Morphological and molecular characterization of Phlebotomus (Larroussius) perniciosus Newstead and Phlebotomus (L.) longicuspis Nitzulescu in Tunisia is reported. Different localities in central and southern Tunisia were sampled. Sand flies were collected by sticky-paper traps and Center for Disease Control traps. For morphological study of males, the copulatory valves (aedeagi) were examined and the number of coxite hairs was recorded. For molecular analysis, the mitochondrial cytochrome b gene and the cytochrome c oxidase I gene were sequenced to investigate the population structure of P. perniciosus and P. longicuspis. The majority of P. perniciosus samples from southern and some specimens from central Tunisia showed single-pointed aedeagi curved at their apicies that were indistinguishable from the P. longicuspis aedeagi and appeared similar to the atypical morph of P. perniciosus, previously described in northern Morocco. The current study gives evidence of a wide distribution of atypical morphs of P. perniciosus in southern and central Tunisia.

Rhipicephalus australis Fuller, the Australian cattle tick, is reinstated and the adults and larvae redescribed from material collected in Australia. This long ignored boophilid was previously known as R. microplus Canestrini for specimens reported in Australia and New Caledonia. The adults of R. australis are easily recognized by a combination of characters, such as the ventro-medial spurs in the palpal segments of the male, and the abundant, plumose, pale white setae on the dorsum of the female. Other details, such as coxal and adanal shields are more variable among different populations and may lead to incorrect determinations. Larvae of R. australis are clearly smaller than those of R. microplus. The use of principal components analysis on body measurements leads to a clear separation of larvae of both taxa. A phylogenetic analysis based on 12S- and 16S-rDNA gene sequences supports the conspecificity of the neotype material on which the reinstatement of the species is proposed, and of the specimens used for previous interspecific crosses. R. australis is now known to be present in Australia, New Caledonia, the island of Borneo, Philippines, Sumatra, Java, New Guinea, Cambodia, and Tahiti. Both R. microplus and R. australis coexist in some countries in southeastern Asia. Given the extreme importance of these ticks for the cattle industry, field data on their distribution in the region are required to know the actual range of these species and to understand the evolution of the group.

Two new species of black flies, Simulium (Gomphostilbia) terengganuense sp. nov. and Simulium (Gomphostilbia) aziruni sp. nov. (Diptera: Simuliidae), are described on the basis of reared adult, pupal, and larval specimens collected from Peninsular Malaysia. Both species are placed in the batoense species-group within the subgenus Gomphostilbia, one of two dominant subgenera of the genus Simulium in Peninsular Malaysia as well as in the Oriental Region. Strikingly, three morphological characteristics that rarely occur in the subgenus Gomphostilbia are found in these two new species: the very narrow female frons and the mushroom-like pupal terminal hooks in S. (G.) terengganuense sp. nov. and the pupal gill composed of an inflated horn-like structure and eight slender filaments in S. (G.) aziruni sp. nov.

Aedes albopictus (Skuse) and Ae. japonicus (Theobald) are important container-inhabiting mosquitoes that transmit disease agents, outcompete native species, and continue to expand their range in the United States. Both species deposit eggs in natural and artificial containers and thrive in peridomestic environments. The goal of our study was to examine the types and characteristics of containers that are most productive for these species in the northeastern United States. In total, 306 containers were sampled in urban, suburban, and rural areas of New Jersey. Multiple biotic and abiotic factors were recorded in an attempt to identify variables associated with the productivity of each species. Based on pupal abundance and density of container types, results showed that tires, trash cans, and planter dishes were the most important containers for Ae. albopictus, while planter dishes were the most important containers for Ae. japonicus. Container color (black and gray), material (rubber), and type (tires) were correlated with species presence for Ae. albopictus and Ae. japonicus. These factors may play a role in the selection of oviposition sites by female mosquitoes or in the survival of their progeny. Differences in species composition and abundance were detected between areas classified as urban, suburban, and rural. In urban and suburban areas, Ae. albopictus was more abundant in container habitats than Ae. japonicus; however, Ae. japonicus was more abundant in rural areas, and when water temperatures were below 14°C Our results suggest many variables can influence the presence of Ae. albopictus and Ae. japonicus in container habitats in northeastern United States.

The Asian tiger mosquito, Aedes albopictus (Skuse), is perhaps the most successful invasive mosquito species in contemporary history. In the United States, Ae. albopictus has spread from its introduction point in southern Texas to as far north as New Jersey (i.e., a span of ≈14° latitude). This species experiences seasonal constraints in activity because of cold temperatures in winter in the northern United States, but is active year-round in the south. We performed a laboratory experiment to examine how life-history traits of Ae. albopictus from four populations (New Jersey [39.4° N], Virginia [38.6° N], North Carolina [35.8° N], Florida [27.6° N]) responded to photoperiod conditions that mimic approaching winter in the north (short static daylength, short diminishing daylength) or relatively benign summer conditions in the south (long daylength), at low and high larval densities. Individuals from northern locations were predicted to exhibit reduced development times and to emerge smaller as adults under short daylength, but be larger and take longer to develop under long daylength. Life-history traits of southern populations were predicted to show less plasticity in response to daylength because of low probability of seasonal mortality in those areas. Males and females responded strongly to photoperiod regardless of geographic location, being generally larger but taking longer to develop under the long daylength compared with short day lengths; adults of both sexes were smaller when reared at low larval densities. Adults also differed in mass and development time among locations, although this effect was independent of density and photoperiod in females but interacted with density in males. Differences between male and female mass and development times was greater in the long photoperiod suggesting differences between the sexes in their reaction to different photoperiods. This work suggests that Ae. albopictus exhibits sex-specific phenotypic plasticity in life-history traits matching variation in important environmental variables.

The Maremma Plain (central Italy) was hyper-endemic for malaria until the mid-20th century, when a national campaign for malaria elimination drastically reduced the presence of the main vector Anopheles labranchiae Falleroni. However, the introduction of rice cultivation over 30 yr ago has led to an increase in the An. labranchiae population and concern over possible malaria reemergence. We studied the impact of anthropogenic environmental changes on the abundance and distribution of An. labranchiae in Maremma, focusing on rice fields, the main breeding sites. Adults and larvae were collected in three main areas with diverse ecological characteristics. Data were collected on human activity, land use, and seasonal climatic and demographic variations. We also interviewed residents and tourists regarding their knowledge of malaria. Our findings showed that the most important environmental changes have occurred along the coast; An. labranchiae foci are present throughout the area, with massive reproduction strictly related to rice cultivation in coastal areas. Although the abundance of this species has drastically decreased over the past 30 yr, it remains high and, together with climatic conditions and the potential introduction of gametocyte carriers, it may represent a threat for the occurrence of autochthonous malaria cases. Our findings suggest the need for the continuous monitoring of An. labranchiae in the study area. In addition to entomological surveillance, more detailed knowledge of human-induced environmental changes is needed, so as to have a more complete database that can be used for vector-control plans and for properly managing emergencies related to autochthonous introduced cases.

Many organisms, such as insects, filarial nematodes, and ticks, contain heritable bacterial endosymbionts that are often closely related to transmissible tickborne pathogens. These intracellular bacteria are sometimes unique to the host species, presumably due to isolation and genetic drift. We used a polymerase chain reaction/electrospray ionization-mass spectrometry assay designed to detect a wide range of vectorborne microorganisms to characterize endosymbiont genetic signatures from Amblyomma americanum (L.), Amblyomma maculatum Koch, Dermacentor andersoni Stiles, Dermacentor occidentalis Marx, Dermacentor variabilis (Say), Ixodes scapularis Say, Ixodes pacificus Cooley & Kohls, Ixodes ricinus (L.), and Rhipicephalus sanguineus (Latreille) ticks collected at various sites and of different stages and both sexes. The assay combines the abilities to simultaneously detect pathogens and closely related endosymbionts and to identify tick species via characterization of their respective unique endosymbionts in a single test.

Weather is important determinant of mosquito abundance that, in turn, influences vectorborne disease dynamics. In temperate regions, transmission generally is seasonal as mosquito abundance and behavior varies with temperature, precipitation, and other meteorological factors. We investigated how such factors affected species-specific mosquito abundance patterns in Saginaw County, MI, during a 17-yr period. Systematic sampling was undertaken at 22 trapping sites from May to September, during 1989–2005, for 19,228 trap-nights and 300,770 mosquitoes in total. Aedes vexans (Meigen), Culex pipiens L. and Culex restuans Theobald, the most abundant species, were analyzed. Weather data included local daily maximum temperature, minimum temperature, total precipitation, and average relative humidity. In addition to standard statistical methods, cross-correlation mapping was used to evaluate temporal associations with various lag periods between weather variables and species-specific mosquito abundances. Overall, the average number of mosquitoes was 4.90 per trap-night for Ae. vexans, 2.12 for Cx. pipiens, and 1.23 for Cx. restuans. Statistical analysis of the considerable temporal variability in species-specific abundances indicated that precipitation and relative humidity 1 wk prior were significantly positively associated with Ae. vexans, whereas elevated maximum temperature had a negative effect during summer. Cx. pipiens abundance was positively influenced by the preceding minimum temperature in the early season but negatively associated with precipitation during summer and with maximum temperature in July and August. Cx. restuans showed the least weather association, with only relative humidity 2–24 d prior being linked positively during late spring—early summer. The recently developed analytical method applied in this study could enhance our understanding of the influences of weather variability on mosquito population dynamics.

Studies on diurnal activity and factors affecting the questing behavior in Dermacentor reticulatus (F.) adults were conducted in an open-type habitat in eastern Poland (51° 15′ N, 22° 36′ E). Observations, for 25 min each, were made at 2-h intervals between 1000 and 1800 hours, during the autumn (from late September to mid-October 2000) and spring (from late April to mid-May 2001) activity peaks. The questing behavior of adult D. reticulatus ticks depended on the time of the day and season. The autumn activity was higher than that in spring, yet both these periods were characterized by a rising trend from the morning to early afternoon with a peak at ≈1400 hours (on average 40 and 30 specimens per one collection, respectively). In spring, the high activity (on average 31.8 specimens/ collection) persisted until late afternoon (1800 hours), while in the autumn period it declined before nightfall (on average 21.4 specimens/collection at 1800 hours). Females predominated over males in both seasonal periods of activity and greater differences in the sex ratio were recorded in the spring. We found a negative correlation between the diurnal activity of the adult ticks and temperature and between the number of active ticks and the length of day both in the autumn and spring. However, no statistically significant correlation was demonstrated between the diurnal activity of adult ticks and humidity. The investigations show that, in addition to environmental factors, tick host-seeking behavior is dependent on the activity of their hosts and biological traits of this species.

Bed bugs (Cimex lectularius L.) are a resurgent pest worldwide and infestations within the United States are increasing at a rapid rate. Because of the physical and psychological discomfort inflicted by their blood feeding habits, and allergies and secondary infections associated with bites, bed bugs are recognized as a significant public health problem. Although bed bug infestations are spreading and becoming more prevalent, we have a poor understanding of their dispersal patterns and sources of infestation. To help fill this gap, we conducted a genetic study of 21 bed bug infestations from the eastern United States, nearly all of which came from single rooms within residences. We genotyped samples comprised of 8–10 individuals per infestation at nine polymorphic microsatellite loci. Despite high genetic diversity across all infestations, with 5–17 alleles per locus (mean = 10.3 alleles per locus), we found low genetic diversity (1–4 alleles per locus) within all but one of the infestations. These results suggest that nearly all the studied infestations were started by a small propagule possibly consisting of a singly mated female and/or her progeny, or a female mated with multiple males that were highly related to her. All infestations were strongly genetically differentiated from each other (mean pairwise F_ST between populations = 0.68) and we did not find strong evidence of a geographic pattern of genetic structure, indicating infestations located in closer proximity to each other were nearly as genetically differentiated as those located hundreds of kilometers away. The high level of genetic diversity across infestations from the eastern United States together with the lack of geographically organized structure is consistent with multiple introductions into the United States from foreign sources.

In all triplicate tests of six plant essential oils and of vanillin mixtures, we corroborated strong insecticidal and repellent activities against adult Aedes aegypti (L.). Essential oils with potent toxic fumigant activities also exhibited repellency. Compared with N, N-diethyl-3-methylbenzamide, 5% of the essential oil concentrations of cassia, lemongrass, lemon eucalyptus, and xanthoxylum oils did not show repellent effects. However, a composition of lemongrass oil, xanthoxylum oil, and vanillin (1:3:1, vol:vol:wt) provided 270 min of complete protection time (CFT) compared with 247.5 min CPT with 15% N, N-diethyl-3-methylbenzamide. The CPT depended on concentration, presence of vanillin, or on both factors. When we applied a mixture of lemongrass oil: xanthoxylum oil: vanillin (1:1:1, vol:vol:wt) to the Viscopearl formulation, or porous cellulose beads, it provided gradual release of volatile compounds, thus showing >90% of repellency for 2 h. The behavioral and electrophysiological approaches we drew upon in our current study demonstrated that plant essential oil mixtures combined with vanillin showed strong and durable repellency to the mosquito. We claim that such combinations of plant essential oils and vanillin found in current study propose a viable commercial product suitable for future application in protecting a person from mosquito bites.

Mendelian crosses were used to study the mode of inheritance of Cry toxin resistance in a Culex quinquefasciatus Say (Diptera: Culicidae) colony (CqAB11A) that evolved insecticide resistance under laboratory selection with a deletion mutant of Bacillus thuringiensis subsp. israelensis de Barjac lacking the Cyt1Aa toxin component but containing its three major Cry toxins, Cry4Aa, Cry4Ba, and Cry11Aa. High levels of resistance were observed to Cry toxins. F1 offspring of reciprocal crosses to a sensitive colony showed intermediate levels of resistance with no maternal effect, indicating autosomal inheritance. Dose—response data for backcross offspring deviated significantly from the monofactorial model when tested with Cry4Aa Cry4Ba Cry11Aa, Cry4Aa Cry4Ba, or Cry11Aa. However, tests with Cry11Ba from B. thuringiensis subsp. jegathesan (Seleena, Lee, Lecadet) fit the monofactorial model. Dominance of F1 offspring was calculated at different concentrations of Cry-toxin suspensions and, as reported for other Cry-resistant Culex, generally decreased as concentration increased. A subset of colony CqAB11A was reared without selection pressure for 18 generations with little change in susceptibility, indicating a highly homozygous population. Consistent with reports for other Cry-resistant Culex, the data show these mosquitoes evolved resistance to B. thuringiensis Cry toxins at multiple loci in response to selection pressure and that cross-resistance to Cry11Ba was conferred by one of those loci.

After the acute infection period, birds persistently infected with West Nile virus (family Flaviviridae, genus Flavivirus, WNV) occasionally shed virus into the bloodstream, but these virions normally are inactivated by neutralizing antibody. The current work tested the hypothesis that these host neutralizing antibodies protect mosquito vectors from WNV infection and reevaluated the minimum WNV infectious dose necessary to infect Culex tarsalis Coquillett. To determine whether host antibodies protect mosquitoes from infection, Cx. tarsalis and Culex stigmatosoma Dyar were fed bloodmeals containing avian blood, WNV, and sera with or without WNV-specific neutralizing antibodies. When viral particles were completely bound by antibody, mosquitoes were protected from infection; however, when incompletely bound, WNV titers as low as 10^2.3 plaque-forming units (pfu)/ml resulted in 5% infection. These data indicated that avian antibodies were protective to mosquito vectors and were not dissociated during digestion. Because recrudescent viremias may not attain the same magnitude as initial acute viremias, Cx. tarsalis vector competence was reevaluated focusing on the fate of low-titered bloodmeals. Females were evaluated for vector competence after ingesting bloodmeals containing 10^2.2, 10^3.4, 10^4.5,10^5.5, or 10^6.5 WNV pfu/ml. Infection increased with bloodmeal titer, with 1% of the mosquitoes ingesting 10^3.4 pfu/ml and 45% of the mosquitoes ingesting 10^6.5 pfu/ml developing disseminated infections. The incomplete neutralization of recrudescent virus may be sufficient to infect a low proportion of competent blood-feeding Culex mosquitoes and perhaps allow persistently infected birds to provide a mechanism for arbovirus overwintering.

West Nile virus (family Flaviviridae, genus Flavivirus, WNV) is now endemic in California across a variety of ecological regions that support a wide diversity of potential avian and mammalian host species. Because different avian hosts have varying competence for WNV, determining the blood-feeding patterns of Culex (Diptera: Culicidae) vectors is a key component in understanding the maintenance and amplification of the virus as well as tangential transmission to humans and horses. We investigated the blood-feeding patterns of Culex tarsalis Coquillett and members of the Culex pipiens L. complex from southern to northern California. Nearly 100 different host species were identified from 1,487 bloodmeals, by using the mitochondrial gene cytochrome c oxidase I (COI). Cx. tarsalis fed on a higher diversity of hosts and more frequently on nonhuman mammals than did the Cx. pipiens complex. Several WNV-competent host species, including house finch and house sparrow, were common bloodmeal sources for both vector species across several biomes and could account for WNV maintenance and amplification in these areas. Highly competent American crow, western scrub-jay and yellow-billed magpie also were fed upon often when available and are likely important as amplifying hosts for WNV in some areas. Neither species fed frequently on humans (Cx. pipiens complex [0.4%], Cx. tarsalis [0.2%]), but with high abundance, both species could serve as both enzootic and bridge vectors for WNV.

The distribution of Aedes (Stegomyia) aegypti (L.), the main vector of dengue viruses (DENV) worldwide, overlaps with Aedes (Gymnometopa) mediovittatus (Coquillett), the Caribbean treehole mosquito, in urban, suburban, and rural areas. Ae. mediovittatus is a competent vector of DENV with high rates of vertical DENV transmission in the laboratory. This study determined whether Ae. mediovittatus feeds on humans and compared its feeding patterns with co-occurring Ae. aegypti in two rural communities of Puerto Rico. Adult mosquitoes were captured for three consecutive days every week from July 2009 to May 2010 using BG-Sentinel traps with skin lures that were placed in the front yard of houses in both communities. Three methods were used to identify the 756 bloodmeals obtained in this study: a multiplex polymerase chain reaction (PCR) for humans and dogs targeting cytochrome b; a PCR targeting the 16S rRNA; and a nested PCR targeting cytochrome b. Ae. mediovittatus fed mostly on humans (45–52%) and dogs (28–32%) but also on cats, cows, horses, rats, pigs, goats, sheep, and chickens. Ae. aegypti fed mostly on humans (76–79%) and dogs (18–21%) but also on cats, horses, and chickens. Our results indicate that Ae. mediovittatus may have a relatively high rate of vector—human contact, which might facilitate virus transmission or harborage in rural areas of Puerto Rico.

Although a variety of disease agents have been reported from bed bugs, the mechanical and biological disease transmission potential of bed bugs remains unelucidated. In this study we assayed survivability of the mildly pathogenic spotted fever group rickettsia, Rickettsia parkeri, in bed bugs after feeding on R. parkeri-infected chicken blood. Two groups of 15 adult bed bugs each were fed on infected or noninfected blood, and two groups of fourth-instar bed bugs also were fed on either infected or noninfected blood. One group of 15 adult bed bugs received no bloodmeal and was included as an additional control. Two weeks postfeeding, two pools of five live bed bugs from each group were surface sterilized, macerated, and placed in Vero cell cultures in an attempt to grow live organism. The remaining five individual bed bugs from each group were dissected, their salivary glands were removed for immunofluorescence assay (IFA) staining, and the remaining body parts were processed for polymerase chain reaction (PCR) analysis. Results indicated that no immature (now molted to fifth instar) bed bugs were positive for R. parkeri by IFA or PCR, indicating that organisms did not survive the molting process. After 4 wk of cell culture, no organisms were seen in cultures from any of the treatment or control groups, nor were any cultures PCR positive. However, two of the adult bed bugs were IFA positive for rickettsia-like organisms, and these two specimens were also PCR positive using R. parkeri-specific primers. These IFA and PCR results indicate that remnants of Rickettsia parkeri (possibly whole organisms) survived in the bugs for 2 wk, but the viability of the organisms in these two specimens could not be determined.

Estimating a mosquito's vector competence, or likelihood of transmitting disease, if it takes an infectious bloodmeal, is an important aspect of predicting when and where outbreaks of infectious diseases will occur. Vector competence can be affected by rearing temperature and interand intraspecific competition experienced by the individual mosquito during its larval development. This research investigates whether a new morphological indicator of larval rearing conditions, wing shape, can be used to distinguish reliably temperature and competitive conditions experienced during larval stages. Aedes albopictus (Skuse) and Aedes aegypti (L.) (Diptera: Culicidae) larvae were reared in low intraspecific, high intraspecific, or high interspecific competition treatments at either 22 or 32°C. The right wing of each dried female was removed and photographed. Nineteen landmarks and 20 semilandmarks were digitized on each wing. Shape variables were calculated using geometric morphometric software. Canonical variate analysis, randomization multivariate analysis of variance, and visualization of landmark movement using deformation grids provided evidence that although semilandmark position was significantly affected by larval competition and temperature for both species, the differences in position did not translate into differences in wing shape, as shown in deformation grids. Two classification procedures yielded success rates of 26 - 49%. Accounting for wing size produced no increase in classification success. There seemed to be a significant relationship between shape and size. These results, particularly the low success rate of classification based on wing shape, show that shape is unlikely to be a reliable indicator of larval rearing competition and temperature conditions for Ae. albopictus and Ae. aegypti.

A naturally occurring mutation was detected within the probe binding region targeting the envelope gene sequence of West Nile virus used in real-time polymerase chain reaction assays to test mosquito pools and other samples. A single C→T transition 6nt from the 5′ end of the 16mer in the envelope gene probe-binding region at genomic position 1,194 reduced assay sensitivity. The mutation first was detected in 2009 and persisted at a low prevalence into 2011. The mutation caused a 0.4% false negative error rate during 2011. These data emphasized the importance of confirmational testing and redundancy in surveillance systems relying on highly specific nucleic acid detection platforms.

In 2009–2011, Monroe County in southern Florida experienced locally acquired and traveler-imported focal dengue outbreaks. Aedes aegypti (L.) is the primary vector of dengue virus (DENV) worldwide, is prevalent in Monroe County, and is the suspected vector in Florida. Ae. albopictus (Skuse) is also known to be an important vector of DENV and this species is ubiquitous in Florida; however, it is not yet established in Monroe County. Florida Ae. aegypti (Key West and Stock Island geographic colonies) and Ae. albopictus (Vero Beach geographic colony) were fed blood containing 3.7 Log₁₀ plaque-forming unit equivalents of DENV serotype 1 isolated from a patient involved in the Key West, FL, outbreak in 2010. Mosquitoes were maintained at extrinsic incubation temperatures of 28 or 30°C for an incubation period of 14 d. Vector competence was assessed using rates of infection (percent with virus-positive bodies), dissemination (percent infected with viruspositive legs), and transmission (percent infected with virus-positive saliva). No significant differences were observed in rates of infection or dissemination between Ae. aegypti or Ae. albopictus at either extrinsic incubation temperature. Transmission was observed only at 28°C in both Ae. aegypti (Key West) and Ae. albopictus. The assessment of local mosquito populations for their DENV vector competence is essential and will aid mosquito control operators interested in pinpointing specific vector populations for control. The extent to which vector competence is affected by seasonal changes in temperature is discussed and provides baseline risk assessment data to mosquito control agencies.

The brown widow spider, Latrodectus geometricus C. L. Koch, has become newly established in southern California during the first decade of the 21st century. Brown widows and egg sacs were collected within the urban Los Angeles Basin using timed searches. We also collected and compared the abundance and distribution of the native western black widow spider, Latrodectus hesperus Chamberlin & Ivie, to brown widows. Brown widows were very common around urban structures especially outside homes, in parks, under playground equipment, in plant nurseries and landscaping areas, greatly outnumbering native western black widows, and were very rare or nonexistent in garages, agricultural crops, and natural areas. Western black widows predominated in xeric habitats and were less prevalent around homes. Neither species was found in the living space of homes. In southern California, envenomation risk exists because brown widows are now common in urban areas and the spiders hide where people place their fingers and exert pressure to move objects (e.g., under the curled lip of potted plants, in the recessed handle of plastic trash bins). Nonetheless, brown widow spider bites are less toxic than those of native western black widow spiders and, hence, if they are displacing black widows, overall widow envenomation risk may actually be lower than before brown widow establishment.

Blood-feeding sources of Culex quinquefasciatus Say (Diptera: Culicidae) collected in the eastern region of the Brazilian state of Santa Catarina were analyzed by precipitin technique. One hundred fifty-four female mosquitoes collected by CDC traps in the Navegantes municipality 13–15 February 2005 reacted to one or more of eight antisera, including chicken, dog, goat, sheep, horse, opossum, human and rodent antisera. One hundred thirty-seven specimens (89%) reacted to only one source, and 17 (11%) specimens reacted to two sources. Among the 137 specimens reacting to only one source, reactions to rodent (50.4%), sheep (5.8%), chicken (5.1%), goat (5.1%), dog (2.2%), horse (3.6%), and human (3.6%) antisera were observed. The analyzed species demonstrated a high degree of opportunistic feeding behavior in relation to host preference. Results are compared with results from similar studies, and the low proportion of reactions to human antisera is discussed.

Considering recent studies confirming an increased risk of contracting Lyme disease near metropolitan Chicago, we surveyed a more comprehensive area to assess whether the geographical distribution and establishment of Ixodes scapularis (Say) populations across northeast Illinois are widespread or limited in occurrence. From May through October 2008 and from April through October 2009, 602 I. scapularis ticks of all three life stages (larva, nymph, adult) were collected from sites in Cook, DuPage, Lake, and McHenry counties in northeast Illinois. The surveys were conducted by drag sampling vegetation in public-access forested areas. I. scapularis comprised 56.4% of ticks collected (n = 1,067) at 17 of 32 survey sites. In addition, four other tick species were incidentally collected: Dermacentor variabilis (Say), Haemaphysalis leporispalustris (Packard), Ixodes dentatus (Marx), and Amblyomma americanum (L.). This study updates the I. scapularis distribution in northeast Illinois. Our random sampling of suitable tick habitats across a large geographic area of the Chicago metropolitan area suggests a widespread human exposure to I. scapularis, and, potentially, to their associated pathogens throughout the region. These results prompt continued monitoring and investigation of the distribution, emergence, and expansion of I. scapularis populations and Borrelia burgdorferi transmission within this heavily populated region of Illinois.

From January to December 2009, 55 Amblyomma dissimile (Koch) ticks removed from iguanas in the municipality of Monteria and 3,114 ticks [458 Amblyomma sp. larvae, 2,636 Rhipicephalus microplus (Canestrini) larvae and 20 Amblyomma sp. nymphs] collected over vegetation in Los Cordobas were included in the study. The ticks were pooled into groups from which DNA was extracted. For initial screening of Rickettsia sp., each pool was analyzed by gltA real-time polymerase chain reaction (PCR). Positive pools were further studied using gltA, ompA, and ompB conventional PCR assays. Sequencing and phylogenetic analysis were also conducted. Rickettsial DNA was found in 28 pools of ticks (16 A. dissimile pools and 12 free-living larvae pools) out of 113 (24.7%) using real-time PCR. The same 28 pools were also positive using conventional PCR assays aimed to amplify gltA, ompA, and ompB. For each gene analyzed, PCR products obtained from 4/28 pools (two pools of A. dissimile, one pool of Amblyomma sp. larvae and one pool of Rh. microplus larvae) were randomly chosen and sequenced twice. Nucleotide sequences generated were identical to each other for each of the rickettsial genes gltA, ompA, and ompB, and showed 99.4, 95.6, and 96.4% identity with those of Rickettsia tamurae. They were deposited in the GenBank database under accession numbers JF905456, JF905458, and JF905457, respectively. In conclusion, we present the first molecular evidence of a novel Rickettsia (Rickettsia sp. strain Colombianensi) infecting A. dissimile ticks collected from iguanas, and also Rh. microplus and unspeciated Amblyomma larvae from vegetation in Colombia.