Search

The first continental European association for veterinary pathologists was founded in 1951 as the Arbeitsgemeinschaft der Veterinärpathologen (AG-Vetpath), bringing together veterinary pathologists from Germany, several European countries, and the United States. Yearly meetings were held in conjunction with the Deutsche Gesellschaft für Pathologie (DGP). Although the majority of DGP members were human pathologists, veterinary pathologists had been using the DGP as a forum for scientific exchange since the early 20th century. Renamed in 1969 as the Europäische Arbeitsgemeinschaft für Veterinärpathologen, and in 1974 as the Europäische Gesellschaft für Veterinärpathologie, the AG-Vetpath finally received its present name, the European Society for Veterinary Pathology (ESVP) in 1994. In parallel, national organizations for veterinary pathologists in European countries have also evolved over the years, the earliest being in Germany with the Fachgruppe Allgemeine Pathologie und pathologische Anatomie of the Deutsche Veterinärmedizinische Gesellschaft (DVG). AG-Vetpath represents the parent organization for further specialty organizations like the Gesellschaft für Toxikologische Pathologie (GTP) or the Arbeitskreis Diagnostische Veterinärpathologie (AKDV). Even the European College of Veterinary Pathologists (ECVP) was founded by members of ESVP.

ZusammenfassungDer Porcine Respiratory Disease Complex (PRDC) beschreibt eine klinische Kondition, die sich in Form einer häufig therapieresistenten Atemwegsinfektion bei Mastschweinen manifestiert. Die multifaktorielle Ätiologie beinhaltet infektiöse und nicht infektiöse Faktoren. Bei Entstehung und Verlauf des PRDC spielen neben Management und Hygiene v. a. virale und bakterielle Erreger eine bedeutende Rolle. Das Virus des Porzinen Reproduktiven und Respiratorischen Syndroms (PRRSV), das porzine Circovirus Typ 2 (PCV2), Influenza-A-Virus (IAV) und Mycoplasma (M.) hyopneumoniae stellen die relevantesten Erreger dar. Das klinische Bild und die zugrundeliegenden pathomorphologischen Veränderungen können je nach Erregerbeteiligung variieren. Die Komplexität des PRDC erschwert die Diagnose und auch die Prävention auf Bestandsebene. Der Übersichtsartikel gibt einen Einblick in die Pathomorphologie, Pathogenese sowie Inter-Erreger-Interaktionen und zielt darauf ab, praktizierende Tierärztinnen und Tierärzte bei der Diagnose, Befundinterpretation und Prävention des PRDC zu unterstützen.

A new gene defect in Fleckvieh calves leads to a syndrome with partial phenotype overlap with bovine hereditary zinc deficiency. A mutation in a gene encoding phospholipase D4 ( PLD4), an endosomal exonuclease, causes the disorder. In mice, PLD4 activity indirectly regulates the Toll-like receptor 9 (TLR9) pathway via degradation of microbial DNA. PLD4 absence thus results in visceral macrophage activation comparable to human macrophage activation syndrome. In this study, disease progression and the role of macrophages in affected calves were monitored clinically, clinicopathologically, and histologically over time. Breeding data identified 73 risk matings of heterozygous carriers resulting in 54 potentially PLD4-deficient calves born on farms. PLD4 status was examined via 5′-exonuclease assay, detecting 6 calves carrying the defect. These were purchased and monitored daily until final necropsy. The calves developed progressive skin lesions starting with small scaling areas terminating in severe crusting dermatitis, especially in areas with mechanical exposure. Histological and immunohistochemical analyses indicated that macrophages with cytoplasmic vacuolation increased considerably in skin sections obtained weekly during the disease course. Macrophage increase correlated with increased dermal lesion severity. Macrophage activation was confirmed by prominent phagocytic activity in the superficial dermis using electron microscopy. Dermal mRNA abundance of CCL2 and CCL3 measured by quantitative polymerase chain reaction verified macrophage activation. Further increase in mRNA of downstream molecule MyD88 and cytokine IL12b connected bovine PLD4 deficiency to increased TLR9 pathway activation. In contrast to human macrophage activation syndrome, the main feature of bovine PLD4 deficiency was local disease in organs with contact to microbial DNA (skin, intestine, lungs).

The porcine respiratory disease complex describes a clinical condition that often manifests as treatment-resistant respiratory disease of growing to finishing pigs. Its multifactorial etiology includes infectious and non-infectious factors. Besides management and hygiene conditions, particularly viral and bacterial pathogens contribute to the development and course of PRDC. The porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), influenza A virus (IAV) andDer Porcine Respiratory Disease Complex (PRDC) beschreibt eine klinische Kondition, die sich in Form einer häufig therapieresistenten Atemwegsinfektion bei Mastschweinen manifestiert. Die multifaktorielle Ätiologie beinhaltet infektiöse und nicht infektiöse Faktoren. Bei Entstehung und Verlauf des PRDC spielen neben Management und Hygiene v. a. virale und bakterielle Erreger eine bedeutende Rolle. Das Virus des Porzinen Reproduktiven und Respiratorischen Syndroms (PRRSV), das porzine Circovirus Typ 2 (PCV2), Influenza-A-Virus (IAV) und

Bone replacement and osteosynthesis require materials which can at least temporarily bear high mechanical loads. Ideally, these materials would eventually degrade and would be replaced by bone deposited from the host organism. To date several metals, notably iron and iron-based alloys have been identified as suitable materials because they combine high strength at medium corrosion rates. However, currently, these materials do not degrade within an appropriate amount of time. Therefore, the aim of the present study is the development of an iron-based degradable sponge-like (i.e. cellular) implant for bone replacement with biomechanically tailored properties. We used a metal powder sintering approach to manufacture a cylindrical cellular implant which in addition contains phosphor as an alloying element. No corrosion inhibiting effects of phosphorus have been found, the degradation rate was not altered. Implant prototypes were tested in an animal model. Bone reaction was investigated at the bone-implant-interface and inside the cellular spaces of the implant. Newly formed bone was growing into the cellular spaces of the implant after 12 months. Signs of implant degradation were detected but after 12 months, no complete degradation could be observed. In conclusion, iron-based open-porous cellular biomaterials seem promising candidates for the development of self-degrading and high load bearing bone replacement materials.

This study describes the pathomorphological alterations of bovine tuberculosis through gross and histopathological examinations, assessment of the distribution of lesions and the demonstration of mycobacteria. Samples from lungs, liver, small intestine, their regional lymph nodes and retropharyngeal lymph nodes were collected from 84 cattle with tuberculosis from the Allgau, Germany. Organs were evaluated grossly, histopathologically and by transmission electron microscopy. Mycobacteria and mycobacterial antigens were demonstrated using acid-fast staining and immunohistochemistry (IHC). Bacteriological tests revealed Mycobacterium caprae in all animals. Gross alterations were classified into five patterns (I to V) with an additional pattern of acute exudative pulmonary inflammation (pattern VI). Histological lesions were classified into four types (1-4) with additional lesions occurring in lungs only. Acid-fast staining revealed a low number of bacteria in all tissues, while IHC showed comparatively more mycobacterial antigens within the lesions and also at their periphery. The alimentary tract (68%) was the main portal of entry followed by an aerosol infection (19%). It was assumed that the observed lesions reflect a continuous primary period of infection; there were no lesions typical of a secondary (post-primary) period, as reported in man and also described in the older literature on bovine tuberculosis. The broad spectrum of changes described formerly was not observed in the present cases and the route of infection and nature of acid fast staining showed differences when compared with previous studies of naturally-occurring bovine tuberculosis.

Acute hemorrhagic diarrhea syndrome (AHDS), formerly named canine hemorrhagic gastroenteritis, is one of the most common causes of acute hemorrhagic diarrhea in dogs, and is characterized by acute onset of diarrhea, vomiting, and hemoconcentration. To date, histologic examinations have been limited to postmortem specimens of only a few dogs with AHDS. Thus, the aim of our study was to describe in detail the distribution, character, and grade of microscopic lesions, and to investigate the etiology of AHDS. Our study comprised 10 dogs with AHDS and 9 control dogs of various breeds, age, and sex. Endoscopic biopsies of the gastrointestinal tract were taken and examined histologically (H&E, Giemsa), immunohistochemically ( Clostridium spp., parvovirus), and bacteriologically. The main findings were acute necrotizing and neutrophilic enterocolitis (9 of 10) with histologic detection of clostridia-like, gram-positive bacteria on the necrotic mucosal surface (9 of 10). Clostridium perfringens isolated from the duodenum was identified as type A (5 of 5) by multiplex PCR (5 of 5). In addition, each of the 5 genotyped isolates encoded the pore-forming toxin netF. Clostridium spp. (not C. perfringens) were cultured from duodenal biopsies in 2 of 9 control dogs. These findings suggest that the pore-forming netF toxin is responsible for the necrotizing lesions in the intestines of a significant proportion of dogs with AHDS. Given that the stomach was not involved in the process, the term “acute hemorrhagic diarrhea syndrome” seems more appropriate than the frequently used term “hemorrhagic gastroenteritis.”

The safety of veterinary vaccines is assessed in clinical trials in Europe. The assessment of the local tissue reaction to vaccination by magnetic resonance imaging (MRI) could reduce the number of animals needed because repeated examinations can be performed in the same animal over time. The present study compared the evaluation of local tissue reactions to vaccination using MRI in live pigs with histopathology of porcine tissue, the current gold standard in regulatory safety testing. Eight piglets each were administered one of two commercial vaccines into marked injection sites. All animals were sedated and scanned repeatedly by MRI using a contrast agent up to day 29 after vaccination. On day 29, the animals were euthanized and underwent a pathological examination. The MRI results were compared with the pathomorphological findings at the injection site by regression analysis. The MR images and the pathological examinations yielded matching results concerning the sizes of the affected tissue volumes or areas. The use of MRI for regulatory safety testing can reduce the number of animals needed to 8 per examination group. The volume of a local reaction and its progression over time can be evaluated and documented. If persistent lesions develop a final pathomorphological examination is needed to identify the kind and local distribution of the reaction.

ZusammenfassungBeschrieben wird ein Fall der septikämischen Salmonellose und des Porzinen Dermatitis-und-Nephropathie-Syndroms (PDNS) mit Nachweis von Salmonella (S.) Choleraesuis var. Kunzendorf und dem porzinen Circovirus Typ 2 (PCV2) bei einem Mastschwein aus einem Bio - betrieb. Auf diesem biologisch wirtschaftenden Schweinemastbetrieb traten 6 Wochen nach dem Einstallen der Tiere vermehrt Todesfälle sowie Durchfall und Husten auf. Das identifizierte Salmonellen-Serovar wurde in Deutschland in den letzten Jahren vor allem bei Wildschweinen nachgewiesen, während der Erreger bei Hausschweinen in Westeuropa keine nennenswerte Rolle mehr spielte. Eine direkte Erregerübertragung von Wildschweinen auf die Hausschweine lässt sich im vorliegenden Fall nicht nachweisen. Vor dem Hintergrund, dass Wildschweine als Erregerreservoir für Tierseuchenerreger eine bedeutende Rolle spielen, unterstreicht dieser Fall jedoch, wie wichtig es ist, epidemiologische Zusammenhänge zu berücksichtigen und die Schweinehaltungshygieneverordnung zur Vermeidung von direkten oder indirekten Kontaktmöglichkeiten zwischen Haus- und Wildschweinen einzuhalten.

The aim of the study was to determine the frequency and age-dependency of adrenal changes in dogs and cats, which were dissected between 2004 and 2006 at the Institute of Veterinary Pathology, LMU Munich. The adrenal glands of 101 dogs and 159 cats were dissected free from adjacent tissue, weighed, lamellated perpendicular to its longitudinal axis and were embedded in paraffin and plastic for histologic inspection. Ageing cats most frequently showed accessory cortical nodules in the capsule, cortex and medulla, pigment deposition and shock bodies. The appearance of accessory cortical nodules and pigment deposition were linked to the age of the cats and that of shock bodies was linked to infectious diseases. Inflammatory alterations were rare and neoplastic changes were not observed. In dogs, the most frequent alterations were accessory cortical nodules in the capsule, cortex and medulla and circulatory disturbances and there was a significant linkage of the appearance of cortical nodules and pigment deposition and age. Metabolic, inflammatory and tumorous diseases were rarely observed in the dogs examined. In summary, cats and dogs show similar incidental alterations of the adrenals. Clinical relevant adrenal lesions such as atrophy or primary neoplasia were rarely diagnosed.

The diagnosis of canine intestinal lymphoma by morphological examination is challenging, especially when endoscopic tissue specimens are used. The utility of detection of antigen receptor gene rearrangement by polymerase chain reaction (PARR) in canine lymphoma has been well established, but its usefulness to distinguish enteritis and intestinal lymphoma remains unclear. In this retrospective study we assessed clonality of 29 primary canine intestinal lymphoma, 14 enteritis and 15 healthy control cases by PARR analysis, using formalin-fixed, paraffin-embedded full-thickness tissue specimens. We could detect monoclonal rearrangements in 22 of 29 canine intestinal lymphomas [76%; 95% confidence interval (CI) 56-90%] and polyclonal rearrangements in all of the enteritis and healthy control cases (100%; CI 88-100%). We revealed a predominance of T-cell phenotype compared to B-cell phenotype (85%; CI 65-96% and 15%; CI 4-35%, respectively). We showed that PARR analysis contributes to differentiation of canine intestinal lymphoma from enteritis and to phenotyping of lymphomas.

PCV2 can be divided into three different genotypes: PCV2a, PCV2b and PCV2c. Since 2004/2005 PCV2b has become the predominant genotype in the domestic pig population worldwide. In the years 2010 and 2012 PCV2b mutant strains (mPCV2), classified as PCV2b-1C strains, were detected in porcine circovirus diseases (PCVD) affected pigs in China and the United States, respectively. Within one year (April 2013-April 2014) newly emerging mPCV2 strains were isolated in seven German pig farms routinely vaccinating against PCV2. Histopathological, clinical and molecular biological findings including in-situ hybridization (ISH) and real-time PCR indicate PCVD in the affected animals. Characterized isolates from five farms were closely related to the PCV2b-1C reference strain BDH (GenBank no. HM038017), whereas strains from two other farms were only 99.1% and 99.0% identical (based on the nucleotide sequence of the complete genome) to mPCV2 strain BDH, respectively.

The objective of the present study was to assess safety and immune responses in gilts after intradermal application of Porcilis® PRRS in two different application sites under field conditions. Forty-four gilts were allocated to one of three groups: Gilts of group 1 (n = 10) served as non-vaccinated controls, gilts of group 2 (n = 17) were vaccinated intradermally in the neck and gilts of group 3 (n = 17) received an intradermal vaccination in the perianal region. Clinical observations, local injection site reactions and histopathologic examination of the injection site were used for safety assessments. Frequency and degree of clinical signs were not significantly different between all three groups. Minor local reactions for both vaccination groups were observed; however, at 6, 7, 8, 9 and 15 days post-vaccination (dpv), the mean injection site reaction score was significantly lower in pigs vaccinated in the perianal region. In histopathologic examination, an extended inflammatory dimension was observed more frequently in pigs vaccinated in the neck. Blood samples were analyzed to quantify the post-vaccination humoral (ELISA and virus neutralization test) and cellular (IFN-γ ELISPOT) immune responses. PRRSV-specific antibodies were present in the serum of all vaccinated animals from 14 dpv onwards, whereas all control pigs remained negative throughout the study. Neutralizing antibody titers were significantly higher in pigs vaccinated in the perianal region at 28 dpv. At 14, 21 and 28 dpv, PRRSV-specific IFN-γ secreting cells were significantly increased in both vaccination groups compared to non-vaccinated gilts. Analysis of mean numbers of PRRSV-specific IFN-γ secreting cells did not result in statistically significant differences between both vaccination groups. The results of this study indicate that the perianal region is a safe alternative application site for intradermal vaccination of gilts with Porcilis PRRS. Furthermore, the intradermal application of Porcilis PRRS induced humoral and cellular immune responses independent of the administration site.

Current knowledge on bovine besnoitiosis, caused by the emerging apicomplexan pathogen Besnoitia besnoiti, is still fragmentary. So far, studies dealing with ultrastructural pathology focused mainly on the easily accessible chronic stage, whereas ultrastructural investigations of tachyzoites were confined to in vitro studies. In the study presented here, the ultrastructural pathology of naturally B. besnoiti-infected cattle in the acute and chronic disease stages and experimentally B. besnoiti-infected mice was monitored. Further, the ultrastructure of tachyzoites and bradyzoites was investigated. Skin samples of two adult Limousin cows and one adult Limousin bull naturally infected with B. besnoiti and liver and skin samples of gamma-interferon knockout mice infected with B. besnoiti were examined in semithin sections stained with toluidine blue and safranin and in ultrathin sections contrasted with uranyl acetate and lead citrate. Samples of vessel walls of the bull and nasal mucosa of one cow were examined by scanning electron microscopy. Few tachyzoites-like endozoites were detected for the first time in bovine skin, and large numbers of tachyzoites were detected in murine skin and liver. Within tissue cysts in bovine skin, numerous bradyzoites were observed displaying signs of degeneration. Tachyzoites had apicomplexan endozoite ultrastructure. B. besnoiti tachyzoites and bradyzoites differed in shape and the number of amylopectin granules. Transmission and scanning electron microscopy confirmed the presence of two different cyst wall layers, and the present results on cyst wall ultrastructure were in accordance with those previously obtained by histological sections.

The anaerobic intestinal spirochaete

Background: Zinc deficiency-like (ZDL) syndrome is an inherited defect of Fleckvieh calves, with striking similarity to bovine hereditary zinc deficiency (BHZD). However, the causative mutation in a phospholipase D4 encoding gene (PLD4) shows no connection to zinc metabolism. Objective: To describe clinical signs, laboratory variables, and pathological findings of ZDL syndrome and their utility to differentiate ZDL from BHZD and infectious diseases with similar phenotype. Animals: Nine hospitalized calves with crusting dermatitis and confirmed mutation in PLD4 and medical records from 25 calves with crusting dermatitis or suspected zinc deficiency. Methods: Prospective and retrospective case series. Results: The 9 calves (age: 5-53 weeks) displayed a moderate to severe crusting dermatitis mainly on the head, ventrum, and joints. Respiratory and digestive tract inflammations were frequently observed. Zinc supplementation did not lead to remission of clinical signs in 4 calves. Laboratory variables revealed slight anemia in 8 calves, hypoalbuminemia in 6 calves, but reduced serum zinc concentrations in only 3 calves. Mucosal erosions/ulcerations were present in 7 calves and thymus atrophy or reduced thymic weights in 8 calves. Histologically, skin lesions were indistinguishable from BHZD. Retrospective analysis of medical records revealed the presence of this phenotype since 1988 and pedigree analysis revealed a common ancestor of several affected calves. Conclusions and Clinical Importance: ZDL syndrome should be suspected in Fleckvieh calves with crusting dermatitis together with diarrhea or respiratory tract inflammations without response to oral zinc supplementation. Definite diagnosis requires molecular genetic confirmation of the PLD4 mutation.

Safety testing is one major part of the licensing procedure for veterinary vaccines and demands a large number of animals. Magnetic resonance imaging (MRI) was tested as an alternative, which may lead to a reduction in numbers of animals required for safety testing, and, correspondingly to a detailed description of the three-dimensional extent of the local tissue reaction repetitively in live pigs. In previous pig studies the following questions arose:To answer these questions the following study was performed by comparing two vaccine groups of suckling piglets (8 animals per group; A and B) with two control groups (4 animals per group; C and D). One control group was injected with a saline solution (C) and the other was only tattoo marked (D). The animals were examined using MRI at days 1, 8, 15, 22, 29, 36, and 43 post vaccination, ending with a final pathomorphologic examination. Pathomorphologic examination confirmed MRI findings. Saline solution does not result in a local tissue reaction as detected after injecting vaccines. Tattoo marking causes no local tissue reaction, neither in MRI nor in pathomorphologic examination. Therefore, MRI can be used as an alternative method for safety testing of vaccines in pigs of different age categories offering repetitive measurements of local tissue reactions. Involved cells might be examined only in a final pathomorphologic examination at the end of the trial on a reduced number of animals.

The pathogenesis of bovine besnoitiosis, a disease of increasing concern within Europe, is still incompletely understood. In this study, disease progression after natural infection with the causative apicomplexan Besnoitia besnoiti was monitored in histological skin sections of 5 individual female cattle over time. High-frequency skin sampling of 2 cattle with mild and 2 with severe acute, subacute, and chronic besnoitiosis, as well as from 1 animal during subclinical disease, enabled documentation from the beginning of the disease. Skin sections were stained with hematoxylin and eosin and Giemsa, periodic acid–Schiff reaction, and anti- Besnoitia immunohistochemistry. In all 4 clinically affected animals, tachyzoite-like endozoites could be detected for the first time by immunohistochemistry, and tissue cyst evolution was monitored. Besnoitiosis-associated lesions were not detected in the animal showing the subclinical course. Because of the inconsistency of the nomenclature of Besnoitia tissue cyst layers in the literature, a new nomenclature for B. besnoiti cyst wall layers is proposed: tissue cysts consist of a hypertrophied host cell with enlarged nuclei, an intracytoplasmic parasitophorous vacuole with bradyzoites, a sometimes vacuolated inner cyst wall, and an outer cyst wall in more developed cysts. Inner and outer cyst walls can be readily distinguished by using special stains. In 1 animal, extracystic B. besnoiti zoites were immunohistochemically detected during the chronic stage. At necropsy, the 2 severely affected cows displayed large numbers of B. besnoiti cysts in a variety of tissues, including the corium of the claws, contributing mainly to the development of chronic laminitis in these 2 cases.

The genus Dermocystidium comprises fungal pathogens of the order Dermocystida in the class Ichthyosporea (http://www.ncbi.nlm.nih.gov/taxonomy). Several different Dermocystidium species have been described, infecting a variety of fish hosts and producing gill infections, skin lesions and visceral disease all over the world (Wildgoose 1995; Pekkarinen & Lotman 2003; Pekkarinen et al. 2003; Feist et al. 2004; Zhang & Wang 2005). To our knowledge, there are two reports on Dermocystidium infections in cardinal tetra, Paracheirodon axelrodi and neon tetra, Paracheirodon innesi (Reichenbach-Klinke 1982; Lewisch 2010). However, these reports did neither include histological and ultrastructural examinations nor molecular genetic investigations to confirm the diagnosis and identify the aetiologic species. In January 2013, increased mortalities of cardinal tetra, P. axelrodi of a 350-L aquarium were reported, occurring after purchase of additional fish. During the clinical examination, most of the fish were swimming in normal active condition, but some were lethargic or displayed a transparent mass on the skin of the head or body. The masses were up to 5 mm in diameter and contained a central, white tubular structure. Similar masses were located on the fins of some fish, but were considerably smaller in these locations. A parasitic infestation of the skin of these fish was excluded via microscopic examination, and normal results were obtained after analysis of the water values and inspection of the pump and filter system. Six cardinal tetra, P. axelrodi and two firehead tetra, Hemigrammus bleheri were submitted for pathological examination in formalin and for molecular genetic examination in ethanol. Only two cardinal tetra displayed skin lesions. Before embedding, one tetra was post-fixed in Davidson’s fixative and sections of glycolmethacrylate/methylmethacrylate-embedded samples were routinely processed for histological examination and stained with haematoxylin and eosin (HE), Giemsa, silver impregnation and periodic acid Schiff (PAS) reaction according to standard protocols. Samples of the mass were also routinely processed for transmission and scanning electron microscopy on a transmission electron microscope (Zeiss EM 10) or on a digital scanning electron microscope (Zeiss DSM 950), respectively. The macroscopic examination of the cardinal tetra revealed a focally extensive, bulging, hemispherical, transparent oedema (4 mm in diameter) of the ventral skin on the head. Central, within Correspondence M C Langenmayer, Institute of Veterinary Pathology at the Centre for Clinical Veterinary Medicine, LMU Munich, Veterin€ arstrasse 13, 80539 Munich, Germany (e-mail: langenmayer@patho.vetmed.uni-muenchen.de) Langenmayer and Lewisch contributed equally to this work.

Background Etiology of hemorrhagic gastroenteritis (HGE) syndrome in dogs is unknown and histopathologic and microbial investigations have only been performed post mortem. Objective To identify characteristic intra vitam endoscopic and histologic mucosal lesions, as well as bacterial species, within the mucosa of dogs with HGE. Animals Ten dogs diagnosed with HGE were included. Eleven dogs with gastroduodenoscopy and different intestinal diseases were used as controls for microbial changes. Dogs pretreated with antibiotics or diagnosed with any disease known to cause bloody diarrhea were excluded from the study. Methods In this prospective study, gastrointestinal biopsies were collected from 10 dogs with HGE. Endoscopic and histologic changes were assessed according to WSAVA guidelines. Biopsies from the stomach, duodenum, ileum, and colon were investigated by histology and by immunohistochemistry for the presence of Clostridium spp. and parvovirus. The first duodenal biopsy taken with a sterile forceps was submitted for bacterial culture. Results Acute mucosal lesions were only found in the intestines, not in the stomach. Clostridium spp., identified as Clostridium perfringens in 6/9 cases, were detected on the small intestinal mucosa in all dogs with HGE, either by culture or immunohistopathology. In the control group, C. perfringens could only be cultured in one of 11 dogs. Conclusions and Clinical Importance The results of this study demonstrate an apparent association between C. perfringens and the occurrence of acute hemorrhagic diarrhea. The term “HGE,” which implies the involvement of the stomach, should be renamed as “acute hemorrhagic diarrhea syndrome.”

Morphological lesions in kidneys and brain are all too often considered diagnostic for confirmation of encephalitozoonosis in rabbits. The current study evaluated the diagnostic value of histology versus other etiological tests, including immunohistochemistry and real-time polymerase chain reaction (PCR) for Encephalitozoon cuniculi infection diagnosis. Samples of brain, heart, lungs, intestine, liver, and kidneys from 81 rabbits were examined for morphological lesions attributed to E. cuniculi infection as well as for the presence of spores and E. cuniculi antigen. Of these, 55 rabbits were tested for E. cuniculi DNA. Histological changes consistent with E. cuniculi infection were seen in 33 rabbits (41%, 33/81) representing 87% (33/38) of all rabbits with confirmed E. cuniculi infection. Brains of these rabbits displayed 6 different types of focal lesions corresponding to the stage of infection and specific tissue response. In 5 rabbits that were tested positive, histology was either inconclusive or inconspicuous. Etiological diagnosis was based on histological spore detection in 16% (6/38) of infected rabbits. Immunohistochemistry was more sensitive (42%, 16/38) than histological spore detection, and real-time PCR proved to be the most sensitive of all investigated methods (30/35, 86% of the examined rabbits with E. cuniculi infection). Encephalitozoon cuniculi infection rarely occurs without characteristic kidney and brain lesions. However, the spectrum of brain changes is wider than previously reported. Based on these findings, confirmation of pathogenic E. cuniculi infection should include standard histology of the predilection sites and a specific etiological assay, preferably real-time PCR.

A gastric biopsy specimen from a 14-year-old Yorkshire terrier was analysed using light microscopy, immunohistochemistry and transmission electron microscopy. The biopsies were obtained from a 3 × 3 × 4 cm mass located at the gastric cardia. Light microscopy revealed solid nests and sheets of tumour cells separated by abundant fibrovascular stroma in all gastric layers. The tumour cells were monomorphic, with eosinophilic cytoplasm and demonstrated palisading at the periphery. Nuclei were round to oval with densely packed chromatin and inconspicuous nucleoli. The tumour cells immunohistochemically stained strongly positive for chromogranin A, synaptophysin, and pan-cytokeratin. Electron microscopy revealed electron dense neurosecretory granules of 100 - 150 nm in diameter. Metastases were found in regional lymph nodes. Gastric neuroendocrine carcinoma was diagnosed, according to the histological, immunohistochemical and electron microscopic features.

In Europe, cystic echinococcosis is rare in horses and is mostly diagnosed at slaughter or postmortem examination. Equine cystic echinococcosis can be caused by various Echinococcus taxa, but only Echinococcus equinus (the "horse strain") is known to produce fertile cysts. In Europe, E. equinus appears to be endemic in Great Britain, Ireland, Spain, and Italy and has sporadically been reported in Belgium and Switzerland. The present report describes the first case of a molecularly confirmed E. equinus infection in a horse foaled and raised in Germany. The 19-year-old mare was presented for examination of inappetence, emaciation, and respiratory symptoms. X-ray radiographs of the thorax showed 2 well-circumscribed tumor-like masses, each approximately 10 cm in diameter in the caudal lung field. The horse was euthanized as its condition rapidly deteriorated. Necropsy revealed 2 thick-walled hydatid cysts, each 7-8 cm in diameter in the lung. The tri-layered cyst walls consisted of an outer adventitial layer, a laminated acellular intermediate layer, and an inner germinal membrane. Grossly, the cysts contained a clear, amber liquid with hydatid sand. Light microscopy of the hydatid sand revealed free protoscoleces, intact and ruptured brood capsules, calcareous corpuscles, and debris. Samples of protoscoleces underwent molecular characterization, and the diagnosis of E. equinus was confirmed by restriction fragment length polymorphism-polymerase chain reaction and sequence analysis of the complete mitochondrial nicotinamide adenine dinucleotide dehydrogenase subunit 1 gene.

Intestinal obstruction resulting from an intramural foreign body-associated pyogranuloma was diagnosed in four dogs. Vomiting and weight loss were the main clinical signs. On physical examination, a mass in the abdomen was detected in three dogs. Abdominal radiography demonstrated the presence of soft tissue opacity in three of the dogs and gas-filled dilated intestinal loops in all four dogs. Abdominal ultrasonography showed hyperkinetic fluid-filled dilated intestinal loops and a hypoechoic small intestinal mass in all the dogs. Exploratory coeliotomy confirmed the presence of a jejunal mass, which was removed by resection and anastomosis in all the dogs. In one of the dogs a linear foreign body was also found cranial to the mass and was removed through a separate enterotomy incision. The lesions were diagnosed as foreign body-associated intestinal pyogranulomas on histological examination. Three dogs recovered without complications, but the fourth showed signs of septic peritonitis four days after surgery and was euthanased at the owner's request. The other three dogs remained disease-free 12 to 42 months after surgery.

The present case reports the detection of Salmonella (S.) Choleraesuis var. Kunzendorf and porcine circovirus type 2 in an organic fattening pig suffering from septicaemic salmonellosis and porcine dermatitis and nephropathy syndrome. Six weeks after pigs had been housed in an organic fattening farm, an increase in mortality, diarrhea and coughing was observed. In recent years, S. choleraesuis var. Kunzendorf has been frequently detected in wild boars in Germany, whereas the same serovar did not play an important role in the Western European domestic pig population. A direct transmission of this serovar from wild boars to domestic pigs in this case could not be proven. However, because wild boars are important reservoirs for the spread of epizootic diseases to the domestic pig population, this case emphasises the importance of taking epidemiological relationships under consideration and to comply with biosecurity measures according to German law (Schweinehaltungshygieneverordnung).

Borrelia persica, a bacterium transmitted by the soft tick Ornithodoros tholozani, causes tick-borne relapsing fever in humans in the Middle East, Central Asia and the Indian peninsula. Immunocompetent C3H/HeOuJ mice were infected intradermally with B. persica at varying doses: 1 x 106, 1 x 104, 1 x 102 and 4 x 100 spirochetes/mouse. Subsequently, blood samples were collected and screened for the presence of B. persica DNA. Spirochetes were detected in all mice infected with 1 x 106, 1 x 104 and 1 x 102 borrelia by real-time PCR targeting the flaB gene of the bacterium. Spirochetemia developed with a one- to two-day delay when 1 x 104 and 1 x 102 borrelia were inoculated. Mice injected with only four organisms were negative in all tests. No clinical signs were observed when infected mice were compared to negative control animals. Organs (heart, spleen, urinary bladder, tarsal joint, skin and brain) were tested for B. persica-specific DNA and cultured for the detection of viable spirochetes. Compiled data show that the target organs of B. persica infections are the brain and the skin. A newly developed serological two-tiered test system (ELISA and western blot) for the detection of murine IgM, IgG and IgA antibody titers against B. persica showed a vigorous antibody response of the mice during infection. In conclusion, the infection model described here for B. persica is a platform for in vivo studies to decipher the so far unexplored survival strategies of this Borrelia species., Author Summary The spirochete Borrelia persica is a tick-borne bacterium that is transmitted by the vector Ornithodoros tholozani to its vertebrate host in the Middle East, Central Asia and the Indian peninsula. Current migration of vast numbers of individuals from this area increases the likelihood that B. persica infections will be introduced into new geographic regions. After infection and distribution by the bloodstream, relapsing fever episodes occur in humans. Since no reliable in vivo tools have existed so far to study this organism, a murine model was established in this study to characterize the infection kinetics in immunocompetent mice. Aspects of the potential infectivity of the laboratory strain and of potential clinical signs, spirochetemia and antibody response as well as organ tropism and histopathological reactions were studied. With the successful infection model presented here, further studies are conceivable in order to gain advanced insights into the pathogenesis of B. persica infection and to characterize in detail the host immune response mounted against the bacterium. We propose that this model could also be used for the development of new rapid diagnostic approaches to initiate or monitor treatment regimes in order to clear or prevent the infection with B. persica.

Objectives Feline infectious peritonitis (FIP) is an important cause of death in the cat population worldwide. The ante-mortem diagnosis of FIP in clinical cases is still challenging. In cats without effusion, a definitive diagnosis can only be achieved post mortem or with invasive methods. The aim of this study was to evaluate the use of a combined reverse transcriptase nested polymerase chain reaction (RT-nPCR) and sequencing approach in the diagnosis of FIP, detecting mutations at two different nucleotide positions within the spike (S) gene. Methods The study population consisted of 64 cats with confirmed FIP and 63 cats in which FIP was initially suspected due to similar clinical or laboratory signs, but that were definitively diagnosed with another disease. Serum/plasma and/or effusion samples of these cats were examined for feline coronavirus (FCoV) RNA by RT-nPCR and, if positive, PCR products were sequenced for nucleotide transitions within the S gene. Results Specificity of RT-nPCR was 100% in all materials (95% confidence interval [CI] in serum/plasma 83.9–100.0; 95% CI in effusion 93.0–100.0). The specificity of the sequencing step could not be determined as none of the cats of the control group tested positive for FCoV RNA. Sensitivity of the ‘combined RT-nPCR and sequencing approach’ was 6.5% (95% CI 0.8–21.4) in serum/plasma and 65.3% (95% CI 50.4–78.3) in effusion. Conclusions and relevance A positive result is highly indicative of the presence of FIP, but as none of the control cats tested positive by RT-nPCR, it was not possible to confirm that the FCoV mutant described can only be found in cats with FIP. Further studies are necessary to evaluate the usefulness of the sequencing step including FCoV-RNA-positive cats with and without FIP. A negative result cannot be used to exclude the disease, especially when only serum/plasma samples are available.

Zusammenfassung Gegenstand und Ziel: Ziel dieser retrospektiven Arbeit war die Prüfung der diagnostischen Validität verschiedener anamnestischer Angaben und klinischer Befunde im Hinblick auf Endokarditis (EK). Material und Methoden: Zur Auswertung gelangten 958 Klinikkarten der Patienten der Klinik für Wiederkäuer aus den Jahren 1997–2004. Die Tiere wurden aufgrund ihres Herzbefundes, der sich aus der klinischen Untersuchung ergab (Herzgeräusch oder kein Herzgeräusch) und aufgrund des Sektionsbefundes (EK oder keine EK) in vier Gruppen eingeteilt. Ergebnisse: Zwischen den an EK erkrankten und nicht erkrankten Tieren bestanden signifikante Unterschiede hinsichtlich Ernährungszustand, Verhalten, Jugularvenen- und Herzbefunden. Außerdem litten EK-Patienten häufiger an Gliedmaßenproblemen. Schlussfolgerung und klinische Relevanz: Aus dem Vorbericht, den Ergebnissen der klinischen Untersuchung sowie den hämatologischen und klinisch-chemischen Befunden lässt sich der Verdacht auf eine Endokarditis zwar erhärten, aber die Sicherung der Diagnose ist nach wie vor in vielen Fällen der pathologischen Untersuchung vorbehalten.

This study documents the characteristics of a large series of spontaneously occurring thymomas in a laboratory colony of European hamsters (Cricetus cricetus). Thymomas are rare organotypic neoplasms originating from the thymic epithelial compartment. Because the hamster thymomas largely resembled their human counterparts, the recent World Health Organization (WHO) classification of human thymic epithelial tumors was used. Forty hamsters of both sexes aged 3-29 months were examined macroscopically and histo- logically. In 22 (55%) of the 40 animals, necropsy revealed enormous whitish masses in the anterior medias- tinum, with a diameter ranging from 0.5 to 4.5 cm and a lobulated structure. The anatomy of the thymus region was normal in the remaining 18 hamsters. Histologically, the tumors presented as thymuslike organoid structures with areas of medullary and cortical differentiation and a predominance of lymphoid cells. A network of epithelial cells in the cortical areas, demonstrated immunohistochemically with a cross-reactive antibody against pancytokeratin, supported the diagnosis of thymoma. Cortical lymphocytes showed positive staining with cross- reacting antibodies against CD3 and terminal deoxynucleotidyl transferase, characteristic of immature T cells. On the basis of these findings, the tumors were classified as B1 thymomas, in some cases with AB or B2 components, according to the new WHO classification for human thymic epithelial tumors. Several possible differential diagnoses must be kept in mind when tumorous masses are found in the an- terior mediastinum. The most frequent neoplasms are lymphomas, followed by thymomas, tumors of ectopic thyroid tissue, and neuroendocrine or mesenchymal tu- mors. Thymomas are rare malignancies both in human and veterinary medicine. They have been described in a wide range of domestic and laboratory animals and occur more frequently in older animals. In humans, thymomas usually present in the fourth to fifth decade but can occur at all ages without sex predisposition. 28

Clinical data from 488 cats (1979-2000) with histopathologically confirmed feline infectious peritonitis (FIP) and 620 comparable controls were evaluated retrospectively to assess the value of several diagnostic tests frequently used in the evaluation of cats with suspected FIP. Diagnostic utility of serum albumin to globulin ratio for the diagnosis of FIP was greater than of the utility of serum total protein and gamma-globulin concentrations. Diagnostic utility of these variables was higher when performed on effusion. On effusion, positive and negative predictive values of Rivalta's test, a test that distinguishes between exudates and transudates (0.86 and 0.97), anti-coronavirus antibody detection (0.90 and 0.79), and immunofluorescence staining of coronavirus antigen in macrophages (1.00 and 0.57) were investigated. The positive and negative predictive values of presence of anti-coronavirus antibodies were 0.44 and 0.90, respectively, antibody concentrations (1:1,600) were 0.94 and 0.88. presence of immune complexes measured by a competitive enzyme-linked immunosorbent assay were 0.67 and 0.84, and detection of viral RNA by serum reverse-transcriptase polymerase chain reaction (RT-PCR) were 0.90 and 0.47. Effusion RT-PCR was performed in 6 cats; it was positive in all 5 cats with FIP and negative in the cat with another disease. Diagnostic assays on the fluid in cats with body effusion had good predictive values. Definitive diagnosis of FIP on the basis of measurement of various variables in serum was not possible. Serum tests can only be used to facilitate the decision for more invasive diagnostic methods.

ZusammenfassungBei Störungen des Allgemeinbefindens, die insbesondere mit respiratorischen und neurologischen Symptomen einhergehen, sind bei allen Spezies auch Systemmykosen in Betracht zu ziehen. In dem vorliegenden Fallbericht wird eine generalisierte Infektion mit Cryptococcus neoformans bei einem Meerschweinchen vorgestellt. Diese in der Tiermedizin eher selten beschriebene Infektionskrankheit hat in der Humanmedizin mit der Verbreitung des humanen Immundefizienzvirus und den konsekutiven opportunistischen Infektionen an Bedeutung zugenommen.

The anaerobic intestinal spirochaete Brachyspira (B.) suanatina was first described in 2007 but since then no further isolates have been reported from pigs. Accordingly, when the species was validly published in 2016, the overall occurrence and clinical relevance in pigs were unknown. In a fattening farm in southern Germany, mucohaemorrhagic diarrhoea was observed in 60 per cent (750 animals) of the finisher pigs. A diagnostic workup including Brachyspira culture, Salmonella culture, Lawsonia intracellularis-specific, B. hyodysenteriae-specific and B. pilosicoli-specific multiplex PCR and postmortem examination of severely affected pigs was performed. Tests for Salmonella species, Lawsonia intracellularis and B. hyodysenteriae were all negative. Gross and microscopic lesions were in agreement with dysentery and spirochaetes could be demonstrated by silver staining in tissue samples of the caecum at the ileal papilla. B. suanatina was cultured from faeces or colon of all (five) animals sampled and identified using nox-RFLP, partial nox-gene-sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). According to the initial report from Scandinavia, B. suanatina can be isolated from birds and cross-species infection could be demonstrated infecting pigs with an avian isolate. Thus outdoor production as in the case presented here and international trade may pose a risk for infection of naive herds.

The reservoirs and the modes of transmission of the most frequent microsporidial species in humans, Enterocytozoon bieneusi , are still unknown. We have examined fecal samples of 26 humans and 350 animals from 37 species to find 18 samples containing this parasite from humans, cats, pigs, cattle, and a llama. Genotypic characterization of the internal transcribed spacer of the rRNA gene resulted in 14 different genotypes, 6 of them previously undescribed. Phylogenetic analysis revealed the lack of a transmission barrier between E. bieneusi from humans and animals (cats, pigs, and cattle). Thus, E. bieneusi appears to be a zoonotic pathogen.

A juvenile dwarf rabbit ( Oryctolagus cuniculus) with clinical signs of dyspnea and suspected ascites was submitted for necropsy. The main macroscopic findings were a watery red pleural effusion and some whitish striated foci in the lungs. In addition, there were multifocal scars in the cortex of the kidneys. The histologic examination of the lungs showed a severe granulomatous pneumonia with detection of acid-fast bacilli, in the kidneys, an interstitial chronic lymphoplasmacellular nephritis with interstitial fibrosis, and in the brain, a multifocal granulomatous and partly necrotizing encephalitis with detection of spores, suggestive of encephalitozoonosis. In the lungs, Mycobacterium genavense was verified by polymerase chain reaction and 16S ribosomal RNA gene sequencing. To our knowledge, this is the first report of an M. genavense infection in a rabbit, with the lungs being the only affected organ. Therefore, an aerogen infection seems to be the most contemplable way of infection.