Carp edema virus (CEV) was initially detected in Japan in the 1970s as the causative agent of koi sleepy disease (KSD) in koi and common carp (Cyprinus carpio L.) populations. CEV has spread all over the world mainly by trading of asymptomatic carrier fish. In Germany KSD was detected for the first time in spring 2009 from samples collected from imported Japanese koi at water temperatures below 10 degrees C. In 2014 large outbreaks of KSD in common carp and koi became evident in Germany. Several unsuccessful investigations were carried out to isolate the agent in permanently growing fish derived cell cultures. For diagnosis, the clinical signs of KSD were difficult to differentiate from the clinical signs related with koi herpesvirus disease (KHVD), except during the advanced stage of disease, when the fish started to lie down ("sleep") on the bottom of the tank. The only successful and reliable diagnostic methods were PCRs and qPCR. However, virus sequences obtained from diseased specimens demonstrated high geographical diversity. Therefore, the original primer pairs, published from Japan in the 1990s, gave negative or uncertain results in detection of CEV in samples from carp and koi in Europe, and new primers and probes had to be designed for PCR and nested PCR as well as for TaqMan and SYBR green based qPCRs. While some of them recognised CEV obtained from koi samples only, a decision was made to use only PCR and qPCR assays that detect all variants of CEV for diagnostic testing. Three molecular assays were chosen to conduct a laboratory comparison test with 10 Animal Health Laboratories all over Germany. The aim was to evaluate and standardise CEV detection with defined samples, thereby making the results directly comparable. The results show that even though all laboratories used different PCR kits and cycler systems, they were able to identify CEV in samples from koi and common carp with given primers and a probe. Only in samples diluted above 1:1000, were the results negative by PCR and qPCR in a few laboratories. Some participating laboratories also identified CEV from koi and carp samples by sequence analysis. These results confirm that accurate and reproducible CEV diagnostic results can be achieved resulting in both an improved control strategy, and the implementation of more effective biosecurity practices, for koi and carp farms. Additionally, it was shown that primers and the probe designed for PCR, nested PCR and qPCR are very sensitive and specific to CEV. The results were reproducible across multiple laboratories that utilised different manufacturers of both PCR kits and machines as well as varying assay conditions.

Koi herpesvirus (KHV) causes KHV disease (KHVD). The virus is highly contagious in carp or koi and can induce a high mortality. Latency and, in some cases, a lack of signs presents a challenge for virus detection. Appropriate immunological detection methods for anti-KHV antibodies have not yet been fully validated for KHV. Therefore, it was developed and validated an enzyme-linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs.

Aim: EBP causes large economic losses during the post weaning period in cattle breeding. Diagnostic procedures in calves include clinical examination and taking of nasal swabs for further microbiological examination. In a first study, exhaled breath analysis using gas chromatography - ion mobility spectrometry (GC-IMS) and measurement of respiratory mechanics by means of impulse oscillometry (IOS) complemented the conventionally used diagnostic procedures. Material and methods: Fourteen calves (age: between 50 and 103 days) suspected to suffer of EBP were included in the study. Results of different diagnostic procedures (clinical examination, bacteriological cultivation, IOS measurements and analysis of exhaled breath samples) were compared and related to each other. Results: Eight calves were diagnosed as infected by Pasteurella multocida. Analysis of exhaled breath by GC-IMS yielded 82 clusters of peaks representing volatile organic compounds (VOC). Peak areas of four clusters (c15, c17, c29 and c37) were significantly different (p < 0.05) between P. multocida infected and non-infected calves. Additionally, indole was detected in breath samples of P. multocida infected calves. Peak areas of three clusters (c20, c22 and c30) had significant (p < 0.05) relations to lung function parameters (Vt, Rex3, Xex7) measured by impulse oscillometry. Conclusion: This first study revealed differences in VOC between P. multocida infected and non-infected calves. However, some VOCs seem to correlate to impaired lung function, independent to the causative pathogen. Further research is necessary to assess the value of novel procedures in diagnosis of respiratory infectious diseases.

Bei der enzootischen Bronchopneumonie (EBP) des Kalbes handelt es sich um eine multikausal bedingte Erkrankung. Die Diagnostik im Stall besteht in der Durchführung einer klinischen Untersuchung und der Entnahme von Nasentupferproben zur Abklärung einer viralen oder bakteriellen Infektion. In dieser Studie sollte überprüft werden, ob Zusammenhänge zwischen flüchtigen organischen Verbindungen (VOC) in der ausgeatmeten Luft und den isolierten Krankheitserregern sowie zu klinischen und anderen Befunden bestehen.

Influences of different culture media and growth surfaces on morpho-functional characteristics of equine endometrial epithelial cells (EEC) in vitro were investigated in order to optimize culture conditions. Therefore, the results were compared to the endometrium in situ. Equine EEC were cultured using different media with varying serum components and additives: foetal bovine serum plus additives (MFBS+A), foetal bovine serum (MFBS), horse serum 1 (MHS1), horse serum 2 (MHS2) and horse serum 3 (MHS3). Uncovered and Matrigel (TM)-covered cell culture inserts, as well as cell culture flasks were used. Cultured cells were examined cytomorphologically (hoemoloun-eosin staining, giemsa staining), cytochemically (alcian blue staining) and immunocytochemically (oestrogen receptor (ER) alpha, progesterone receptor, proliferating cell nuclear antigen, inhibin-alpha, cytokeratin 8, 18, 19, vimentin, a-actin, desmin, transforming growth factor (TGF)-alpha, -beta 1, -beta 2, -beta 3). Further, tissue samples of the uteri from which the cells were isolated were examined histopathologically (hoemalaun-eosin staining), histochemically (alcian blue staining) and immunohistochemically, using the above-mentioned antibodies. Regardless of the used media and the state of the endometrial cycle at point of isolation, two morphologically different cell types and three growth patterns were found. None of these facts led to apparent differences concerning immunolabelling. Whereas growth surfaces, duration of culture and state of endometrial cycle at point of isolation did not cause any obvious influences on the immunolabelling of EEC in vitro concerning the utilized antibodies, the culture media did. Especially cells grown in MFBS+A exhibited characteristics comparable to uterine glandular epithelia in situ concerning cytokeratin 8, 18, 19, TGF-beta 1, -beta 3 and ER alpha.

A new method of in vitro culture of equine endometrial epithelial and stromal cells is described. After dissection, the endometrial tissue was dissociated in collagenase type II, sieved through nylon gauze and the different cell types were separated by centrifugation and differential adhesion. Cells were seeded at a density of 1x10(4) viable cells per cm(2) on several surfaces and cultured at 37 degrees C in humidified atmosphere. For both cell types the same serum-supplemented culture medium was utilized whereby confluent monolayer evolved after 10 to 14 days of incubation. Confluent cultures were dissociated using 0.25% trypsin-EDTA (stromal cells) or alfazyme (epithelial cells) and passaged up to 19 times. Further on, a successful cryoconservation-protocol could be established. With it, we created the precondition to establish a co-culture system for equine endometrial epithelial and stromal cells with the long-term objective to study growth characteristics, effects of steroid hormones and mutual influences between the cell populations, as well as potential pathogenetic factors and etiological influences on equine endometrosis.

This case report describes clinical signs, treatment and pathological findings of a 15-year-old pony more with acute necrotising pancreatitis which was presented due to continuous severe abdominal pain, anorexia, tachycardia (88/min), tachypnoea (68/min), increased body temperature (40,8 degrees C), abdominal distension, absence of abdominal sounds, anuria and no defecation. Increased peritoneal fluid and distended intestinal loops were seen sonographically. Biochemical analysis of venous blood resulted in hyperlipasaemia (>6000 U/l), hypertriglyceridaemio (29,75 mmol/l), hyperglycaemia (40,79 mmol/l) and elevated activities of GLDH- (993,3 U/I), AST- (2663,1 U/l), LDH- (2830 U/I) and GGT (458,8 U/l). Serum insulin was normal (12,1 mu U/ml). Peritoneal fluid was haemorrhagic and had increased activity of lipase (>36000 U/l). The patient was treated with crystalloid (4 ml NaC1 7,5%/kg BW and 100 ml NaCI 0,9%/kg BW) as well as colloidal infusions (10 ml hydroxyl ethyl starch 10%/kg BW) and with Flunixin meglumine (1,1 mg/kg BW iv.) as analgesic. Due to bad prognosis of general condition the pony was euthanized. Acute necrotising pancreatitis was diagnosed based on the pathologic-anatomical and -histological findings. The pancreas showed extended necrosis with extensive destruction of organ-specific structures and multifocal necrosis in the pancreatic region and mesentery fat. Acute pancreatitis often is an incidental finding during routine necropsy. Clinicians should consider pancreotitis as a differential diagnosis in horses presented with abdominal pain, fever and increased enzyme activity. Especially in horses with hypertriglyceridaemia and hyperglycaemia it is advisable to analyse concentrations of lipase as well as amylase in blood and peritoneal fluid.

This study aimed to characterize the composition and distribution of the extracellular matrix (ECM) components in normal canine mitral valves (MV) and in chronic heart valve disease (CVD). MV of 50 dogs (normal (n=9), mild (n=13), moderate (n=17), severe (n=11) CVD) were investigated macroscopically, histologically (H.-E., picrosirius red) and immunohistochemically (collagen I, III, IV, V, VI, elastin, laminin, fibronectin, heparan sulphate). In normal MV, ECM components were expressed in a typical layered pattern. In mild CVD, basement membrane components (laminin, collagen IV, fibronectin) were increased. Advanced CVD was characterized by myxomatous nodular lesions displaying a marginal and a central region comprised mainly of collagen I, VI and fibronectin in the former and collagen I and III in the latter. Collagen IV and laminin appeared multifocally in marked CVD. In conclusion, not only an accumulation of proteoglycans, but also a distinctly altered expression of basement membrane components, and collagens characterizes CVD.

Over a five-year-period, five horses underwent unilateral thyroidectomy. Individual data including medical history, physical, laboratory, endoscopic, and ultrasonographic findings, surgical technique, histopathological findings, complications, and outcome are described. Five horses, 14 to 18 years of age had an enlarged unilateral thyroid mass. On ultrasonogrophy the enlarged thyroids had either a heterogeneous (3 horses) or homogeneous (2 horses) appearance and were histopothologically characterized as thyroid carcinoma (in 4 horses). After the surgery no one of the horses had ipsilateral laryngeal hemiplegia. No recrudescence or metastases were observed 15 months to five years after surgery. Complications like ipsilateral laryngeal hemiplegia can be avoided by correct surgical techniques. Statements about the malignancy of such tumors by ultrasonographic examination are not appropriate.

Viral diseases represent precursors and primary factors of losses in horse breeding. While the main target of vaccination is the protection against Equine Herpes Virus (EHV) infection of type I and 4 there is only little attention paid to type 2. There have been several reports of EHV-2-outbreaks during the last three to four decades and the infection rate is assumed to be very high. During spring-time 2004 and 2008 a total of 13 foals of the year from different geographic regions of Germany have been diagnosed positive regarding EHV-2. Common features of clinical history have been progressive respiratory problems with dyspnoea and serous to mucopurulent discharge for about two weeks. Histopathologically varying stages of purulent to necrotizing pneumonia with intralesional detection of bacteria could be observed as well as a diffuse alveolar damage in other less affected regions. Although Clinical features are predominantly nonspecific, virological or serological investigations may give the chance to make the right decisions concerning an appropriate therapy to reduce losses in foal breeding.

Up to now, the caryophyllid cestode Atractolytocestus huronensis Anthony, 1958, a parasite of common carp, has attracted little attention in Germany. Based on recent publications from the Czech Republic and Hungary, it appears probable that this cestode may be increasingly common in Germany. There is a strong connection between the occurrence of A. huronensis and imports of common carp from the Czech Republic and southern Germany. Although in most cases no clinical alterations in parasitized carp have been observed, care should be taken to avoid further dissemination and to prevent possible losses in commercial pond farming.

In contrast to intestinal coccidia (Eimeria sp.), cyst-forming coccidia of the genus Sarcocystis have only rarely been described as a cause of disease in pet birds. Intra vitam diagnosis is often difficult to achieve due to the role of the bird as an intermediate host without exogenous stages. The course of the disease is commonly peracute and can lead to the sudden death of the animal without cyst formation in the muscle. Animals that survive the acute stage form tissue cysts which can grow up to several millimeters. The source of infection are the sporocysts in the faeces of the final (carnivorous) host. For the differential diagnosis Toxoplasma- or Leukozytozoon-infections have to be considered. The therapy of Sarcocystis-infections in usually restricted to surgical removal of cysts.