Severe Acute Respiratory Syndrome Coronavirus 2 Vasculopathy in a Syrian Golden Hamster Model.

Clinical evidence of vascular dysfunction and hypercoagulability as well as pulmonary vascular damage and microthrombosis are frequently reported in severe cases of human coronavirus disease 2019 (COVID-19). Syrian golden hamsters recapitulate histopathologic pulmonary vascular lesions reported in patients with COVID-19. Herein, special staining techniques and transmission electron microscopy further define vascular pathologies in a Syrian golden hamster model of human COVID-19. The results show that regions of active pulmonary inflammation in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection are characterized by ultrastructural evidence of endothelial damage with platelet marginalization and both perivascular and subendothelial macrophage infiltration. SARS-CoV-2 antigen/RNA was not detectable within affected blood vessels. Taken together, these findings suggest that the prominent microscopic vascular lesions in SARS-CoV-2-inoculated hamsters likely occur due to endothelial damage followed by platelet and macrophage infiltration.

California sea lion (Zalophus californianus) lymph-node explant reveals involvement and possible transcriptional regulation of SLAM and nectin-4 during phocine distemper virus infection.

Phocine distemper virus (PDV) is a significant cause of mortality for phocid seals; however, the susceptibility of otariids to this virus is poorly understood. The authors used a lymph-node explant culture system from California sea lions (Zalophus californianus, CSL) to investigate: (1) the role of signaling lymphocyte activation molecule (SLAM) and nectin-4 in PDV infection and their cellular expression patterns, (2) if PDV induces transcriptional regulation of cell-entry receptors, and (3) the involvement of apoptosis in PDV infection. PDV replicated in the lymph-node explants with peak replication 3 days post-infection (dpi), but the replication was not sustained 4 to 5 dpi. The PDV+ cells co-localized SLAM and nectin-4. These cells expressed IBA1, indicating a histiocytic lineage. Comparison of receptor expression between infected and mock-infected lymph nodes suggested transcriptional downregulation of both receptors during the initial stage of infection and upregulation during the late stage of infection, but the values lack of statistical significance. Cleaved caspase-3+ cells were slightly increased in the infected lymph nodes compared with the mock-infected lymph node from 1 to 4 dpi, but without statistical significance, and a few apoptotic cells co-expressed PDV. The results suggest that lymph-node explants might be an important model to study PDV pathogenesis. CSLs have the potential to be infected with PDV, as they express both cell-entry receptors in histiocytes. The lack of statistical significance in the PDV replication, transcriptional regulation of viral receptors, and changes in apoptosis suggest that although CSL might be infected by PDV, they might be less susceptible than phocid species.

Characterization of Pseudogymnoascus destructans conidial adherence to extracellular matrix: Association with fungal secreted proteases and identification of candidate extracellular matrix binding proteins.

Pseudogymnoascus destructans is the etiological agent of white-nose syndrome (WNS), a fungal skin infection of hibernating bats. Pathophysiology of the disease involves disruption of bat metabolism and hibernation patterns, which subsequently causes premature emergence and mortality. However, information on the mechanism(s) and virulence factors of P. destructans infection is minimally known. Typically, fungal adherence to host cells and extracellular matrix (ECM) is the critical first step of the infection. It allows pathogenic fungi to establish colonization and provides an entry for invasion in host tissues. In this study, we characterized P. destructans conidial adherence to laminin and fibronectin. We found that P. destructans conidia adhered to laminin and fibronectin in a dose-dependent, time-dependent and saturable manner. We also observed changes in the gene expression of secreted proteases, in response to ECM exposure. However, the interaction between fungal conidia and ECM was not specific, nor was it facilitated by enzymatic activity of secreted proteases. We therefore further investigated other P. destructans proteins that recognized ECM and found glyceraldehyde-3-phosphate dehydrogenase and elongation factor 1-alpha among the candidate proteins. Our results demonstrate that P. destructans may use conidial surface proteins to recognize laminin and fibronectin and facilitate conidial adhesion to ECM. In addition, other non-specific interactions may contribute to the conidial adherence to ECM. However, the ECM binding protein candidates identified in this study highlight additional potential fungal virulence factors worth investigating in the P. destructans mechanism of infection in future studies.

Distribution of canine distemper virus and nectin-4 in raccoon (Procyon lotor) skin.

Raccoons (Procyon lotor) are abundant in urban/wildland interfaces and are key sources of canine distemper virus (CDV) outbreaks in domestic, zoo, and free-ranging wildlife species. CDV is pantropic, which provides multiple potential routes of transmission (urine, respiratory secretions, feces), but the specific role of skin as a target of infection, as a diagnostic sample, or as a potential source of environmental persistence and transmission is unknown. We have characterized the distribution of CDV and its known receptor, nectin-4, in skin samples of 36 raccoons. Even with skin samples that were grossly and histologically normal, immunohistochemistry of skin was useful in the diagnosis of CDV infection, which was found in both epithelium and endothelium. Nectin-4 was codistributed with cellular targets of viral infection. Skin secretions, shed keratinocytes, and hair of CDV infected raccoons are all potential environmental fomites.

Evaluation of Atherosclerotic Lesions and Risk Factors of Atherosclerosis in Raptors in Northern California.

Atherosclerosis is a chronic inflammatory disease that has been reported to affect the cardiovascular system of many avian species. However, atherosclerosis in raptor species has not been fully evaluated. The aim of this study was to histologically characterize central and peripheral atherosclerotic lesions in raptors that were submitted to a pathology service in Northern California from 1986 to 2013. We also evaluated risk factors, including age, gender, origin, and avian family. Atherosclerotic lesions were categorized as minimal, mild, moderate, or severe, based on the severity of the lesions and their distribution within the arterial wall. Among the central arteries, lesions were determined to be of greater severity in the aorta than in the pulmonary artery. More than 50% of the peripheral arteries were affected, including 53.1% (17/32) myocardial, 52% (13/25) coronary, 62.9% (22/35) arteries in the kidney, 52.2% (12/23) gonadal and 51.7% (15/29) splenic arteries; however, hepatic and pulmonary arteries were uncommonly affected. Atherosclerosis was diagnosed in 17 raptor species representing 4 families: Accipitridae, Cathartidae, Falconidae, and Strigidae. The overall prevalence (95% CI) of atherosclerosis in raptors was 2.3% (36/1574; range, 1.63%-3.19%) with the Falconidae having the highest prevalence at 7.4% (9/122; range 3.64%-13.93%) and with 0% detected in the Tytonidae and Pandionidae families. A multiple logistic regression model that jointly accounted for differences in risk by family, age, and gender found that the risk in Accipitridae was significantly less than that of Falconidae, that adult raptors were at greater risk of atherosclerotic lesions than juveniles were, and that females were more frequently affected than males were.

Granulomatous Inflammation of the Muzzle in White-Tailed Deer (Odocoileus virginianus) and Mule Deer (Odocoileus hemionus) Associated With Mannheimia granulomatis.

Since 2002, reports of deer with swollen muzzles from throughout the United States have resulted in significant interest by wildlife biologists and wildlife enthusiasts. The condition was identified in 25 white-tailed deer (Odocoileus virginianus) and 2 mule deer (O. hemionus). Microscopic lesions consisted of severe, granulomatous or pyogranulomatous inflammation of the muzzle, nasal planum, and upper lip, as well as similar but less severe inflammation of the hard palate. Lymphadenitis of regional lymph nodes was common and granulomatous pneumonia was present in one individual. Splendore-Hoeppli material was typical in the center of inflammatory foci. Other than the single instance of pneumonia, systemic disease was not evident. Various bacterial species were isolated in culture, most of which were not morphologically consistent with the colonies of small, gram-negative bacteria observed in the center of the granulomas. Amplification and sequencing of the bacterial 16S rRNA gene from tissues of affected deer resulted in the identification of Mannheimia granulomatis. Laser capture microdissection was used to confirm that the colonies in the inflammatory foci were M. granulomatis. The cases described here are reminiscent of a bovine disease in Brazil and Argentina, locally called lechiguana. Although the inflammation of lechiguana is mostly truncal, the microscopic lesions are very similar and are also attributed to M. granulomatis. It is unclear if this is an emerging infectious disease of deer, or if it is a sporadic, uncommon condition that has only recently been recognized.

Evaluation of transanal minimally invasive surgery for submucosal rectal resection in cadaveric canine specimens.

OBJECTIVE: To evaluate the feasibility of transanal minimally invasive surgery (TAMIS) for submucosal rectal resection in large breed dogs. STUDY DESIGN: Cadaveric study. SAMPLE POPULATION: Canine cadavers (n = 6) weighing between 37.5 and 60 kg. METHODS: Dogs were positioned in sternal recumbency. After rectal cleansing, a transanal access platform was placed in the rectum, and a pneumorectum was established. An area of ventral rectal wall approximately 2 × 2 cm was resected in a submucosal plane by using laparoscopic instruments and submitted for histopathological evaluation. The rectal wall defect was closed with a single-layer continuous suture pattern with barbed suture. Postoperatively, the rectum was removed en bloc and evaluated for suture or surgical penetration of the serosal surface. RESULTS: Submucosal rectal resection was successfully completed by using TAMIS in all dogs. The median length of resected specimens after fixation was 24.5 mm (range 9.8-26.5). In two of six dogs, suture was macroscopically visible on the serosal surface, but no dogs had evidence of iatrogenic full-thickness surgical penetration of the rectum. The median distance from the aborad extent of the suture closure line to the anocutaneous junction was 35 mm (range, 35-105). CONCLUSION: Submucosal resection of the canine rectal wall was feasible in large breed dogs by using TAMIS. No evidence of full-thickness penetration of the rectal wall was seen in these cadaveric specimens. CLINICAL SIGNIFICANCE: Transanal minimally invasive surgery may provide an alternative minimally invasive approach for resection for benign adenomatous rectal polyps in large breed dogs that might otherwise require a rectal pull-through.

Evolution and Cryo-electron Microscopy Capsid Structure of a North American Bat Adenovirus and Its Relationship to Other Mastadenoviruses.

Since the first description of adenoviruses in bats in 2006, a number of micro- and megabat species in Europe, Africa, and Asia have been shown to carry a wide diversity of adenoviruses. Here, we report on the evolutionary, biological, and structural characterization of a novel bat adenovirus (BtAdV) recovered from a Rafinesque's big-eared bat (Corynorhinus rafinesquii) in Kentucky, USA, which is the first adenovirus isolated from North American bats. This virus (BtAdV 250-A) exhibits a close phylogenetic relationship with Canine mastadenovirus A (CAdV A), as previously observed with other BtAdVs. To further investigate the relationships between BtAdVs and CAdVs, we conducted mass spectrometric analysis and single-particle cryo-electron microscopy reconstructions of the BtAdV 250-A capsid and also analyzed the in vitro host ranges of both viruses. Our results demonstrate that BtAdV 250-A represents a new mastadenovirus species that, in contrast to CAdV, has a unique capsid morphology that contains more prominent extensions of protein IX and can replicate efficiently in a phylogenetically diverse range of species. These findings, in addition to the recognition that both the genetic diversity of BtAdVs and the number of different bat species from disparate geographic regions infected with BtAdVs appears to be extensive, tentatively suggest that bats may have served as a potential reservoir for the cross-species transfer of adenoviruses to other hosts, as theorized for CAdV. IMPORTANCE: Although many adenoviruses are host specific and likely codiverged with their hosts over millions of years, other adenoviruses appear to have emerged through successful cross-species transmission events on more recent time scales. The wide geographic distribution and genetic diversity of adenoviruses in bats and their close phylogenetic relationship to Canine mastadenovirus A (CAdV A) has raised important questions about how CAdV A, and possibly other mammalian adenoviruses, may have emerged. Although most adenoviruses tend to cause limited disease in their natural hosts, CAdV A is unusual in that it may cause high morbidity and sometimes fatal infections in immunocompetent hosts and is thus an important pathogen of carnivores. Here, we performed a comparative evolutionary and structural study of representative bat and canine adenoviruses to better understand the relationship between these two viral groups.

Differential Disease Susceptibilities in Experimentally Reptarenavirus-Infected Boa Constrictors and Ball Pythons.

Inclusion body disease (IBD) is an infectious disease originally described in captive snakes. It has traditionally been diagnosed by the presence of large eosinophilic cytoplasmic inclusions and is associated with neurological, gastrointestinal, and lymphoproliferative disorders. Previously, we identified and established a culture system for a novel lineage of arenaviruses isolated from boa constrictors diagnosed with IBD. Although ample circumstantial evidence suggested that these viruses, now known as reptarenaviruses, cause IBD, there has been no formal demonstration of disease causality since their discovery. We therefore conducted a long-term challenge experiment to test the hypothesis that reptarenaviruses cause IBD. We infected boa constrictors and ball pythons by cardiac injection of purified virus. We monitored the progression of viral growth in tissues, blood, and environmental samples. Infection produced dramatically different disease outcomes in snakes of the two species. Ball pythons infected with Golden Gate virus (GoGV) and with another reptarenavirus displayed severe neurological signs within 2 months, and viral replication was detected only in central nervous system tissues. In contrast, GoGV-infected boa constrictors remained free of clinical signs for 2 years, despite high viral loads and the accumulation of large intracellular inclusions in multiple tissues, including the brain. Inflammation was associated with infection in ball pythons but not in boa constrictors. Thus, reptarenavirus infection produces inclusions and inclusion body disease, although inclusions per se are neither necessarily associated with nor required for disease. Although the natural distribution of reptarenaviruses has yet to be described, the different outcomes of infection may reflect differences in geographical origin.IMPORTANCE New DNA sequencing technologies have made it easier than ever to identify the sequences of microorganisms in diseased tissues, i.e., to identify organisms that appear to cause disease, but to be certain that a candidate pathogen actually causes disease, it is necessary to provide additional evidence of causality. We have done this to demonstrate that reptarenaviruses cause inclusion body disease (IBD), a serious transmissible disease of snakes. We infected boa constrictors and ball pythons with purified reptarenavirus. Ball pythons fell ill within 2 months of infection and displayed signs of neurological disease typical of IBD. In contrast, boa constrictors remained healthy over 2 years, despite high levels of virus throughout their bodies. This difference matches previous reports that pythons are more susceptible to IBD than boas and could reflect the possibility that boas are natural hosts of these viruses in the wild.

Congenital Ocular Abnormalities in Free-Ranging White-Tailed Deer.

Congenital ocular abnormalities in cervids have been previously reported as individual cases from various regions of the United States and include microphthalmia, anophthalmia, congenital cataracts, dermoids, and colobomata. A common underlying cause for these abnormalities, such as nutritional deficiencies, environmental toxin exposures, or genetic mutations, has not been established. This retrospective study summarized and compared cases of suspected congenital ocular abnormalities in free-ranging white-tailed deer ( Odocoileus virginianus) submitted to the Southeastern Cooperative Wildlife Disease Study (SCWDS) in Athens, Georgia, to the preexisting literature. Of 3645 accessions of white-tailed deer submitted to SCWDS, 15 qualifying case records were found. An additional 15 cases were reported previously in the literature. Conditions described in SCWDS cases included microphthalmia (8/15), congenital cataracts (3/15), anophthalmia (2/15), colobomata (1/15), anterior segment dysgenesis (1/15), ectopic lacrimal gland tissue (1/15), and congenital blindness with corneal opacity (1/15). Most (11/15; 73%) of the SCWDS cases were male fawns with an average age of 4 months at presentation, consistent with previously described cases. Most animals had bilateral abnormalities with few extraocular congenital abnormalities, also consistent with existing reports. Cases were variably tested for various infectious agents at the time of submission; 2 cases were seropositive for bluetongue virus. Spatiotemporal clustering of cases was not evident. This study provided a concise and systematic summary of known existing cases of congenital ocular defects in fawns but did not identify a cause.

Cyclic avian mass mortality in the northeastern United States is associated with a novel orthomyxovirus.

UNLABELLED: Since 1998, cyclic mortality events in common eiders (Somateria mollissima), numbering in the hundreds to thousands of dead birds, have been documented along the coast of Cape Cod, MA, USA. Although longitudinal disease investigations have uncovered potential contributing factors responsible for these outbreaks, detecting a primary etiological agent has proven enigmatic. Here, we identify a novel orthomyxovirus, tentatively named Wellfleet Bay virus (WFBV), as a potential causative agent of these outbreaks. Genomic analysis of WFBV revealed that it is most closely related to members of the Quaranjavirus genus within the family Orthomyxoviridae. Similar to other members of the genus, WFBV contains an alphabaculovirus gp64-like glycoprotein that was demonstrated to have fusion activity; this also tentatively suggests that ticks (and/or insects) may vector the virus in nature. However, in addition to the six RNA segments encoding the prototypical structural proteins identified in other quaranjaviruses, a previously unknown RNA segment (segment 7) encoding a novel protein designated VP7 was discovered in WFBV. Although WFBV shows low to moderate levels of sequence similarity to Quaranfil virus and Johnston Atoll virus, the original members of the Quaranjavirus genus, additional antigenic and genetic analyses demonstrated that it is closely related to the recently identified Cygnet River virus (CyRV) from South Australia, suggesting that WFBV and CyRV may be geographic variants of the same virus. Although the identification of WFBV in part may resolve the enigma of these mass mortality events, the details of the ecology and epidemiology of the virus remain to be determined. IMPORTANCE: The emergence or reemergence of viral pathogens resulting in large-scale outbreaks of disease in humans and/or animals is one of the most important challenges facing biomedicine. For example, understanding how orthomyxoviruses such as novel influenza A virus reassortants and/or mutants emerge to cause epidemic or pandemic disease is at the forefront of current global health concerns. Here, we describe the emergence of a novel orthomyxovirus, Wellfleet Bay virus (WFBV), which has been associated with cyclic large-scale bird die-offs in the northeastern United States. This initial characterization study provides a foundation for further research into the evolution, epidemiology, and ecology of newly emerging orthomyxoviruses, such as WFBV, and their potential impacts on animal and/or human health.

Choroidal Vasculopathy and Retinal Detachment in a Bald Eagle ( Haliaeetus leucocephalus ) With Lead Toxicosis.

A subadult male bald eagle ( Haliaeetus leucocephalus ) was presented for severe depression and weakness. Physical examination findings included depressed mentation, dehydration, sternal recumbency, poor body condition, and bilateral, whole-head, horizontal nystagmus. A heavy-metal panel was performed, and blood lead levels were 6.1 ppm. Treatment for lead poisoning was initiated, including subcutaneous fluids and parenteral calcium-disodium ethylenediaminetetraacetic acid, ceftiofur, and meloxicam. Ophthalmic examination findings included absent menace response, absent dazzle reflex, slow and incomplete direct pupillary light reflex, mild anterior uveitis, incipient cataracts, multifocal retinal tears, and retinal separation in both eyes. Because of poor prognosis for vision and release to the wild, the eagle was euthanatized. No lesions were observed on gross postmortem examination. Histologically, extensive myocardial necrosis and multisystemic arteriolar vasculopathy were identified. The eyes were examined after tissue processing, and the vasculopathy extended into the choriocapillaris and was associated with a secondary, bilateral, exudative, retinal detachment. This is the first report in avian species characterizing the histopathologic ocular lesions of lead poisoning.

Antleroma in a free-ranging white-tailed deer (Odocoileus virginianus).

A 2-year-old male free-ranging white-tailed deer (Odocoileus virginianus) was diagnosed with bilateral expansile tumors of antler origin. The deer was found dead by a landowner in High Springs, Florida. Two roughly spherical, multilobular, broad-based, bony, velvet-covered masses originated from each antler pedicle. These masses replaced or displaced many of the bones and soft tissues of the skull and extended through the left cribriform plate and the right petrous temporal bone, compressing portions of the brain. Microscopically, the masses closely resembled normal-growing antler, containing all the elements thereof but with areas of necrosis and hemorrhage suggestive of ischemi or trauma. Tumorlike outgrowths termed antleromas have been described in free-ranging and captive cervids and typically are associated with disruptions in the seasonal rise and fall of circulating testosterone necessary for normal antler growth, casting, and regeneration.

A preliminary report on the contact-independent antagonism of Pseudogymnoascus destructans by Rhodococcus rhodochrous strain DAP96253.

BACKGROUND: The recently-identified causative agent of White-Nose Syndrome (WNS), Pseudogymnoascus destructans, has been responsible for the mortality of an estimated 5.5 million North American bats since its emergence in 2006. A primary focus of the National Response Plan, established by multiple state, federal and tribal agencies in 2011, was the identification of biological control options for WNS. In an effort to identify potential biological control options for WNS, multiply induced cells of Rhodococcus rhodochrous strain DAP96253 was screened for anti-P. destructans activity. RESULTS: Conidia and mycelial plugs of P. destructans were exposed to induced R. rhodochrous in a closed air-space at 15°C, 7°C and 4°C and were evaluated for contact-independent inhibition of conidia germination and mycelial extension with positive results. Additionally, in situ application methods for induced R. rhodochrous, such as fixed-cell catalyst and fermentation cell-paste in non-growth conditions, were screened with positive results. R. rhodochrous was assayed for ex vivo activity via exposure to bat tissue explants inoculated with P. destructans conidia. Induced R. rhodochrous completely inhibited growth from conidia at 15°C and had a strong fungistatic effect at 4°C. Induced R. rhodochrous inhibited P. destructans growth from conidia when cultured in a shared air-space with bat tissue explants inoculated with P. destructans conidia. CONCLUSION: The identification of inducible biological agents with contact-independent anti- P. destructans activity is a major milestone in the development of viable biological control options for in situ application and provides the first example of contact-independent antagonism of this devastating wildlife pathogen.

Demographic Risk Factors Vary in the Invasion Front of Chronic Wasting Disease in West Virginia, USA.

After detecting chronic wasting disease (CWD) in white-tailed deer (Odocoileus virginianus) in Hampshire County, West Virginia, USA, in 2005, we investigated the change of CWD apparent prevalence and potential factors influencing infection risk during the invasion front. Over eight sampling years (2006-2012 and 2017) during a 12-yr period within a 101-km2-area monitoring zone, we sampled and tested a total of 853 deer for CWD by ELISA and immunohistochemistry. Bayesian logistic regression of risk factors included collection year, age class, sex, and adjusted body weight (weight after accounting for sex, age, kidney fat index, and number of fetuses). In the whole-herd model (n=634), collection year, age, and adjusted body weight were associated with increased odds of CWD, whereas an age-weight interaction had a negative relationship. We found that males drove the positive associations with age and adjusted body weight, whereas females were responsible for the negative interaction effect. These findings suggest potential behavioral and physiological mechanisms related to sex that may influence CWD exposure. Older males exhibited higher CWD prevalence, aligning with previous studies. Notably, the novel finding of adjusted body weight as a risk factor in males warrants further investigation, and this study highlights the need for future research on social behavior and its role in CWD transmission within white-tailed deer populations.

Trichomonad Disease in Wild Turkeys (Meleagris gallopavo): Pathology and Molecular Characterization of Histomonas, Tetratrichomonas, Tritrichomonas, and Simplicimonas spp.

The Wild Turkey (Meleagris gallopavo) is a geographically widespread, popular game bird and conservation icon in North America. Following successful reestablishment in parts of the US and Canada, regional declines have fueled population health concerns; therefore, understanding mortality causes and spatiotemporal patterns of disease is important to uncover potential ongoing and future health risks. Histomonosis, caused by the trichomonad Histomonas meleagridis, is a well-established and potentially fatal disease in Wild Turkeys; however, its prevalence and potential population health impacts remain poorly understood. Moreover, molecular tools recently have allowed for the detection of additional trichomonads that similarly can cause fatal disease in Wild Turkeys. We describe and compare disease due to H. meleagridis with that of Tetratrichomonas gallinarum, Tritrichomonas sp., and Simplicimonas sp. in Wild Turkeys in the southeastern US. Among 857 Wild Turkeys evaluated postmortem from 2002 to 2023, 34 (4.0%) were diagnosed with trichomonad disease, often assumed to be histomonosis prior to molecular testing. However, among 25 debilitating to fatal trichomonad disease cases for which etiologies were confirmed by PCR from 2015 to 2023, H. meleagridis was detected in 16/25 (64.0%), T. gallinarum in 6/25 (24.0%), Tritrichomonas sp. in 2/25 (8.0%) and Simplicimonas sp. in 1/25 (4.0%). These turkeys had similar clinical manifestations, and although lesion patterns varied to some extent, liver and/or intestinal tract was most commonly affected. Coinfections were common among all turkeys with trichomonad disease from 2015 to 2023 (21/25, 84.0%) and included viruses (lymphoproliferative disease virus, avian poxvirus), bacteria (Streptococcus gallolyticus, Listeria monocytogenes, Escherichia coli), and other protozoa (Sarcocystis sp., Haemoproteus sp.). Our results highlight the importance of molecular diagnostic testing in determining etiologies of trichomonad disease in Wild Turkeys. Further evaluation of the epidemiology and pathogenesis of trichomonad disease and its varied etiologies in Wild Turkeys and other birds is warranted to better understand risk factors and potential health impacts.

Validation of a Field-Portable, Handheld Real-Time PCR System for Detecting Pseudogymnoascus destructans, the Causative Agent of White-Nose Syndrome in Bats.

White-nose syndrome (WNS), caused by the fungus Pseudogymnoascus destructans, has decimated bat populations across North America. Despite ongoing management programs, WNS continues to expand into new populations, including in US states previously thought to be free from the pathogen and disease. This expansion highlights a growing need for surveillance tools that can be used to enhance existing monitoring programs and support the early detection of P. destructans in new areas. We evaluated the feasibility of using a handheld, field-portable, real-time (quantitative) PCR (qPCR) thermocycler known as the Biomeme two3 and the associated field-based nucleic acid extraction kit and assay reagents for the detection of P. destructans in little brown bats (Myotis lucifugus). Results from the field-based protocol using the Biomeme platform were compared with those from a commonly used laboratory-based qPCR protocol. When using dilutions of known conidia concentrations, the lowest detectable concentration with the laboratory-based approach was 108.8 conidia/mL, compared with 1,087.5 conidia/mL (10 times higher, i.e., one fewer 10× dilution) using the field-based approach. Further comparisons using field samples suggest a high level of concordance between the two protocols, with positive and negative agreements of 98.2% and 100% respectively. The cycle threshold values were marginally higher for most samples using the field-based protocol. These results are an important step in establishing and validating a rapid, field-assessable detection platform for P. destructans, which is urgently needed to improve the surveillance and monitoring capacity for WNS and support on-the-ground management and response efforts.

Role of multiple hosts in the cross-species transmission and emergence of a pandemic parvovirus.

Understanding the mechanisms of cross-species virus transmission is critical to anticipating emerging infectious diseases. Canine parvovirus type 2 (CPV-2) emerged as a variant of a feline parvovirus when it acquired mutations that allowed binding to the canine transferrin receptor type 1 (TfR). However, CPV-2 was soon replaced by a variant virus (CPV-2a) that differed in antigenicity and receptor binding. Here we show that the emergence of CPV involved an additional host range variant virus that has circulated undetected in raccoons for at least 24 years, with transfers to and from dogs. Raccoon virus capsids showed little binding to the canine TfR, showed little infection of canine cells, and had altered antigenic structures. Remarkably, in capsid protein (VP2) phylogenies, most raccoon viruses fell as evolutionary intermediates between the CPV-2 and CPV-2a strains, suggesting that passage through raccoons assisted in the evolution of CPV-2a. This highlights the potential role of alternative hosts in viral emergence.

Infestation of two brown-headed cowbirds (Molothrus ater) with the hunchback mite (Harpirhynchus quasimodo).

A recently described North American mite, Harpirhynchus quasimodo (Acariformes: Harpirhynchidae), was identified in a wild brown-headed cowbird (Molothrus ater) from Tennessee and another from Arkansas, U. S. A., during 2010 and 2011, respectively. Consistent with a single previous report of H. quasimodo infestation, both cowbirds had multiple skin masses composed of epidermal cysts filled with harpirhynchid mites, keratin, and cellular debris. Although very little is currently known about this avian mite, these three reports suggest that H. quasimodo infestation is a rare source of overt disease in free-ranging brown-headed cowbirds from the southeastern United States; however, further research and surveillance efforts are needed.