Clinical, pathologic, and immunohistochemical features of pituitary tumors in 4 chinchillas.

Pituitary tumors are rare in chinchillas. This report describes the clinical, gross, histologic, and immunohistochemical characteristics of pituitary tumors in 4 chinchillas. The affected chinchillas were females between 4 and 18 years of age. Clinically, neurologic signs were most commonly reported and included depression, obtundation, seizure, head-pressing, ataxia, and possible blindness. Computed tomography scanning of 2 chinchillas revealed solitary intracranial extra-axial masses in the region of the pituitary gland. Two pituitary tumors were confined to the pars distalis; the other 2 invaded the brain. Based on their microscopic appearances and lack of distant metastases, all 4 tumors were diagnosed as pituitary adenomas. Immunohistochemically, all pituitary adenomas were weakly to strongly positive for growth hormone, most consistent with the diagnosis of somatotropic pituitary adenomas. To the authors' knowledge, this is the first detailed report of the clinical, pathologic, and immunohistochemical features of pituitary tumors in chinchillas.

VITAMIN C DEFICIENCY-ASSOCIATED LESIONS IN A COLONY OF COMMON VAMPIRE BATS (DESMODUS ROTUNDUS) IN A ZOO FACILITY.

The Milwaukee County Zoo has housed common vampire bats (Desmodus rotundus) since 1973. The bats are fed defibrinated cow's blood supplemented with a liquid pediatric multivitamin. From July 2013 to May 2014, multiple deaths occurred in colony bats, including five juveniles with multiple bone fractures and failure of endochondral ossification, three adults with cerebellar necrosis, and one adult with subcutaneous hemorrhage. In November 2013, an adult bat developed a nonhealing left wing hematoma and eventually succumbed 9 mo later. A postmortem examination revealed multifocal extensive necrohemorrhagic and suppurative ulcerative dermatitis with no underlying cause determined. From July to December 2014, five of nine adult bats in the colony developed similar hematomas along with gingival bleeding. One euthanized bat had a serum ascorbic acid level of 0.08 mg/dl and marked generalized subcutaneous hemorrhage. A therapeutic trial was initiated in which two bats received defibrinated cow's blood supplemented only with oral vitamin C, 100 mg/kg PO q24h for 3 d, and then 50 mg/kg PO q24h. Two other bats received nonsupplemented defibrinated cow's blood and were given vitamin K 3.3 mg/kg SC q12h for 3 d, and then 3.3 mg/kg SC q24h for 7 d. The bats supplemented with vitamin C improved, supporting a diagnosis of vitamin C deficiency. All bats were subsequently supplemented with vitamin C leading to resolution of all lesions within 10 d to 2 mo. Vitamin C is necessary for collagen synthesis, which is required for proper wound healing, capillary and cartilage strength, osteoid production, and pial membrane formation of the cerebellum. Several bat species cannot synthesize vitamin C and require a dietary source. This is the first report of vitamin C deficiency in a colony of vampire bats leading to severe chronic subcutaneous hemorrhage, bone fragility, microfractures, cerebellar necrosis, and death.

DIFFUSE IDIOPATHIC SKELETAL HYPEROSTOSIS WITH SECONDARY DYSPHAGIA IN A BLACK-HANDED SPIDER MONKEY (ATELES GEOFFROYI).

A 32-yr-old male black-handed spider monkey (Ateles geoffroyi) with marked kyphosis and reduced spinal range of motion developed intermittent regurgitation, which was managed with an acid reducer. Diffuse idiopathic skeletal hyperostosis (DISH) was suspected in this animal due to radiographically evident ossification of the anterior longitudinal ligament. At repeat radiographic evaluation 1.5 yr later, due to weight loss and increased frequency of regurgitation, the cervical spine was deviated ventrally and appeared to be impinging on the thoracic inlet. The spider monkey was humanely euthanized due to poor prognosis, and the presumptive diagnosis of DISH was confirmed via postmortem computed tomography and necropsy. DISH has not been reported in black-handed spider monkeys, and secondary dysphagia, an uncommon but recognized consequence in humans, has not been reported in a nonhuman primate. Earlier recognition of this possibly underreported disease process may increase treatment options and effectiveness of intervention.

A Survey of Prescribers' Attitudes, Knowledge, Comfort, and Fear of Consequences Related to an Opioid Overdose Education and Naloxone Distribution Program.

CONTEXT: The alarming increase in opioid-related fatalities has placed the United States in the midst of an opioid epidemic. In an effort to reduce opioid-related fatalities, each Veterans Affairs (VA) hospital is responsible for development of an OEND (Overdose Education and Naloxone Distribution) program to enhance naloxone distribution. OBJECTIVE: To assess attitudes, knowledge, comfort level, and fear of consequences at the prescriber level related to the OEND program at VA Connecticut Healthcare System (VACHS). DESIGN: Anonymous surveys were administered to VACHS prescribers to assess attitude, knowledge, comfort level, and fear of consequences using a 5-point Likert scale. SETTING: VACHS. PARTICIPANTS: VACHS prescribers (primary care physicians/internists, psychiatrists, physician assistants/advanced practice registered nurses, pharmacists) in primary care/medicine, mental health, and substance use disorder (SUD). METHODS: Differences across 4 domains were analyzed using analysis of variance testing, and pairwise post hoc analysis was conducted using Tukey's HSD for all significant findings. Regression modeling was conducted to evaluate correlations between each group and survey responses. MAIN OUTCOME: Identify differences and potential barriers to program implementation across professions and practice specialty. RESULTS: While primary care/medicine, mental health, and SUD prescribers had positive attitudes toward dispensing naloxone, SUD clinicians reported being more comfortable, more knowledgeable, and less fearful of consequences. In general, psychiatrists reported to be more knowledgeable and comfortable prescribing naloxone. In addition, prescribers who received naloxone training reported themselves to be more comfortable, more knowledgeable, and less fearful of the consequences of dispensing naloxone. CONCLUSIONS: Results indicate that barriers to OEND implementation continue to exist, and not all clinicians or practice specialties at VACHS are comfortable with dispensing naloxone. A targeted approach to training health care clinicians may lead to increased acceptance of naloxone dispensing. Additional research is needed to elicit the best method of improved acceptance of naloxone to expand naloxone access.

MYOSITIS OSSIFICANS TRAUMATICA IN A VAMPIRE BAT (DESMODUS ROTUNDUS).

A 15-yr-old sexually intact female vampire bat ( Desmodus rotundus ) was diagnosed with myositis ossificans traumatica of the abdominal wall. The bat presented with a large ulcerated firm mass along the abdomen. Radiographs and cytology were performed, followed by surgical exploration. The mass was determined to be nonresectable and the bat was euthanized. Histopathology showed severe necrotizing, degenerative, and pyogranulomatous myositis with osseous and cartilaginous metaplasia, fibrosis, and ulceration, which were consistent with myositis ossificans traumatica. Myositis ossificans traumatica is commonly associated with previous trauma to skeletal muscle. Two years prior, this bat had an emergency Caesarian section at this site, which was postulated to elicit a marked tissue response leading to this condition. Myositis ossificans traumatica is infrequently reported in humans, dogs, cats, pigs, and horses. To the author's knowledge, this is the first report of this condition in a bat.

"TERATOCARCINOMATOSIS" IN AN EMU (DROMAIUS NOVAEHOLLANDIAE).

An adult female emu (Dromaius novaehollandiae) with no previous abnormal clinical signs was found dead in its stall. A postmortem examination revealed carcinomatosis of unknown origin. Histopathology identified the masses as teratomas with malignant transformation by the presence of poorly differentiated tissue and dissemination throughout the coelomic cavity. We propose a new term, "teratocarcinomatosis," for this finding. This is the first case of a reproductive tumor described in an emu and the first case of such marked spread of malignant teratomas in a bird.

Virulence and immunity orchestrated by the global gene regulator sigL in Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis causes Johne's disease in ruminants, a chronic enteric disease responsible for severe economic losses in the dairy industry. Global gene regulators, including sigma factors are important in regulating mycobacterial virulence. However, the biological significance of such regulators in M. avium subsp. paratuberculosis rremains elusive. To better decipher the role of sigma factors in M. avium subsp. paratuberculosis pathogenesis, we targeted a key sigma factor gene, sigL, activated in mycobacterium-infected macrophages. We interrogated an M. avium subsp. paratuberculosis ΔsigL mutant against a selected list of stressors that mimic the host microenvironments. Our data showed that sigL was important in maintaining bacterial survival under such stress conditions. Survival levels further reflected the inability of the ΔsigL mutant to persist inside the macrophage microenvironments. Additionally, mouse infection studies suggested a substantial role for sigL in M. avium subsp. paratuberculosis virulence, as indicated by the significant attenuation of the ΔsigL-deficient mutant compared to the parental strain. More importantly, when the sigL mutant was tested for its vaccine potential, protective immunity was generated in a vaccine/challenge model of murine paratuberculosis. Overall, our study highlights critical role of sigL in the pathogenesis and immunity of M. avium subsp. paratuberculosis infection, a potential role that could be shared by similar proteins in other intracellular pathogens.

Bacterial load and inflammation in fetal tissues is not dependent on IL-17a or IL-22 in 10-14 day pregnant mice infected with Listeria monocytogenes.

In this study, we first assessed the effect of intragastric infection of pregnant mice with Listeria monocytogenes on relative expression of select genes associated with T cell subsets. Relative gene expression was moderately increased in placental tissues for IFNγ, IL-4, IL-17a, IL-22, CD3, and FoxP3. To assess the roles of IL-17a and IL-22 in resistance to listeriosis during pregnancy, we compared the severity of maternal and fetal infection in IL-17a((-/-)), IL-22((-/-)), and IL-17a((-/-))/IL-22((-/-)) mice with that of wild type C57BL/6 mice. Intragastric infection with modest numbers of bacterial cells (10(5) CFU) caused reproducible maternal and fetal infection in all four mouse strains. We recovered greater numbers of CFU from the bloodstream of pregnant IL-22((-/-)) mice than pregnant wild type mice. Otherwise we found no significant difference in bacterial load in maternal or fetal tissues (spleen, liver, fetoplacental units) from pregnant IL-17a((-/-)), IL-22((-/-)), or IL-17a((-/-))/IL-22((-/-)) or wild type mice. Nor did we observe histopathologic differences in severity of inflammation in maternal or fetal tissues from the various groups of mice. Although IL-17a and IL-22 are up-regulated in placental tissue, our study suggests that antibacterial resistance and the host inflammatory response are not dependent on IL-17a or IL-22 during infection of mice with L. monocytogenes at 10-14 days of gestation.

Five observations of a third morphologically distinct feline Demodex mite.

BACKGROUND: Feline demodicosis is caused by infestation with Demodex cati and/or Demodex gatoi. These two mites have distinctive morphological appearances. OBJECTIVES: To describe five observations of a morphologically distinct feline Demodex mite in 10 cats. All cats were in or adopted from an animal shelter. RESULTS: A mite with blunted ends longer than D. gatoi but shorter than D. cati was observed in one or multiple cats from animal shelters. Mean mite size was 139 ± 4.5 µm (n = 41 mites). Similar features among the cases included a history of recent recovery or presence of concurrent illness at the time of diagnosis. Pruritus was variable. Hairs were easily epilated in large amounts, and mites were found on either skin scrapings or hair trichograms; mites were most commonly found on the proximal third of hairs examined via trichogram. CLINICAL IMPORTANCE: Reports of this mite are uncommon but when present tend to be in cats with concurrent illnesses. Awareness of this mite morphology will hopefully provide other investigators with specimens for molecular testing to determine whether this is a third species or a variant of D. gatoi or D. cati.

Sternal osteosarcoma in a blue crane (Anthropoides paradiseus).

A previously healthy, adult male blue crane (Anthropoides paradiseus) became peracutely distressed and ataxic. Examination revealed hemoptysis and a large mass associated with the cranial aspect of the sternum (keel). The patient died before treatment could be initiated. Gross necropsy revealed a large, hemorrhagic mass originating from the cranial aspect of the keel and extending into the cranial coelomic cavity. Histopathology confirmed the mass as an osteosarcoma. This is the first known report of an osteosarcoma in any crane species.

Characterization of a novel heat shock protein (Hsp22.5) involved in the pathogenesis of Mycobacterium tuberculosis.

Tuberculosis is a worldwide health problem, given that one-third of the world's population is currently infected with Mycobacterium tuberculosis. Understanding the regulation of virulence on the molecular level will provide a better understanding of how M. tuberculosis can establish chronic infection. Using in vivo microarray analysis (IVMA), we previously identified a group of genes that are activated in BALB/c mouse lungs compared to in vitro cultures, including the rv0990c gene. Our analysis indicated that this gene is a member of the heat shock regulon and was activated under other stress conditions, including survival in macrophages or during the late phase of chronic tuberculosis in the murine lungs. Deletion of rv0990c from the genome of M. tuberculosis strain H37Rv affected the transcriptional profiles of many genes (n = 382) and operons involved in mycobacterial survival, including the dormancy regulon, ATP synthesis, respiration, protein synthesis, and lipid metabolism. Comparison of the proteomes of the mutant to those of the wild-type strain further confirmed the differential expression of 15 proteins, especially those involved in the heat shock response (e.g., DnaK and GrpE). Finally, the rv0990c mutant strain showed survival equivalent to that of the isogenic wild-type strain during active tuberculosis in guinea pigs, despite showing significant attenuation in BALB/c mice during the chronic phase of the disease. Overall, we suggest that rv0990c encodes a heat shock protein that plays an important role in mycobacterial virulence. Hence, we renamed rv0990c heat shock protein 22.5 (hsp22.5), reflecting its molecular mass.

CtpV: a putative copper exporter required for full virulence of Mycobacterium tuberculosis.

Copper is a required micronutrient that is also toxic at excess concentrations. Currently, little is known about the role of copper in interactions between bacterial pathogens and their human hosts. In this study, we elucidate a mechanism for copper homeostasis in the human pathogen Mycobacterium tuberculosis via characterization of a putative copper exporter, CtpV. CtpV was shown to be required by M. tuberculosis to maintain resistance to copper toxicity. Furthermore, the deletion of ctpV resulted in a 98-gene transcriptional response, which elucidates the increased stress experienced by the bacteria in the absence of this detoxification mechanism. Interestingly, although the ΔctpV mutant survives close to the wild-type levels in both murine and guinea pig models of tuberculosis, animals infected with the ΔctpV mutant displayed decreased lung damage, and mutant-infected mice had a reduced immune response to the bacteria as well as a significant increase in survival time relative to mice infected with wild-type M. tuberculosis. Overall, our study provides the first evidence for a connection between bacterial copper response and the virulence of M. tuberculosis, supporting the hypothesis that copper response could be important to intracellular pathogens, in general.

Pregnancy reduces the genetic resistance of C57BL/6 mice to Listeria monocytogenes infection by intragastric inoculation.

In this study, we compared genetically resistant C57BL/6 and susceptible A/J mice for their resistance to Listeria monocytogenes infection during pregnancy. Intragastric infection with modest numbers of bacterial cells (10(5) CFU) caused reproducible fetal infection and abortion in both mouse strains. Bioluminescence imaging demonstrated dissemination of L. monocytogenes cells from maternal to fetal organs within 3 days of intragastric infection. Although non-pregnant C57BL/6 mice were significantly more resistant to infection than non-pregnant A/J mice, C57BL/6 and A/J mice had similar microbial loads (CFU) in maternal and fetal tissues during pregnancy. Inflammation and necrosis, however, were more severe in A/J mice as evaluated by semi-quantitative histopathology. Although the microbial load in fetal tissues was similar for all fetuses within a single uterus, inflammation and necrosis varied among individual fetuses and placentas. We also noted that the uterus is a target for L. monocytogenes infection in non-pregnant mice.

Disseminated histoplasmosis in a Bengal tiger (Panthera tigris).

Disseminated infection with Histoplasma capsulatum was diagnosed in a 7-yr-old female Bengal tiger (Panthera tigris). Clinical signs were nonspecific with the exception of brief periods of tachypnea for 5 days prior to death. H. capsulatum organisms were found in the lungs, tracheobronchial lymph nodes, and liver. Diagnosis was confirmed by tracheal wash, urine H. capsulatum enzyme immunoassay, and necropsy results. This report represents the first published account of disseminated histoplasmosis in a tiger.

Hydrocephalus in a yellow-headed Amazon parrot (Amazona ochrocephala oratrix).

A 37-year-old female yellow-headed Amazon parrot (Amazona ochrocephala oratrix) was presented after a 4-month-period behavior change and intermittent episodes of obtunded mentation. Clinical findings on physical examination included ataxia, a weak grasp, and reluctance to move. Results of magnetic resonance imaging were consistent with severe hydrocephalus without evidence of cerebrospinal fluid obstruction. The bird was treated with tapering dosages of prednisolone over a 4-month period, during which time the episodes did not occur. Discontinuation of treatment was attempted several times but resulted in relapse. After 3.5 years of maintenance treatment with prednisolone, the bird was presented subsequent to a 5-hour episode of obtunded mentation and worsening neurologic signs. Despite increasing the dose of prednisolone and providing additional supportive care, the bird's condition worsened, and euthanasia was elected. Necropsy findings included severe hydrocephalus with significant loss of right cerebral parenchyma and no evidence of cerebrospinal fluid obstruction. Histologic examination of the remaining cerebral parenchyma revealed a moderate, multifocal, cellular infiltrate; encephalomalacia; fibrosis; and hemosiderosis in tissue adjacent to the distended ventricles. Other findings included hepatic vacuolar degeneration. Diagnostic imaging and postmortem findings were consistent with a diagnosis of hydrocephalus ex vacuo. To our knowledge, this is the first report of hydrocephalus in an Amazon parrot as well as the first report of hydrocephalus in any avian species associated with long-term follow-up and prolonged corticosteroid treatment.

Granular cell tumor in a lumbar spinal nerve of a dog.

A 2-year-old Great Dane dog with a 2.5-week history of progressive paraparesis was presented to the Veterinary Medical Teaching Hospital at the University of Wisconsin-Madison. Neurologic examination revealed nonambulatory paraparesis with reduced to absent withdrawal hind-limb reflexes and lumbar pain. Magnetic resonance imaging and gross pathology confirmed a larger regional lumbar mass and a second smaller extradural mass within the spinal canal. The left lumbar mass was associated with extensive hemorrhage; dissection showed a dark-red, soft, well-circumscribed mass measuring 2 cm x 1.5 cm x 0.5 cm within the left fourth lumbar spinal nerve. Histopathological evaluation with immunohistochemistry revealed sheets of round to polygonal cells with diffuse granular cytoplasm demonstrating diastase-resistant periodic acid-Schiff reactivity and positive immunoexpression of S100 and neuron-specific enolase. The smaller extradural mass within the spinal canal exhibited similar morphology. Based on gross, histological, and immunohistochemical evidence, the masses were diagnosed as granular cell tumor.

Proventricular dilatation disease associated with Avian bornavirus in a scarlet macaw (Ara macao).

A case of proventricular dilatation disease is described in a scarlet macaw (Ara macao) from clinical presentation to diagnosis with molecular methods. The initial clinical signs were depression progressing to head pressing over several days. A leukocytosis with toxic heterophil changes, hypoalbuminemia, and increased serum activity of aspartate aminotransferase and creatine kinase were present. Lead and zinc assays were within reference ranges, and results from Chlamydophila and polyomavirus testing were negative. Contrast-enhanced fluoroscopy revealed normal gastrointestinal transit times and motility as well as the presence of 2 small metallic foreign bodies in the ventriculus. The macaw was treated with antimicrobials, analgesics, vitamins E and B complex, force-feeding, and fluid administration with little improvement. Euthanasia was elected, and histologic examination of brain tissue revealed a perivascular lymphoplasmacytic infiltration, while the lungs had evidence of a fungal pneumonia. Tissue samples from the brain and proventriculus tested positive for the presence of Avian bornavirus genotype 2, while serology confirmed Avian bornavirus infection.

Treatment of feline gastrointestinal small-cell lymphoma with chlorambucil and glucocorticoids.

Gastrointestinal (GI) lymphoma is the most frequently diagnosed form of lymphoma in the cat and is categorized into two distinct forms based on the size of neoplastic lymphocytes. Treatments for both large- and small-cell GI lymphoma have been described previously; however, multiple chemotherapy protocols were used, a minimal amount of histopathological characterization was provided, and, in most studies, the majority of diagnoses were obtained via endoscopic pinch biopsies. Twenty-eight cats (24 with full-thickness intestinal biopsies) were diagnosed with small-cell GI lymphoma and treated with a combination of chlorambucil and glucocorticoids. The majority of cases were strongly CD3+, and many displayed epitheliotropism. The overall clinical response rate was 96%, with a median clinical remission duration of 786 days. Follow-up identified seven cats with relapsed disease-all of which were treated with a rescue protocol of cyclophosphamide and glucocorticoids; the response rate was 100%, and four of the 28 cats were diagnosed with a second malignancy.

A Protective Vaccine against Johne's Disease in Cattle.

Johne's disease (JD) caused by Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) is a chronic infection characterized by the development of granulomatous enteritis in wild and domesticated ruminants. It is one of the most significant livestock diseases not only in the USA but also globally, accounting for USD 200-500 million losses annually for the USA alone with potential link to cases of Crohn's disease in humans. Developing safe and protective vaccines is of a paramount importance for JD control in dairy cows. The current study evaluated the safety, immunity and protective efficacy of a novel live attenuated vaccine (LAV) candidate with and without an adjuvant in comparison to an inactivated vaccine. Results indicated that the LAV, irrespective of the adjuvant presence, induced robust T cell immune responses indicated by proinflammatory cytokine production such as IFN-γ, IFN-α, TNF-α and IL-17 as well as strong response to intradermal skin test against M. paratuberculosis antigens. Furthermore, the LAV was safe with minimal tissue pathology. Finally, calves vaccinated with adjuvanted LAV did not shed M. paratuberculosis post-challenge, a much-desired characteristic of an effective vaccine against JD. Together, this data suggests a strong potential of testing LAV in field trials to curb JD in dairy herds.

A single dose polyanhydride-based nanovaccine against paratuberculosis infection.

Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) causes Johne's disease in ruminants and is characterized by chronic gastroenteritis leading to heavy economic losses to the dairy industry worldwide. The currently available vaccine (inactivated bacterin in oil base) is not effective in preventing pathogen shedding and is rarely used to control Johne's disease in dairy herds. To develop a better vaccine that can prevent the spread of Johne's disease, we utilized polyanhydride nanoparticles (PAN) to encapsulate mycobacterial antigens composed of whole cell lysate (PAN-Lysate) and culture filtrate (PAN-Cf) of M. paratuberculosis. These nanoparticle-based vaccines (i.e., nanovaccines) were well tolerated in mice causing no inflammatory lesions at the site of injection. Immunological assays demonstrated a substantial increase in the levels of antigen-specific T cell responses post-vaccination in the PAN-Cf vaccinated group as indicated by high percentages of triple cytokine (IFN-γ, IL-2, TNF-α) producing CD8+ T cells. Following challenge, animals vaccinated with PAN-Cf continued to produce significant levels of double (IFN-γ, TNF-α) and single cytokine (IFN-γ) secreting CD8+ T cells compared with animals vaccinated with an inactivated vaccine. A significant reduction in bacterial load was observed in multiple organs of animals vaccinated with PAN-Cf, which is a clear indication of protection. Overall, the use of polyanhydride nanovaccines resulted in development of protective and sustained immunity against Johne's disease, an approach that could be applied to counter other intracellular pathogens.