Tissue and cellular tropism of Eptesicus fuscus gammaherpesvirus in big brown bats, potential role of pulmonary intravascular macrophages.

Gammaherpesviruses (γHVs) are recognized as important pathogens in humans but their relationship with other animal hosts, especially wildlife species, is less well characterized. Our objectives were to examine natural Eptesicus fuscus gammaherpesvirus (EfHV) infections in their host, the big brown bat (Eptesicus fuscus), and determine whether infection is associated with disease. In tissue samples from 132 individual big brown bats, EfHV DNA was detected by polymerase chain reaction in 41 bats. Tissues from 59 of these cases, including 17 from bats with detectable EfHV genomes, were analyzed. An EfHV isolate was obtained from one of the cases, and electron micrographs and whole genome sequencing were used to confirm that this was a unique isolate of EfHV. Although several bats exhibited various lesions, we did not establish EfHV infection as a cause. Latent infection, defined as RNAScope probe binding to viral latency-associated nuclear antigen in the absence of viral envelope glycoprotein probe binding, was found within cells of the lymphoid tissues. These cells also had colocalization of the B-cell probe targeting CD20 mRNA. Probe binding for both latency-associated nuclear antigen and a viral glycoprotein was observed in individual cells dispersed throughout the alveolar capillaries of the lung, which had characteristics of pulmonary intravascular macrophages. Cells with a similar distribution in bat lungs expressed major histocompatibility class II, a marker for antigen presenting cells, and the existence of pulmonary intravascular macrophages in bats was confirmed with transmission electron microscopy. The importance of this cell type in γHVs infections warrants further investigation.

Isolation, characterization and prevalence of a novel Gammaherpesvirus in Eptesicus fuscus, the North American big brown bat.

Little is known about the relationship of Gammaherpesviruses with their bat hosts. Gammaherpesviruses are of interest because of their long-term infection of lymphoid cells and their potential to cause cancer. Here, we report the characterization of a novel bat herpesvirus isolated from a big brown bat (Eptesicus fuscus) in Canada. The genome of the virus, tentatively named Eptesicus fuscus herpesvirus (EfHV), is 166,748 base pairs. Phylogenetically EfHV is a member of Gammaherpesvirinae, in which it belongs to the Genus Rhadinovirus and is closely related to other bat Gammaherpesviruses. In contrast to other known Gammaherpesviruses, the EfHV genome contains coding sequences similar to those of class I and II host major histocompatibility antigens. The virus is capable of infecting and replicating in human, monkey, cat and pig cell lines. Although we detected EfHV in 20 of 28 big brown bats tested, these bats lacked neutralizing antibodies against the virus.

Activation of innate immune-response genes in little brown bats (Myotis lucifugus) infected with the fungus Pseudogymnoascus destructans.

Recently bats have been associated with the emergence of diseases, both as reservoirs for several new viral diseases in humans and other animals and, in the northern Americas, as hosts for a devastating fungal disease that threatens to drive several bat species to regional extinction. However, despite these catastrophic events little Information is available on bat defences or how they interact with their pathogens. Even less is known about the response of bats to infection during torpor or long-term hibernation. Using tissue samples collected at the termination of an experiment to explore the pathogenesis of White Nose Syndrome in Little Brown Bats, we determined if hibernating bats infected with the fungus Pseudogymnoascus destructans could respond to infection by activating genes responsible for innate immune and stress responses. Lesions due to fungal infection and, in some cases, secondary bacterial infections, were restricted to the skin. However, we were unable to obtain sufficient amounts of RNA from these sites. We therefore examined lungs for response at an epithelial surface not linked to the primary site of infection. We found that bats responded to infection with a significant increase in lungs of transcripts for Cathelicidin (an anti-microbial peptide) as well as the immune modulators tumor necrosis factor alpha and interleukins 10 and 23. In conclusion, hibernating bats can respond to experimental P. destructans infection by activating expression of innate immune response genes.

Targeting the detection of chronic wasting disease using the hunter harvest during early phases of an outbreak in Saskatchewan, Canada.

Chronic wasting disease (CWD) is a fatal disease of North American cervids that was first detected in a wild, hunter-shot deer in Saskatchewan along the border with Alberta in Canada in 2000. Spatially explicit models for assessing factors affecting disease detection are needed to guide surveillance and control programs. Spatio-temporal patterns in CWD prevalence can be complicated by variation in individual infection probability and sampling biases. We assessed hunter harvest data of mule deer (Odocoileus hemionus) and white-tailed deer (Odocoileus virginianus) during the early phases of an outbreak in Saskatchewan (i.e., 2002-2007) for targeting the detection of CWD by defining (1) where to look, and (2) how much effort to use. First, we accounted for known demographic heterogeneities in infection to model the probability, P(E), that a harvested deer was infected with CWD given characteristics of the harvest location. Second, in areas where infected deer were harvested we modelled the probability, P(D), of the hunter harvest re-detecting CWD within sample units of varying size (9-54 km(2)) given the demographics of harvested deer and time since first detection in the study area. Heterogeneities in host infection were consistent with those reported elsewhere: mule deer 3.7 times >white-tailed deer, males 1.8 times>females, and quadratically related to age in both sexes. P(E) increased with number of years since the first detection in our study area (2002) and proximity to known disease sources, and also varied with distance to the South Saskatchewan River and small creek drainages, terrain ruggedness, and extent of agriculture lands within a 3 km radius of the harvest. The majority (75%) of new CWD-positive deer from our sample were found within 20 km of infected deer harvested in the previous year, while approximately 10% were greater than 40 km. P(D) modelled at 18 km(2) was best supported, but for all scales, P(D) depended on the number of harvested deer and time since the first infected deer was harvested. Within an 18 km(2) sampling unit, there was an 80% probability of detecting a CWD-positive deer with 16 harvested deer five years after the initial infected harvest. Identifying where and how much to sample to detect CWD can improve targeted surveillance programs early in the outbreak of the disease when based on hunter harvest.

Survival of radio-marked mallards in relation to management of avian botulism.

Avian botulism outbreaks are frequently perpetuated by type C toxin produced by Clostridium botulinum proliferating in decomposing bird carcasses and consumption of toxic maggots from these carcasses by healthy birds. Therefore, removing bird carcasses has been advocated for disease management because availability of toxic maggots should be reduced, increasing duck survival. However, this management is expensive, and its effect on waterfowl mortality under field conditions is unknown. We radio-marked 419 molting mallards on 11 lakes in western Canada during July-August 1999-2001 and monitored them for 30 days, testing whether survival was higher on lakes with carcass removal. Botulism occurred on 10 lakes. On five carcass removal lakes, greater-than-normal effort was made to conduct early, thorough surveillance and immediately remove carcasses; on six nonremoval lakes, no carcasses were removed. In 1999, estimated 30-day survival probabilities ranged from 0.149 (95% CI=0.065-0.304) on one large lake with carcass removal to 0.466 (95% CI=0.270-0.674) and 0.618 (95% CI=0.443-0.767) on two nonremoval lakes. As a result, we conducted work on smaller wetlands thereafter, reasoning that any management benefit would be easier to detect. In 2000, estimated 30-day survival probabilities were 0.313 (95% CI=0.143-0.556) and 0.794 (95% CI=0.609-0.905) on two carcass removal lakes versus 0.525 (95% CI=0.362-0.682) and 0.743 (95% CI=0.564-0.866) on two nonremoval lakes. In 2001, botulism was detected on two nonremoval lakes where survival probabilities were 0.845 (95% CI=0.630-0.946) and 0.942 (95% CI=0.778-0.987), and on one removal lake where survival probability was 1.0 (95% CI=0.99-1.0), but not detected on the other removal lake where no marked birds died from botulism (1.0, 95% CI=0.99-1.0). Survival tended to be higher on lakes with lower carcass density, but when data were organized by carcass removal versus nonremoval, mallard survival was not consistently greater on lakes where carcasses were removed.

Sheep-associated malignant catarrhal fever in free-ranging moose (Alces alces) in Saskatchewan, Canada.

Malignant catarrhal fever (MCF) is a sporadic disease of artiodactyls caused by several viruses in the Gammaherpesvirinae. We report two cases of MCF in free-living moose (Alces alces) from Saskatchewan. One was a thin, dehydrated, adult male found recumbent in 2006. At necropsy, ulcers were found in the intestine, bladder, and corneas. Microscopically, there was lymphocytic vasculitis and perivasculitis in many organs with infrequent fibrinoid necrosis. Ovine herpes virus-2 (OHV-2) was identified by polymerase chain reaction. A segment of the herpesviral DNA polymerase gene was 99% identical to published OHV-2 sequences. During a retrospective search of earlier cases, a female moose with lymphoplasmacytic meningoencephalitis examined in 2003 was identified and OHV-2 was amplified from paraffin-embedded tissues from this animal. We believe this to be the first description of MCF in free-ranging moose in North America. Infection requires contact with infected sheep or goats, and MCF in moose may become more prevalent as moose distribution continues to expand into agricultural prairie.