Endogenous retroviruses of sheep: a model system for understanding physiological adaptation to an evolving ruminant genome

Endogenous retroviruses (ERVs) are present in the genome of all vertebrates and are remnants of ancient exogenous retroviral infections of the host germline transmitted vertically from generation to generation. Sheep betaretroviruses offer a unique model system to study the complex interaction between retroviruses and their host. The sheep genome contains 27 endogenous betaretroviruses (enJSRVs) related to the exogenous and pathogenic Jaagsiekte sheep retrovirus (JSRV), the causative agent of a transmissible lung cancer in sheep. The enJSRVs can protect their host against JSRV infection by blocking early and late steps of the JSRV replication cycle. In the female reproductive tract, enJSRVs are specifically expressed in the uterine luminal and glandular epithelia as well as in the conceptus (embryo and associated extraembryonic membranes) trophectoderm and in utero loss-of-function experiments found the enJSRVs envelope (env) to be essential for conceptus elongation and trophectoderm growth and development. Collectively, available evidence in sheep and other mammals indicate that ERVs coevolved with their hosts for millions of years and were positively selected for biological roles in genome plasticity and evolution, protection of the host against infection of related pathogenic and exogenous retroviruses, and placental development.     

Stomach wall evaluation using helical hydro-computed tomography

In helical hydro-computed tomography (helical hydro-CT), water is used as a neutral luminal contrast medium together with intravenous iodine contrast medium for the diagnosis and staging of human gastric neoplasia. We evaluated the feasibility of helical hydro-CT in 11 healthy animals (nine dogs and two cats). Adequate uniform gastric distension was obtained with 30 ml water/kg body weight. Fourteen client-owned dogs and four cats with suspected or diagnosed gastric neoplasia then underwent helical hydro-CT followed by intravenous contrast medium administration. Focal thickening with moderate contrast enhancement was found in 10 dogs and 3 cats. The extent of the lesion was assessed easily in all these patients. Three dogs and one cat had a normal stomach wall. One dog had multifocal thickening of the antrum but no histopathologic diagnosis was made. Helical hydro-CT, followed by intravenous contrast medium administration, is a simple technique for assessing the stomach wall.     

Laparoscopic sterilization of Sardinia donkeys using an endoscopic stapler

OBJECTIVE: To describe a laparoscopic technique for sterilization of the male donkey using an endoscopic linear stapler (ELS). STUDY DESIGN: Clinical report. ANIMALS: Male Sardinia donkeys (n=6), aged 24-36 months. METHODS: Laparoscopic sterilization was performed under standing sedation or recumbent under general anesthesia. An endoscopic linear stapler was used to transect, and control hemorrhage of, the spermatic cord. Serum testosterone concentrations were measured preoperatively and at 3, 6, and 12 months after sterilization. One testis from each of 2 donkeys was collected at 12 months for histologic examination. RESULTS: Sterilization was successfully achieved. The only minor intraoperative complication experienced was hemorrhage from 1 spermatic cord stump, which was re-stapled. No other major short- or long-term complications occurred. Serum testosterone concentrations were < 15 ng/dL at 3, 6, and 12 months after surgery in all donkeys. Testicular atrophy without evidence of revascularization was observed in the 2 testes examined histologically. CONCLUSIONS: Sterilization of descended testes by intraabdominal laparoscopic transection of the spermatic cord is a simple practical method, with minimal intraoperative or postoperative complications. Testicular revascularization did not occur. CLINICAL RELEVANCE: Laparoscopic sterilization in donkey can be performed using an endoscopic linear stapler as an alternative to scrotal castration.     

Computed tomography-guided fine-needle aspiration and tissue-core biopsy of bone lesions in small animals

In humans, free-hand computed tomography (CT)-guided biopsy is an accurate method to obtain a tissue sample. There are only a few reports of this technique in veterinary medicine. In the present study, 21 dogs and two cats underwent a free-hand CT-guided tissue-core biopsy (17 animals) or fine-needle aspiration (six animals) of a bone lesion. Two out of 17 tissue-core samples were also cultured. All 17 tissue-core biopsy samples were diagnostic (accuracy of 100%). Five out of six aspirates were diagnostic (accuracy of 83.3%). The overall accuracy was 95.7%. In one aspirate, cytologic quality was insufficient containing only blood. No major complications were encountered. Fourteen neoplastic, two infectious and six benign lesions were diagnosed. CT examination after intravenous contrast medium added useful information to avoid large vessels and to biopsy-viable tissue. Free-hand CT-guided tissue-core biopsy and aspiration appears to be a safe and very accurate procedure for use in the diagnosis of bone-associated diseases in small animals.     

Fatal necrotizing fasciitis and myositis in a captive common bottlenose dolphin (Tursiops truncatus) associated with Streptococcus agalactiae.

A common bottlenose dolphin (Tursiops truncatus) was presented for necropsy after acute onset of gastrointestinal signs and cutaneous lesions that rapidly progressed to death. Gross and microscopic findings were characterized by locally extensive severe necrohemorrhagic fasciitis and cellulitis, and severe necrotizing myositis in the head and dorsocranial thorax, with numerous disseminated gram-positive cocci. Streptococcus agalactiae was isolated from the lesions and from visceral organs (liver and lung), and it was identified by standard microbiology techniques. This communication is the first report of necrotizing fasciitis in a marine mammal associated with S. agalactiae.     

Iron content in vegetative and reproductive organs of nectarine trees in calcareous soils during the development of chlorosis

We investigated for 2 years (1995–1996) the time course development of chlorosis and the variation of iron (Fe) content in vegetative and reproductive organs in two nectarine orchards planted with cv Spring Red and cv Stark Redgold on calcareous soils of the Po valley (Italy) with the final aim to evaluate possible tools for the early prognosis of Fe chlorosis and a more efficient fertilization management. Due to the withdrawal of Fe supply, floral Fe concentration significantly decreased in 1996 as compared with 1995 in cv Spring Red, but not in Stark Redgold. Correlation coefficients between Fe and chlorophyll (Chl) from the same leaves were always higher when Fe was considered as amount present per leaf or per unit of leaf area than as leaf dry weight. The fact that chlorotic and green leaves had similar Fe concentration could be explained by an overestimate of Fe in the chlorotic leaf as a consequence of a reduction of its size. However, the decrease of Chl concentration between 60 and 120 days after full bloom (DAFB) occurred while leaf Fe content generally increased, indicating that even during chlorosis development leaves were supplied with some iron. We therefore suggest that the development of chlorosis was associated with an inactivation of Fe in the leaf apoplast. In 1995, regardless the cultivar, floral Fe concentration and leaf Chl were never correlated. In 1996 floral Fe concentration was linearly related to leaf Chl recorded 60 and 120 DAFB in cv Spring Red only. Floral Fe concentration at full bloom 1996, regardless the variety, was linearly related to leaf Chl determined in spring of the previous year, suggesting that flower Fe concentration might be used for assessing the storage of iron during the previous season.     

Characterization of the in vitro core surface proteome of Mycoplasma mycoides subsp. mycoides, the causative agent of contagious bovine pleuropneumonia

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides (Mmm) is a severe cattle disease, present in many countries in sub-Saharan Africa. The development of improved diagnostic tests and vaccines for CBPP control remains a research priority. Polyacrylamide gel electrophoresis and mass spectrometry were used to characterize the Triton X-114 soluble proteome of nine Mmm strains isolated from Europe or Africa. Of a total of 250 proteins detected, 67 were present in all strains investigated. Of these, 44 were predicted to be lipoproteins or cytoplasmic membrane-associated proteins and are thus likely to be members of the core in vitro surface membrane-associated proteome of Mmm. Moreover, the presence of all identified proteins in other ruminant Mycoplasma pathogens were investigated. Two proteins of the core proteome were identified only in other cattle pathogens of the genus Mycoplasma pointing towards a role in host–pathogen interactions. The data generated will facilitate the identification and prioritization of candidate Mycoplasma antigens for improved control measures, as it is likely that surface-exposed membrane proteins will include those that are involved in host–pathogen interactions.     

COMPUTED TOMOGRAPHIC CHARACTERISTICS OF EOSINOPHILIC PULMONARY GRANULOMATOSIS IN FIVE DOGS

Canine pulmonary eosinophilic granulomatosis is a rare inflammatory pulmonary disease characterized by formation of eosinophilic granulomas that tend to obliterate the normal pulmonary architecture. The purpose of this retrospective study was to describe the CT characteristics of confirmed idiopathic pulmonary eosinophilic granulomatosis in a group of dogs. Five dogs met inclusion criteria. All patients were young adult dogs of variable breeds. No dog had concurrent occult heartworm disease. Computed tomographic characteristics most commonly included pulmonary masses and nodules of variable size, and lesions were most commonly located in the caudal lung lobes. Four dogs had large pulmonary masses with or without additional nodules and one dog had nodular lesions disseminated throughout the entire lung parenchyma. All large eosinophilic granulomas were smoothly margined, heterogeneous pulmonary masses displaying heterogeneous contrast enhancement. A honeycomb‐like enhancement pattern was observed in all but one mass and consisted of multiple hyperattenuating rims delineating central hypoattenuating areas, suggestive of bronchiectatic lung with peripheral enhancing airway walls and fluid‐filled, necrotic bronchial lumen. One dog had evidence of tracheobronchial lymphadenopathy. Findings indicated that canine eosinophilic pulmonary granulomatosis should be included as a differential diagnosis for dogs with CT characteristics of multiple pulmonary masses and/or nodules in caudal lung lobes, and a honeycomb‐like enhancement pattern in masses after intravenous administration of iodinated contrast medium.     

Fork reversal and ssDNA accumulation at stalled replication forks owing to checkpoint defects

Checkpoint-mediated control of replicating chromosomes is essential for preventing cancer. In yeast, Rad53 kinase protects stalled replication forks from pathological rearrangements. To characterize the mechanisms controlling fork integrity, we analyzed replication intermediates formed in response to replication blocks using electron microscopy. At the forks, wild-type cells accumulate short single-stranded regions, which likely causes checkpoint activation, whereas rad53 mutants exhibit extensive single-stranded gaps and hemi-replicated intermediates, consistent with a lagging-strand synthesis defect. Further, rad53 cells accumulate Holliday junctions through fork reversal. We speculate that, in checkpoint mutants, abnormal replication intermediates begin to form because of uncoordinated replication and are further processed by unscheduled recombination pathways, causing genome instability.