Molecular Detection of Avian Influenza Virus from Sediment Samples in Waterfowl Habitats on the Delmarva Peninsula, United States.

Avian influenza viruses (AIV) affect many species of birds including waterfowl and may persist in sediment in aquatic habitats. Sediment samples were collected from two areas representative of prime migration and overwintering waterfowl habitat in Dorchester County, Maryland in the fall and winter of 2013-2014. Samples were screened for the presence of AIV via reverse transcriptase-quantitative PCR targeting the matrix gene. Although 13.6% of sediment samples were positive for the AIV matrix gene across all collection dates and locations, differences in detection were noted with location and collection season. Percentage of AIV-positive sediment samples recovered corresponded to trends in waterfowl abundance at collection sites both temporally and spatially. These findings provide further support for the assertion that the presence of AIV in the aquatic environment is likely affected by the total number, site-specific density, and array of waterfowl species.

Isolation and characterization of new genetic types of Toxoplasma gondii and prevalence of Trichinella murrelli from black bear (Ursus americanus).

Black bears (Ursus americanus) are hosts for two important zoonotic parasites, Toxoplasma gondii and Trichinella spp. and bears are hunted for human consumption in the USA. Little is known of the genetic diversity of T. gondii circulating in wildlife. In the present study, antibodies to T. gondii were found in juice from tongues of 17 (25.7%) of 66 wild black bear from Maryland during the hunting season of 2010 and 2011. Antibodies to T. gondii were assessed by the modified agglutination test. Tongues of 17 seropositive bears were bioassayed in mice and viable T. gondii was isolated from three samples. These three T. gondii isolates (TgBbMd1-3) were further propagated in cell culture and DNA isolated from culture-derived tachyzoites was characterized using 11 PCR-RFLP markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Results revealed three genotypes. TgBbMd1 is a Type 12 strain (ToxoDB PCR-RFLP genotype #4) and TgBbMd2 is ToxoDB PCR-RFLP genotype #216, and TgBbMd3 is a Type II clonal strain (ToxoDB PCR-RFLP genotype #1). The isolate TgBbMd2 was highly virulent for outbred Swiss Webster mice; all infected mice died of acute toxoplasmosis. Results indicate that mouse virulent strains of T. gondii are circulating in wildlife in the USA. These 66 tongues in addition to tongues collected during hunts in previous years were further investigated for the presence of muscle larvae of Trichinella spp. Tongues from 40 bears in 2005, 41 in 2006, 51 in 2007, 56 in 2008, 68 in 2009, 67 in 2010, and 66 in 2011 were subjected to digestion with pepsin/HCl and microscopic examination. Two bears were infected with Trichinella spp.; one in 2008 and one in 2009. Genotyping of collected muscle larvae revealed that the infecting species in both cases was Trichinella murrelli.

Potential of low pathogenicity avian influenza viruses of wild bird origin to establish experimental infections in turkeys and chickens.

The potential of low pathogenicity (LP) avian influenza virus (AIV) isolates of wild bird origin to establish infection in commercial turkeys and broiler chickens was studied. Isolates, representing subtypes H5N1, H7N3, H6N2, and H3N6, were recovered in 2005 and 2006 from waterfowl and shorebirds in the Delmarva Peninsula region of the east coast of the United States. The LP AIV isolates were not pathogenic for 2-wk-old meat-type turkeys and broiler chickens. No mortality, clinical signs, or gross lesions were observed following intratracheal and conjunctival sac routes of exposures with 10(6.0) EID50 (embryo infectious dose) per bird. Isolates resulting in an established infection based on virus isolation were: A/mallard/Maryland/1159/ 2006 (H5N1) in the upper respiratory tract of turkeys; A/mallard/Delaware/418/2005 (H7N3) in the upper respiratory and intestinal tracts of turkeys and chickens; and A/shorebird-environment/Delaware/251/2005 (H3N6) in the upper respiratory and intestinal tracts of chickens. Infections were also confirmed by production of AIV-specific serum antibodies detected by hemagglutination inhibition.