Leah Colton

West Nile virus-infected mosquitoes, Louisiana, 2002.

Culex quinquefasciatus was identified as probable vector.

Host-Seeking Heights, Host-Seeking Activity Patterns, and West Nile Virus Infection Rates for Members of the Culex pipiens Complex at Different Habitat Types Within the Hybrid Zone, Shelby County, TN, 2002 (Diptera: Culicidae)

Abstract Host-seeking heights, host-seeking activity patterns, and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) infection rates were assessed for members of the Culex pipiens complex from July to December 2002, by using chicken-baited can traps (CT) at four ecologically different sites in Shelby County, TN. Host-seeking height was assessed by CT placed at elevations of 3.1, 4.6, and 7.6 m during one 24-h period per month. Host-seeking activity was assessed by paired CT placed at an elevation of 4.6 m. Can traps were sampled at one 10-h daytime interval and at seven 2-h intervals during the evening, night, and morning. Cx. pipiens complex mosquitoes accounted for 87.1% of collected mosquitoes. Culex (Melanoconion) erraticus (Dyar & Knab) accounted for 11.9% of specimens. The average number of Cx. pipiens complex mosquitoes collected per 24-h CT period from July to September was lowest at a rural middle income site (1.7), intermediate at an urban middle income site (11.3), and highest at an urban low income site (47.4). Can traps at the forested site failed to collect Cx. pipiens complex mosquitoes. From July to September at urban sites, Culex pipiens pipiens L. was the rarest of the three complex members accounting for 11.1–25.6% of specimens. At the rural site, Culex pipiens quinquefasciatus Say was the rarest member of the complex. Cx. p. pipiens was not collected after September. Mean abundance of Cx. pipiens complex mosquitoes was higher in traps at 7.6 m than in traps at 4.6 m. Abundances at 3.1 m were intermediate and not significantly different from abundances at the other heights. Initiation of host-seeking activity was associated with the end of civil twilight and activity occurred over an extended nighttime period lasting 8–10 h. All 11 WNV-positive mosquitoes were Cx. pipiens complex mosquitoes collected from urban sites in traps placed at elevations of 4.6 and 7.6 m. Infection rates were marginally nonsignificant by height. Infection rates, host-seeking heights, and activity patterns were not significantly different among members of the Cx. pipiens complex.

Quantification of West Nile virus in vector mosquito saliva.

ABSTRACT Saliva was collected from 4 species of mosquitoes intrathoracically inoculated with West Nile virus (WNV). The amount of infectious virus in the saliva was quantified by plaque assay and the number of WNV genomic equivalents (GE) was measured by reverse transcriptase–polymerase chain reaction. Ochlerotatus triseriatus had the greatest mean amount of infectious virus per saliva collection, followed by Aedes albopictus, Culex pipiens, and Cx. quinquefasciatus. The mean GE/saliva collection was also greatest in Oc. triseriatus, followed by Cx. quinquefasciatus, Cx. pipiens, and Ae. albopictus. The variance of log GE/saliva collection for Ae. albopictus was significantly lower than the variance for the other 3 species. This study provides a basis for comparing this component of vector competence and for determining the amounts of virus inoculated into vertebrates in experimental host competence studies.

Oviposition Activity Patterns and West Nile Virus Infection Rates for Members of the Culex pipiens Complex at Different Habitat Types within the Hybrid Zone, Shelby County, TN, 2002 (Diptera: Culicidae)

Abstract Oviposition activity and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) infection rates were assessed for members of the Culex pipiens complex from July through December 2002 by using gravid traps placed at four ecologically different sites in the southern portion of the hybrid zone in Shelby County, TN. Molecular assays identified three members of the Cx. pipiens complex: Cx. pipiens pipiens L., Cx. p. quinquefasciatus Say, and Cx. p. pipiens–Cx. p. quinquefasciatus hybrids (hybrids). The Cx. pipiens complex accounted for 90% of mosquitoes collected in gravid traps. All 285 WNV-positive mosquitoes were Culex mosquitoes, and 277 (97%) were Cx. pipiens complex mosquitoes. Infection rates among members of the Cx. pipiens complex were not significantly different. Infection rates were significantly higher at two urban sites than at a rural site, and WNV was not detected at a forested site. At urban sites, abundances of members of the Cx. pipiens complex corresponded to a simple latitude model of the hybrid zone. Cx. p. quinquefasciatus was most abundant (46.4%), followed by hybrids (34.1%) and Cx. p. pipiens (19.5%). The relative abundances at a rural site were reversed with Cx. p. pipiens (48.4%) being most abundant. This demonstrates that spatial habitat variation may profoundly influence the distribution of members of the Cx. pipiens complex within the hybrid zone. Members of the Cx. pipiens complex did not display different oviposition patterns. However, oviposition patterns assessed hourly at urban and rural sites were significantly different. At urban sites, oviposition activity of Cx. pipiens complex mosquitoes was bimodal with an evening peak associated with sunset and a morning peak associated with sunrise. At the rural site, the evening peak was pronounced and the morning peak weak and similar to nighttime activity.

QUANTIFICATION OF WEST NILE VIRUS IN THE SALIVA OF CULEX SPECIES COLLECTED FROM THE SOUTHERN UNITED STATES

ABSTRACT Culex quinquefasciatus, Cx. restuans, Cx. pipiens complex, and Cx. nigripalpus were collected as larvae or egg rafts from the southern USA. Adult female mosquitoes were intrathoracically inoculated with ~1,000 plaque-forming units of West Nile virus (WNV) and saliva was collected from them 5 days later. The amount of infectious WNV in the saliva samples was quantified by plaque assay and WNV RNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). More than 90% of the mosquitoes had either infectious virus or viral RNA in their saliva. The RT-PCR assay detected a greater percent of samples with WNV RNA than the plaque assay detected infectious virus. Pairwise comparisons revealed 6 significant differences between the 7 groups surveyed. The Cx. nigripalpus secreted lower mean amounts of WNV than 3 other groups, and a difference was found between early- and late-season Cx. quinquefasciatus collected in Louisiana.

Molecular Detection and Identification of Bartonella Species in Rat Fleas from Northeastern Thailand

The presence of Bartonella species in Xenopsylla cheopis fleas collected from Rattus spp. (R. exulans, R. norvegicus, and R. rattus) in Khon Kaen Province, Thailand was investigated. One hundred ninety-three fleas obtained from 62 rats, were screened by polymerase chain reaction using primers specific for the 16S-23S intergenic spacer region, and the presence of Bartonella DNA was confirmed by using the citrate synthase gene. Bartonella DNA was detected in 59.1% (114 of 193) of fleas examined. Sequencing demonstrated the presence of Bartonella spp. similar to B. elizabethae, B. rattimassiliensis, B. rochalimae, and B. tribocorum in the samples tested with a cutoff for sequence similarity ≥ 96% and 4 clustered together with the closest match with B. grahamii (95.5% identity). If X. cheopis proves to be a competent vector of these species, our results suggest that humans and animals residing in this area may be at risk for infection by several zoonotic Bartonella species.

Ecology of Potential West Nile Virus Vectors in Southeastern Louisiana: Enzootic Transmission in the Relative Absence of Culex quinquefasciatus

A study of West Nile virus (WNV) ecology was conducted in St. Tammany Parish, Louisiana, from 2002 to 2004. Mosquitoes were collected weekly throughout the year using Centers for Disease Control and Prevention (CDC) light traps placed at 1.5 and 6 m above the ground and gravid traps. A total of 379,466 mosquitoes was collected. WNV was identified in 32 pools of mosquitoes comprising four species; 23 positive pools were from Culex nigripalpus collected during 2003. Significantly more positive pools were obtained from Cx. nigripalpus collected in traps placed at 6 m than 1.5 m that year, but abundance did not differ by trap height. In contrast, Cx. nigripalpus abundance was significantly greater in traps placed at 6 m in 2002 and 2004. Annual temporal variation in Cx. nigripalpus peak seasonal abundance has important implications for WNV transmission in Louisiana. One WNV-positive pool, from Cx. erraticus, was collected during the winter of 2004, showing year-round transmission. The potential roles of additional mosquito species in WNV transmission in southeastern Louisiana are discussed.

Experimental infection of Swiss webster mice with four rat bartonella strains: host specificity, bacteremia kinetics, dose dependent response, and histopathology.

Groups of Swiss Webster outbred mice were each inoculated with one of four bartonella strains originally isolated from Rattus spp. at doses ranging from 10(1) to 10(7) bacteria per mouse. One strain, Rn1691yn (Bartonella coopersplainensis-like), infected mice and produced bacteremias at levels up to 10(5)bacteria/ml of blood and from 3 to 8 weeks duration. A dose dependent response was also observed with differing proportions of mice bacteremic following inoculation at different doses. In addition weeks-to-months long lags in bacteremia manifestation occurred following lower dose exposures. The possibility of bacterial transmission from bacteremic mice to uninfected cagemates was assessed and no naïve mice became infected from contacts with infected mice. Finally, a subset of bacteremic mice inoculated with high doses of Rn1691yn were examined histopathologically and multifocal, granulomatous lesions were detected in both liver and kidneys. The host specificity and infectivity of the strains is discussed in relation to their potential for zoonotic transmission to incidental hosts.

Experimental infection of three laboratory mouse stocks with a shrew origin Bartonella elizabethae strain: an evaluation of bacterial host switching potential

Background : Bartonella elizabethae has been reported as a causative agent of human illnesses and strains of this bacterium are commonly isolated from commensal small mammals in Asia. Methods : Since the zoonotic potential of a pathogen is often related to its host switching ability, we explored the capacity of a B. elizabethae strain to host switch by subcutaneously inoculating groups of Swiss Webster, BALB/c, and C57BL/6 mice with the bacteria at a range of doses. Results : A low number of mice in each of the three groups showed susceptibility to infection at high doses (105 and 106 bacteria), and developed bacteremias of 6–8 weeks duration. Conclusion: The capacity of this B. elizabethae strain to switch hosts can have important public health consequences for humans in areas of Asia where many small mammal populations have high bartonellae infection prevalences and live as commensals with humans.

Human isolates of Bartonella tamiae induce pathology in experimentally inoculated immunocompetent mice

BackgroundBartonella tamiae, a newly described bacterial species, was isolated from the blood of three hospitalized patients in Thailand. These patients presented with headache, myalgia, anemia, and mild liver function abnormalities. Since B. tamiae was presumed to be the cause of their illness, these isolates were inoculated into immunocompetent mice to determine their relative pathogenicity in inducing manifestations of disease and pathology similar to that observed in humans.MethodsThree groups of four Swiss Webster female mice aged 15-18 months were each inoculated with 106-7 colony forming units of one of three B. tamiae isolates [Th239, Th307, and Th339]. A mouse from each experimental group was sampled at 3, 4, 5 and 6 weeks post-inoculation. Two saline inoculated age-matched controls were included in the study. Samples collected at necropsy were evaluated for the presence of B. tamiae DNA, and tissues were formalin-fixed, stained with hematoxylin and eosin, and examined for histopathology.ResultsFollowing inoculation with B. tamiae, mice developed ulcerative skin lesions and subcutaneous masses on the lateral thorax, as well as axillary and inguinal lymphadenopathy. B. tamiae DNA was found in subcutaneous masses, lymph node, and liver of inoculated mice. Histopathological changes were observed in tissues of inoculated mice, and severity of lesions correlated with the isolate inoculated, with the most severe pathology induced by B. tamiae Th239. Mice inoculated with Th239 and Th339 demonstrated myocarditis, lymphadenitis with associated vascular necrosis, and granulomatous hepatitis and nephritis with associated hepatocellular and renal necrosis. Mice inoculated with Th307 developed a deep dermatitis and granulomas within the kidneys.ConclusionsThe three isolates of B. tamiae evaluated in this study induce disease in immunocompetent Swiss Webster mice up to 6 weeks after inoculation. The human patients from whom these isolates were obtained had clinical presentations consistent with the multi-organ pathology observed in mice in this study. This mouse model for B. tamiae induced disease not only strengthens the causal link between this pathogen and clinical illness in humans, but provides a model to further study the pathological processes induced by these bacteria.