Morphological details are provided for the dorsal and ventral surfaces of both extremities and the micropylar area of eggs of Haemagogus (Haemagogus) capricornii Lutz, captured in the Biological Reserve of Tinguá, State of Rio de Janeiro, Brazil. The eggs were observed by scanning electron microscopy with a morphometrical analysis of the main structures. The outer chorionic cells on the ventral surface were extremely regular, such as those observed in Hg. equinus and Hg. janthinomys. The tubercles present differences in form, size, and distribution. Filaments to attach to the substrate were observed in this species.

The Afrotropical Funestus and the Oriental-African Minimus Groups of the Myzomyia Series of Anopheles subgenus Cellia are considered distinct only because of their geographical separation. For the first time, a phylogenetic study was done on six Oriental and four Afrotropical species based on the comparison of ribosomal (ITS2, D3) and mitochondrial (COI) nucleotide sequences. Both maximum parsimony (MP) and maximum likelihood (ML) analyses revealed that the groups form a monophyletic assemblage containing four clades. The inclusion of Afrotropical An. leesoni with the Oriental species was confirmed, whereas An. rivulorum, also an Afrotropical species, was placed in a basal position relative to the African and Oriental species. The biogeography of the Afrotropical and Oriental species was examined in relation to the phylogeny and estimates of divergence time. Divergence events correspond to periods of major tectonic movement as well as periods of great aridity or humidity.

Correct classification of the insect vector is central to the study of arboviral disease. A simple molecular method for identification of the main vectors of the mosquito-borne viruses, dengue, yellow fever, and Rift Valley fever in Senegal, West Africa, was developed. We present a system in which the five mosquito species (Diptera: Culicidae) responsible for the majority of flaviviral disease transmission in Senegal can be reliably identified using small amounts of DNA coextracted during flaviviral screening procedures, via an easy amplification of the mitochondrial gene cytochrome oxidase c subunit I or II (COI or COII, respectively). We observed that despite very similar morphology, the two cryptic disease vector species Aedes furcifer Edwards and Aedes taylori Edwards are highly divergent at the molecular level. This sequence variation was used as a basis for the development of a polymerase chain reaction-restriction fragment-length polymorphism system for the differentiation of the two species. We also present the first investigation of the phylogeny of the culicine mosquitoes based on all COI and COII sequences currently available. There seems to be very low intraspecific variation in both genes, whereas interspecific variation is high. As a consequence, COI and COII are ideal candidates for the molecular identification of disease vectors to species level, whereas deeper divergences remain equivocal by using these genes. This system provides a new technique for the accurate identification of culicine disease vectors in West Africa and provides a basis for the expansion of such methods into the study of a range of diseases.

The intraspecific variability of Triatoma dimidiata Latreille, a major vector of Chagas disease, was studied in four departments of Guatemala. Insects were collected from either domestic and sylvatic habitats, and their cuticular hydrocarbon pattern and head morphology were analyzed using ordination and classification techniques. A significant discrimination was obtained both with morphometric and hydrocarbon analyses. Insects from northern departments were easily differentiated from southern conspecifics. Distinctive hydrocarbon pattern and head shape were detected for insects collected from caves in the north central region of the country, posing concern about their taxonomic status.

Ixodes ricinus L. is the principal European vector of Borrelia burgdorferi sensu lato, the causative agent of Lyme borreliosis. Subtractive hybridization was used to isolate tick genes that were induced in whole ticks after blood meals on uninfected and B. burgdorferi-infected guinea pigs. Novel cDNA clones with similarity to cytochrome c oxidase, salivary secreted protein, actin, and a cysteine protease propeptide were induced after a blood meal. Novel cDNA clones with similarity to thioredoxin peroxidases, dolichyl-phosphate β-glucosyltransferase, glutathione S-transferase, defensin, ML domain-containing protein, and von Willebrand factor were induced after B. burgdorferi infection. Virtual Northern analysis was used to verify that these genes were differentially expressed in ticks after a pathogen-infected blood meal and to detect their tissues of expression. The characterization of genes that are induced after an infected blood meal is essential for gaining an understanding of the molecular mechanisms that underlie vector-pathogen interactions.

The lesser mealworm, Alphitobius diaperinus (Panzer), is a carrier of Campylobacter spp. in poultry facilities; however, the beetle’s importance in the epidemiology of campylobacteriosis is not known. A series of laboratory experiments were designed to test the vector and reservoir competence of the lesser mealworm for Campylobacter jejuni. In the first experiment, C. jejuni was swabbed onto the outer surface of adult and larval beetles to determine how long bacteria can survive on the beetles’ exterior. Next, adult and larval mealworms were allowed to drink from a solution containing C. jejuni and the duration of internal carriage was monitored. For the third experiment, beetles drank from a Campylobacter suspension and the duration of fecal shedding of bacteria was determined. In the last experiment, 3-d-old chickens were fed either one or 10 infected beetles, and cloacal swabs were tested periodically for Campylobacter. C. jejuni was detected on the exterior of larval beetles for 12 h, from the interior of larvae for 72 h, and from the feces of larvae for 12 h after exposure. Ninety percent of the birds that consumed a single adult or larval beetles became Campylobacter-positive, whereas 100% of the birds that consumed 10 adults or larvae became positive. These experiments demonstrated that the lesser mealworm could acquire and harbor Campylobacter from an environmental source. We found that the lesser mealworm was capable of passing viable bacteria to chickens that consumed the beetle. The beetle should be included in attempts to maintain Campylobacter-free poultry facilities.

Two Trypanosoma cruzi-derived cloning vectors, pTREX-n and pBs:CalB1/CUB01, were used to drive the expression of green fluorescent protein (GFP) and DsRed in Trypanosoma rangeli Tejera, 1920, and Trypanosoma cruzi Chagas, 1909, isolates, respectively. Regardless of the species, group, or strain, parasites harboring the transfected constructs as either episomes or stable chromosomal integrations showed high-level expression of fluorescent proteins. Tagged flagellates of both species were used to experimentally infect Rhodnius prolixus Stal, 1953. In infected bugs, single or mixed infections of T. cruzi and T. rangeli displayed the typical cycle of each species, with no apparent interspecies interactions. In addition, infection of kidney monkey cells (LLC-MK2) with GFP-T. cruzi showed that the parasite retained its fluorescent tag while carrying out its life cycle within cultured cells. The use of GFP-tagged parasites as a tool for biological studies in experimental hosts is discussed, as is the application of this method for copopulation studies of same-host parasites.

Since first discovered in the New York City area in 1999, West Nile virus (WNV) has become established over much of the continental United States and has been responsible for >10,000 cases of severe disease and 400 human fatalities, as well as thousands of fatal infections in horses. To develop appropriate surveillance and control strategies, the identification of which mosquito species are competent vectors and how various factors influence their ability to transmit this virus must be determined. Therefore, we evaluated numerous mosquito species for their ability to transmit WNV under laboratory conditions. This report contains data for several mosquito species not reported previously, as well as a summary of transmission data compiled from previously reported studies. Mosquitoes were allowed to feed on chickens infected with WNV isolated from a crow that died during the 1999 outbreak in New York City. These mosquitoes were tested ≈2 wk later to determine infection, dissemination, and transmission rates. All Culex species tested were competent vectors in the laboratory and varied from highly efficient vectors (e.g., Culex tarsalis Coquillett) to moderately efficient ones (e.g., Culex nigripalpus Theobald). Nearly all of the Culex species tested could serve as efficient enzootic or amplifying vectors for WNV. Several container-breeding Aedes and Ochlerotatus species were highly efficient vectors under laboratory conditions, but because of their feeding preferences, would probably not be involved in the maintenance of WNV in nature. However, they would be potential bridge vectors between the avian–Culex cycle and mammalian hosts. In contrast, most of the surface pool-breeding Aedes and Ochlerotatus species tested were relatively inefficient vectors under laboratory conditions and would probably not play a significant role in transmitting WNV in nature. In determining the potential for a mosquito species to become involved in transmitting WNV, it is necessary to consider not only its laboratory vector competence but also its abundance, host-feeding preference, involvement with other viruses with similar transmission cycles, and whether WNV has been isolated from this species under natural conditions.

In 1985–2002, surveillance for bovine arboviruses was conducted in Kagoshima, located in the most southern part of the main islands of Japan and known to be an area where bovine arboviral diseases have frequently been epidemic. Culicoides biting midges were collected in a cowshed by light traps. A total of 456,300 Culicoides biting midges representing 13 species were collected, and a portion of each pool of midges were tested for virus isolation. Overall, 85 isolates of six different viruses were obtained from the collected midges. The isolated viruses included two Orthobunyaviruses, Akabane and Aino viruses; three Orbiviruses, Chuzan, D’Aguliar, and Ibaraki viruses; and one unclassified virus, a bunyavirus-like virus. The viruses were most frequently isolated from Culicoides oxystoma Kieffer (85.9% of 85 isolates). Isolations of all viruses except for the bunyavirus-like virus were made from this species. Our data indicated that C. oxystoma is a potential vector for bovine arboviruses in southern Japan.

Entomological surveys were conducted in five rural communities (138 domiciliary units [DUs]) in the southern Andes of Ecuador. Adobe walls and ceramic tile roofs were predominant construction materials. A 35% house infestation rate with Panstrongylus chinai (Del Ponte, 1929) (0.7%), Panstrongylus rufotuberculatus (Champion, 1899) (0.7%), Rhodnius ecuadoriensis (Lent & León, 1958) (27%), and/or Triatoma carrioni (Larrousse, 1926) (7%) was found. Adults and nymphs of R. ecuadoriensis and T. carrioni were found in intradomiciliary and peridomiciliary areas. Breeding triatomine colonies were present in 85% of infested DUs, and the average insect crowding was 52 ± 113 triatomine bugs per infested house. T. cruzi-like organisms were found by microscopic examination in the feces or hindgut but not the salivary glands of 4% of examined R. ecuadoriensis and 12% T. carrioni. Serological tests detected a general anti-T. cruzi antibody seroprevalence of 3.9% (n = 1136). Only 2% of individuals had heard of Chagas disease, and although triatomines were reported as a major nuisance by the population they were not considered vectors of disease. Additional baseline field research is needed for the design and implementation of a Chagas disease control program in the region.

In California, Ixodes pacificus Cooley & Kohls nymphs have been implicated as the primary bridging vectors to humans of the spirochetal bacterium causing Lyme disease (Borrelia burgdorferi). Because the nymphs typically do not ascend emergent vegetation, risk of human exposure is minimal in grasslands, chaparral, and woodland-grass. Instead, woodlands with a ground cover dominated by leaf litter (hereinafter referred to as woodland-leaf) have emerged as a primary risk habitat for exposure to B. burgdorferi-infected nymphs. As a means of differentiating woodland-leaf habitats from others with minimal risk (e.g., chaparral, grassland, and woodland-grass), we constructed a maximum likelihood model of these habitat types within a 7,711-ha area in southeastern Mendocino County based on the normalized difference vegetation index derived from Landsat 5 Thematic Mapper imagery (based on a 30 by 30-m pixel size) over four seasons. The overall accuracy of the model to discriminate woodland-leaf, woodland-grass, open grassland, and chaparral was 83.85% (Kappa coefficient of 0.78). Validation of the accuracy of the model to classify woodland-leaf yielded high values both for producer accuracy (93.33% of validated woodland-leaf pixels correctly classified by the model) and user accuracy (96.55% of model-classified validation pixels correctly categorized as woodland-leaf). Woodland-leaf habitats were found to be highly aggregated within the examined area. In conclusion, our model successfully used remotely sensed data as a predictor of habitats where humans are at risk for Lyme disease in the far-western United States.

The prevalence of ‘Ehrlichia walkeri’ in Ixodes ricinus in northern Italy was studied. A total of 456 ticks were collected from seven places around Belluno, and 292 living ticks were examined using polymerase chain reaction (PCR). Nineteen ticks were positive for gltA of ‘E. walkeri’, and the prevalence rate for each place ranged from 0 to 12.8%. The disparity of prevalence rates among areas examined suggests that ‘E. walkeri’ is not widespread in ticks and requires an animal host for its maintenance in nature.

Eggs of Liosarcophaga dux (Thomson) (Diptera: Sarcophagidae) are elongated and slightly bean-shaped, measuring ≈1.5 mm in length. Each is covered externally by an eggshell comprised of polygonal patterns. In this study, scanning and transmission electron microscopy were used to examine the eggshell of this species of flesh fly for the first time. No plastron region or median area was detected. Ultrathin sectioning of the eggshell revealed multiple layers in the shell that could be observed using transmission electron microscopy.

Head lice are very common and mainly affect children between 3 and 12 yr old. Little is known about the way nits, the eggs of the head louse, are attached to the hair. In this report, an objective measurement procedure for the ease with which nits can be removed is presented. The first peak force, associated with the start of nit movement, and the average and maximal force during the sliding of the nit were measured. The three force variables correlated with the length of the cylinder by which the nit was attached to the hair. A negative correlation was found between the maximum force exerted and the distance of the nit from the scalp. The method described in this report can be used to determine the in vitro efficacy of various products to remove nits.