Disseminated protothecosis with central nervous system involvement in a dog in New Zealand.

CASE HISTORY: A 1-year-old Border Terrier presented with acute onset of neurological signs and neck pain. CLINICAL FINDINGS: Severe generalised ataxia, muscle tremors and cranial nerve deficits were noted. Multifocal brain lesions were suspected based on neurological examination. Computed tomography revealed an abdominal mass and cerebellar herniation through the foramen magnum. LABORATORY AND PATHOLOGICAL FINDINGS: Cytological and histopathological analysis of the abdominal mass revealed necrotising and granulomatous lymphadenitis with intralesional algal elements most consistent with Prototheca spp.. Culture of a sample from the mesenteric lymph node confirmed the presence of Prototheca spp. which was identified as P. bovis based on sequencing of a DNA fragment amplified by PCR. Following inadequate response to symptomatic therapy and poor prognosis, the dog was subjected to euthanasia. Histopathological evaluation of the central nervous system lesions, identified granulomatous meningitis and ventriculitis with the presence of intralesional algae. DIAGNOSIS: Disseminated protothecosis with granulomatous meningitis and ventriculitis caused by Prototheca bovis (formerly P. zopfii gen. 2). CLINICAL RELEVANCE: This is the first case report of disseminated protothecosis with central nervous system involvement in a dog in New Zealand.

A Comprehensive Review on Crustaceans' Immune System With a Focus on Freshwater Crayfish in Relation to Crayfish Plague Disease.

Freshwater crayfish immunity has received great attention due to the need for urgent conservation. This concern has increased the understanding of the cellular and humoral defense systems, although the regulatory mechanisms involved in these processes need updating. There are, however, aspects of the immune response that require clarification and integration. The particular issues addressed in this review include an overall description of the oomycete Aphanomyces astaci, the causative agent of the pandemic plague disease, which affects freshwater crayfish, and an overview of crustaceans' immunity with a focus on freshwater crayfish. It includes a classification system of hemocyte sub-types, the molecular factors involved in hematopoiesis and the differential role of the hemocyte subpopulations in cell-mediated responses, including hemocyte infiltration, inflammation, encapsulation and the link with the extracellular trap cell death pathway (ETosis). In addition, other topics discussed include the identity and functions of hyaline cells, the generation of neoplasia, and the emerging topic of the role of sessile hemocytes in peripheral immunity. Finally, attention is paid to the molecular execution of the immune response, from recognition by the pattern recognition receptors (PRRs), the role of the signaling network in propagating and maintaining the immune signals, to the effector elements such as the putative function of the Down syndrome adhesion molecules (Dscam) in innate immune memory.

Interferon therapies in small animals.

Interferons (IFNs) are cytokines that play an important role in the immune response of animals and humans. A number of studies reviewed here have evaluated the use of human, canine and feline IFNs as treatments for infectious, inflammatory and neoplastic disease in dogs and cats. Recombinant canine IFN-γ is deemed an efficacious therapy for canine atopic dermatitis. Recombinant feline IFN-ω is effective against canine parvoviral enteritis and has also been recommended for canine atopic dermatitis. Based on limited evidence, recombinant canine IFN-α could be a topical treatment option for dogs with gingivitis and keratoconjunctivitis sicca. Conclusive evidence is lacking for other diseases and large randomised controlled trials are needed before IFNs can be recommended for other indications.

Patterns and processes of pathogen exposure in gray wolves across North America.

The presence of many pathogens varies in a predictable manner with latitude, with infections decreasing from the equator towards the poles. We investigated the geographic trends of pathogens infecting a widely distributed carnivore: the gray wolf (Canis lupus). Specifically, we investigated which variables best explain and predict geographic trends in seroprevalence across North American wolf populations and the implications of the underlying mechanisms. We compiled a large serological dataset of nearly 2000 wolves from 17 study areas, spanning 80° longitude and 50° latitude. Generalized linear mixed models were constructed to predict the probability of seropositivity of four important pathogens: canine adenovirus, herpesvirus, parvovirus, and distemper virus-and two parasites: Neospora caninum and Toxoplasma gondii. Canine adenovirus and herpesvirus were the most widely distributed pathogens, whereas N. caninum was relatively uncommon. Canine parvovirus and distemper had high annual variation, with western populations experiencing more frequent outbreaks than eastern populations. Seroprevalence of all infections increased as wolves aged, and denser wolf populations had a greater risk of exposure. Probability of exposure was positively correlated with human density, suggesting that dogs and synanthropic animals may be important pathogen reservoirs. Pathogen exposure did not appear to follow a latitudinal gradient, with the exception of N. caninum. Instead, clustered study areas were more similar: wolves from the Great Lakes region had lower odds of exposure to the viruses, but higher odds of exposure to N. caninum and T. gondii; the opposite was true for wolves from the central Rocky Mountains. Overall, mechanistic predictors were more informative of seroprevalence trends than latitude and longitude. Individual host characteristics as well as inherent features of ecosystems determined pathogen exposure risk on a large scale. This work emphasizes the importance of biogeographic wildlife surveillance, and we expound upon avenues of future research of cross-species transmission, spillover, and spatial variation in pathogen infection.

[Disseminated protothecosis with ulcerative granulomatous colitis in a Rhodesian Ridgeback from Germany]. / Ulzerative granulomatöse Kolitis durch Prototheca spp. bei einem Rhodesian Ridgeback in Deutschland.

A 10-month-old male Rhodesian Ridgeback was presented to the Clinic of Small Animal Medicine, LMU, Germany, with a 6-month history of chronic diarrhea and hematochezia. The dog lived in Germany and had never traveled abroad. Complete blood count and serum biochemistry performed by the referring veterinarian revealed neutrophilia, hyperkalemia, and hyponatremia, with a basal cortisol of 4.3 µg/dl, which excluded hypoadrenocorticism. Since antibiotic treatment had not resulted in any improvement, a 2 week course of prednisolone administration had been initiated, leading to a marked deterioration of intestinal signs and a significant weight loss of 6 kg. At the time of referral, the patient was markedly emaciated, dehydrated, hypovolemic and had a rectal temperature of 39.6 °C. Abdominal ultrasound showed a thickened and irregular colonic wall. On colonoscopy, an irregular colonic mucosa with ulcerations was observed. Histopathologic examination revealed an ulcerative granulomatous colitis, and on Periodic acid-Schiff reaction (PAS) numerous organisms consistent with Prototheca spp. were identified. Prototheca zopfii infection was confirmed by culture and MALDI-TOF MS. In order to test for an underlying immunodeficiency, immunoglobulin levels in serum were determined. IgM was decreased, while IgG and IgA levels were within the reference interval. Due to deterioration of general condition, grave prognosis and costs of a treatment trial, the patient was euthanized one week later, and necropsy was performed. Prototheca spp. were detected on histopathologic examination in the lymphnodes, however not in the eyes or the central nervous system. Protothecosis should be considered an differential diagnosis in dogs with chronic diarrhea and ulcerative granulomatous colitis even in dogs living in Germany. Histopathologic examination of colonic biopsies with special stains such as PAS is recommended in every dog with signs of chronic large bowel disease in order to avoid missing this rare infectious disease.

Cytologic, histologic, microbiologic, and electron microscopic characterization of a canine Prototheca wickerhamii infection.

An adult dog was presented for chronic cough and a recent development of ulcerated, erythematous nares with nasal discharge. Cytology of enlarged peripheral lymph nodes revealed many intracellular and extracellular organisms. These round or rarely oval organisms measured approximately 5-9 µm in diameter and frequently contained several globular structures, ranging from deeply basophilic to magenta. A thin, clear halo was present. Smaller 1-2 µm, magenta forms were also observed. Fungal culture yielded small, wet, raised, irregularly shaped, white to pale tan colonies. Microbiologic staining of cultured material revealed features suggestive of algae. Histopathology of the lymph nodes revealed marked granulomatous inflammation with intralesional algal organisms suggestive of Prototheca. Electron microscopic findings were also consistent with protothecosis. Polymerase chain reaction, followed by direct DNA sequencing, identified the organism as Prototheca wickerhamii. A brief literature review discussing protothecosis in veterinary medicine is included.

Emergence of Fungal-Like Organisms: Prototheca.

The genus Prototheca consists of achlorophyllic algae that are ubiquitous in the environment and animal intestines. However, this organism has forfeited its photosynthetic ability and switched to parasitism. In 1894, Krüger described two microorganisms isolated in Germany from mucous flux of Tilia and Ulmus spp., namely Prototheca moriformis and P. zopfii. Based on their yeast-like colony morphology, Krüger classified these organisms as fungi. The genus is now included within the class Trebouxiophyceae, order Chlorellales, and family Chlorellaceae. Historically, protothecosis and infections caused by green algae have been studied in the field of medical mycology. Prototheca spp. have been found to colonize human skin, fingernails, the respiratory tract, and digestive system. Although human infection by Prototheca is considered rare, an increase in infections has been noted among immunosuppressed patients, those on corticosteroid treatment, or both. Moreover, the first human outbreak of protothecal algaemia and sepsis was recently reported in a tertiary care chemotherapy oncology unit in 2018. Prototheca is also a causative pathogen of bovine disease. Prototheca zopfii and P. blaschkeae are associated with bovine mastitis, which causes a reduction in milk production and secretion of thin, watery milk containing white flakes. Economic losses are incurred either directly via reduced milk production and premature culling of affected animals or indirectly as a result of treatment and veterinary care expenses. Thus, knowledge of this fungal-like pathogen is essential in human and veterinary medicine. In this mini-review, I briefly introduce human and animal protothecoses.

Cumulative Effects of Thermal and Fisheries Stressors Reveal Sex-Specific Effects on Infection Development and Early Mortality of Adult Coho Salmon (Oncorhynchus kisutch).

Multiple stressors are commonly encountered by wild animals, but their cumulative effects are poorly understood, especially regarding infection development. We conducted a holding study with repeated gill and blood sampling to characterize the effects of cumulative stressors on infection development in adult coho salmon. Treatments included chronic thermal stress (15°C vs. 10°C) and acute gill net entanglement with an air exposure (simulating fisheries bycatch release). The potential loadings of 35 infectious agents and the expression of 17 host immune genes were quantified using high-throughput quantitative polymerase chain reaction, while host physiology was characterized with chemical analysis of blood. Temporal increases in infectious agent richness and loads were concurrent with decreased expression of immune genes in fish sampled in the river. In the laboratory, mortality was minimal in cool water regardless of fishery treatment (<15%). Elevated water temperature under laboratory conditions increased mortality of males and females (8% and 28% mortality, respectively, delayed by >1 wk) and enhanced mortality associated with handling and biopsy (∼40% both sexes). Experimental gillnetting at high temperature further enhanced female mortality (73%). Fish held at high temperature demonstrated heavier infectious agent loads, osmoregulatory impairment, suppressed female maturation, and upregulation of inflammatory and extracellular immune genes. At high temperature, heavy Parvicapsula minibicornis loads were associated with premature mortality. Females exhibited physiological impairment from both stressors after 1 wk, and infection burdens correlated poorly with immune gene regulation compared with males. Cumulative effects of multiple stressors on female mortality are likely a function of physiological impairment and enhanced infections at high temperature.

Effects of aloe-emodin on innate immunity, antioxidant and immune cytokines mechanisms in the head kidney leucocytes of Labeo rohita against Aphanomyces invadans.

The effect of aloe-emodin incorporated diets on innate immune response, disease resistance, pro and/or anti-inflammatory cytokine gene transcription in Labeo rohita against Aphanomyces invadans is reported for the first time. In healthy and infected groups fed with 5 mg aloe-emodin enriched diet the white blood cell (WBC) count increased significantly (p > 0.05) after 6th week. In both groups fed with any enriched diet the biochemical parameters such as albumin, globulin, and albumin/globulin ratio did not vary significantly; however with 5 mg aloe-emodin diet the albumin and globulin levels increased significantly (p > 0.05) after 6th week. The serum phagocytic activity (PA), respiratory burst activity (RBA), serum complement C3 (CC3), and lysozyme activity (LA) did not increase with any diet between weeks 2 and 4, whereas with 5 mg aloe-emodin diet increased significantly in both groups after 6th week. The pro-inflammatory cytokines such as IL-1ß, IL-8, TNF-α, and iNOS significantly modulated the expression in both groups on being fed with 5 mg aloe-emodin incorporation diet on 8th week. Healthy fish fed with any aloe-emodin diet did not suffer mortality. However, the infected fish fed with 1, 5, and 10 mg kg-1 aloe-emodin diets registered 5%, 10%, and 15% mortality. The present study indicates that healthy and infected L. rohita exhibited enhanced innate immune response, disease resistance, pro and/or anti-inflamatory cytokine gene transcription levels against A. invadans.

The use of absorbable staples for skin closure after tibial plateau leveling osteotomy.

OBJECTIVE: To compare the use of stainless steel staples with absorbable staples for closure of skin incisions in dogs undergoing tibial plateau leveling osteotomy (TPLO). STUDY DESIGN: Prospective study. SAMPLE POPULATION: Client-owned dogs (n = 80). METHODS: With client consent, dogs were randomly assigned a staple type (stainless steel or absorbable) immediately prior to closure of a TPLO skin incision. Incisions were compared for length, staple type and number, and an inflammation-infection score 2 weeks after surgery. RESULTS: Overall, 18.8% of incisions were diagnosed with inflammation or infection. No difference was found between inflammation-infection scores, incision length, number of staples used, or general anesthetic time between the 2 staple groups. However, wound closure was faster with stainless steel staples (22.50 seconds; range, 11-180) by approximately 30 seconds compared with absorbable staples (56.50 seconds; range, 18-190; P < .001). Time taken to close the incision correlated negatively with the number of occasions that absorbable staples were used (P = .01). CONCLUSION: Absorbable skin staples were successfully used to close skin incisions after TPLO and were not associated with an increased level of inflammation or infection in our clinical setting. CLINICAL SIGNIFICANCE: Absorbable staples may be considered to close surgical wounds when subsequent suture removal would be impractical, without specific concerns over inflammation or infection of the wound.

Short communication: Evaluation of the efficacy of novel disbudding methods for dairy calves.

The objectives of this study were to (1) evaluate whether cryoablation or the administration of clove oil was as efficacious as cautery disbudding at preventing horn growth, and (2) evaluate whether the efficacy of cautery disbudding is affected by removing or leaving the horn bud tissue intact after disbudding of dairy calves. At approximately 4 d of age (4.0 ± 0.88 d of age, mean ± SD), 265 dairy heifer calves from 3 dairy farms (farm 1: n = 129 calves; farm 2: n = 109 calves; farm 3: n = 27 calves) were disbudded. Each calf had 1 of 4 treatments randomly assigned to each horn bud: (1) clove oil (0.5 mL) administered subcutaneously under the horn bud (CLOV, n = 135 buds); (2) a liquid nitrogen-filled probe applied to the horn bud area for 30 s (CRYO, n = 134 buds); (3) cautery disbudding using an electric hot-iron and the horn bud removed (BUDOFF, n = 130); or (4) cautery disbudding and the horn bud tissue left intact (BUDON, n = 131). Calves were assessed for signs of infection at the disbudding site frequently within the first month after disbudding. At approximately 6 mo of age (6 ± 2.2 mo, mean ± SD) calves were assessed for scur or horn growth. The disbudding procedures were considered successful if no scur or horn development was observed. Within the first month, 12% of disbudding wounds showed some indication of infection, such as pus, exudate, or swelling; of the infected buds, 52% were associated with the BUDON treatment, 27% with CLOV, 25% with BUDOFF, and 2% with CRYO treatments. At 6 mo of age, BUDOFF was the most effective method of preventing horn growth and CRYO was the least efficacious [mean percentage of success: BUDOFF: 100% (95% CI: 97.7-100.0); CRYO: 1% (95% CI: 0.2-5.3)]. Injecting clove oil under the horn bud was 87% (95% CI: 80.6-92.5) successful. Not removing the horn bud tissue after cautery disbudding reduced the efficacy of this method by 9% [91% success (95% CI: 83.8-95.7)]; moreover, this method was associated with more infection at the site of disbudding. It appears as though the clove oil treatment could be used as an alternative to cautery disbudding of dairy calves; however, further research is needed to evaluate the tissue damage and associated pain caused by clove oil and to refine this technique (i.e., administration methods to improve efficacy) before it could be considered an alternative to cautery.

Estimating the value of infectious or noninfectious foot disorder prevention strategies within dairy farms, as influenced by foot disorder incidence rates and prevention effectiveness.

A farm-level stochastic simulation model was adapted to estimate the value of implementing foot disorder prevention on a dairy farm by calculating the return on investment associated with implementation of foot disorder prevention. Two potential strategies for foot disorder prevention were tested: strategy 1 was prevention focused on reducing infectious foot disorders (i.e., digital dermatitis) in the model, and strategy 2 was prevention focused on reducing noninfectious foot disorders (i.e., sole ulcer and white line disease) in the model. For each strategy, we evaluated the effect of foot disorder incidence on the value of prevention by setting the incidence of foot disorders at 3 levels. For strategy 1, the level of digital dermatitis incidence without prevention in all parities was 20, 40, or 60%, and the incidence level of the noninfectious foot disorders in the model were held constant. For strategy 2, levels of sole ulcer and white line disease incidence without prevention in parity ≥3 cows were 5, 15, or 25%, and the incidence level of the infectious foot disorders included in the model were held constant; the incidence levels of noninfectious foot disorders in younger cows were adjusted to be lower. Overall, 6 scenarios were run, 1 for each prevention strategy × foot disorder incidence rate combination. To evaluate how the effectiveness of each prevention strategy would influence the investment value, the effectiveness of prevention could vary from a prevention risk ratio (RR) of 0.0 (100% reduction in disorder incidence) to 1.0 (0% reduction in disorder incidence). When implementing strategy 1, the return on prevention investment per cow-year (mean ± standard deviation) when prevention effectiveness was low (prevention RR = 0.91 to 1.0) and the digital dermatitis incidence rate was originally 20, 40, or 60% was $0.6 ± 0.4, $1.2 ± 0.9, and $1.8 ± 1.3, respectively. In comparison, the return on prevention investment per cow-year when prevention effectiveness was high (prevention RR = 0.00 to 0.09) and the digital dermatitis incidence rate was originally 20, 40, or 60% was $12.2 ± 3.0, $24.4 ± 6.0, and $36.5 ± 9.0, respectively. When implementing strategy 2, the return on prevention investment per cow-year when prevention effectiveness was low and noninfectious foot disorder incidence rates were originally 5, 15, or 25% in parity ≥3 cows was $0.6 ± 0.4, $1.9 ± 1.1, and $3.2 ± 1.9, respectively. In comparison, the return on prevention investment per cow-year when prevention effectiveness was high and noninfectious foot disorder incidence rates were originally 5, 15, or 25% in parity ≥3 cows was $12.4 ± 1.5, $37.3 ± 4.6, and $62.2 ± 7.6, respectively. The return on investment for foot disorder prevention would depend on the cost of the prevention strategy and the other benefits associated with the selected prevention strategy. This model could be used as a decision support tool to help identify the amount that could be paid to implement a selected prevention strategy.

Perinatal immuno/inflammatory responses in the presence or absence of bovine fetal infection.

BACKGROUND: It is known that the bovine fetus can mount an immune and inflammatory reaction to infection, but it is not known whether there is a contemporaneous maternal response. Nor is it known whether the response of calves which die perinatally, with or without infection, differs from that of live perinates. Hence, the objective of this study was to determine if acute phase reactant and immunoglobulin concentrations differed between calves (and their dams) in three groups: live calves (CC; n = 21) and dead calves with (PM INF+; n = 22) or without (PM INF-; n = 89) in utero infection. In calf plasma, serum amyloid A, haptoglobin, immunoglobulins M, G1 and G2 and interleukin-6 were measured. In dam serum, SAA and Hp was measured and in amniotic and abomasal fluid, IL-6 was measured. RESULTS: Live calves had higher plasma concentrations of SAA and IL-6 than dead calves with (PM INF+) or without (PM INF-) in utero infection. Calves in the PM INF-, but not PM INF+ group, had higher Hp concentrations than calves in the CC group. Calves in the PM INF+ group had higher IgG1 concentrations than calves in the PM INF- and CC groups. Except for higher IgG1 and IgG2 concentrations, biomarker values did not differ significantly between dead calves with or without in utero infection. Live calves had higher IL-6 concentrations in abomasal fluid compared to PM INF- calves. There were no significant differences in blood biomarker concentrations between dams of the three groups of calves. Amniotic fluid IL-6 concentrations were higher from the dams of control calves than the dams of uninfected calves. CONCLUSIONS: Differences in biomarkers (higher Hp and IgG1; lower SAA and IL-6) between perinatal mortalities and live perinates probably reflect differences between these two groups in age at sampling (SAA and IL-6) and in utero infection (IgG1). Out of the six analytes measured in calves, only IgG1 and IgG2 were biomarkers of (chronic) in utero infection.

A Case of Prototheca zopfii Genotype 1 Infection in a Dog (Canis lupus familiaris).

Protothecosis is a rare disease caused by environmental algae of the genus Prototheca. These are saprophytic, non-photosynthetic, aerobic, colorless algae that belong to the Chlorellaceae family. Seven different species have been described. Prototheca zopfii genotype 2 and P. wickerhamii are most commonly involved in pathogenic infections in humans and animals. The objective of this work is to describe, for the first time, a case of protothecosis caused by P. zopfii genotype 1 in a dog. The dog, a 4-year-old mix bred male, was presented to a veterinary clinic in Montevideo, Uruguay, with multiple skin nodules, one of which was excised by surgical biopsy. The sample was examined histologically and processed by PCR, DNA sequencing, and restriction fragments length polymorphisms for the detection and genotyping of P. zopfii. In addition, transmission electron microscopy and scanning electron microscopy were performed. Histology showed severe ulcerative granulomatous dermatitis and panniculitis with myriads of pleomorphic algae. Algal cells were 4-17 µm in size, with an amphophilic, 2-4-µm-thick wall frequently surrounded by a clear halo, contained flocculant material and a deeply basophilic nucleus, and internal septae with daughter cells (endospores) consistent with endosporulation. Ultrastructurally, algal cells/endospores at different stages of development were found within parasitophorous vacuoles in macrophages. Prototheca zopfii genotype 1 was identified by molecular testing, confirming the etiologic diagnosis of protothecosis.

Effect of immunization of rohu Labeo rohita with inactivated germinated zoospores in providing protection against Aphanomyces invadans.

Infection with Aphanomyces invadans is one of the most destructive diseases of freshwater fishes. Indian major carps, the dominant cultured species in the Indian sub-continent are highly susceptible to this disease. Till date, there is no effective treatment for control of this disease and immunization can be one of the strategies to reduce disease-related losses. In the present study, inactivated germinated zoospores of A. invadans were evaluated as antigen in conjunction with and without adjuvant Montanide™ ISA 763 A VG, for assessing their efficacy in rendering protection against A. invadans infection. For the experiment, rohu Labeo rohita, (n = 160, 74 ±â€¯12 g) were divided into 4 groups (C, A, G and GA) with 40 fish in each group. The fish in groups i.e., C, A, G and GA were injected intraperitoneally with PBS, adjuvant emulsified with PBS, inactivated germinated zoospores, and inactivated germinated zoospores emulsified with adjuvant, respectively. After 21 days of immunization, the fish were given a booster dose as above. After 7 days of the booster dose, the fish were challenged with zoospores of A. invadans to determine the relative percent survival (RPS). The results revealed that all the fish in C, A and G group succumbed to infection (0% RPS), although there was delayed mortality in fish from A and G groups in comparison to the C group. However, the fish in GA group showed significantly higher (P < 0.05) protection (66.7% RPS). In addition, following booster immunization (before challenge), the antibody level in the GA group was significantly higher (P < 0.05) than the control group. In western blotting, sera from G and GA groups showed reactivity with peptides of about 54 KDa. Following challenge (on 14th day), the antibody level as well as total antiprotease activity in fish of all the groups was significantly decreased in comparison to pre-challenge, except fish in GA group not exhibiting any gross lesions. In addition, sera of surviving fish of GA group showed significant inhibition of germination of zoospores and germlings growth in comparison to other groups (P < 0.05). Further, histopathological examination of the muscle tissue revealed that, in fish of GA group without any gross lesions, there were well developed granulomas and extensive mononuclear cell infiltration restricted to the site of injection, whereas in other groups, there was extensive myonecrosis with proliferating hyphae. These preliminary findings indicate that inactivated germinated zoospores of A. invadans in combination with adjuvant could stimulate good immune response and confer remarkable protection in rohu.

Problems that can arise in cattle before and after turnout.

This article has been prepared by Arthur Otter of the APHA Cattle Expert Group.

Prevalence and disease associations in feline thrombocytopenia: a retrospective study of 194 cases.

OBJECTIVES: To assess the prevalence of thrombocytopenia in a referral population of cats in the UK, to identify disease processes associated with thrombocytopenia and to assess the proportion of thrombocytopenic cats that tested positive for feline leukaemia virus or feline immunodeficiency virus. MATERIALS AND METHODS: Retrospective analysis of medical records at a UK referral hospital. Cats were grouped by mechanism of thrombocytopenia and disease process (where known). RESULTS: Prevalence of thrombocytopenia was 5·9%. The most common disease processes associated with thrombocytopenia were haematological or infectious disease and neoplasia; 11% of thrombocytopenic cats tested were positive for feline leukaemia virus, which is lower than reported previously. Cats presenting with unexplained haemorrhage had significantly lower platelet counts than other thrombocytopenic cats. Primary immune-mediated thrombocytopenia was less commonly diagnosed than in dogs and associated with the most severe platelet depletion in this study. CLINICAL SIGNIFICANCE: Thrombocytopenia in cats may be more prevalent than previously reported and severe thrombocytopenia may be associated with spontaneous haemorrhage. Severe thrombocytopenia in cats appears less commonly immune-mediated than in dogs. Thrombocytopenia did not appear to be associated with retroviral infections.

In vitro investigations on extracellular proteins secreted by Aphanomyces invadans, the causative agent of epizootic ulcerative syndrome.

BACKGROUND: Proteases produced by many microorganisms, including oomycetes, are crucial for their growth and development. They may also play a critical role in disease manifestation. Epizootic ulcerative syndrome is one of the most destructive fish diseases known. It is caused by the oomycete Aphanomyces invadans and leads to mass mortalities of cultured and wild fish in many countries. The areas of concern are Australia, China, Japan, South and Southeast Asian countries and the USA. Extracellular proteases produced by this oomycete are believed to trigger EUS pathogenesis in fish. To address this activity, we collected the extracellular products (ECP) of A. invadans and identified the secreted proteins using SDS-PAGE and mass spectrometery. A. invadans was cultivated in liquid Glucose-Peptone-Yeats media. The culture media was ultra-filtered through 10 kDa filters and analysed using SDS-PAGE. Three prominent protein bands from the SDS gel were excised and identified by mass spectrometery. Furthermore, we assessed their proteolytic effect on casein and immunoglobulin M (IgM) of rainbow trout (Oncorhynchus mykiss) and giant gourami (Osphronemus goramy). Antiprotease activity of the fish serum was also investigated. RESULTS: BLASTp analysis revealed that the prominent secreted proteins were proteases, mainly of the serine and cysteine types. Proteins containing fascin-like domain and bromodomain were also identified. We could demonstrate that the secreted proteases showed proteolytic activity against the casein and the IgM of both fish species. The anti-protease activity experiment showed that the percent inhibition of the common carp serum was 94.2% while that of rainbow trout and giant gourami serum was 7.7 and 12.9%, respectively. CONCLUSIONS: The identified proteases, especially serine proteases, could be the potential virulence factors in A. invadans and, hence, are candidates for further functional and host-pathogen interaction studies. The role of identified structural proteins in A. invadans also needs to be investigated further.

Spontaneous and Experimentally Induced Pathologies in the Naked Mole Rat (Heterocephalus glaber).

The naked mole rat (Heterocephalus glaber, Rüppell, 1842) is a unique eusocial rodent with unusually long lifespan. Therefore, the study of spontaneous and experimentally induced pathologies in these animals is one of the most important tasks of gerontology. Various infections, noninfectious pathologies (including age-dependent changes), and tumors have been described in the naked mole rat. The most frequent pathologies are traumas (bite wounds), purulent and septic complications of traumatic injuries, renal tubular calcinosis, chronic progressive nephropathy, hepatic hemosiderosis, testicular interstitial cell hyperplasia, calcinosis cutis, cardiomyopathy, and dysbiosis-related infectious lesions of the digestive system. However, the summarized data on pathology (including tumor incidence) and on the causes of mortality are insufficient. There are only few publications about the results of experiments where pathologies were induced in the naked mole rat. All these problems could be subjects for promising future studies without which adequate studies on mechanisms providing the long lifespan of the naked mole rat are impossible, as well as the elucidation of causes of tumor resistance of this species.