. In anticipation of a country-wide deployment of Pyrethroid-treated nets for control of mosquito-borne diseases in Nigeria, this study assessed susceptibility of Culex quinquefasciatus Say. (Diptera: Culicidae) to Pyrethroids in Owhelogbo, Ejeme and Oria-Abraka communities in Delta State, Niger-Delta, Nigeria. Three to five day old Cx. quinquefasciatus were exposed to Deltamethrin (0.05%), Permethrin (0.75%), and Alphacypermethrin (0.05%) using World Health Organization bioassay method. Polymerase chain reaction (PCR) was employed in characterization of species and knockdown mutation. Results revealed that Cx. quinquefasciatus were generally susceptible (98-100%) to Deltamethrin, Permethrin, and Alphacypermethrin in the three communities with the exception of Owhelogbo where resistance to Deltamethrin (97%) was suspected. Knockdown time to Deltamethrin (11.51, 11.23, and 12.68 min), Permethrin (28.75, 13.26, and 14.49 min), and Alphacypermethrin (15.07, 12.50, and 13.03 min) were considerably low for Owhelogbo, Ejeme, and Oria-Abraka Cx. quinquefasciatus populations, respectively. Species identification result showed that all amplified samples were Cx. quinquefasciatus; however, no kdr allele was found in the three populations. Deployment of pyrethroid-treated nets for control of mosquito-borne diseases in Niger-Delta region of Nigeria is capable of reducing burden of diseases transmitted by Cx. quinquefasciatus as well as addressing nuisance value of the vector; however, caution must be entertained so as not to increase selection pressure thereby aiding resistance development.

The Original Scientific Description of the Lone Star Tick (Amblyomma americanum, Acari: Ixodidae) and Implications for the Species’ Past and Future Geographic Distributions

Amblyomma americanum L. is an important vector in North America originally described by Linnaeus based on Pehr Kalm’s 1754 report. While Kalm’s ‘Travels into North America’ is well known, his 1754 report remains obscure. Some authors were skeptical that Kalm referred to A. americanum because he encountered them at sites farther north outside of the species’ range. However, the details in 1754 report leave no doubt that Kalm described lone star ticks. In this historical review, we provide support for Kalm’s identification using a modern translation of his 1754 report and other sources. We also delineate distributional changes of lone star ticks from the pre-colonization era to the present and interpret them in the context of large-scale anthropogenic changes in the landscape. In this framework, the lone star tick’s current northward expansion is a recolonization of their former range. Extensive deforestation and extirpation of their principal host species, white-tailed deer, led to A. americanum’s disappearance from the northern parts of its range by the 20th century. Subsequent recolonization by second-growth forest and increases in white-tailed deer populations by the mid-20th century is now allowing A. americanum to reclaim its former range. These changes in the land appear to be the driving force behind A. americanum’s present expansion. Understanding this species’ history and the factors contributing to its current expansion will enable better predictions about its future distribution and potential to transmit human pathogens.

Trypanosomatids in Phlebotomine Sand Flies (Diptera: Phlebotominae) From Anthropic and Sinantropic Landscapes in a Rural Settlement in the Brazilian Amazon

Trypanosomatids (Kinetoplastida:Trypanosomatidae) protozoa are a diverse group of obligate parasites. The genera Trypanosoma and Leishmania are the most studied because of their medical importance. This work aims to evaluate the effects of anthropization processes on the composition of the phlebotomine sand fly fauna and the natural infection by Trypanosomatids, with emphasis on Leishmania. At all 3,186 sand flies were collected, distributed in 13 genera and 52 species, being Ny. umbratilis the most abundant species. There was no difference in the diversity between canopy and soil environments. The species abundance and richness were higher in the forest environment while species diversity and evenness were highest in the forest edge. The ITS1 region was used by PCR-RFLP to identify the fragment profiles of Leishmania species, followed by genetic sequencing. Here were analyzed 100 pools of female sand flies, being six positive for DNA parasite. PCR-RFLP fragment patterns similar to Endotrypanum sp. were observed in Nyssomyia anduzei, Psychodopygus amazonensis and Lutzomyia gomezi, and those fragments similar to Leishmania (Leishmania) amazonensis were observed in Bichromomyia flaviscutellata. ITS1 sequencing confirmed the presence of Leishmania sp. in Bi. flaviscutellata, and Leishmania (Viannia) naiffi in Ny. anduzei, Psychodopygus amazonensis, and Lu. gomezi. This is the first record of Lu. gomezi and Ps. amazonensis infection by L. naiffi in the State of Amazonas. These results show the trypanosomatid infection in sandflies from different landscapes in a rural settlement, and the finding of species infected with L.(V.) naiffi suggest that they can develop a role in the transmission cycle of leishmaniasis.

Forensically Relevant Flesh Flies (Diptera, Sarcophagidae, Sarcophaginae) of Southern Brazil

Flesh flies comprise a large fly family distributed worldwide that has great importance for forensic entomology. A robust and updated checklist of Sarcophaginae flies recorded in Southern Brazil is presented, based on material collected in the Rio Grande do Sul state and on a literature review. The forensic importance (high, moderate, or none) of the recorded flesh flies for estimating the postmortem interval (PMI) or inferring cases of neglect was determined based on their ecological habits. A total of 2,831 specimens representing 38 taxa were collected from three studies. Oxysarcodexia was the most abundant and species-rich genus in all three studies. Dexosarcophaga carvalhoi (Lopes) was registered for the first time in Southern Brazil. The checklist for Southern Brazil comprises 91 species distributed in 19 genera, with Oxysarcodexia, Peckia, and Lepidodexia representing the richest genera. Considering the importance of species for PMI estimation, 10, 42, and 39 species were classified as having high, moderate, or no forensic relevance, respectively. Moreover, five myiasis-causing sarcophagine flies recorded in Southern Brazil were considered to have the potential to reveal cases of neglect or mistreatment. This study revealed that Microcerella halli (Engel) and Peckia (Euboettcheria) australis (Townsend) are the most important species for PMI estimation in Southern Brazil, as they are often found breeding in corpses in this region.

Spinosyns Delivered in Sugar Meals to Aedes aegypti and Aedes albopictus (Diptera: Culicidae): Acute Toxicity and Subacute Effects on Survival, Fecundity, and Fertility

Sugar is an essential source of nutrition for adult mosquitoes to acquire energy. Toxic sugar bait (TSB) provides a promising method for mosquito control by incorporating toxins into artificial sources of sugar (i.e., toxic baits) presented to wild populations. Spinosyns comprise a family of bacterial secondary metabolites with a unique mode of action against the insect nervous system, an appealing environmental safety profile, and potential for incorporation into sugar baits. This research evaluated acute and subacute effects of spinosad (spinosyns A and D) and spinetoram (spinosyns J and L) in sugar meals on survival, fecundity, and fertility of Aedes aegypti and Aedes albopictus. Acute toxicity of spinosyns doubled from 24 to 48 h of assessment, revealing a relatively slow and cumulative action of the formulated spinosyns. Median lethal concentrations at 48 h were lower for spinetoram than for spinosad, lower for Ae. albopictus than Ae. aegypti, and lower for males than females. When exposed to subacute LC50 concentrations of spinosad and spinetoram for 24 h, survival of males and females of both species was diminished compared with controls, fecundity of females was increased, but fertility as measured by hatch rate of eggs was decreased. The formulations may have increased the nutritive value of the sugar meals thereby boosting fecundity, while toxifying embryos, reducing fertility. The inclusion of subacute effects of spinosyns allows assessment of the broader consequences of TSB for adult mosquito control.

Detection of Trypansoma cruzi in Kissing Bugs (Hemiptera: Reduviidae: Triatominae) Collected Across Oklahoma

Trypanosoma cruzi is the causative agent of Chagas disease in humans and dogs in the Americas. Transmission predominantly occurs via the feces of infected kissing bugs (Hemiptera: family Reduviidae; subfamily Triatominae) contaminating bite site wounds or mucous membranes. To better understand Chagas disease entomologic risk in Oklahoma, kissing bugs collected from within the state were tested for T. cruzi DNA. Data including county of insect collection, species and instar, and specific locations where specimens were found were collated. Triatomines were also tested by PCR to potentially identify DNA of vertebrate species on which specimens had recently fed. In total, 110 kissing bugs from 22 counties were tested. All triatomines were identified as Triatoma sanguisuga nymphs or adults, with the exception of one possible T. lecticularia adult. Trypanosoma cruzi DNA was detected in 22 (20%) triatomines from 12 counties spanning the state. The majority of T. cruzi PCR positive kissing bugs were found inside homes or associated structures (i.e., garages, porches). Vertebrate DNA was identified in 27 (24.5%) triatomines, with human DNA detected in 25 (92.6%) of these specimens, and canine and raccoon DNA detected in one specimen each (3.7%). Two specimens tested positive for both T. cruzi and human DNA and one specimen tested positive for both T. cruzi and raccoon DNA. Results from this study indicate that kissing bugs carrying T. cruzi are widespread in Oklahoma, that positive kissing bugs infest homes and associated structures, and that human-vector, canine-vector, and wildlife-vector contact all occur within the state.

Daily Variation in Sampled Densities of Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) Nymphs at a Single Site—Implications for Assessing Acarological Risk

The public health challenge posed by tick-borne disease (TBD) has increased efforts to characterize the spatial and temporal distribution of ticks and associated pathogens to better focus tick control strategies and personal protection measures. We describe variability in nymphal Ixodes scapularis Say and Amblyomma americanum (L.) density derived from daily drag sampling at a single location in New Jersey over 4 yr and explore how observed differences in daily collections might affect the estimation of acarological risk. We found significant variability in the density of host-seeking nymphs that could suggest substantially different rates of human-tick encounters depending on sampling date, habitat, and ambient weather conditions. The spatial and temporal variability in the distribution of 2 sympatric tick species with different host preferences and questing strategies, suggests that to produce results that are comparable among sites across the area sampled, surveillance efforts may be limited to shorter collection seasons, fewer sites or less sampling effort (fewer plots or fewer visits) per site, and a geographic scope that minimizes the potential temporal and spatial biases indicated here. Our results illustrate that evaluation of models of tick distribution or relative acarological risk based on surveillance data requires a full description of the diversity of habitats sampled and the conditions under which sampling is performed. The array of factors that affect tick host-seeking and that could bias interpretation of sampling results emphasizes the need to standardize sampling protocols and for more caution when interpreting tick sampling data collected over large temporal and spatial scales.

Diptera (Dryomyzidae, Fanniidae, Muscidae, Piophilidae) Associated With Pig Carcasses in a Forest Habitat of Poland: Sex-Related Patterns of Visitation and Effectiveness of Sampling Methods

Our knowledge on many necrophilous dipterans was limited for a long time mainly due to taxonomic issues. Therefore, findings on the sex-related associations with pig carcasses in Dryomyzidae, Fanniidae, Muscidae, and Piophilidae are presented. Moreover, the effectiveness of pitfall traps and sweep nets in fly sampling were compared, since the correct method of collecting insects is critical for the development and application of entomological tools for forensic purposes. The trapped numbers of all species differed significantly in respect to both sexes. The sex ratios were strongly female-biased in most species of Muscidae and in Fanniidae. More females were also collected in Piophilidae, but the proportion of males was higher when compared to muscids. Within Dryomyzidae, the sex ratio differed according to species. Only Dryomyza flaveola (Fabricius, 1794) (Diptera: Dryomyzidae) and Muscina pascuorum (Meigen, 1826) (Diptera: Muscidae) had male-biased sex ratios. Differences in residency patterns between sexes in Stearibia nigriceps (Meigen, 1826) (Diptera: Piophilidae), Liopiophila varipes (Meigen, 1830) (Diptera: Piophilidae), and D. flaveola were statistically insignificant. All studied species were recorded for each sampling method. The effect of the sampling method on the number of individuals was statistically significant for all dipterans but Piophilidae. Pitfall traps outcompeted the aerial sweep net in representing Dryomyzidae and Muscidae, whereas both methods were almost equally effective in the collection of Piophilidae. A varied effectiveness of sampling methods was observed in Fanniidae. The sampling method significantly affected the determination of the appearance time of flies on carcasses, but the effect of species and sampling method was insignificant. Implications for forensic entomology are discussed.