Isolation and genetic characterization of Toxoplasma gondii from mute swan (Cygnus olor) from the USA.

Little is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study, antibodies to T. gondii were determined in serum samples from 632 mute swans (Cygnus olor) collected from different areas of the USA. Sera were tested by T. gondii modified agglutination test; 54 (8.5%) of 632 samples were seropositive with titers of 25 in 28 sera, 50 in 22 sera, 100 in three samples, and 200 or higher in one swan. Hearts from 14 seropositive swans were bioassayed in mice and viable T. gondii (designated TgSwanUs1-3) were isolated from the hearts of three. These three T. gondii isolates 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 of genotyping revealed that two strains (TgSwanUs1, TgSwanUs2) were Type III (ToxoDB PCR-RFLP genotype #2), and TgSwanUs3 was a new genotype designated here as ToxoDB PCR-RFLP genotype #216. Pathogenicity of oocysts derived from these three strains was determined in Swiss Webster (SW) outbred mice. All mice infected with oocysts and tachyzoites of the atypical isolate (TgSwanUs3) died of acute toxoplasmosis, irrespective of the dose. Oocysts of the remaining two isolates were less pathogenic but differed from each other; 10 oocysts of the TgSwanUs1 killed all inoculated mice whereas 1 million oocysts of the TgSwanUs2 were needed to kill all infected SW mice. Isolation of T. gondii from mute swan indicates that the local waters were contaminated by T. gondii oocysts, and that mouse T. gondii virulent strains are circulating in wildlife. Mute swan is a new host record for T. gondii.

Leukocyte response to eastern equine encephalomyelitis virus in a wild passerine bird.

Leukocyte counts are frequently used to assess the immunologic status of animals; however, few studies have directly looked at the predictive value of leukocyte counts and an animal's ability to respond to an infection with a pathogen. Understanding how an animal's leukocyte profile is altered by an active infection can assist with interpretation of leukocyte profiles in animals for which infection status is not known. In this study we examine the leukocyte counts of gray catbirds (Dumetella carolinensis) infected with eastern equine encephalomyelitis virus (EEEV). Blood smears were collected from infected catbirds on -4, 2, 5, and 14 days postinoculation (dpi) with EEEV, and from a corresponding uninfected control group, to monitor leukocyte counts. Although we found that preinfection leukocyte counts were not a reliable predictive of a catbird's viremia, we did find that infected catbirds exhibited significant hematologic changes in response to EEEV infection. We observed a significant drop in all subpopulations of leukocytes (i.e., lymphocytes, monocytes, and granulocytes) following infection. Lymphocytes and granulocytes still had not recovered to preinfection levels at 14 dpi. Uninfected catbirds also exhibited statistically significant changes in leukocyte counts, but this was due to a slight increase at 14 dpi and was not considered biologically relevant. Studies such as this can provide important information for field ecoimmunologists that use leukocyte counts to assess immunocompetence in free-living animals.

Prevalence of avian paramyxovirus 1 and avian influenza virus in double-crested Cormorants (Phalacrocorax auritus) in eastern North America.

Although it is well established that wild birds, such as cormorants, carry virulent avian paramyxovirus serotype 1 (APMV-1; causative agent of Newcastle disease) and avian influenza virus (AIV), the prevalence of these viruses among Double-crested Cormorants (Phalacrocorax auritus) in the Great Lakes region of North America has not been rigorously studied. We determined the prevalences of APMV-1 and AIV in Double-crested Cormorants from the interior population of eastern North America. From 2009 to 2011, oropharyngeal and cloacal swabs and serum samples were collected from 1,957 individual Double-crested Cormorants, ranging from chicks to breeding adults, on breeding colony sites in Michigan, Wisconsin, and Mississippi, USA, and Ontario, Canada, as well as on the wintering grounds of migratory populations in Mississippi, USA. Prevalence of antibodies to APMV-1 in after-hatch year birds was consistently high across all three years, ranging from 86.3% to 91.6%. Antibody prevalences in chicks were much lower: 1.7, 15.3, and 16.4% in 2009, 2010, and 2011, respectively. Virulent APMV-1 was detected in six chicks sampled in 2010 in Ontario, Canada. Only one adult was positive for AIV-specific antibodies and five individuals were positive for AIV matrix protein, but the latter were negative for H5 and H7 AIV subtypes. We provide further evidence that Double-crested Cormorants play an important role in the maintenance and circulation of APMV-1 in the wild, but are unlikely to be involved in the circulation of AIV.