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.

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.

N-dihydrogalactochitosan reduces mortality in a lethal mouse model of SARS-CoV-2.

The rapid emergence and global dissemination of SARS-CoV-2 that causes COVID-19 continues to cause an unprecedented global health burden resulting in nearly 7 million deaths. While multiple vaccine countermeasures have been approved for emergency use, additional treatments are still needed due to sluggish vaccine rollout, vaccine hesitancy, and inefficient vaccine-mediated protection. Immunoadjuvant compounds delivered intranasally can guide non-specific innate immune responses during the critical early stages of viral replication, reducing morbidity and mortality. N-dihydrogalactochitosan (GC) is a novel mucoadhesive immunostimulatory polymer of ß-0-4-linked N-acetylglucosamine that is solubilized by the conjugation of galactose glycans with current applications as a cancer immunotherapeutic. We tested GC as a potential countermeasure for COVID-19. GC was well-tolerated and did not produce histopathologic lesions in the mouse lung. GC administered intranasally before and after SARS-CoV-2 exposure diminished morbidity and mortality in humanized ACE2 receptor expressing mice by up to 75% and reduced infectious virus levels in the upper airway. Fluorescent labeling of GC shows that it is confined to the lumen or superficial mucosa of the nasal cavity, without involvement of adjacent or deeper tissues. Our findings demonstrate a new application for soluble immunoadjuvants such as GC for preventing disease associated with SARS-CoV-2 and may be particularly attractive to persons who are needle-averse.

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.

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.

Lung and lymph node explants to study the interaction between host cells and canine distemper virus.

Canine distemper virus (CDV, family Paramyxoviridae) is a widely known fatal disease in unvaccinated dogs and wild carnivores. The virus enters via the respiratory tract and rapidly spreads to the lymphoid organs. To investigate viral entry into these tissues, a dog tissue explant model was developed for lung and lymph nodes. Canine lung explants were cultured with CDV for three days. During this time CDV antigens were visible on alveolar cells, which were CD163-positive and SLAM-positive (signaling lymphocytic activation molecule), demonstrating that they were macrophages. The lymph node explants were maintained for five days. During this time the viral replication increased progressively by each day post infection and syncytia were observed by day three, post exposure. The microscopic distribution of CDV-positive cells in the lymph nodes, including the syncytia, and co-expression of CD163 and SLAM, demonstrated that they were macrophages. These findings suggest that alveolar macrophages are the first cells in the lung to become infected during CDV infection, and lymph node explants showed similar replication rates and virus-cell interactions as seen in experimental live animals. This demonstrates the utility of canine respiratory and lymphoid explant model to evaluate cell entry and viral replication of CDV and other morbilliviruses in dogs or other susceptible carnivores.

Zika virus persistence in the male macaque reproductive tract.

Zika virus (ZIKV) is unique among mosquito-borne flaviviruses in that it is also vertically and sexually transmitted by humans. The male reproductive tract is thought to be a ZIKV reservoir; however, the reported magnitude and duration of viral persistence in male genital tissues vary widely in humans and non-human primate models. ZIKV tissue and cellular tropism and potential effects on male fertility also remain unclear. The objective of this study was to resolve these questions by analyzing archived genital tissues from 51 ZIKV-inoculated male macaques and correlating data on plasma viral kinetics, tissue tropism, and ZIKV-induced pathological changes in the reproductive tract. We hypothesized that ZIKV would persist in the male macaque genital tract for longer than there was detectable viremia, where it would localize to germ and epithelial cells and associate with lesions. We detected ZIKV RNA and infectious virus in testis, epididymis, seminal vesicle, and prostate gland. In contrast to prepubertal males, sexually mature macaques were significantly more likely to harbor persistent ZIKV RNA or infectious virus somewhere in the genital tract, with detection as late as 60 days post-inoculation. ZIKV RNA localized primarily to testicular stem cells/sperm precursors and epithelial cells, including Sertoli cells, epididymal duct epithelium, and glandular epithelia of the seminal vesicle and prostate gland. ZIKV infection was associated with microscopic evidence of inflammation in the epididymis and prostate gland of sexually mature males, pathologies that were absent in uninfected controls, which could have significant effects on male fertility. The findings from this study increase our understanding of persistent ZIKV infection which can inform risk of sexual transmission during assisted reproductive therapies as well as potential impacts on male fertility.

Listeriosis with viral coinfections in 8 gray foxes, 8 wild turkeys, and 2 young cervids in the southeastern United States.

Listeria monocytogenes is a bacterium that can cause disease in many species, including humans, livestock, and wildlife. Increased interactions via shared habitats may promote pathogen transmission among these groups. Our objectives were to evaluate the Southeastern Cooperative Wildlife Disease Study diagnostic data to characterize and compare L. monocytogenes-induced lesions and comorbidities in gray foxes and wild turkeys, and to describe cases of listeriosis in 2 cervids. From 1991-2020, 8 gray foxes, 8 wild turkeys, a neonatal elk, and a white-tailed deer fawn from several eastern states in the United States were diagnosed with listeriosis. All 8 foxes had hepatitis and/or hepatic necrosis with intralesional gram-positive bacilli, and concurrent canine distemper virus (CDV) infection; 2 of the foxes had been vaccinated recently for CDV. L. monocytogenes was cultured from the liver (6 of 8) or lung (2 of 8) of foxes. Lesions in wild turkeys included hepatocellular necrosis (3 of 8), heterophilic hepatitis (1 of 8), heterophilic granulomas (1 of 8), intrasinusoidal gram-positive bacilli without hepatic lesions (1 of 8), granulomatous dermatitis (1 of 8), and/or granulomatous myocarditis (2 of 8). Lymphoproliferative disease viral DNA was detected in 5 of 6 turkeys tested; reticuloendotheliosis viral DNA was detected in 2 of 3 turkeys tested. Both cervids had systemic listeriosis, with L. monocytogenes isolated from liver. Immunohistochemistry for Listeria spp. on select cases revealed immunolabeling in affected organs. Listeriosis was thus established as a cause of morbidity and mortality in 3 wildlife species, which often suffered from concurrent infections and likely immunosuppression.

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.

Plant pathogens provide clues to the potential origin of bat white-nose syndrome Pseudogymnoascus destructans.

White-nose syndrome has killed millions of bats, yet both the origins and infection strategy of the causative fungus, Pseudogymnoascus destructans, remain elusive. We provide evidence for a novel hypothesis that P. destructans emerged from plant-associated fungi and retained invasion strategies affiliated with fungal pathogens of plants. We demonstrate that P. destructans invades bat skin in successive biotrophic and necrotrophic stages (hemibiotrophic infection), a mechanism previously only described in plant fungal pathogens. Further, the convergence of hyphae at hair follicles suggests nutrient tropism. Tropism, biotrophy, and necrotrophy are often associated with structures termed appressoria in plant fungal pathogens; the penetrating hyphae produced by P. destructans resemble appressoria. Finally, we conducted a phylogenomic analysis of a taxonomically diverse collection of fungi. Despite gaps in genetic sampling of prehistoric and contemporary fungal species, we estimate an 88% probability the ancestral state of the clade containing P. destructans was a plant-associated fungus.

Distinctive Gross Presentation in Free-Ranging White-Tailed Deer (Odocoileus virginianus) with Rabies.

The white-tailed deer (Odocoileus virginianus) is a popular game species in North America and often lives in close proximity to humans and domestic animals. Deer with neurologic signs are of high interest to the general public and wildlife managers because of disease and safety concerns. Our aim was to describe diagnostic findings from free-ranging white-tailed deer diagnosed with rabies from across the eastern US from 2000 to 2021, with emphasis on gross lesions in the skin and soft tissue overlying the skull. We reviewed diagnostic reports of white-tailed deer cases submitted to the Southeastern Cooperative Wildlife Disease Study for those diagnosed with rabies from 2000 to 2021. Rabies virus infection was confirmed by immunohistochemistry or fluorescent antibody test of brain, or both. Nine adult deer from five states were diagnosed with rabies, including seven (78%) females and two (22%) males. Three (33%) deer were found dead, and six (67%) were humanely dispatched for abnormal behavior. Six deer heads were examined grossly and had lesions, including forehead or periorbital alopecia, cutaneous erythema, abrasions and ulcers, and subcutaneous edema. Histologic examination was performed for eight of nine cases, all of which had intraneuronal eosinophilic inclusion (Negri) bodies in cerebrum, cerebellum, or both. Most (6/8; 75%) had perivascular lymphoplasmacytic encephalitis. Rabies should be considered a differential diagnosis in deer with this pattern of head lesions, suggestive of head rubbing or head pressing.

Prevalence and genetic characterization of a Babesia microti-like species in the North American river otter (Lontra canadensis).

A 4.5-month-old, male, North American river otter (Lontra canadensis) from Athens-Clarke County, Georgia, USA being temporarily housed at a rehabilitation facility, presented with a three-day history of lethargy, anorexia, and severe anemia. Antemortem blood smears revealed intraerythrocytic piroplasms. Supportive care and antiparasitic treatments were initiated, but the animal died three days following presentation. Gross necropsy revealed yellow discoloration of all adipose tissue throughout the carcass and a mildly enlarged, diffusely yellow to pale orange liver. Microscopically, moderate, centrilobular hepatocellular degeneration and necrosis were observed, consistent with hypoxia secondary to apparent hemolytic anemia. Piroplasms were frequently observed in red blood cells in histologic sections. The nearly full-length 18S rRNA gene sequence (1588 bp) was identical to a previously described piroplasm from North American river otters from North Carolina. Phylogenetically, based on the 18S rRNA gene sequence, the otter Babesia sp. was in a sister group with a clade that included several strains of Babesia microti-like species including Babesia sp. from badgers (Meles meles), Babesia vulpes, and Babesia sp. from raccoons (Procyon lotor). To better understand the distribution and genetic variability of this Babesia species, otters from four states in the eastern U.S. and California were tested. Overall, 30 of 57 (53%) otters were positive for Babesia sp. None of four otters from California were positive, but prevalences in eastern states were generally high, 5/9 (55%) in Georgia, 7/14 (50%) in South Carolina, 10/17 (59%) in North Carolina, and 8/13 (62%) in Pennsylvania). Partial 18S rRNA gene sequences from all populations were identical to the clinical case sequence. No Babesia sensu stricto infections were detected. There were six unique COI sequences (937 bp) detected in 18 positive otters. The most common lineage (A) was detected in 12 of 18 (67%) samples from Georgia, North Carolina, South Carolina, and Pennsylvania. Lineage B was found in two otters and the remaining lineage types were found in single otters. These six lineages were 99-99.8% similar to each other and were < 88% similar to related parasites such as B. vulpes, B. microti-like species of raccoons, B. microti, and B. rodhaini. Phylogenetically, the Babesia sp. of otters grouped together in a well-supported clade separate from a sister group including B. vulpes from fox (Vulpes vulpes) and domestic dogs. In conclusion, this report demonstrates that this piroplasm is a potential pathogen of North American river otters and the parasite is widespread in otter populations in the eastern United States.

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.

TEMPORAL AND SPATIAL PATTERNS IN CANINE DISTEMPER VIRUS CASES IN WILDLIFE DIAGNOSED AT THE SOUTHEASTERN COOPERATIVE WILDLIFE DISEASE STUDY, 1975-2019.

Canine distemper is a high-impact disease of many mammal species and has caused substantial carnivore population declines. Analysis was conducted on passive surveillance data of canine distemper (CDV)-positive wild mammal cases submitted to the Southeastern Cooperative Wildlife Disease Study, Athens, Georgia, US, between January 1975 and December 2019. Overall, 964 cases from 17 states were CDV positive, including 646 raccoons (Procyon lotor), 254 gray foxes (Urocyon cinereoargenteus), 33 striped skunks (Mephitis mephitis), 18 coyotes (Canis latrans), four red foxes (Vulpes vulpes), three gray wolves (Canis lupus), three American black bears (Ursus americanus), two American mink (Mustela vison), and one long-tailed weasel (Mustela frenata). Raccoon and gray fox case data from the state of Georgia (n=441) were selected for further analysis. Autoregressive integrated moving average models were developed predicting raccoon and gray fox case numbers. The best-performing model for gray foxes used numbers of gray fox CDV cases from the previous 2 mo and of raccoon cases in the present month to predict the numbers of gray fox cases in the present month. The best-performing model for raccoon prediction used numbers of raccoon CDV cases from the previous month and of gray fox cases in the present month and previous 2 mo to predict numbers of raccoon cases in the present month. Temporal trends existed in CDV cases for both species, with cases more likely to occur during the breeding season. Spatial clustering of cases was more likely to occur in areas of medium to high human population density; fewer cases occurred in both the most densely populated and sparsely populated areas. This pattern was most prominent for raccoons, which may correspond to high transmission rates in suburban areas, where raccoon population densities are probably highest, possibly because of a combination of suitable habitat and supplemental resources.

Facial distortion due to chronic inflammation of unknown cause in a cat.

CASE SUMMARY: An 8-year-old neutered male indoor cat was presented for evaluation of a year-long history of swelling over the bridge of the nose that extended from the subcutaneous tissue of the right upper eyelid to the dorsum of the skull. Intermittent regression of the mass lesion was reported with antibiotic or corticosteroid therapy; however, progressive swelling, malaise and hiding behavior persisted. CT revealed an aggressive osteolytic mass lesion in the right and left nasal cavities and extending into the frontal sinuses. Rhinoscopy using a 2.8 mm rigid telescope revealed somewhat normal-appearing turbinates rostrally and ventrally on the left side, with turbinate destruction on the right. After obtaining a biopsy from the right side of the nasal cavity, thick material filling the entire nasal cavity was visible caudally and was extracted endoscopically from a rostral approach. Surgical biopsy of the dorsal nasal bridge resulted in protrusion of inspissated material from the incision site. Rhinoscopic exploration revealed that the material extended into both frontal sinuses. Following extensive debridement and medical therapy, marked resolution of facial asymmetry was achieved. RELEVANCE AND NOVEL INFORMATION: Facial distortion is often considered suggestive of a neoplastic process; however, it can also be seen with fungal and mycobacterial infections, and, in this case, an inflammatory condition of unknown etiology. In this cat, aggressive intervention and debridement of necrotic debris resulted in substantial bony remodeling of the skull and return to normal activity levels.

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.

A Baylisascaris Outbreak in Fox Squirrels (Sciurus niger) and Subsequent Detection of Francisella tularensis in Kansas, USA.

Thorough epidemiologic investigations of wildlife mortality events are often challenging, in part because of the dynamic variables involved. In May 2011, six fox squirrels (Sciurus niger) in Clinton State Park, Kansas, US were euthanized after exhibiting clinical signs of neurologic disease. Postmortem examination of two squirrels revealed that these individuals died of Baylisascaris larva migrans, which resulted in meningoencephalitis and variable pneumonia and myocarditis. Fecal flotation of raccoon (Procyon lotor) feces collected in the area revealed Baylisascaris sp. ova, presumably Baylisascaris procyonis, in one of nine samples. Additional fox squirrel carcasses were submitted for diagnostic evaluation from eastern Kansas for 1 yr following the Baylisascaris sp. outbreak. This monitoring unexpectedly resulted in the detection of Francisella tularensis, the zoonotic pathogen that causes tularemia, in two fox squirrels. The increased attention to fox squirrel mortalities prompted by the outbreak of Baylisascaris sp. larva migrans revealed cases of tularemia that may not have been otherwise detected. Although F. tularensis is endemic in Kansas, the current distribution and prevalence of B. procyonis in raccoons and other hosts in Kansas are poorly understood. This yearlong mortality investigation illustrated the importance of wildlife health monitoring as a means of assessing public health risks, especially during unusual wildlife mortality events.