Vet. Sci., Volume 6, Issue 4 (December 2019) – 29 articles

Superficial corneal ulcers that fail to heal within a normal time period and are refractory to conventional therapy in dogs are common in veterinary practice. Different etiologies can lead to this result, including spontaneous chronic corneal epithelial defects (SCCEDs) and ulcerative keratitis associated with bullous keratopathy. Thus, there is an urgent need to find new therapeutic approaches such as matrix therapy replacement. To determine the efficacy of a new ophthalmic treatment (Clerapliq^®) for SCCEDs and ulcerative keratitis associated with bullous keratopathy, a total of 11 dogs referred to the clinic because of nonhealing erosive ulcers after a classic primary treatment were enrolled to get this new treatment. Dogs underwent ophthalmic exams and 7 dogs (10 eyes) were diagnosed with superficial ulceration and 4 dogs (5 eyes) with bullous keratopathy due to endothelial dystrophy/degeneration. They received eye drops of Clerapliq^® every 3 days until recovery. The results showed that the corneas with recurrences of the ulcers were resolved predominantly by using Clerapliq^® every 3 days in 83.3% of the cases during a period of treatment ranging between 6 to 35 days. Therefore, this new approach using matrix therapy regenerating technology in treating superficial ulcers and bullous keratopathy in dogs can be successfully considered as an adjunctive therapy. Full article

The objective of this study was to evaluate if administration of clove oil prevents scur/horn growth in dairy cattle long term. At approximately 4 days of age, calves had one of four treatments assigned to each horn bud: (1) clove oil administered subcutaneously under the horn bud (CLOV, n = 132); (2) cautery disbudded and the horn bud removed (BUDOFF, n = 126); (3) cautery disbudded and the horn bud tissue left intact (BUDON, n = 129); (4) a liquid nitrogen filled probe applied to the horn bud area (CRYO, n = 131). At approximately 16 months of age, all cattle were checked for scur or horn development. A sub-set of scurs/horns from the CLOV cattle were removed to evaluate tissue and structural development. In total, 5% of CLOV buds developed into horns and 63% into scurs; 10% of the scurs looked like normally developed horns but they were not attached to the skull. Cautery disbudding prevented scur and horn development in cattle when the horn bud tissue was removed, but some scur growth was observed in the BUDON treatment. CRYO was 100% ineffective at preventing scur/horn growth. Injecting clove oil under the horn bud appeared to delay horn development, but not prevent it, when administered to 4 day old dairy calves. Full article

Incidence of canine mammary carcinoma is two times higher than the rate of human breast cancer. Mammary tumors are the most common type of cancer in intact female dogs and account for about half of all neoplasms in these dogs. Well-established models of breast cancer have shown that neoplastic cells often have a loss of intercellular communication, particularly gap junction proteins. Thus, the objective of this study is to explore the aspect of gap junction intercellular communication in canine mammary carcinoma, non-cancerous (CMEC) and cancerous (CMT12, CMT27, and CF41.Mg) cells, and patient-derived tumors. Both non-cancerous and cancerous mammary cells express connexins 26 and 43 using immunofluorescence; however, the level of expression is significantly different in quantitative analysis using western blot in which connexin 43 in both CMT12 and CMT27 is significantly decreased compared to CMEC. Furthermore, a decrease of gap junction capacity in CMT12 and CMT27 was observed compared to CMEC. Immunostaining of CMT27-xenograft tumors revealed positive Cx26 and negative Cx43 expression. Similarly, immunostaining of spontaneous canine mammary tumors revealed that Cx26 is present in all tumors while Cx43 is present in 25% of tumors. Overall, the study provides for the first time that a differential pattern of connexin expression exists between non-cancerous and cancerous mammary cells in dogs. This study will pave the path for further in vitro work of connexins in comparative canine models and possibly allow for novel therapeutics to be developed. Full article

Background—The study aims to assess the efficacy of a permethrin 54.5%-fipronil 6.1%-based spot-on solution in dogs naturally infested with Neotrombicula in an open-label controlled study. Methods—Ten naturally infested dogs received one drop per affected site on day (D) 0, and on D14, the rest of the pipette was applied on the skin between the shoulders. Five dogs served as sentinels. Parasite score (0–3), skin lesions (0–4), and investigator pruritus scale (0–4) were assessed on D0, D1, D14, and D28. Results—No treated dogs developed adverse reactions. Parasite score of sentinel dogs was maintained between 1.8 (D0, D1, and D28) and 2.2 (D14). In treated dogs, D0 parasite score was 2.4. It was significantly reduced from D1 (0.5; p < 0.002) to D28 (0.1; p < 0.002). The lesion score was 2.9 on D0 and D1; it was significantly reduced on D14 (0.6; p < 0.002) and D28 (0.1; p < 0.002). Similarly, investigator pruritus scale (D0, 2.2) scores significantly decreased on D14 (0.4; p < 0.004) and D28 (0.2; p < 0.002). Conclusions—The combination permethrin-fipronil appears to be well-tolerated, rapidly and durably effective in the control of localized canine harvest mite infestation. Full article

Biological properties of the African swine fever (ASF) virus isolates originating from various regions of the Russian Federation (2013–2018) were studied in a series of experimental infections. Comparative analysis allowed us to establish the differences in the key characteristics of the infection, such us the duration of the incubation periods, disease, and the onset of death. The incubation period averaged 4.1 days, varying from 1 to 13 days. An average duration of the disease was 6.3 days and varied from 0 to 18 days. Overall case fatality was 94.5%, and antibodies were detected only in 19.3% of the animals. The biological properties of isolates Odintsovo 02/14 and Lipetsk 12/16 were significantly different from others. For this two, the presence of antibodies to the virus was detected in 71.4% and 75% of animals respectively and mortality levels were of 87.5% and 50%. Full article

The single intradermal comparative cervical tuberculin (SICCT) test is the primary test for ante-mortem diagnosis of bovine tuberculosis (TB) in England and Wales. When an animal is first classified as an inconclusive reactor (IR) using this test, it is not subject to compulsory slaughter, but it must be isolated from the rest of the herd. To understand the risk posed by these animals, a case-control study was conducted to measure the association between IR status of animals and the odds of them becoming a reactor to the SICCT at a subsequent test. The study included all animals from herds in which only IR animals were found at the first whole herd test in 2012 and used data from subsequent tests up until the end of 2016. Separate mixed-effects logistic regression models were developed to examine the relationship between IR status and subsequent reactor status for each risk area of England and for Wales, adjusting for other explanatory variables. The odds of an animal becoming a subsequent reactor during the study period were greater for IR animals than for negative animals in the high-risk area (odds ratio (OR): 6.85 (5.98–7.86)) and edge area (OR: 8.79 (5.92–13.04)) of England and in Wales (OR: 6.87 (5.75–8.22)). In the low-risk area of England, the odds were 23 times greater, although the confidence interval around this estimate was larger due to the smaller sample size (11–48, p < 0.001). These findings support the need to explore differential controls for IR animals to reduce the spread of TB, and they highlight the importance of area-specific policies. Full article

Mycobacterium avium subspecies paratuberculosis (MAP) is endemic in the Dutch dairy goat population causing economic loss, and negatively influencing welfare. Moreover, there are concerns about a potential zoonotic risk. Therefore the industry’s objectives are to decrease MAP prevalence, limit economic losses as well as reduce the concentration of MAP in (bulk) milk. To diminish within-farm spread of infection, vaccination, age dependent group housing with separation of newborns from adults, as well as rearing on artificial or treated colostrum and milk replacers are implemented. However, the importance of MAP contaminated colostrum and milk as a route of infection in dairy goat herds is unknown. Therefore the aim of this study was to detect the presence of MAP DNA in colostrum and milk from dairy goats in infected herds. A convenience sample of 120 colostrum samples and 202 milk samples from MAP infected dairy goat herds were tested by IS900 real-time Polymerase Chain Reaction (PCR) for MAP DNA. Furthermore, 22 colostrum samples and 27 post mortem milk samples of goats with clinical signs consistent with paratuberculosis from known infected herds were tested. The majority of samples were from goats vaccinated against MAP. Positive or doubtful PCR results were obtained in none of the 120 and two of the 22 colostrum samples, and in eight of the 202 and four of the 27 milk samples Negative PCR results were obtained in the remaining 140 (99%) colostrum samples and 217 (95%) milk samples. Full article

Background: Peste des petits ruminants virus (PPRV) is the causative agent of PPR, which can cause an acute, highly contagious and fatal disease of sheep and goats, resulting in significant economic losses for commercial animal husbandry due to its high mortality and morbidity. As professional antigen-presenting cells, dendritic cells (DCs) play a unique role in innate immunity. This study aimed to gain a deeper understanding of the transcriptional response of bone marrow-derived dendritic cells (BMDCs) stimulated with PPRV. Results: Transcriptional profiling was performed using RNA sequencing. Herein, we reported that compared to untreatedBMDCs, 4492 differentially expressed genes (DEGs) were identified following PPRV stimulation, out of these DEGs 2311 were upregulated and 2181 were downregulated, respectively. A total of three gene ontology (GO) term clusters of biological process, cell component and molecular function were significantly enriched in 963 GO terms in the PPRV-stimulated BMDCs. These GO clusters were related to inflammatory response, cell division and vacuole, anchoring junction, positive regulation of cellular component and nucleoside binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of DEGs were enriched in a chemokine signaling pathway, protein processing in endoplasmic reticulum, cell cycle and mTOR signaling pathway. Additionally, identified DEGs of BMDCs were further validated by qRT-PCR and the results were in accordance with the change of the genes. This study suggested the effects of PPRV stimulation on the maturation and function of BMDCs. Conclusion: We found that the dramatic BMDCs transcriptome changes triggered were predominantly related to an inflammatory response and chemokine signaling pathway. Full article

African swine fever (ASF) is one of the most threatening diseases for the pig farming sector worldwide. As an effective vaccine is lacking, strict application of control measures is the only way to fight the disease in both industrial farms and backyard holdings. With generally low biosecurity standards, the latter are at particular risk for disease introduction and offer challenging conditions for disease control. In the following case report, we describe the overall course of an ASF outbreak in a Bulgarian backyard farm and the implemented control measures. Farm facilities and available data have been investigated to estimate the possible source, spread and time point of virus introduction. Contact with contaminated fomites entering the stable via human activities was regarded to be the most likely introduction route. The slow disease spread within the farm contributes to the hypothesis of a moderate contagiosity. As no further ASF outbreaks have been detected in domestic pig farms in the region, it could be demonstrated that successful disease control in small-scale farms can be reached. Thus, the report contributes to a better understanding of ASF in the backyard sector. Full article

Exposure of neonates to Mycobacterium avium subsp. paratuberculosis (MAP) via infected dams is the primary mode of transmission of Johne’s disease. Little is known about the impacts of feeding colostrum and supplemental vitamins on the gut microbiome in calves exposed to MAP. In the present study, calves were assigned at birth to one of six treatment groups: (1) Colostrum deprived (CD), no vitamins; (2) colostrum replacer (CR), no vitamins; (3) CR, vitamin A; (4) CR, vitamin D₃; (5) CR, vitamin E; (6) CR, vitamins A, D₃, E, with five calves per treatment in a 14-day study. All calves were orally inoculated with MAP on days 1 and 3 of the study. Differences due to vitamin supplementation were not significant but treatment groups CR-A, CR-E, and CR-ADE had higher numbers of MAP-positive tissues overall. Shannon diversity indices demonstrated regional differences in microbial communities, primarily Proteobacteria, Bacteroidetes, and Firmicutes, between the ileum, cecum, and spiral colon of all calves. CD calves exhibited increased richness compared with CR calves in the cecum and spiral colon and harbored increased Proteobacteria and decreased Bacteroidetes in the mucosa compared with the lumen for all three tissues. Overall, supplementation with vitamins did not appear to influence gut microbiome or impact MAP infection. Feeding of colostrum influenced gut microbiome and resulted in fewer incidences of dysbiosis. Full article

Marek’s disease virus (MDV) and the reticuloendotheliosis virus (REV) are two of the primary oncogenic viruses that significantly affect chickens. In Brazil, there have been no previous published reports on the presence of field REV alone or in coinfection. This retrospective study analyzes samples from a case of lymphoproliferative lesions from a backyard chicken flock. MDV and REV were detected by PCR and classified as MDV1 and REV3, respectively, through sequencing and phylogenetic analysis based on the glycoprotein B (gB) genes for MDV and the polymerase (pol) and envelope (env) genes for REV. Real-time PCR reactions were performed for MDV to rule out the presence of the Rispens vaccine strain. This is the first report of the presence of REV in coinfection with a MDV clinical case in Brazil and the first molecular characterization of REV in South America. This study highlights the importance of molecular diagnosis for REV and MDV in poultry. In addition, this study highlights the distribution of these two viruses worldwide and the latent risk of them solely or in coinfection to this part of the world. Full article

Accurate diagnosis of mycobacterial infections, such as bovine tuberculosis and paratuberculosis, remains challenging. Available direct diagnostic tests aimed at detecting the pathogen are highly specific but lack sensitivity, depending on the stage of infection and the prevalence of infection in a population. The sensitivity of indirect diagnostic assays that measure the host immune response to infection is similarly affected by disease characteristics. The choice of antigen used to detect a host response to infection has a critical impact on test sensitivity and specificity. Many indirect tests rely on crude antigen preparations and cell-free extracts, of which the production is poorly standardized. Moreover, these preparations contain ample uncharacterized cross-reactive compounds. To enhance serological test specificity, existing assays depend on the pre-treatment of samples and a relatively high cut-off value, that in turn influences test sensitivity. Research therefore focuses on the identification of more specific, defined antigens to improve diagnostics. In the current study, we extracted phosphatidylinositol mannosides (PIMs) and investigated their potential use in antibody-based tests. Our results demonstrate that specific IgG class antibodies are generated against PIMs in cows, but this is unrelated to tuberculosis or paratuberculosis infection status, making these antigens unsuitable for diagnostic applications. In addition, we demonstrate that PIMs are widely present in crude antigen preparations and in serum pre-absorption buffer. Our results indicate that PIMs are cross-reactive compounds with immunodominant B cell epitopes that could impair serological test specificity. Full article

Osteochondral defects are a common problem in both human medicine and veterinary practice although with important limits concerning the cartilaginous tissue regeneration. Interest in the subchondral bone has grown, as it is now considered a key element in the osteochondral defect healing. The aim of this work was to generate and to evaluate the architecture of three cell-free scaffolds made of collagen, magnesium/hydroxyapatite and collagen hydroxyapatite/wollastonite to be implanted in a sheep animal model. Scaffolds were designed in a bilayer configuration and a novel “Honey” configuration, where columns of hydroxyapatite were inserted within the collagen matrix. The use of different types of scaffolds allowed us to identify the best scaffold in terms of integration and tissue regeneration. The animals included were divided into four groups: three were treated using different types of scaffold while one was left untreated and represented the control group. Evaluations were made at 3 months through CT analysis. The novel “Honey” configuration of the scaffold with hydroxyapatite seems to allow for a better reparative process, although we are still far from obtaining a complete restoration of the defect at this time point of follow-up. Full article

European badgers are a wildlife reservoir of bovine tuberculosis in parts of Great Britain. Accurate diagnosis of tuberculosis in badgers is important for the development of strategies for the control of the disease. Sensitive serological tests for badger TB are needed for reasons such as cost and simplicity. Assay of mucosal IgA could be useful for diagnosing respiratory pathogens such as Mycobacterium bovis and for monitoring the response to mucosal vaccination. To develop an IgA assay, we purified secretory IgA from badger bile, identifying secretory component (SC), heavy chain (HC) and light chain (LC), at 66, 46 and 27 Kda, respectively, on the basis of size comparison with other species. Monoclonal antibodies (mAbs) were generated to purified IgA. We selected two for ELISA development. The detection limit of the IgA-specific mAbs was found to be approximately 20 ng/mL when titrated against purified badger bile. One monoclonal antibody specific for badger IgA was used to detect IgA in serum and tracheal aspirate with specificity to an immunodominant antigen of M. bovis. An M. bovis infection dose-dependent IgA response was observed in experimentally infected badgers. IgA was also detected by immunohistochemistry in the lungs of bTB-infected badgers. With further characterisation, these represent new reagents for the study of the IgA response in badgers. Full article

Antigens extracted using ethanol (EtOH) and incorporated in the EtOH vortex ELISA (EVELISA) test have previously shown high specificity and sensitivity for detecting Mycobacterium avium subspecies paratuberculosis (Map) and M. bovis infections in cattle. The objective of this study is to define the components present in the EtOH extract. We show that this extract is composed of lipid, carbohydrate, and proteins on the surface of the bacilli, and that EtOH removes the outer layer structure of Map which comprise these elements. To identify proteins, polyclonal antibodies to the EtOH prep were produced and used to screen a Map genomic expression library. Seven overlapping clones were identified with a single open reading frame, MAP_0585, common to all. MAP_0585, which encodes a hypothetical protein, was recombinantly produced and used to demonstrate strong reactivity in sera from hyperimmunized rabbits, but this protein is not strongly immunogenic in cattle with Johne’s disease. A panel of monoclonal antibodies was used to determine the presence of additional proteins in the EtOH extract. These antibodies demonstrated that a well-known antigen, termed MPB83, is present in M. bovis EtOH extracts and a fatty acid desaturase (MAP_2698c) is present in Map EtOH extracts, while lipoarabinomannan was common to both. The lipid and carbohydrate components of the extract were analyzed using thin layer chromatography and lectin binding, respectively. Lectin biding and protease treatment of the EtOH extract suggest the antigenic component is carbohydrate and not protein. These results give further insight into this important antigen prep for detecting mycobacterial diseases of cattle. Full article

In this review, information was summarized on endoparasites found in six non-domesticated neotropical animals. These mammals have the potential to be domesticated. The animals included three rodents, agouti (Dasyprocta leporina), lappe (Agouti paca), and capybara (Hydrochoerus hydrochaeris); a marsupial, manicou (Didelphis marsupialis insularis); and an artiodactyl, the collared peccary (Tayassu tajacu/Peccari tajacu) and a ruminant (the red brocket deer, Mazama americana). While there are many descriptions of the parasites present, the majority of publications failed to note the effect of them on the animals. Most information is available on endoparasites of capybara, while the endoparasites of the red brocket deer were the least reported. The manicou was reported to have had the most number of endoparasites, 44 species of parasites were reported, while there were only 24 endoparasites reported in the lappe. The most common parasites found in these neotropical animals were Paraspidodera uncinata, Strongyloides spp., Eimeria spp., Moniezia benedeni, Trichuris spp., Physocephalus spp., and Giardia spp. A large majority of the studies concluded that these animals were reservoirs for parasites that could affect domesticated livestock. Endoparasites of zoonotic significance were Echinoccocus spp., Trichuris spp., Giardia spp., and Cryptosporidium spp. Full article

Several novel porcine pestiviruses that are linked to disease outbreaks in commercial pig farms were discovered during recent years. Bungowannah pestivirus (BuPV; new species Pestivirus F) causes sudden death in young pigs, but has only ever been isolated in the Australian region Bungowannah. Atypical porcine pestivirus (APPV; new species Pestivirus K) on the other hand has been found in multiple countries worldwide and is potentially linked to congenital tremor, a disease that causes considerable production problems in pig farms. To assess the seroprevalences of both viruses in German commercial farms during the years 2009/10 and 2018, two approaches were selected. Antibodies against Pestivirus F were detected by a traditional in-house indirect immunofluorescence test against the culture-grown virus isolate, while for the detection of Pestivirus K-specific antibodies, a newly developed test system utilizing a chimeric construct of bovine viral diarrhea virus 1 (BVDV-1; species Pestivirus A) containing the E1 and E2 encoding sequences of APPV was established. A total of 1115 samples originating from 122 farms located in seven German federal states were investigated. Antibodies against Bungowannah virus could not be detected, confirming the absence of this virus in other regions than the initially affected Australian pig farm complex. In contrast, antibodies against APPV were highly prevalent throughout Germany at both investigated time points. The seroprevalence at the state level fluctuated to some degree, but the overall percentage remained stable, as is to be expected for an endemic pestivirus lacking any form of control measures. Full article

The presence of a wildlife reservoir for Mycobacterium bovis complicates the eradication of bovine tuberculosis (BTB) from domestic cattle populations. For the BTB eradication program in Fiji, there is concern about the small Indian mongoose (Herpestes auropunctatus), which is overabundant and in direct contact with cattle. Consequently, a survey of mongooses trapped on three BTB affected dairy farms led to necropsy of 85 mongooses during January–February 2017. Thirty (35%) mongooses had gross pathological changes including possible granulomas detected at necropsy, and tissues from these animals were taken for histopathological examination. Granulomatous lesions were present in 53% of animals examined histopathologically but acid-fast bacilli were not observed and the majority of lesions in lung and kidney were associated with the nematodes Pulmostrongylus herpestis and Capillaria sp., respectively. Nevertheless, assuming test sensitivity of 35% for the current study, from this sample of 85 mongooses it can be concluded with 95% confidence that if present in the mongoose population susceptible to trapping, M. bovis prevalence was ≤10%. The prevalence of intercurrent lesions raised concerns about gross pathology as a screening test for M. bovis infection in mongooses in Fiji, and therefore pathogen detection methods such as bacterial culture and direct tissue PCR are recommended for future surveys. These are needed to completely rule out the mongoose as a reservoir host for M. bovis in Fiji. Full article

Immune system recognize and fight back foreign microorganisms and inner modifications that lead to deficient cell and tissue functions. During a dog’s life, the immune system needs to adapt to different physiological conditions, assuring surveillance and protection in a careful and controlled way. Pregnancy alters normal homeostasis, requiring a balance between immunity and tolerance. The embryos and fetus should be protected from infections, while the female dog must tolerate the growing of semi-allografts in her uterus. After birth, newborn puppies are at great risk of developing infectious diseases, because their immune system is in development and immune memory is absent. Passive transfer of immunity through colostrum is fundamental for puppy survival in the first weeks of life, but hampers the development of an active immune response to vaccination. At the end of life, dogs experience a decline in the structure and functional competence of the immune system, compromising the immune responses to novel antigenic challenges, such as infections and vaccines. Therefore, the current article reviews the general processes related to the development of the dog´s immune system, providing an overview of immune activity throughout the dog’s life and its implications in canine health, and highlighting priority research goals. Full article

Enzyme linked immunosorbent assays (ELISAs) have been developed for the detection of antibodies against contagious caprine pleuropneumonia (CCPP), the causative agent of which is Mycoplasma capricolum subsp. Capripneumoniae (Mccp). The currently available commercial CCPP competitive ELISA (CCPP cELISA) kit produced and supplied by IDEXX Company (Westbrook, Maine, United States) is relatively expensive for most African laboratories. To address this issue and provide a variety of choices, a sensitive and specific blocking-ELISA (b-ELISA) test to detect antibodies against CCPP was developed. We describe the newly developed CCPP blocking-ELISA based on the blocking of an epitope of a monoclonal antibody (Mccp-25) by a positive serum sample against the Mccp protein coated on a plate. The Percentage Inhibition (PI) cut-off value for the CCPP b-ELISA was set at 50 using 466 CCPP negative and 84 CCPP positive small ruminant sera. Of the negative sera, 307 were obtained from the Botswana National Veterinary Laboratory (BNVL) and 159 from the Friedrich-Loeffler-Institute (FLI) Germany. The 84 positive sera samples came from experimentally vaccinated goats at the AU-PANVAC facility in Debre-Zeit, Ethiopia. The relative diagnostic sensitivity and specificity of the CCPP b-ELISA was 93% and 88%, respectively. This test result indicated good correlation with that of the commercial CCPP cELISA by IDEXX Company (Westbrook, Maine, United States) with a Cohen’s κ agreement of κ agreement of 0.85. The newly developed CCPP b-ELISA will be useful in the detection of antibodies for the diagnosis CCPP and for sero-surveillance during vaccination campaigns. Full article

Molecular typing of bacterial isolates provides a powerful approach for distinguishing Mycobacterium bovis (M. bovis) genotypes. It is known that M. bovis strain virulence plays a role in prevalence and spread of the disease, suggesting that strain virulence and prevailing genotypes are associated. However, it is not well understood whether strain virulence correlates with particular genotypes. In this study, we assessed the in vitro intracellular growth of 18 M. bovis isolates in bovine macrophages as an indicator of bacterial virulence and sought a relationship with the genotype identified by spoligotyping. We found 14 different spoligotypes—11 were already known and three spoligotypes had never been reported before. We identified 2 clusters that were phylogenetically related, containing 10 and 6 strains, respectively, and 2 orphan strains. Intracellular growth and phagocytic rates of 18 M. bovis strains were heterogeneous. Our results suggest that M. bovis intracellular growth and phagocytosis are independent of the bacterial lineage identified by spoligotyping. Full article

Mycobacterium avium subspecies paratuberculosis (Map) causes chronic granulomatous disease in cattle and ruminant livestock, causing substantial economic losses. Current vaccines delay clinical signs but cannot train the immune system to fully eradicate latent Map. During latency, Map uses host defenses, cage-like macrophage clusters called granuloma, as incubators for months or years. We used an in vitro model to investigate the early coordination of macrophages into granuloma upon Map infection over ten days. We found that at multiplicities of infection (MOI; Map:macrophages) of 1:2 and below, the macrophages readily form clusters and evolve pro-inflammatory cytokines in keeping with a cell-mediated immune response. At higher MOIs, viability of host macrophages is negatively impacted. At 1:4 MOI, we quantified viable Map in our model and confirmed that intracellular Map reproduced over the first five days of infection. Host cells expressed Type 1-specific cytokines, and Map-infected macrophages displayed reduced motility compared to Map-exposed, uninfected macrophages, suggesting an important role for uninfected macrophages in the early aggregative response. Reported is the first in vitro JD granuloma model capturing Map and macrophage viability, size distribution of resulting clusters, motility of monocyte-derived macrophages, and cytokine response during clustering, allowing quantitative analysis of multiple parameters of the Map-specific granulomatous response. Full article

The ventricular filling velocities during diastole and the influence of isoflurane anesthesia on these blood flow velocities of the racing pigeon (n = 43) are evaluated by pulsed-wave (PW) Doppler sonography. Sonographic examination demonstrates an early passive ventricular (E wave) and late active (A wave) ventricular filling. The results indicate differences between the two heart ventricles. Especially, the E wave velocity of the right heart is significantly lower than in the left heart, which is explained by the crescent-shaped cavity of the right ventricle around the left ventricle. The faster active filling velocities are significantly influenced by heart rate in conscious birds. Anesthesia with isoflurane leads to a significant decrease of the diastolic blood flow velocities, and the A wave velocities of both ventricles are especially influenced. Anesthesia with isoflurane induces a high incidence of insufficiencies of the left atrioventricular valve in the preejection period. These observations indicate that a contraction of the left ventricle myocardium is important for a complete valvular closure and for normal functioning of this heart valve. The effective closure of the right atrioventricular muscle valve in anesthetized pigeons supports the observation of the fast innervation of this muscle valve via a direct connection to the right atrium. Full article

Wilson’s disease (WD), an autosomal recessive disorder, results in copper accumulation in the liver as a consequence of mutations in the gene ATPase copper transporting beta (ATP7B). The disease is characterized by chronic hepatitis, eventually resulting in liver cirrhosis. Recent studies have shown that dysregulation of nuclear receptors (NR) by high hepatic copper levels is an important event in the pathogenesis of liver disease in WD. Intracellular trafficking of ATP7B is mediated by COMMD1 and, in Bedlington terriers, a mutation in the COMMD1 gene results in high hepatic copper levels. Here, we demonstrate a reduced Farnesoid X nuclear receptor (FXR)-activity in liver biopsies of COMMD1-deficient dogs with copper toxicosis, a unique large animal model of WD. FXR-induced target genes, small heterodimer partner (SHP), and apolipoprotein E (ApoE) were down-regulated in liver samples from COMMD1-deficient dogs with hepatic copper accumulation. In contrast, the relative mRNA levels of the two CYP-enzymes (reduced by FXR activity) was similar in both groups. These data are in line with the previously observed reduced FXR activity in livers of ATP7B−/− mice and WD patients. Therefore, these data further corroborate on the importance of the COMMD1-deficient dogs as a large animal model for WD. Full article

A black bear of 29-year-old (Ursus americanus) died unexpectedly in captivity without any gross lesions or clinical signs. We identified a firm, lobulated, yellowish tan, and well-circumscribed mass embedded inside the testicular tissue at the time of necropsy. The tumor sections exhibited soft necrotic and hemorrhagic areas beneath its capsule. Histologically, the tumor comprised Sertoli cells arranged in tubules and solid sheets supported by prominent fibrous connective tissues. The Sertoli cells were positive for vimentin and ER-β expression, whereas it showed negative staining for inhibin-α, cytokeratin 19, and S-100. To the best of our knowledge, this is the rare case report of testicular Sertoli cell tumor in black bear. Full article

This communication presents a successful story of an attempt to treat and manage a case of canine melioidosis, a severe tropical disease caused by Burkholderia pseudomallei. A 10-year-old dog was trapped with barbed wires, causing an infected wound around its neck and back, which was later diagnosed as severe melioidosis. The dog was treated based on a modified human protocol. Intravenous meropenem injections (20 mg/kg twice daily) were given for 14 days to prevent death from sepsis prior to treatment with oral sulfamethoxazole-trimethoprim (25 mg/kg twice daily) for 20 weeks to eliminate the bacteria. Canine melioidosis is an unusual infection in dogs, even in Thailand where melioidosis is highly endemic. This successful case management was solely based on proper diagnosis and appropriate treatments. Full article

Infectious diseases, particularly of wildlife, are intrinsically linked to human and domestic animal health. Reports of sarcoptic mange in black bears (Ursus americanus) are increasing in multiple states in the USA and while the reason is unknown, mange in other species has been associated with immunosuppression from multiple causes. Serum from bears across Pennsylvania were collected to determine the seroprevalence of five pathogens important for animal and/or human health: Canine distemper virus (CDV), canine parvovirus (CPV), canine adenovirus-1 (CAV), Toxoplasma gondii, and Trichinella sp. from bears with sarcoptic mange as well as bears that were clinically normal. Several of these pathogens, particularly canine distemper virus, are associated with immunosuppression and secondary infections in other hosts. In addition to describing the seroprevalence and relating these findings to data from other regions, statistics were performed to determine if antibodies to any of these pathogens were associated with mange in bears. The overall seroprevalence to these pathogens was as follows: CDV 7.1% (17/240), CPV 16% (15/94), CAV 6.9% (6/87), Toxoplasma gondii 64.9% (194/299), and Trichinella spiralis 3.2% (7/220). While there was no association between mange and antibodies to these pathogens, infection with one or more of these pathogens has implications for bears, other wildlife, domestic animal, and human health. Full article