Seasonal or pathological findings? Morphofunctional characteristics of the equine endometrium during the autumn and spring transition.

The deep anoestrous phase in winter is part of the anovulatory season in mares and is bordered by the autumn and spring transitional periods (ATP/STP). To define an annual time span for effective prognostic biopsy sampling, the aim of this study was to provide a morphofunctional characterization of the endometrium during ATP and STP. To outline both transitional periods, endometrial specimens were taken in September, October and November (n = 76) as well as February, March and April (n = 184) with the requirement of a detailed clinical documentation. Tissue samples were examined histologically with special emphasis on the functional endometrial morphology. Additionally, an immunohistochemical evaluation was performed on selected specimens regarding the expression of oestrogen receptor α, progesterone receptor and Ki67-antigen. An absent to low endometrial activity was ascertained in more than 60% of all specimens from late October onwards, whereas a comparably lacking or low activity in STP was observed until early April. Approximately 30% (ATP) to 22% (STP) of all samples exhibited a predominantly "irregular" endometrial differentiation. During the transitional periods, the clinically evaluated ovarian status (transrectal palpation, transrectal ultrasonography and/or serum progesterone and oestrogen analyses) and the endometrial functional morphology were in accordance with approximately 70% of all cases. The expression of steroid hormone receptors and Ki67-antigen was generally low. Given that endometrial maldifferentiations were frequently found during ATP and STP, its occurence might display a characteristic and physiological feature of the transitional periods. Regarding the functional endometrial morphology, a diagnostic biopsy sampling should therefore be performed between late April and before September.

Establishment and characterization of a coculture system of equine endometrial epithelial and stromal cells.

To investigate the equine endometrium as close to the in vivo situation as possible, we established a coculture system for epithelial and stromal cells (ECs/SCs). ECs and SCs were isolated from nine endometrial tissue specimens. ECs obtained as glandular formations were cultivated on one side of the semipermeable membrane of a Millicell® insert. After 2 days, SCs (2 × 104 cells/membrane) were seeded onto the other side of the same membrane. During cocultivation, the low serum containing culture medium (Theuß et al., 2010) was supplemented with different concentrations and combinations of 17ß-estradiol (2.0-3.0 pg/ml medium) and progesterone (0.5-15.0 ng/ml medium). Once the cocultures formed continuous cell layers as determined by phase-contrast microscopy, the membranes were fixed and processed for light microscopical examination. Cytokeratin 19, steroid hormone receptors and the uterine proteins uteroglobin and calbindinD9k were detected using immunocytochemistry to determine the degree of culture purity and functional cellular differentiation. The culture purity of the EC layer averaged ≥95%. Uteroglobin and calbindinD9k were consistently expressed in ECs, while hormone receptors were predominantly absent in both cell populations. An explicit cytomorphological epithelial differentiation with formation of round-oval to polygonal cell forms was encountered in ≤50% of all ECs and independent of supplemented steroids. Based on the findings altogether, and despite the partly absent congruence to the in situ prerequisites, we established a standardized and reproducible coculture system, which offers a basic approach for studies of physiologic and pathophysiologic issues in the mare.

[Pathology of South American Camelids: a retrospective study of necropsies at the Institute of Veterinary Pathology, University of Leipzig, Germany]. / Sektionsbefunde von Neuweltkameliden. Eine retrospektive Analyse des Sektionsgutes des Leipziger Instituts für Veterinär-Pathologie.

OBJECTIVE: The number of South American Camelids (New World Camelids) housed in Germany has increased in the recent years. While these species were formerly kept solely in zoological gardens, ever more private and commercial livestock is being established. Compared to indigenous livestock animals, they bear some distinctive differences, particularly in terms of digestive tract anatomy and physiology. Therefore, it is of considerable interest for veterinarians working with South American Camelids to obtain knowledge about the distinguishing features of these animals and the typical diseases affecting them in Germany. MATERIAL AND METHODS: For this purpose, the necropsy reports, including the anamnestic data, and their diagnostic usefulness, from 1995 to 2012 were studied retrospectively. RESULTS: Du- ring this period, a total of 233 New World Camelids were examined (195 alpacas and 38 llamas). Anamnestic data of diagnostic usefulness regarding the cause of disease were only submitted in a limited number of cases, because most of the animals died without specific symptoms. The following were the most frequent pathological findings: enteritis (n = 91), gastritis (n = 76), cachexia (n = 73), pneumonia (n = 30), stomatitis (n = 27), azotaemia (n = 22) and anaemia (n = 9). An endoparasitosis occurred in 107 cases and was considered the predominant cause of enteritis. CONCLUSION: As with indigenous ruminants, llamas and alpacas primarily suffered from diseases of the digestive and respiratory tracts. Other organ systems were affected to a lesser extent. CLINICAL RELEVANCE: Even in cases with severe alterations in the affected organs, South American Camelids do not show or show too late diagnostically indicative clinical symptoms. Therefore, a detailed clinical examination of these animals is important.

[Cowpox virus infection in an alpaca (Vicugna pacos) - clinical symptoms, laboratory diagnostic findings and pathological changes]. / Kuhpockenvirusinfektion bei einem Alpaka (Vicugna pacos) - klinische Symptomatik, Diagnostik und pathologische Befunde.

Orthopoxvirus infections appear to be rare in South American Camelids, because only a few cases have been reported in the literature. Based on a generalized infection with cowpox virus in an alpaca, the clinical symptoms, laboratory diagnostic findings and the pathological changes are described. The case history showed a long treatment because of chronic skin lesions. The main clinical symptom was miliary papules over the entire skin. Furthermore, a bilateral mucopurulent conjunctivitis occurred as well as excessive salivation due to a severe erosive-ulcerative stomatitis. Although the animal received intensive treatment, it died 8 days after admission to the clinic. During necropsy, an erosive-ulcerative laryngitis as well as a necrotising pneumonia and lymphadenitis were observed. Histopathological examination of representative organ samples led to the diagnosis of a suspected orthopoxvirus infection. Electron microscopy and quantitative polymerase chain reaction (qPCR) of tissue samples confirmed this diagnosis. The virus could be isolated in tissue culture and a PCR with subsequent nucleotide sequencing identified cowpox virus as the causative agent for this generalised infection.

The conclusiveness of less-invasive imaging techniques (computer tomography, X-ray) with regard to their identification of bone diseases in a primate model (Callithrix jacchus).

BACKGROUND: Although common marmosets seem to be appropriate animal models to examine bone diseases, no data about the conclusiveness of less-invasive techniques are available. Therefore, the aim was to combine different techniques to analyse changes in bone metabolism of common marmosets with bone diseases. METHODS: Five monkeys were examined by X-ray, computer tomography (CT), histology and immunohistochemistry (IHC). RESULTS: Monkeys with lowest bone mineral density (BMD) showed increased bone marrow, decreased cancellous bone and decreased contrast in X-ray. Highest alkaline phosphatase (AP)-levels were detected in bones with low elastic modulus. Expression of osteopontin (OPN), osteocalcin (OC) and runt-related transcriptions factor 2 (RUNX 2) was detected in bones with high modulus. No expression was present in bones with lower modulus. Collagen type I and V were found in every bone. CONCLUSIONS: In conclusion, CT, X-ray and AP are useful techniques to detect bone diseases in common marmosets. These observations could be confirmed by IHC.

Disseminated infection with Mycobacterium genavense in a grizzled giant squirrel (Ratufa macroura) associated with the isolation of an unknown Mycobacterium.

Disseminated infection with Mycobacterium genavense was diagnosed in an adult grizzled giant squirrel (Ratufa macroura). Microscopical examination showed granulomatous inflammation in the brain, kidneys, lungs and maxilla with intracellular acid-fast bacilli. M. genavense and a novel species of Mycobacterium (proposed name 'Mycobacterium lipsiensis') were identified.