Comparative sequence analyses of La Crosse virus strain isolated from patient with fatal encephalitis, Tennessee, USA.

We characterized a La Crosse virus (LACV) isolate from the brain of a child who died of encephalitis-associated complications in eastern Tennessee, USA, during summer 2012. We compared the isolate with LACV sequences from mosquitoes collected near the child's home just after his postmortem diagnosis. In addition, we conducted phylogenetic analyses of these and other sequences derived from LACV strains representing varied temporal, geographic, and ecologic origins. Consistent with historical findings, results of these analyses indicate that a limited range of LACV lineage I genotypes is associated with severe clinical outcomes.

Population Dynamics of Aedes aegypti and Aedes albopictus in Two Rural Villages in Southern Mexico: Baseline Data for an Evaluation of the Sterile Insect Technique.

Indoor and outdoor ovitraps were placed in 15 randomly selected houses in two rural villages in Chiapas, southern Mexico. In addition, ovitraps were placed in five transects surrounding each village, with three traps per transect, one at the edge, one at 50 m, and another at 100 m from the edge of the village. All traps were inspected weekly. A transect with eight traps along a road between the two villages was also included. Population fluctuations of Aedes aegypti and Ae. albopictus were examined during 2016-2018 by counting egg numbers. A higher number of Aedes spp. eggs was recorded at Hidalgo village with 257,712 eggs (60.9%), of which 58.1% were present in outdoor ovitraps and 41.9% in indoor ovitraps, compared with 165,623 eggs (39.1%) collected in the village of Río Florido, 49.0% in outdoor and 51.0% in indoor ovitraps. A total of 84,047 eggs was collected from ovitraps placed along transects around Río Florido, compared to 67,542 eggs recorded from transects around Hidalgo. Fluctuations in egg counts were associated with annual variation in precipitation, with 2.3 to 3.2-fold more eggs collected from ovitraps placed in houses and 4.8 to 5.1-fold more eggs in ovitraps from the surrounding transects during the rainy season than in the dry season, respectively. Aedes aegypti was the dominant species during the dry season and at the start of the rainy season in both villages. Aedes albopictus populations were lower for most of the dry season, but increased during the rainy season and predominated at the end of the rainy season in both villages. Aedes albopictus was also the dominant species in the zones surrounding both villages. The numbers of eggs collected from intradomiciliary ovitraps were strongly correlated with the numbers of eggs in peridomiciliary ovitraps in both Río Florido (R2 adj = 0.92) and Hidalgo (R2 adj = 0.94), suggesting that peridomiciliary sampling could provide an accurate estimate of intradomiciliary oviposition by Aedes spp. in future studies in these villages. We conclude that the feasibility of sterile insect technique (SIT)-based program of vector control could be evaluated in the isolated Ae. aegypti populations in the rural villages of our baseline study.

Vector competence of eastern and western forms of Psorophora columbiae (Diptera: Culicidae) mosquitoes for enzootic and epizootic Venezuelan equine encephalitis virus.

Venezuelan equine encephalitis virus (VEEV) continues to circulate enzootically in Mexico with the potential to re-emerge and cause disease in equines and humans in North America. We infected two geographically distinct mosquito populations of eastern Psorophora columbiae form columbiae (Chiapas, Mexico and Texas, United States) and one mosquito population of western Psorophora columbiae form toltecum (California, United States) with epizootic and enzootic IE VEEV and epizootic IAB VEEV. We detected no differences between epizootic and enzootic IE viruses in their ability to infect any of the mosquito populations analyzed, which suggested that neither species selects for epizootic IE viruses. Psorophora columbiae f. columbiae (Texas) were significantly less susceptible to infection by epizootic IE than Ps. columbiae f. columbiae (Mexico). Psorophora columbiae f. toltecum populations were more susceptible than Ps. columbiae f. columbiae populations to epizootic IE and IAB viruses.

Gonotrophic cycle estimate for Culex quinquefasciatus in Mérida, Yucatán, México.

West Nile virus (WNV) has been present in the Yucatán State, México, since 2002. Culex quinquefasciatus, one of the main vectors of WNV transmission in the United States, is also common in Mexico and may be a key vector of WNV transmission t o humans in t he Yucatán. The aim of this study was to determine the length of the gonotrophic cycle and the survival rates of Cx. quinquefasciatus from Mérida, Yucatán, during the rainy versus the dry season. Mosquitoes were collected during 25-day periods in October (rainy season) and in April (dry season), and captured females were classified by abdominal appearance (freshly fed, late-stage fed, half gravid, and subgravid). To determine the age structure as nulliparous and parous females and to calculate the gonotrophic cycle through a time series and the mosquito survival, we used Davidson formulae. Also, vitellogenesis analysis to monitor egg maturity was conducted during both seasons. Cross-correlation data suggested a similar length of the gonotrophic cycle (4 days) in both seasons. Oogenic development required a minimum of 72 h in each season. However, survival of the mosquito population collected in the rainy season was significantly higher (0.91) with a mean temperature of 28 +/- 1.57 degrees C than was survival in the dry season (0.78) with a mean temperature of 29 +/- 1.10 degrees C. Survival, although higher during the rainy season, did not influence the length of the gonotrophic cycle of Cx. quinquefasciatus in Yucatán.

Host selection and gonotrophic cycle length of Anopheles punctimacula in southern Mexico.

The host preference, survival rates, and length of the gonotrophic cycle of Anopheles punctimacula was investigated in southern México. Mosquitoes were collected in 15-day separate experiments during the rainy and dry seasons. Daily changes in the parous-nulliparous ratio were recorded and the gonotrophic cycle length was estimated by a time series analysis. Anopheles punctimacula was most abundant during the dry season and preferred animals to humans. The daily survival rate in mosquitoes collected in animal traps was 0.96 (parity rate = 0.86; gonotrophic cycle = 4 days). The length of gonotrophic cycle of 4 days was estimated on the base of a high correlation coefficient value appearing every 4 days. The minimum time estimated for developing mature eggs after blood feeding was 72 h. The proportion of mosquitoes living enough to transmit Plasmodium vivax malaria during the dry season was 0.35.

Biological variation in two Anopheles vestitipennis populations with different feeding preferences in southern Mexico.

The lengths of gonotrophic cycle and egg development and survival rate were studied in Anopheles vestitipennis collected in horse and human-baited traps in southern Mexico. The gonotrophic cycle duration was estimated using cross-correlation analysis, whereas the survival rate was assessed using a vertical method. Daily changes of parity rates gave significant correlation indices at 3 and 4 days in the zoophilic and anthropophilic populations, respectively. The minimum time required to develop mature eggs after blood feeding was 54 and 60 h, and the survival rate was 0.93 and 0.88 in zoophilic and anthropophilic female mosquito populations, respectively. These biological differences provide additional support for the existence of subpopulations with distinctive feeding preferences within An. vestitipennis in southern Mexico.

Cemeteries are effective sites for monitoring la crosse virus (LACv) and these environments may play a role in LACv infection.

La Crosse encephalitis (LAC) is the leading arboviral disease among children, and was previously limited to the upper Midwest. In 2012 nine pediatric cases of LAC occurred in eastern Tennessee, including one fatal case. In an attempt to identify sites near an active LACv infection and describe the abundance and distribution of potential LACv vectors near a fatal LAC case in the Appalachian region, we initiated an end of season study using a combination of questing and oviposition mosquito traps placed at forty-nine sites consisting of cemeteries and houses within 16 radial kilometers of two pediatric infections. LACv was isolated from three Aedes triseriatus pools collected from cemeteries and spatial clustering analysis identified clusters of Ae. triseriatus and Ae. albopictus populations that overlapped in the same area as the 2012 LACv cases. Results indicate cemeteries are effective sites for monitoring LACv. The role of cemeteries and specific environmental features will be the focus of future investigations.

Historical inability to control Aedes aegypti as a main contributor of fast dispersal of chikungunya outbreaks in Latin America.

The arrival of chikungunya fever (CHIKF) in Latin American countries has been expected to trigger epidemics and challenge health systems. Historically considered as dengue-endemic countries, abundant Aedes aegypti populations make this region highly vulnerable to chikungunya virus (CHIKV) circulation. This review describes the current dengue and CHIKF epidemiological situations, as well as the role of uncontrolled Ae. aegypti and Aedes albopictus vectors in spreading the emerging CHIKV. Comments are included relating to the vector competence of both species and failures of surveillance and vector control measures. Dengue endemicity is a reflection of these abundant and persistent Aedes populations that are now spreading CHIKV in the Americas. This article forms part of a symposium in Antiviral Research on "Chikungunya discovers the New World."

Serologic survey of domestic animals for zoonotic arbovirus infections in the Lacandón Forest region of Chiapas, Mexico.

A serologic survey in domestic animals (birds and mammals) was conducted in four communities located in the Lacandón Forest region of northeastern Chiapas, Mexico, during June 29 to July 1, 2001, with the objective to identify zoonotic arboviruses circulating in this area. We collected 202 serum samples from healthy domestic chickens, geese, ducks, turkeys, horses and cattle. The samples were tested by plaque-reduction neutralization test for antibodies to selected mosquito-borne flaviviruses (family Flaviviridae), including St. Louis encephalitis (SLE), Rocio (ROC), Ilheus (ILH), Bussuquara (BSQ), and West Nile (WN) viruses, and selected alphaviruses (family Togaviridae), including Western equine encephalitis (WEE), Eastern equine encephalomyelitis (EEE), and Venezuelan equine encephalitis (VEE) viruses. Neutralizing antibodies to SLE virus were detected in two (8%) of 26 turkeys, 15 (23%) of 66 cattle, and three (60%) of five horses. Antibodies to VEE virus were detected in 29 (45%) of 65 cattle. Because some of these animals were as young as 2 months old, we demonstrated recent activity of these two viruses. Sub-typing of the VEE antibody responses indicated that the etiologic agents of these infections belonged to the IE variety of VEE, which has been reported from other regions of Chiapas. WN virus-neutralizing antibodies were detected in a single cattle specimen (PRNT(90) = 1:80) that also circulated SLE virus-neutralizing antibodies (PRNT(90) = 1:20), suggesting that WN virus may have been introduced into the region. We also detected weak neutralizing activity to BSQ virus in four cattle and a chicken specimen, suggesting the presence of this or a closely related virus in Mexico. There was no evidence for transmission of the other viruses (ROC, ILH, EEE, WEE) in the study area.

Fine structure of the eggs of Anopheles (Anopheles) punctimacula.

The egg of Anopheles (Anopheles) punctimacula is described from scanning electron micrographs. Eggs of An. punctimacula are boat shaped, with lateral floats extending 70% of the length of the egg. Plastronlike polyhedral chorionic cells with distinctive boundaries and round tubercles in the cell field cover the dorsal, lateral, and ventral surfaces. Narrow decks enclosing a field of irregular jagged tubercles and 2-4 lobed tubercles are present at both egg poles.

Mark-recapture studies of host selection by Anopheles (Anopheles) vestitipennis.

We present herein the results of a series of mark-recapture experiments with female Anopheles vestitipennis. Theses experiments used human and animal hosts to assess the degree of anthropophily of field-caught specimens, originally collected on either host, and of their offspring. Fidelity of mosquitoes to particular hosts was estimated by recapturing marked host-seeking mosquitoes returning for a 2nd blood meal. Results indicated that mosquitoes seeking animal hosts were more faithful (80.48%; 33 of 41) in returning to their original host than were those seeking human hosts (63%; 49 of 78).

Diversity of mosquitoes and the aquatic insects associated with their oviposition sites along the Pacific coast of Mexico.

BACKGROUND: The abundance, richness and diversity of mosquitoes and aquatic insects associated with their oviposition sites were surveyed along eight states of the Pacific coast of Mexico. Diversity was estimated using the Shannon index (H'), similarity measures and cluster analysis. METHODS: Oviposition sites were sampled during 2-3 months per year, over a three year period. Field collected larvae and pupae were reared and identified to species following adult emergence. Aquatic insects present at oviposition sites were also collected, counted and identified to species or genus. RESULTS: In total, 15 genera and 74 species of mosquitoes were identified: Anopheles pseudopunctipennis, An. albimanus and Aedes aegypti were the most abundant and widely-distributed species, representing 47% of total mosquito individuals sampled. New species records for certain states are reported. Anopheline diversity was lowest in Sinaloa state (H' = 0.54) and highest in Chiapas (H' = 1.61) and Michoacán (H' = 1.56), whereas culicid diversity was lowest in Michoacán (H' = 1.93), Colima (H' = 1.95), Sinaloa (H' = 1.99) and Jalisco (H' = 2.01) and highest in Chiapas (H' = 2.66). In total, 10 orders, 57 families, 166 genera and 247 species of aquatic insects were identified in samples. Aquatic insect diversity was highest in Chiapas, Oaxaca and Michoacán (H' = 3.60-3.75). Mosquito larval/pupal abundance was not correlated with that of predatory Coleoptera and Hemiptera. CONCLUSION: This represents the first update on the diversity and geographic distribution of the mosquitoes and aquatic insects of Mexico in over five decades. This information has been cataloged in Mexico's National Biodiversity Information System (SNIB-CONABIO) for public inspection.

Antibodies to West Nile virus in wild and farmed crocodiles in southeastern Mexico.

Surveillance for evidence of West Nile virus (WNV) infection in Morelet's crocodiles (Crocodylus moreletii) was conducted in Campeche State, Mexico, in 2007. Sera from 62 crocodiles (32 free-ranging and 30 captive) were assayed for antibodies to WNV by epitope-blocking enzyme-linked immunosorbent assay. Antibodies to WNV were detected in 13 (41%) wild and nine (30%) captive crocodiles, and the overall antibody prevalence was 35%. Although evidence of WNV infection in captive crocodiles has been reported in Mexico, we provide the first evidence of WNV exposure in wild crocodiles in Mexico.

Serologic evidence of flavivirus infection in bats in the Yucatan Peninsula of Mexico.

We captured 140 bats of seven species in Merida City in the Yucatan Peninsula of Mexico in 2010. Serum was collected from each bat and assayed by plaque reduction neutralization test (PRNT) using six flaviviruses: West Nile virus, St. Louis encephalitis virus, and dengue viruses 1-4. Flavivirus-specific antibodies were detected in 26 bats (19%). The antibody-positive bats belonged to three species: the Pallas's long-tongued bat (Glossophaga soricina), Jamaican fruit bat (Artibeus jamaicensis), and great fruit-eating bat (Artibeus lituratus), and their flavivirus antibody prevalences were 33%, 24%, and 9%, respectively. The PRNT titers were usually highest for dengue virus 2 or dengue virus 4, but none of the titers exceeded 80. These data could indicate that most of the antibody-positive bats had been infected with dengue virus. However, because all titers were low, it is possible that the bats had been infected with another (perhaps unrecognized) flavivirus not included in the PRNT analysis, possibly a virus more closely related to dengue virus than to other flaviviruses. Each serum sample was assayed for flavivirus RNA by reverse transcription PCR, but all were negative.

West Nile virus activity in mosquitoes and domestic animals in Chiapas, México.

Prior to 2006, West Nile virus (WNV) had not been definitively detected in Chiapas, the southernmost state of Mexico, although it circulates elsewhere in Mexico and Central America. We collected over 30,000 mosquitoes and blood-sampled 351 domestic animals in Chiapas in search for evidence of current or recent transmission of WNV. Two mosquito pools tested positive for WNV RNA and 17 domestic animals tested positive for specific WNV-neutralizing antibodies, including young animals (<1 year old) in four of five sampled locations. The two WNV-positive mosquito pools were collected on the Pacific coastal plain of Chiapas in June, 2006, and included a pool of Culex nigripalpus, a suspected vector of WNV, and a pool of Cx. interrogator. The sequence of a 537-nucleotide portion of a cDNA amplicon derived from the WNV NS5 gene from the Cx. interrogator pool contained a single silent nucleotide substitution when compared to WNV strain NY99.