AN OUTBREAK OF FELINE INFECTIOUS PERITONITIS IN THREE RELATED SAND CATS (FELIS MARGARITA) IN HUMAN CARE.

Feline infectious peritonitis (FIP) is a systemic disease in felid species caused by infection with mutated forms of feline coronavirus (FCoV), and outbreaks can devastate exotic felid populations in human care. Feline infectious peritonitis was diagnosed in three of four related juvenile sand cats (Felis margarita) from a single institution over a 6-wk period. Case 1 was a 7-mon-old male found deceased with no premonitory signs. Case 2, an 8-mon-old male (littermate to Case 1), and Case 3, a 6-mon-old male (from a different litter with identical parentage), were evaluated for lethargy and anorexia 1 mon after Case 1. Both exhibited transient anisocoria and progressive lethargy, anorexia, and dehydration despite antibiotic and supportive treatment. Approximately 1 wk after initial presentation, Case 2 was humanely euthanized, and Case 3 was found deceased. Necropsy findings included intrathoracic and/or intra-abdominal lymphadenopathy (3/3 cases), bicavitary effusion (2/3), multifocal tan hepatic and intestinal nodules (1/3), and multifocal yellow renal nodules (1/3). Histologically, all cats had severe pyogranulomatous vasculitis in multiple organs, and the presence of FCoV antigen was confirmed using immunohistochemical staining. Next-generation sequencing of the virus from Case 3's affected kidney demonstrated ∼93% homology to the UG-FH8 virus, a serotype 1 feline alphacoronavirus isolated from Denmark. Future research will focus on comparative viral genomic sequencing with the goals of identifying potential sources of FCoV infection and identifying features that may have contributed to the development of FIP in this species.

DEVELOPMENT AND VALIDATION OF A NOVEL DUPLEX PROBE-HYBRIDIZATION QUANTITATIVE PCR FOR LYMPHOMA-ASSOCIATED MIROUNGINE GAMMAHERPESVIRUS 3 IN NORTHERN ELEPHANT SEALS (MIROUNGA ANGUSTIROSTRIS).

Recently, a novel gammaherpesvirus, miroungine gammaherpesvirus 3 (MirGHV3), was described in two juvenile elephant seals (Mirounga angustirostris) with diffuse large B-cell lymphoma. We developed and validated a quantitative (q)PCR for rapid detection of MirGHV3 and investigated its potential association with lymphoma. We developed a duplex probe-hybridization qPCR with MirGHV3 DNA polymerase (pol) as the target gene. Each primer-probe combination was cross-validated against the others. Interference was not seen when they were run in the same well as a duplex assay. Twenty-three samples from seven northern elephant seals were tested using the duplex assay. Viral DNA was detected by the assay in 9 of 9 (100%) tissues affected by lymphoma and in 6 of 14 (43%) samples from tissues unaffected by lymphoma. There was a strong correlation between viral copies detected with each of the assays (P=0.0002). Viral load was significantly higher in tissues affected by lymphoma than in those unaffected (P<0.0001). Excluding the virus-negative samples, viral load was still significantly higher in tissues affected by lymphoma than in those unaffected (P=0.0004). This is consistent with a potential role of MirGHV3 in oncogenesis in northern elephant seals, although more studies are needed to determine this definitively. The qPCR developed has utility for further investigations of MirGHV3.

Diffuse large B cell lymphoma and a novel gammaherpesvirus in northern elephant seals Mirounga angustirostris.

Two emaciated male northern elephant seal (NES) Mirounga angustirostris pups were admitted to The Marine Mammal Center (Sausalito, California, USA) and treated for malnutrition. Complete blood counts showed a progressive moderate to marked leukocytosis characterized by a predominance of large monomorphic mononuclear cells of probable lymphoid origin, frequently with flower-shaped nuclei. Both seals were euthanized due to suspected lymphoid neoplasia. At necropsy, most lymph nodes in both pups were markedly enlarged, some with distinct white nodules, the spleens were diffusely enlarged, and the intestinal mucosae were thickened. Histopathologic features consistent with disseminated large cell lymphoma were identified to varying degrees of severity in lymph nodes, bone marrow, liver, tonsils, spleen, liver, intestines, kidneys, lower urinary tract, and several other organs. Immunohistochemical staining of neoplastic cells was most consistent with B lymphocyte origin, with most cells staining positively for Pax 5 and CD20 with admixed small CD3-positive T lymphocytes and CD204-positive macrophages. PCR and sequencing identified a novel gammaherpesvirus, herein called miroungine gammaherpesvirus 3, from affected tissues. This virus is in a clade outside of named genera that utilize hosts in the suborder Caniformia. The present study is the first description of diffuse large B cell lymphoma with leukemic manifestation and concomitant detection of a novel gammaherpesvirus in free-living NESs. Further research regarding the prevalence of this new gammaherpesvirus and its associated pathogenesis in this species is indicated.

Gut microbiota and age shape susceptibility to clostridial enteritis in lorikeets under human care.

BACKGROUND: Enteritis is a common cause of morbidity and mortality in lorikeets that can be challenging to diagnose and treat. In this study, we examine gut microbiota in two lorikeet flocks with enteritis (Columbus Zoo and Aquarium-CZA; Denver Zoo-DZ). Since 2012, the CZA flock has experienced repeated outbreaks of enteritis despite extensive diet, husbandry, and clinical modifications. In 2018, both CZA and DZ observed a spike in enteritis. Recent research has revealed that the gut microbiota can influence susceptibility to enteropathogens. We hypothesized that a dysbiosis, or alteration in the gut microbial community, was making some lorikeets more susceptible to enteritis, and our goal was to characterize this dysbiosis and determine the features that predicted susceptibility. RESULTS: We employed 16S rRNA sequencing to characterize the cloacal microbiota in lorikeets (CZA n = 67, DZ n = 24) over time. We compared the microbiota of healthy lorikeets, to lorikeets with enteritis, and lorikeets susceptible to enteritis, with "susceptible" being defined as healthy birds that subsequently developed enteritis. Based on sequencing data, culture, and toxin gene detection in intestinal contents, we identified Clostridium perfringens type A (CZA and DZ) and C. colinum (CZA only) at increased relative abundances in birds with enteritis. Histopathology and immunohistochemistry further identified the presence of gram-positive bacilli and C. perfringens, respectively, in the necrotizing intestinal lesions. Finally, using Random Forests and LASSO models, we identified several features (young age and the presence of Rhodococcus fascians and Pseudomonas umsongensis) associated with susceptibility to clostridial enteritis. CONCLUSIONS: We identified C. perfringens type A and C. colinum associated with lorikeet necrohemorrhagic enteritis at CZA and DZ. Susceptibility testing of isolates lead to an updated clinical treatment plan which ultimately resolved the outbreaks at both institutions. This work provides a foundation for understanding gut microbiota features that are permissive to clostridial colonization and host factors (e.g. age, prior infection) that shape responses to infection.

Nonsteroidal anti-inflammatory drugs restore immune function to respiratory syncytial virus in geriatric cotton rats (Sigmodon hispidus).

Respiratory syncytial virus (RSV) infection is not only a childhood disease, but also a serious health risk for the elderly. We investigated in cotton rats how age affected viral clearance, immune responses, and whether pharmacological intervention was beneficial. Our results demonstrated that in geriatric animals, virus grew to similar titers, but with delayed clearance, compared to adult animals. After primary infection with RSV, geriatric animals were susceptible to secondary infection and results indicated a defective humoral immune response. Depletion of cytotoxic T lymphocytes (CTL) during primary infection delayed clearance, indicating the necessary role of CTL. Pharmacological intervention through nonsteroidal anti-inflammatory ibuprofen resulted in faster viral clearance and complete protection after immunization. In addition, the CTL response in the presence of ibuprofen seemed to be restored. It appears that in geriatric animals, immune functions are not as effective as in adult animals and that anti-inflammatory therapy may restore effective immune function.

Consistency and strength of grapheme-color associations are separable aspects of synesthetic experience.

Consistency of synesthetic associations over time is a widely used test of synesthesia. Since many studies suggest that consistency is not a completely reliable feature, we compared the consistency and strength of synesthetes' grapheme-color associations. Consistency was measured by scores on the Synesthesia Battery and by the Euclidean distance in color space for the specific graphemes tested for each participant. Strength was measured by congruency magnitudes on the Implicit Association Test. The strength of associations was substantially greater for synesthetes than non-synesthetes, suggesting that this is a novel, objective marker of synesthesia. Although, intuitively, strong associations should also be consistent, consistency and strength were uncorrelated, indicating that they are likely independent, at least for grapheme-color synesthesia. These findings have implications for our understanding of synesthesia and for estimates of its prevalence since synesthetes who experience strong, but inconsistent, associations may not be identified by tests that focus solely on consistency.

Immunogenicity and inflammatory properties of respiratory syncytial virus attachment G protein in cotton rats.

Human respiratory syncytial virus (RSV) is a leading cause of lower respiratory tract infection in infants and young children worldwide. The attachment (G) protein of RSV is synthesized by infected cells in both a membrane bound (mG) and secreted form (sG) and uses a CX3C motif for binding to its cellular receptor. Cell culture and mouse studies suggest that the G protein mimics the cytokine CX3CL1 by binding to CX3CR1 on immune cells, which is thought to cause increased pulmonary inflammation in vivo. However, because these studies have used RSV lacking its G protein gene or blockade of the G protein with a G protein specific monoclonal antibody, the observed reduction in inflammation may be due to reduced virus replication and spread, and not to a direct role for G protein as a viral chemokine. In order to more directly determine the influence of the soluble and the membrane-bound forms of G protein on the immune system independent of its attachment function for the virion, we expressed the G protein in cotton rat lungs using adeno-associated virus (AAV), a vector system which does not itself induce inflammation. We found no increase in pulmonary inflammation as determined by histology and bronchoalveolar lavage after inoculation of AAVs expressing the membrane bound G protein, the secreted G protein or the complete G protein gene which expresses both forms. The long-term low-level expression of AAV-G did, however, result in the induction of non-neutralizing antibodies, CD8 T cells and partial protection from challenge with RSV. Complete protection was accomplished through co-immunization with AAV-G and an AAV expressing cotton rat interferon α.

Cotton Rat Placenta Anatomy and Fc Receptor Expression and Their Roles in Maternal Antibody Transfer.

Respiratory syncytial virus (RSV) is the leading cause of bronchiolitis and viral pneumonia in infants and young children worldwide. Currently no vaccine is available to prevent RSV infection, but virus-neutralizing monoclonal antibodies can be given prophylactically, emphasizing the protective potential of antibodies. One concept of RSV vaccinology is mothers' immunization to induce high antibody titers, leading to passive transfer of high levels of maternal antibody to the fetus through the placenta and to the neonate through colostrum. Cotton rats are an excellent small animal model for RSV infection and have been used to test maternal immunization. To mechanistically understand antibody transfer in the cotton rat model, we characterized the cotton rat placenta and Fc receptor localization. Placentas from cotton rats at midgestation (approximately day 14) and at late gestation (approximately day 25) and neonatal (younger than 1 wk) gastrointestinal tracts were collected for light microscopy, immunohistochemistry, and transmission electron microscopy. The cotton rat placenta is hemotrichorial and has 5 distinct layers: decidua, junctional zone, labyrinth, chorionic plate, and yolk sac. Consistent with the transfer of maternal antibodies, the majority of the Fc receptors are present in the yolk sac endoderm and fetal capillary endothelium of the chorionic plate, involving 10% of the cells within the labyrinth. In addition, Fc receptors are present on duodenal and jejunal enterocytes in cotton rats, similar to humans, mice, and rats. These findings provide the structural basis for the pre- and postnatal transfer of maternal antibodies described in cotton rats.

Pulmonary function analysis in cotton rats after respiratory syncytial virus infection.

The cotton rat (Sigmodon hispidus) is an excellent small animal model for human respiratory viral infections such as human respiratory syncytial virus (RSV) and human metapneumovirus (HMPV). These respiratory viral infections, as well as other pulmonary inflammatory diseases such as asthma, are associated with lung mechanic disturbances. So far, the pathophysiological effects of viral infection and allergy on cotton rat lungs have not been measured, although this information might be an important tool to determine the efficacy of vaccine and drug candidates. To characterize pulmonary function in the cotton rat, we established forced oscillation technique in uninfected, RSV infected and HDM sensitized cotton rats, and characterized pulmonary inflammation, mucus production, pulmonary edema, and oxygenation. There was a gender difference after RSV infection, with females demonstrating airway hyper-responsiveness while males did not. Female cotton rats 2dpi had a mild increase in pulmonary edema (wet: dry weight ratios). At day 4 post infection, female cotton rats demonstrated mild pulmonary inflammation, no increase in mucus production or reduction in oxygenation. Pulmonary function was not significantly impaired after RSV infection. In contrast, cotton rats sensitized to HDM demonstrated airway hyper-responsiveness with a significant increase in pulmonary inflammation, increase in baseline tissue damping, and a decrease in baseline pulmonary compliance. In summary, we established baseline data for forced oscillation technique and other respiratory measures in the cotton rat and used it to analyze respiratory diseases in cotton rats.

Patterns of Osteopontin Expression in Abusive Head Trauma Compared with Other Causes of Pediatric Traumatic Brain Injury.

OBJECTIVE: To examine levels of plasma osteopontin (OPN), a recently described neuroinflammatory biomarker, in children with abusive head trauma (AHT) compared with children with other types of traumatic brain injury (TBI). STUDY DESIGN: The study cohort comprised children aged <4 years diagnosed with TBI and seen in the intensive care unit in a tertiary children's hospital. Patients were classified as having confirmed or suspected AHT or TBI by other mechanisms (eg, motor vehicle accidents), as identified by a Child Protection Team clinician. Serial blood samples were collected at admission and at 24, 48, and 72 hours after admission. Levels of OPN were compared across groups. RESULTS: Of 77 patients identified, 24 had confirmed AHT, 12 had suspected AHT, and 41 had TBI. There were no differences in the Glasgow Coma Scale score between the patients with confirmed AHT and those with suspected AHT and those with TBI (median score, 4.5 vs 4 and 7; P = .39). At admission to the emergency department, OPN levels were significantly higher in children with confirmed AHT compared with the other 2 groups (mean confirmed AHT, 471.5 ng/mL; median suspected AHT, 322.3 ng/mL; mean TBI, 278.0 ng/mL; P = .03). Furthermore, the adjusted mean trajectory levels of OPN were significantly higher in the confirmed AHT group compared with the other 2 groups across all subsequent time points (P = <.01). CONCLUSIONS: OPN is significantly elevated in children with confirmed AHT compared with those with suspected AHT and those with other types of TBI. OPN expression may help identify children with suspected AHT to aid resource stratification and triage of appropriate interventions for children who are potential victims of abuse.

Distemper Encephalomyelitis Presenting with Lower Motor Neuron Signs in a Young Dog.

A 4 mo old spayed female mixed-breed dog was presented for focal lower motor neuron signs of the right forelimb and marked hyperesthesia on axillary palpation. Her signs progressed rapidly over the following days to diffuse lower motor neuron signs in all limbs and a seizure. MRI demonstrated a focal, slightly right-sided, 2.5 cm region of noncontrast-enhancing T2 hyperintensity and T1 isointensity at C4-C5 spinal cord segments. Imaging of the brain was unremarkable. The dog was euthanized as a result of poor prognosis. Polymerase chain reaction on cerebrospinal fluid and immunohistochemistry of brain tissue were both positive for canine distemper virus. This report documents an atypical presentation of canine distemper encephalomyelitis causing lower motor neuron signs and hyperesthesia.

SYSTEMIC AMYLOIDOSIS IN A POPULATION OF PRONGHORN ANTELOPE (ANTILOCAPRA AMERICANA).

Thirteen pronghorn antelope (Antilocapra americana) from a single captive herd at the Columbus Zoo and Aquarium underwent complete or partial necropsies between 1997 and 2016. Ten of the 13 animals had histologic evidence of amyloidosis resulting in a 77% prevalence. Histologic and ultrastructural changes were characterized in an attempt to determine the underlying cause of the amyloid. Amyloid detection was performed through histologic examination of hemotoxylin and eosin and Congo red-stained microscopic slides for all 13 animals. Transmission electron microscopy and mass spectrometry was performed on renal tissue from two animals. Pedigree analysis and retrospective investigation into the clinical histories was performed. Histologically, 9/10 animals had amyloid present in the kidneys, 8/10 in the liver, 9/10 in the spleen, 4/10 in the gastrointestinal tract, 3/10 in the adrenal glands, and 2/10 in the thyroid glands. Transmission electron microscopy demonstrated glomerular deposits consistent with amyloid. Mass spectrometry performed on renal specimens from two animals revealed the presence of serum amyloid A. Eight of the 10 animals diagnosed with systemic amyloidosis had a clinical history of haemonchosis (elevated fecal strongyle count), 5/10 were diagnosed with pneumonia postmortem, and 7/10 had postmortem findings consistent with negative energy balance. Serum amyloid A, and ß and γ globulin levels were evaluated in four cases of amyloidosis, and all were within normal ranges for healthy domestic cattle. It was possible that the herd's amyloidosis was associated with a hereditary defect that could be exacerbated by chronic inflammation. However, there was no significant association between the mean degree of relatedness and presence of amyloidosis. In conclusion, systemic amyloidosis in this captive population of pronghorn is common. It is likely reactive and secondary to underlying chronic inflammation caused by haemonchosis and/or pneumonia.

Synesthesia strengthens sound-symbolic cross-modal correspondences.

Synesthesia is a phenomenon in which an experience in one domain is accompanied by an involuntary secondary experience in another, unrelated domain; in classical synesthesia, these associations are arbitrary and idiosyncratic. Cross-modal correspondences refer to universal associations between seemingly unrelated sensory features, e.g., auditory pitch and visual size. Some argue that these phenomena form a continuum, with classical synesthesia being an exaggeration of universal cross-modal correspondences, whereas others contend that the two are quite different, since cross-modal correspondences are non-arbitrary, non-idiosyncratic, and do not involve secondary experiences. Here, we used the implicit association test to compare synesthetes' and non-synesthetes' sensitivity to cross-modal correspondences. We tested the associations between auditory pitch and visual elevation, auditory pitch and visual size, and sound-symbolic correspondences between auditory pseudowords and visual shapes. Synesthetes were more sensitive than non-synesthetes to cross-modal correspondences involving sound-symbolic, but not low-level sensory, associations. We conclude that synesthesia heightens universally experienced cross-modal correspondences, but only when these involve sound symbolism. This is only partly consistent with the idea of a continuum between synesthesia and cross-modal correspondences, but accords with the idea that synesthesia is a high-level, post-perceptual phenomenon, with spillover of the abilities of synesthetes into domains outside their synesthesias. To our knowledge, this is the first demonstration that synesthetes, relative to non-synesthetes, experience stronger cross-modal correspondences outside their synesthetic domains.

The functional significance of the last 5 residues of the C-terminus of cardiac troponin I.

The C-terminal region of cardiac troponin I (cTnI) is known to be important in cardiac function, as removal of the last 17 C-terminal residues of human cTnI has been associated with myocardial stunning. To investigate the C-terminal region of cTnI, three C-terminal deletion mutations in human cTnI were generated: Δ1 (deletion of residue 210), Δ3 (deletion of residues 208-210), and Δ5 (deletion of residues 206-210). Mammalian two-hybrid studies showed that the interactions between cTnI mutants and cardiac troponin C (cTnC) or cardiac troponin T (cTnT) were impaired in Δ3 and Δ5 mutants when compared to wild-type cTnI. Troponin complexes containing 2-[4'-(iodoacetamido) anilino] naphthalene-6-sulfonic acid (IAANS) labeled cTnC showed that the troponin complex containing cTnI Δ5 had a small increase in Ca(2+) affinity (P < 0.05); while the cTnI Δ1- and Δ3 troponin complexes showed no difference in Ca(2+) affinity when compared to wild-type troponin. In vitro motility assays showed that all truncation mutants had increased Ca(2+) dependent motility relative to wild-type cTnI. These results suggest that the last 5 C-terminal residues of cTnI influence the binding of cTnI with cTnC and cTnT and affect the Ca(2+) dependence of filament sliding, and demonstrate the importance of this region of cTnI.

Emotion regulation difficulties in anorexia nervosa before and after inpatient weight restoration.

OBJECTIVE: This study examined: (1) changes in emotion regulation difficulties in underweight inpatients with anorexia nervosa (AN) following weight restoration, (2) differences in emotion regulation between AN subtypes at acute and weight-restored stages of illness. METHOD: Repeated measure analyses of variance examined changes in scores on the Difficulties in Emotion Regulation Scale (DERS; Gratz and Roemer, J Psychopathol Behav Assess, 26, 41-54, 2004) and other clinical variables in a group of inpatients with AN from hospital admission (N = 65) to weight restoration (N = 51). Correlations between BMI and DERS scores at both time points were examined. Emotion regulation difficulties were compared between individuals with AN, restricting type (AN-R) and AN, binge/purge type (AN-BP) at both time points using multivariate analysis of covariance. RESULTS: All clinical variables, except for the DERS, significantly improved with weight restoration (p < .001). There were no associations between BMI and DERS prior to or after weight restoration and AN subtypes did not significantly differ in emotion regulation difficulties. DISCUSSION: Unlike other clinical variables, emotion regulation difficulties in AN did not improve with weight restoration. In addition, both subtypes of AN appear to have similar difficulties with emotion regulation. The treatment of AN might be enhanced by focusing on improving emotion regulation abilities.

The evolution of intergroup bias: perceptions and attitudes in rhesus macaques.

Social psychologists have learned a great deal about the nature of intergroup conflict and the attitudinal and cognitive processes that enable it. Less is known about where these processes come from in the first place. In particular, do our strategies for dealing with other groups emerge in the absence of human-specific experiences? One profitable way to answer this question has involved administering tests that are conceptual equivalents of those used with adult humans in other species, thereby exploring the continuity or discontinuity of psychological processes. We examined intergroup preferences in a nonhuman species, the rhesus macaque (Macaca mulatta). We found the first evidence that a nonhuman species automatically distinguishes the faces of members of its own social group from those in other groups and displays greater vigilance toward outgroup members (Experiments 1-3). In addition, we observed that macaques spontaneously associate novel objects with specific social groups and display greater vigilance to objects associated with outgroup members (Experiments 4-5). Finally, we developed a looking time procedure-the Looking Time Implicit Association Test, which resembles the Implicit Association Test (Greenwald & Banaji, 1995)-and we discovered that macaques, like humans, automatically evaluate ingroup members positively and outgroup members negatively (Experiments 6-7). These field studies represent the first controlled experiments to examine the presence of intergroup attitudes in a nonhuman species. As such, these studies suggest that the architecture of the mind that enables the formation of these biases may be rooted in phylogenetically ancient mechanisms.

Helping behaviour and regard for others in capuchin monkeys (Cebus apella).

Altruism is an evolutionary puzzle. To date, much debate has focused on whether helping others without regard to oneself is a uniquely human behaviour, with a variety of empirical studies demonstrating a lack of altruistic behaviour in chimpanzees even when the demands of behaving altruistically seem minimal. By contrast, a recent experiment has demonstrated that chimpanzees will help a human experimenter to obtain an out-of-reach object, irrespective of whether or not they are offered a reward for doing so, suggesting that the cognitions underlying altruistic behaviour may be highly sensitive to situational demands. Here, we examine the cognitive demands of other-regarding behaviour by testing the conditions under which primates more distantly related to humans--capuchin monkeys--help an experimenter to obtain an out-of-reach object. Like chimpanzees, capuchin monkeys helped human experimenters even in the absence of a reward, but capuchins systematically failed to take into account the perspective of others when they stood to obtain food for themselves. These results suggest an important role for perspective taking and inhibition in altruistic behaviour and seem to reflect a significant evolutionary development in the roots of altruism, and specifically in other-regarding behaviour, between the divergence of New World monkeys and apes.