Antonia Dibernardo

West Nile virus outbreak in North American owls, Ontario, 2002.

Susceptibility of North American owls to WNV is associated with native breeding range.

The prevalence of Borrelia miyamotoi infection, and co-infections with other Borrelia spp. in Ixodes scapularis ticks collected in Canada

BackgroundBlacklegged ticks, Ixodes scapularis are vectors of the tick-borne pathogens Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti. Recently, the I. scapularis-borne bacterium Borrelia miyamotoi has been linked to human illness in North America. The range of this tick is expanding in Canada which may increase the potential for human exposure to these agents.MethodsIn this study, 4938 I. scapularis ticks collected in 2012 were tested following a newly developed PCR-based testing protocol to determine the prevalence of infection with B. miyamotoi and other pathogens in I. scapularis in Canada.ResultsBorrelia miyamotoi was detected in blacklegged ticks from all provinces except Newfoundland, although the infection prevalence was low (<1%). There was significant variation among provinces in the prevalence of infection of ticks with B. burgdorferi and A. phagocytophilum, but not with B. miyamotoi.ConclusionsGiven the widespread distribution of B. miyamotoi, infection due to this agent should be considered in patients who have been exposed to blacklegged ticks in Canada.

Rapid Antigen-Capture Assay To Detect West Nile Virus in Dead Corvids

The utility of the VecTest antigen-capture assay to detect West Nile virus (WNV) in field-collected dead corvids was evaluated in Manitoba and Ontario, Canada, in 2001 and 2002. Swabs were taken from the oropharynx, cloaca, or both of 109 American Crows, 31 Blue Jays, 6 Common Ravens, and 4 Black-billed Magpies from Manitoba, and 255 American Crows and 28 Blue Jays from Ontario. The sensitivity and specificity of the antigen-capture assay were greatest for samples from American Crows; oropharyngeal swabs were more sensitive than cloacal swabs, and interlaboratory variation in the results was minimal. The sensitivity and specificity of the VecTest using oropharyngeal swabs from crows were 83.9% and 93.6%, respectively, for Manitoba samples and 83.3% and 95.8%, respectively, for Ontario birds. The VecTest antigen-capture assay on oropharyngeal secretions from crows is a reliable and rapid diagnostic test that appears suitable for incorporation into a WNV surveillance program.

EVALUATION OF COMMERCIAL ASSAYS FOR DETECTING WEST NILE VIRUS ANTIGEN

ABSTRACT Two commercially available West Nile virus (WNV) detection assays (RAMP® WNV test, Response Biomedical Corp., Burnaby, British Columbia, Canada; and VecTest™ WNV antigen assay, Medical Analysis Systems, Inc., Camarillo, CA) were compared for sensitivity, specificity, and ability to detect WNV in field-collected mosquito pools. Serially diluted stock seed WNV and St. Louis encephalitis virus (SLEV) were used to determine sensitivity and specificity. The RAMP WNV test detected WNV at concentrations as low as 3.17 log10 plaque-forming units per milliliter (PFU/ml), whereas the VecTest assay detected WNV at concentrations as low as 5.17 log10 PFU/ml. Neither test cross-reacted with SLEV. A WNV-specific reverse transcriptase polymerase chain reaction was used to identify positives among field-collected mosquito pools. The RAMP WNV test detected 94% of positive pools and the VecTest assay detected 65% of the positive field-collected pools. Despite these differences, both assays have characteristics that make them useful in WNV surveillance programs.

West Nile virus infection in the eastern loggerhead shrike (Lanius ludovicianus migrans): Pathology, epidemiology, and immunization

An outbreak of West Nile virus (WNV) infection occurred at a captive breeding facility for the endangered eastern loggerhead shrike (Lanius ludovicianus migrans) in August 2002. Within 10 d, five birds died; two were found dead, and the others died shortly after showing neurologic signs. West Nile virus was detected in all organs examined using immunohistochemistry, and its viral genome was amplified from brain and kidney samples using reverse transcription–polymerase chain reaction. None of the remaining birds in the colony had antibodies against WNV, which suggests a mortality rate of 100%. After vaccination with a commercial equine WNV vaccine 31 of 37 (84%) of the birds had WNV neutralizing antibodies.

The First case of Locally Acquired Tick-Borne Babesia Microti Infection in Canada

Infection with Babesia microti can cause severe illness, particularly among asplenic individuals. Blacklegged and Western blacklegged ticks (Ixodes scapularis and Ixodes pacificus, respectively) are the vector through which B microti is transmitted. The distribution of these ticks in Canada has increased over the past several years. In this article, the authors present the first case of babesiosis in Canada that was not diagnosed following travel to an area in which the disease is endemic.

Blood collected on filter paper for wildlife serology: evaluating storage and temperature challenges of field collections.

Abstract Filter-paper (FP) blood sampling can facilitate wildlife research and expand disease surveillance. Previous work indicated that Nobuto FP samples from caribou and reindeer (Rangifer tarandus subspecies) had comparable sensitivity and specificity to serum samples (≥80% for both) in competitive enzyme-linked immunosorbent assays (cELISAs) for Brucella spp., Neospora caninum, and West Nile virus. The same sensitivity and specificity criteria were met in indirect ELISAs for Brucella spp., bovine herpesvirus type 1 (BHV-1), parainfluenza virus type 3 (PI-3), and bovine respiratory syncytial virus (BRSV), with adjusted FP thresholds used for PI-3 and BRSV. Comparable sensitivity and specificity values to serum were also observed for FP in virus neutralization (VN) assays for bovine viral diarrhea virus types I and II; however, reduced sensitivity is a potential limitation of FP samples in protocols that require undiluted serum (i.e., VN and N. caninum cELISA). We evaluated the performance of FP samples from reindeer and caribou in these nine assays after simulating potential challenges of high-latitude field collections: 1) different durations of storage and 2) different processing/storage regimes involving freezing or drying. Sample pairs (serum and FP) were collected from reindeer and caribou populations in 2007–10 and were tested in duplicate. Comparable performance to serum was defined as sensitivity and specificity ≥80%. In the storage experiments, FP performance was determined after 2 mo of storage dry at room temperature, and after two longer periods (variable depending on assay; up to 2 yr). After 1 yr, compared to frozen serum stored for the same period, sensitivity was ≥88% for all but two assays (68% BHV-1; 75% PI-3), and specificity remained >90%. A limited trial evaluated the effect of freezing FP samples as opposed to drying them for storage. There were no observed detrimental effects of freezing on FP sample performance, but rigorous investigation is warranted.

Dengue seroprevalence, seroconversion and risk factors in Dhaka, Bangladesh.

Background Dengue virus (DENV) activity has been reported in Dhaka, Bangladesh since the early 1960s with the greatest burden of dengue fever and dengue hemorrhagic fever cases observed in 2000. Since this time, the intensity of dengue activity has varied from year to year, and its determining factors remained relatively unknown. In light of such gaps in knowledge, the main objectives of this study were to determine the magnitude of seroprevalence and seroconversion among the surveyed population, and establish the individual/household level risk factors for the presence of DENV antibodies among all age groups of target populations in the city of Dhaka. Methodology/Principal findings Considering the lack of fine scale investigations on the factors driving dengue activity in Bangladesh, a prospective cohort study involving serological surveys was undertaken with participant interviews and blood donation across the city of Dhaka in 2012. Study participants were recruited from 12 of 90 wards and blood samples were collected during both the pre-monsoon (n = 1125) and post-monsoon (n = 600) seasons of 2012. The findings revealed that the seroprevalence in all pre-monsoon samples was 80.0% (900/1125) while the seropositivity in the pre-monsoon samples that had paired post-monsoon samples was 83.3% (503/600). Of the 97 paired samples that were negative at the pre-monsoon time point, 56 were positive at the post-monsoon time point. This resulted in a seroprevalence of 93.2% (559/600) among individuals tested during the post-monsoon period. Seroprevalence trended higher with age with children exhibiting a lower seropositivity as compared to adults. Results from this study also indicated that DENV strains were the only flaviviruses circulating in Dhaka in 2012. A multivariate analysis revealed that age, possession of indoor potted plants, and types of mosquito control measures were significant factors associated with DENV seroprevalence; while attendance in public/mass gatherings, and use of mosquito control measures were significantly associated with DENV seroconversion after adjusting for all other variables. Conclusions/Significance Our study suggests that there is a high level of endemic dengue virus circulation in the city of Dhaka which has resulted in significant DENV seroprevalence among its residents. Seropositivity increased with age, however, a substantial proportion of children are at risk for DENV infections. Our serological analysis also documents considerable DENV seroconversion among study participants which indicates that a large proportion of the population in the city of Dhaka were newly exposed to DENV during the study period (pre-and post-monsoon 2012). High levels of seroconversion suggest that there was an intense circulation of DENV in 2012 and this may have resulted in a significant risk for viral associated illness. Findings of our study further indicated that home-based interventions, such as removing indoor potted plants and increased bed net use, in addition to vector control measures in public parks, would reduce exposure to DENV and further decrease risk of viral associated disease.

BLOOD COLLECTED ON FILTER PAPER FOR WILDLIFE SEROLOGY: DETECTING ANTIBODIES TO NEOSPORA CANINUM, WEST NILE VIRUS, AND FIVE BOVINE VIRUSES IN REINDEER

Abstract We compared Nobuto filter paper (FP) whole-blood samples to serum for detecting antibodies to seven pathogens in reindeer (Rangifer tarandus tarandus). Serum and FP samples were collected from captive reindeer in 2008–2009. Sample pairs (serum and FP eluates) were assayed in duplicate at diagnostic laboratories with the use of competitive enzyme-linked immunosorbent assays (cELISAs) for Neospora caninum and West Nile virus (WNV); indirect ELISA (iELISAs) for bovine herpesvirus type 1 (BHV-1), parainfluenza virus type 3 (PI-3), and bovine respiratory syncytial virus (BRSV); and virus neutralization (VN) for bovine viral diarrhea virus (BVDV) types I and II. Assay thresholds were evidence-based values employed by each laboratory. Comparable performance to serum was defined as FP sensitivity and specificity ≥80%. Filter-paper specificity estimates ranged from 92% in the cELISAs for N. caninum and WNV to 98% in the iELISAs for PI-3 and BRSV. Sensitivity was >85% for five tests (most ≥95%) but was insufficient (71–82%) for the PI-3 and BRSV iELISAs. Lowering the threshold for FP samples in these two ELISAs raised sensitivity to ≥87% and reduced specificity slightly (≥90% in three of the four test runs). Sample size limited the precision of some performance estimates. Based on the criteria of sensitivity and specificity ≥80%, and using adjusted FP thresholds for PI-3 and BRSV, FP sensitivity and specificity were comparable to serum in all seven assays. A potential limitation of FP is reduced sensitivity in tests that require undiluted serum (i.e., N. caninum cELISA and BVDV VNs). Possible toxicity to the assay cell layer in VN requires investigation. Results suggested that cELISA is superior to iELISA for detecting antibodies in FP samples from reindeer and other Rangifer tarandus subspecies. Our findings expand the potential utility of FP sampling from wildlife.

Two Anaplasma phagocytophilum Strains in Ixodes scapularis Ticks, Canada

We developed PCR-based assays to distinguish a human pathogenic strain of Anaplasma phagocytophilum, Ap-ha, from Ap-variant 1, a strain not associated with human infection. The assays were validated on A. phagocytophilum-infected blacklegged ticks (Ixodes scapularis) collected in Canada. The relative prevalence of these 2 strains in I. scapularis ticks differed among geographic regions.