Of rodents and ruminants: a comparison of small noncoding RNA requirements in mouse and bovine reproduction

Ruminants are major producers of meat and milk, thus managing their reproductive potential is a key element in cost-effective, safe, and efficient food production. Of particular concern, defects in male germ cells and female germ cells may lead to significantly reduced live births relative to fertilization. However, the underlying molecular drivers of these defects are unclear. Small noncoding RNAs, such as piRNAs and miRNAs, are known to be important regulators of germ-cell physiology in mouse (the best-studied mammalian model organism) and emerging evidence suggests that this is also the case in a range of ruminant species, in particular bovine. Similarities exist between mouse and bovids, especially in the case of meiotic and postmeiotic male germ cells. However, fundamental differences in small RNA abundance and metabolism between these species have been observed in the female germ cell, differences that likely have profound impacts on their physiology. Further, parentally derived small noncoding RNAs are known to influence early embryos and significant species-specific differences in germ-cell born small noncoding RNAs have been observed. These findings demonstrate the mouse to be an imperfect model for understanding germ-cell small noncoding RNA biology in ruminants and highlight the need to increase research efforts in this underappreciated aspect of animal reproduction.     

Retracted: Mycobacterium bovis BCG Danish Strain 1331 isolated from a periarticular lesion in a domestic cat

A 7-year-old male neutered domestic shorthair outdoor cat was referred for chronic left forelimb lameness, which had been treated with intra-articular injections of triamcinolone acetonide. A soft tissue swelling around the elbow joint, extending from the distal humerus to the proximal ulna, was surgically explored and biopsy samples obtained. Mycobacterium bovis was cultured from samples from the soft tissue and bone. The mycobacteria from the media were killed and the DNA extracted and tested on a multiplex real-time PCR for the absence of specific genes and the presence of mycobacterial genus markers. The PCR revealed bacillus Calmette-Guérin Danish Strain 1331; this was also isolated from the prescapular lymph node, muscle and bone, obtained at post mortem examination. Badgers had been vaccinated with the bacillus Calmette-Guérin vaccine SSI (Statens Serum Institute) in the area where the cat lived, in the spring and autumn of the previous year. To the authors' knowledge, this is the first report of infection with M. bovis bacillus Calmette-Guérin Danish Strain 1331 in a domestic cat, potentially associated with annual vaccination of badgers in the proximity of the cat's home.     

Characterization of primary pulmonary adenosquamous carcinoma‐associated pleural effusion

A 10‐year‐old, female spayed Shih Tzu was presented due to weight loss, increased respiratory effort and lethargy, determined to be secondary to a congenital para‐esophageal diaphragmatic defect with partial herniation of the stomach and spleen. Four days following reduction surgery of the displaced abdominal organs thoracic effusion developed. Thoracic fluid evaluation revealed a cell‐rich, protein‐poor modified transudate with neutrophils, reactive mesothelial cells, and atypical epitheloid cells which occasionally appeared to be keratinizing, consistent with neoplastic exfoliation. Thoracic effusion recurred 2 days later, with similar characteristics as the initial sample. Computed tomography (CT) indicated consolidation and displacement of the right middle and accessory lung lobes. Exploratory thoracic surgery demonstrated a thickened, hyperemic right middle lung lobe, and thickened pericardial diaphragmatic ligament. Histologic evaluation of these tissues identified a primary pulmonary adenosquamous carcinoma with intravascular and pleural invasion. Based on these cytologic, histologic, and clinical findings, we conclude that primary pulmonary carcinomas may involve superficial thoracic structures and exfoliate into a thoracic effusion.     

Diagnostic use of cytologic examination of bone marrow from dogs with thrombocytopenia: 58 cases (1994-2004)

OBJECTIVE: To determine the diagnostic use of cytologic examination of bone marrow from dogs with thrombocytopenia. DESIGN: Retrospective case series. ANIMALS: 58 dogs with thrombocytopenia. PROCEDURES: Medical records were searched and reviewed for dogs with thrombocytopenia. Dogs that had thrombocytopenia and cytologic examination of bone marrow were included in the study. Dogs with other hematologic abnormalities, with a previous diagnosis of hematopoietic neoplasia, or that had previous treatment with cytotoxic drugs were excluded. Bone marrow cytologic findings were reviewed. Results were compared between dogs with severe thrombocytopenia (< 20,000 platelets/microL) and dogs with mild to moderate thrombocytopenia (20,000 to 200,000 platelets/microL). RESULTS: 58 dogs met the inclusion criteria. Of 55 dogs with diagnostic bone marrow aspirates, 36 had severe thrombocytopenia. Cytologic evaluation of bone marrow did not reveal substantial nonmegakaryocytic bone marrow abnormalities or result in a definitive diagnosis in any of these dogs. Nineteen dogs with mild to moderate thrombocytopenia had diagnostic bone marrow aspirates. Bone marrow cytologic findings revealed nonmegakaryocytic abnormalities in 4 of these dogs. Significantly fewer dogs with severe thrombocytopenia had abnormalities identified on cytologic examination of bone marrow, compared with dogs with mild to moderate thrombocytopenia. CONCLUSIONS AND CLINICAL RELEVANCE: Cytologic examination of bone marrow is unlikely to provide specific diagnostic or prognostic information in dogs with severe thrombocytopenia.     

Multiplex real-time PCR for the detection and differentiation of equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4)

A multiplex real-time PCR was designed to detect and differentiate equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB) labelled with 6-carboxy-fluorescein (FAM) and VIC for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaherpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.     

Veterinary pathology in the United Kingdom: past, present, and future

This article presents a historical perspective on veterinary anatomic pathology in the United Kingdom from the late nineteenth century to the present. Prior to World War II, the specialty was a rather general one that also included bacteriology and parasitology and was only slightly affected by strong Germanic developments in cell and tissue pathology. The few notable figures of this era include John McFadyean, Sidney Gaiger, and J.R.M Innes. The specialty developed strongly in the second half of the twentieth century, led by a small number of individuals, and was greatly aided by the development of specialist colleges and residency training. Key individuals of this era include W.F. Blakemore, Ernest Cotchin, R.J.M. Franklin, W.F.H. Jarrett, A.R. Jennings, and A.C. Palmer. A remarkable feature of this period has been the increased employment of veterinary pathologists in biomedical industry and in private diagnostic laboratories. While standards of pathology practice have benefited from the college initiatives, there are major financial constraints on the availability of funded training posts in the United Kingdom, and there remain considerable shortages in the supply of pathologists trained to contemporary standards. The acknowledged professional and scientific importance of veterinary pathology needs to be translated into effective financial support for the training that underpins competence in this specialty. Further developments seem likely to be dominated by advances in the technology of tissue handling, applications of molecular biology to pathology, and greater use of telepathology in teaching, in quality assurance, and in continuing professional development.