Two reassortant H16 influenza A viruses were isolated from gulls in California. Seven of the eight segments were most closely related to H16 and H13 isolates from eastern North America and Iceland. Of note is a C-terminal truncation of the nonstructural 1 (NS1) protein in one of the isolates that is usually found in swine H1N1 virus.

Background

Babesia conradae is an intraerythrocytic piroplasm infecting dogs in the southern United States. Ticks have been suspected, but unproven, as vectors. We identified B. conradae and other blood-borne pathogens in 2 kennels of sighthounds with a history of coyote fighting.

Objectives

To examine clinicopathologic abnormalities associated with B. conradae infection, risk factors for infection, and the prevalence of coinfections with other blood-borne pathogens.

Animals

Fifty-five Greyhounds and Greyhound mixes METHODS: Blood samples were collected from each dog for CBC, serum biochemistry panel, conventional and real-time PCR assays (Babesia spp., hemoplasmas, Ehrlichia canis, Bartonella spp., Anaplasma spp., and Rickettsia spp.), vector-borne pathogen ELISA, and immunofluorescent serology and culture for Bartonella spp and Francisella tularensis sero-agglutination test. Associations between B. conradae infection and coyote fighting, age and laboratory abnormalities were investigated.

Results

Twenty-nine dogs were PCR-positive for B. conradae. Of these, 16 were PCR-positive for other vector-borne organisms including Mycoplasma haemocanis, "Candidatus Mycoplasma haematoparvum," E. canis, and a Hepatozoon felis-like organism. Twelve of the 20 dogs tested for seroreactivity to Bartonella spp. antigens were positive, but none were seropositive for tularemia. Infection with B. conradae was associated with a history of aggressive interactions with coyotes; lower hematocrit, leukocyte count, MCHC, platelet count and serum albumin concentration; and higher MCV, MPV, and serum globulin concentration.

Conclusions and clinical importance

Babesia conradae infection should be considered in dogs with anemia, leukopenia, thrombocytopenia, hypoalbuminemia and hyperglobulinemia. As with B. gibsoni, aggressive interactions with other canids may play a role in B. conradae transmission.

Influenza A viruses (IAVs) utilize sialylated host glycans as ligands for binding and infection. The glycan-binding preference of IAV hemagglutinin (HA) is an important determinant of host specificity. Propagation of IAV in embryonated chicken eggs and cultured mammalian cells yields viruses with amino acid substitutions in the HA that can alter the binding specificity. Therefore, it is important to determine the binding specificity of IAV directly in primary samples since it reflects the actual tropism of virus in nature. We developed a novel platform for analysis of IAV binding specificity in samples that contain very low virus titers. This platform consists of a high-density flexible glycan display on magnetic beads, which promotes multivalent interactions with the viral HA. Glycan-bound virus is detected by quantifying the viral neuraminidase activity via a fluorogenic reporter, 2'-(4-methylumbelliferyl)-α-d-N-acetylneuraminic acid. This method eliminates the need for labeling the virus and significantly enhances the sensitivity of detection.

We report the complete genome sequence of a reassortant H14N2 avian influenza virus isolated in 2011 from a northern shoveler in California. This introduced Eurasian subtype acquired seven segments from North American viruses and circulated in the Pacific Flyway 1 year after its detection in the Mississippi Flyway.

A high prevalence and diversity of avian influenza (AI) viruses were detected in a population of wild mallards sampled during summer 2011 in California, providing an opportunity to compare results obtained before and after virus culture. We tested cloacal swab samples prior to culture by matrix real-time PCR, and by amplifying and sequencing a 640bp portion of the hemagglutinin (HA) gene. Each sample was also inoculated into embryonated chicken eggs, and full genome sequences were determined for cultured viruses. While low matrix Ct values were a good predictor of virus isolation from eggs, samples with high or undetectable Ct values also yielded isolates. Furthermore, a single passage in eggs altered the occurrence and detection of viral strains, and mixed infections (different HA subtypes) were detected less frequently after culture. There is no gold standard or perfect reference comparison for surveillance of unknown viruses, and true negatives are difficult to distinguish from false negatives. This study showed that sequencing samples prior to culture increases the detection of mixed infections and enhances the identification of viral strains and sequences that may have changed or even disappeared during culture.