Organochlorine contaminants in wild mink from the lower Great Lakes basin, Canada, 1998-2006.

Organochlorine contaminants were measured in livers of wild mink (Neovison vison) trapped in the lower Great Lakes basin from 1998 to 2006. To assess exposure and potential risk in mink feeding on Great Lakes biota, concentrations of contaminants were compared in mink trapped within 7.8 km of the shoreline as well as at inland sites (i.e., 8-40 km). Overall, significant spatial variation in mean hepatic concentrations of sum PCBs and seven other organochlorines was found in mink from 13 Great Lakes sites, many of which are within the Great Lakes Areas of Concern. Mean sum PCB concentrations, on a lipid weight basis, ranged from 2 µg/g in mink from inland Lake Ontario sites to 44 µg/g in mink from western Lake Erie. Concentrations of other organochlorines in mink were generally low. Mink from western Lake Erie had the highest mean cumulative organochlorine burdens dominated largely by PCBs. A significant age effect was found with 1-year-old mink having significantly higher PCB burdens than mink less than 1 year in age. With respect to published PCB threshold effect concentrations, some mink exceeded those associated with effects on reproduction and survival as well as the presence of jaw lesions. This was most consistently found in western Lake Erie where the health of populations of wild mink may be adversely affected and where no mink 2 years of age or older were collected.

Mercury and other heavy metals in free-ranging mink of the lower Great Lakes basin, Canada, 1998-2006.

Mercury concentrations were examined in livers of free-ranging mink (Neovison vison) trapped in the lower Great Lakes basin from 1998 to 2006. Significant geographic variation in total mercury levels was found in mink from 13 Great Lakes locations, many of which are within Great Lakes Areas of Concern (AOCs). Total mercury levels in mink from these locations were generally low to moderate, with means (±SE) ranging from 0.99 (±0.36) µg/g dry weight in mink from inland Lake Erie to 7.31 (±1.52) µg/g in mink from Walpole Island in the St. Clair River AOC. Overall, mercury exposure was highest in mink trapped in locations associated with large riverine wetlands. Total mercury concentrations were negatively related to the percentage of mercury present as methylmercury and positively related to selenium concentrations, consistent with increasing demethylation of methylmercury with increasing total mercury concentrations. One-year old mink had significantly higher levels of mercury and selenium than mink under 1 year but similar to mink 2 years of age and older. Mercury levels were below those associated with mercury toxicity. Concentrations of cadmium, lead and arsenic were low and largely associated with environmental background levels. Significant age and sex effects were found for cadmium. Lead levels were significantly lower in mink infected by the lung nematode parasite compared to uninfected animals. Further studies of biomagnification of methylmercury in wetland environments, where top predators such as mink may be an increased risk of exposure, are warranted.

Development of a real-time PCR assay using SYBR Green chemistry for monitoring Marek's disease virus genome load in feather tips.

Feather follicles of birds infected with Marek's disease virus (MDV) serve as the sole source of infectious virus particles. The present study was aimed at developing a SYBR Green real-time PCR assay to detect and quantify MDV loads in feather tips targeting meq gene of the virus. The assay had a dynamic range of 8 logs, mean inter- and intra-assay coefficient variation (CV) of <5% and minimum detection limit of 15 MDV genome copies when plasmid DNA was used as the template. The sensitivity of the assay was compared with that of the conventional PCR technique and found to be 2.5-10 times more sensitive than the conventional PCR technique. The assay was validated using feather tip DNA preparations derived from chickens infected with 250 plaque forming units (PFU) of RB1B strain of MDV and sampled on days 7, 14, 21 and 28 post-infection (p.i.) along with uninfected chickens. MDV genome was quantifiable in feather tips of infected birds by day 7 p.i. and the number of MDV copies peaked by day 14 p.i., but then gradually decreased by day 28. This reliable real-time PCR assay may be used for monitoring MDV genome loads in tissues of experimentally or naturally infected birds.

Driving forces behind the evolution of the Aleutian mink disease parvovirus in the context of intensive farming.

Aleutian mink disease virus (AMDV) causes plasmacytosis, an immune complex-associated syndrome that affects wild and farmed mink. The virus can also infect other small mammals (e.g., ferrets, skunks, ermines, and raccoons), but the disease in these hosts has been studied less. In 2007, a mink plasmacytosis outbreak began on the Island of Newfoundland, and the virus has been endemic in farms since then. In this study, we evaluated the molecular epidemiology of AMDV in farmed and wild animals of Newfoundland since before the beginning of the outbreak and investigated the epidemic in a global context by studying AMDV worldwide, thereby examining its diffusion and phylogeography. Furthermore, AMDV evolution was examined in the context of intensive farming, where host population dynamics strongly influence viral evolution. Partial NS1 sequences and several complete genomes were obtained from Newfoundland viruses and analyzed along with numerous sequences from other locations worldwide that were either obtained as part of this study or from public databases. We observed very high viral diversity within Newfoundland and within single farms, where high rates of co-infection, recombinant viruses and polymorphisms were observed within single infected individuals. Worldwide, we documented a partial geographic distribution of strains, where viruses from different countries co-exist within clades but form country-specific subclades. Finally, we observed the occurrence of recombination and the predominance of negative selection pressure on AMDV proteins. A surprisingly low number of immunoepitopic sites were under diversifying pressure, possibly because AMDV gains no benefit by escaping the immune response as viral entry into target cells is mediated through interactions with antibodies, which therefore contribute to cell infection. In conclusion, the high prevalence of AMDV in farms facilitates the establishment of co-infections that can favor the occurrence of recombination and enhance viral diversity. Viruses are then exchanged between different farms and countries and can be introduced into the wild, with the rapidly evolving viruses producing many parallel lineages.

Cytokine gene expression patterns associated with immunization against Marek's disease in chickens.

The present study explored the immunological correlates of protection mediated by a live bivalent vaccine consisting of herpesvirus of turkeys (HVT) and SB-1 against infection with the RB1B strain of Marek's disease virus (MDV). Compared to unvaccinated infected chickens, vaccinated protected birds had lower expression of interleukin (IL)-6, IL-10 and IL-18 genes in spleen. However, there was no difference between these two groups of birds in the expression of interferon (IFN)-gamma, IL-4, IL-12 and inducible nitric oxide synthase (iNOS) genes on day 21 post-infection. Furthermore, protection was associated with lower MDV genome load in spleen but not in feather tips, suggesting that vaccination had little or no effect on curtailing virus transmission. In conclusion, vaccination with a bivalent MD vaccine was associated with distinct cytokine expression patterns in spleen and modulation of cytokine responses by the vaccine may play a role in mediation of protection.

Diagnosing abuse: a systematic review of torn frenum and other intra-oral injuries.

INTRODUCTION: A torn labial frenum is widely regarded as pathognomonic of abuse. METHODS: We systematically reviewed the evidence for this, and to define other intra-oral injuries found in physical abuse. Nine studies documented abusive torn labial frena in 27 children and 24 [corrected] were fatally abused: 22 were less than 5 years old. Only a direct blow to the face was substantiated as a mechanism of injury. RESULTS: Two studies noted accidentally torn labial frena, both from intubation. Abusive intra-oral injuries were widely distributed to the lips, gums, tongue and palate and included fractures, intrusion and extraction of the dentition, bites and contusions. CONCLUSIONS: Current literature does not support the diagnosis of abuse based on a torn labial frenum in isolation. The intra-oral hard and soft tissue should be examined in all suspected abuse cases, and a dental opinion sought where abnormalities are found.

Temporal and spatial trends in chlorinated hydrocarbon concentrations of mink in Canadian Lakes Erie and St. Clair.

Mink (Mustela vison) carcasses were collected from local commercial trappers from fall 1998 to spring 2003 from tributaries and marshes within 4.8 km from the shores of Lakes Erie and St. Clair, including the Wheatley Harbour and St. Clair River Areas of Concern (AOCs), as well as from inland sites (8 to 40 km from shore). Liver concentrations, on a lipid weight basis, of chlorinated hydrocarbons were measured and compared among sites and to tissue concentrations of mink from two previous collections from similar sites over the past 25 years. Mink from the western Lake Erie sites, which included the Wheatley Harbour AOC, had significantly higher concentrations of sum PCBs and PCB Aroclors than those from the St. Clair corridor or inland sites, with concentrations from the rest of Lake Erie being intermediate. Dieldrin concentrations were also significantly higher in western Lake Erie than many other sites, and those of oxychlordane and mirex also tended to be higher (0.05 < p < 0.1). There were no differences in contaminant concentrations between the St. Clair River AOC and the downstream non-AOC Lake St. Clair site, with the exception of slightly higher levels of octachlorostyrene (OCS). Concentrations of PCBs and other chlorinated hydrocarbons in mink showed a general decrease over the past two decades. In contrast, PCBs and some organochlorine pesticides tended to increase, significantly so with oxychlordane, in western Lake Erie mink over the same time period. DDE declined at all sites. Currently, mink liver PCB concentrations are within the range associated with reproductive impairment, as determined from captive mink studies, in 11.7% of all animals collected from the Lakes Erie and St. Clair basin overall, but in almost 40% of individuals from western Lake Erie.

Pathology and tissue distribution of West Nile virus in North American owls (family: Strigidae).

This study describes the macroscopic and microscopic lesions and the viral antigen distribution in 82 owls (Family: Strigidae) of 11 North American and one Eurasian species that died following natural West Nile virus infection. The range of lesions seen was greater than that previously reported for owls, and involved more organs. Two patterns of antigen distribution were identified: one that involved the blood and all major organs; and a second where antigen was sparse, localized, and absent from the blood. The first pattern was associated with species of northern natural breeding range, while the second was seen in owls of a more southern distribution and appeared to be associated with a more prolonged course of illness. Further differences in lesion and antigen distribution appeared to be either species related or individual. The findings underline the complexity and variability of West Nile virus pathology within birds of a relatively narrow taxonomic group.

Thyroid function in nestling tree swallows and eastern bluebirds exposed to non-persistent pesticides and p, p'-DDE in apple orchards of southern Ontario, Canada.

This study examined the associated effects of pesticides and persistent residues of p,p'-DDE on thyroid function in 16-day-old tree swallows (Tachycineta bicolor) and 12-day-old eastern bluebirds (Sialia sialis). Apple orchards sprayed with pesticides in current use and reference sites were chosen for study in southern Ontario, Canada, during 2000-2001. We assessed thyroid hormone concentrations (plasma and muscle thyroxine [T4], and triiodothyronine [T3]) and the response to a challenge with thyroid stimulating hormone (TSH) as well as parameters of thyroid histology. Individual nests were exposed to as many as seven individual pesticide applications and up to five pesticide mixtures. Concentrations of p,p'-DDE in eggs from both species were highest in orchard sites, and ranged from 0.05 to 5.44 mg/kg wet weight for tree swallows, and from 0.17 to 95.4 mg/kg for eastern bluebirds. In 2000, tree swallows from sprayed orchards had elevated plasma T4 concentrations, and thyroid glands with collapsed follicles, hypertrophic epithelia, and higher follicular epithelial cells relative to chicks from reference sites. Plasma T4 concentration was positively correlated with the total number of pesticide mixtures applied during egg incubation through chick rearing. In 2001, basal plasma T3 concentration in tree swallow chicks did not differ between orchard and reference sites, though there were differences among orchards; T3 was positively correlated with total number of individual sprays applied during egg incubation through chick rearing. Tree swallows challenged with TSH showed no significant difference in thyroid hormone concentration or follicular epithelial cell height between exposure groups; however, a significantly greater proportion of chicks from reference sites were found to have thyroids with focal or diffuse hypertrophic epithelia relative to orchard chicks. In 2001, bluebirds from sprayed orchards had significantly higher T3 concentrations in response to TSH challenge; this response was positively correlated with the total number of pesticides applied during egg incubation through chick rearing.

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

From July to September 2002, an outbreak of West Nile virus (WNV) caused a high number of deaths in captive owls at the Owl Foundation, Vineland, Ontario, Canada. Peak death rates occurred in mid-August, and the epidemiologic curve resembled that of corvids in the surrounding Niagara region. The outbreak occurred in the midst of a louse fly (Icosta americana, family Hippoboscidae) infestation. Of the flies tested, 16 (88.9 %) of 18 contained WNV RNA. Species with northern native breeding range and birds >1 year of age were at significantly higher risk for WNV-related deaths. Species with northern native breeding range and of medium-to-large body size were at significantly higher risk for exposure to WNV. Taxonomic relations (at the subfamily level) did not significantly affect exposure to WNV or WNV-related deaths. Northern native breeding range and medium-to-large body size were associated with earlier death within the outbreak period. Of the survivors, 69 (75.8 %) of 91 were seropositive for WNV.

Detecting West Nile virus in owls and raptors by an antigen-capture assay.

We evaluated a rapid antigen-capture assay (VecTest) for detection of West Nile virus in oropharyngeal and cloacal swabs, collected at necropsy from owls (N = 93) and raptors (N = 27). Sensitivity was 93.5%-95.2% for northern owl species but <42.9% for all other species. Specificity was 100% for owls and 85.7% for raptors.