Acute phase protein levels in dogs with mast cell tumours and sarcomas

The acute phase proteins (APP) form part of a non-specific host response to inflammation. They may be induced by a range of different causes, including infection, inflammation, cancer and trauma. As they form part of the earliest response to such insults, they have potential for early identification of disease. In people, APP levels have been shown to correlate both with the extent of disease and also the prognosis in several forms of neoplasia, including prostate, oesophageal and colorectal cancer. As such, they can be used as prognostic and monitoring tools. To date, similar studies in veterinary patients have been limited, largely retrospective in nature and many are non-specific for tumour type. The purpose of this study was to evaluate a panel of four APPs in dogs with naturally occurring mast cell tumours (MCTs) and sarcomas to identify in the first instance whether increased levels of individual APPs, or identifiable combinations of APPs, was linked with the presence of disease. In the patients with MCTs, C-reactive protein (CRP) and α-1 acid glycoprotein levels increased, with a concurrent drop in serum amyloid A levels. In the sarcoma patients, CRP, α-1 acid glycoprotein and haptoglobin were increased. These findings suggest that specific solid tumour types in dogs may be associated with specific changes in APP profiles.     

Inflammatory polyps of the middle ear in 5 dogs

OBJECTIVE: To describe inflammatory polyps of the middle ear in 5 dogs. STUDY DESIGN: Case series. ANIMALS: Five dogs with ear disease. METHODS: Medical records (1995-2001) were reviewed to identify dogs with inflammatory polyps of the middle ear. Signalment, clinical signs, ancillary diagnostic procedures, treatment, postoperative complications, and outcome were recorded. Owners and referring veterinarians were contacted to document outcome. RESULTS: Dogs with inflammatory polyps of the middle ear were male and aged 4 to 13 years. Two dogs had bilateral polyps, whereas 3 had unilateral polyps. The most common clinical presentation was otitis externa and media, with radiographic evidence of otitis media. Polyps were treated by ventral bulla osteotomy (VBO) in 1 dog and total ear canal ablation with lateral bulla osteotomy (TECA-LBO) in 4 dogs. Polyps consisted of a fibrovascular stroma infiltrated with neutrophils, macrophages, lymphocytes, and plasma cells. The overlying epithelium was frequently ulcerated. Immediate postoperative complications included a seroma after VBO (1 dog) and transient unilateral facial nerve paralysis after bilateral TECA-LBO (1 dog). No recurrence occurred within 9 to 69 months. CONCLUSIONS: Unilateral or bilateral, inflammatory polyps can occur in the middle ear of dogs in association with otitis externa and media. No recurrence occurred after surgical removal of the polyps. CLINICAL RELEVANCE: Inflammatory polyps of the middle ear in dogs can be a cause of otitis externa/media. Surgical removal of aural polyps has a good prognosis.     

Barrier pressure at the gastroesophageal junction in anesthetized dogs.

OBJECTIVE: To evaluate the effect of body position on barrier pressure at the gastroesophageal junction in anesthetized Greyhounds and to assess alterations in barrier pressure following gastropexy. ANIMALS: 8 adult Greyhounds. PROCEDURE: Barrier pressure at the gastroesophageal junction was measured by fast (1 cm/s) and slow (1 cm/10 s) withdrawal of a subminiature strain gauge transducer through the gastroesophageal junction in 8 anesthetized dogs. The effect of body position was measured. Each dog then was placed in right-lateral recumbency, and gastropexy was performed in the left flank. Additional measurements were obtained 1, 5, 10, 20, and 30 minutes after gastropexy. RESULTS: Barrier pressure for dogs positioned in sternal recumbency (mean +/- SEM, 1.1 +/- 0.53 mm Hg) was significantly less than for dogs positioned in right lateral or left lateral recumbency. Following gastropexy, there was a steady increase in barrier pressure. Thirty minutes after gastropexy, barrier pressure was significantly higher (13.36 +/- 3.46 mm Hg), compared with the value before surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Barrier pressure in anesthetized dogs is highly variable and influenced by body position. This is most likely the result of anatomic interrelationships between the diaphragm, stomach, and terminal portion of the esophagus. Gastropexy also increases barrier pressure in the immediate postoperative period, which may be clinically relevant in terms of understanding how resolution of gastroesophageal reflux disease associated with hiatal hernia may be affected by gastropexy combined with hernia reduction.     

Pneumonyssoides species infestation in two Pekingese dogs in the UK

Two male, neutered, Pekingese dogs aged four years and 12 years were presented for acute-onset nasal pruritus and sneezing following a visit to a beach in northern Scotland. Routine nasal investigations revealed the presence of the canine nasal mite Pneumonyssoides both by direct visualisation and histopathologically. Resolution of clinical signs was observed following selamectin treatment. To the authors' knowledge, this report describes the first cases of Pneumonyssoides infestation in non-travelled UK dogs.     

Developmental Capabilities of Embryos Produced In Vitro from Prepubertal Lamb Oocytes

Breeding from prepubertal females, known as juvenile in vitro embryo transfer (JIVET), reduces the generation interval and increases the rate of genetic gain in animal breeding programs. While the birth of the first lambs from prepubertal ewes occurred nearly 30 years ago; and there is considerable interest in the commercialization of this technology, its efficiency remains too low. The advent of in vitro production (IVP) of embryo resulted in the more widespread use of JIVET. Morphologic and metabolic differences coupled with reduced in vitro and in vivo development of oocytes derived from prepubertal animals have been reported. Research has been undertaken to optimize donor selection and hormone stimulation methods in an attempt to reduce the variability and increase the proportion of donors responding to hormone stimulation and increase oocyte developmental competence. Yet, this variation persists and the development of oocytes and embryos from prepubertal animals remains reduced when compared with adults. Recent improvements to JIVET, resulting from a modified hormone stimulation regime, have eliminated the failure of donors to respond to hormone stimulation, and increased both the number and developmental competence of oocytes harvested from very young prepubertal lambs. This increased efficiency has facilitated the incorporation of other reproductive technologies such as sperm sexing with JIVET, resulting in the birth of lambs of a pre-determined sex from prepubertal lambs. Increased rates of genetic gain in sheep breeding programs can be achieved by combining sexed sperm with oocytes obtained from lambs as young as 3–4 weeks of age. Continued increases in the efficiency of JIVET resulting from further improvements to hormone stimulation regimes and an increased understanding of the differences between oocytes from adult and prepubertal animals will result in the commercialization of this technology.     

Proposal for a unified nomenclature for target‐site mutations associated with resistance to fungicides

Evolved resistance to fungicides is a major problem limiting our ability to control agricultural, medical and veterinary pathogens and is frequently associated with substitutions in the amino acid sequence of the target protein. The convention for describing amino acid substitutions is to cite the wild‐type amino acid, the codon number and the new amino acid, using the one‐letter amino acid code. It has frequently been observed that orthologous amino acid mutations have been selected in different species by fungicides from the same mode of action class, but the amino acids have different numbers. These differences in numbering arise from the different lengths of the proteins in each species. The purpose of the present paper is to propose a system for unifying the labelling of amino acids in fungicide target proteins. To do this we have produced alignments between fungicide target proteins of relevant species fitted to a well‐studied ‘archetype’ species. Orthologous amino acids in all species are then assigned numerical ‘labels’ based on the position of the amino acid in the archetype protein. © 2016 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Prevalence of the AMHR2 mutation in Miniature Schnauzers and genetic investigation of a Belgian Malinois with persistent Müllerian duct syndrome

Persistent Müllerian duct syndrome (PMDS) is a sex‐limited disorder in which males develop portions of the female reproductive tract. Important consequences of PMDS are cryptorchidism and its sequelae of infertility and increased risk of testicular cancer. Anti‐Müllerian hormone (AMH) and its receptor (AMHR2) induce the regression of the Müllerian ducts in male embryos. In Miniature Schnauzer dogs, the genetic basis has been identified as an autosomal recessive nonsense mutation in AMHR2, but the allele frequency of the mutation is unknown. Thus, the primary objective of this study was to estimate the prevalence of the AMHR2 mutation in North American Miniature Schnauzers, in order to ascertain the value of genetic testing in this breed. An additional objective was to determine whether mutations in AMH or AMHR2 were responsible for PMDS in a Belgian Malinois; this would aid development of a genetic test for the Belgian Malinois breed. Genomic DNA from 216 Miniature Schnauzers (including one known PMDS case) was genotyped for the AMHR2 mutation, and DNA from a single PMDS‐affected Belgian Malinois was sequenced for all coding exons of AMH and AMHR2. The Miniature Schnauzer cohort had an AMHR2 mutation allele frequency of 0.16 and a carrier genotypic frequency of 0.27. The genetic basis for PMDS in the Belgian Malinois was not determined, as no coding or splicing mutations were identified in either AMH or AMHR2. These findings support a benefit to AMHR2 mutation testing Miniature Schnauzers used for breeding or with cryptorchidism.     

Chlamydiaceae in riverine buffalo (Bubalus bubalis) and cows (Bos taurus) in Egypt with and without signs of reproductive disease

Abstract

AIMS: To obtain information and compare the prevalence of Chlamydiaceae in riverine buffalo (Bubalus bubalis) and cows (Bos taurus) in Egypt with and without clinical signs of reproductive disease.

METHODS: Vaginal swabs and blood samples were collected from animals attending Governmental Veterinary Clinics without (buffalo n=39, cows n=20) and with (buffalo n=63, cows n=53) signs of reproductive disease. Serum samples were tested for antibodies to Chlamydiaceae using complement fixation testing (CFT). Vaginal swabs were tested for Chlamydiaceae following inoculation into Vero cells and 6-day-old embryonated chicken eggs, using modified Giménez and immunoperoxidase staining, PCR analyses targeting the omp2 gene, and Restriction Fragment Length Polymorphism PCR (RFLP-PCR) for species identification.

RESULTS: Antibodies to Chlamydiaceae were detected in 30/39 (77%) and 50/63 (79%) buffalo without and with signs of reproductive disease, respectively, and 10/20 (50%) and 39/53 (74%) of cows with and without signs of reproductive disease, respectively. Positive samples from PCR analysis were identified in 31/39 (79%) and 37/63 (59%) buffalo without and with signs of reproductive disease, respectively, and 12/20 (60%) and 46/53 (89%) of cows without and with signs of reproductive disease, respectively. Using RFLP-PCR, 57/68 (84%) of samples from buffalo, and 47/58 (81%) from cows, were identified as Chlamydophila psittaci and the reminder as Cp. abortus. From the CFT and PCR results there was no significant difference in the prevalence of positive samples between species, or between animals without or with signs of reproductive disease.

CONCLUSION: The presence of anti-Chlamydiaceae antibodies in 77% of the animals with signs of reproductive disease and the detection of Chlamydiaceae in 72% of vaginal swabs of the animals suggest a pathogenic role by Chlamydiaceae in riverine buffalo and cows. The main Chlamydiaceae found in the genital tract of cattle in Egypt were Cp. psittaci and Cp. abortus.

CLINICAL RELEVANCE: Chlamydophila spp. should be included in diagnostic algorithms for reproductive disorders, in order to assess the real burden of Chlamydophila associated disease in buffalo and cattle and to evaluate the potential value of vaccines.     

Pregnancy status determination in mares using a rapid lateral flow test for measuring serum oestrone sulphate

AIMS: To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point.

METHODS: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visible detection reagent. Concentrations of the coating antigen and OS monoclonal antibody were optimised so that the working range would allow pregnancy status to be determined from a visual end-point. The test was run by adding 0.1 ml serum to the sample well of a plastic cassette encasing the test membrane. As the serum migrated along the membrane, a test dot and control line were generated on it within 5–10 min. The intensity of the test dot was inversely proportional to the concentration of OS in the serum sample being tested. Results were compared with those from a validated OS enzyme immunoassay (EIA) and subsequent foaling or return to oestrus of the mares.

RESULTS: Serum samples with OS concentrations <10 ng/ml, indicative of non-pregnancy in mares >100 days post-mating, generated a test end-point consisting of a highly visible test dot and control line, whereas serum OS concentrations >50 ng/ml, indicative of pregnancy, generated a control line only. The test correctly identified 384/389 (98.7%) non-pregnant mares tested, and 303/312 (97.1%) pregnant mares tested that were >100 days post-mating. The lateral flow test devices were stable for at least 12 months when stored at 4°C, sealed in aluminium pouches with desiccant.

CONCLUSION: This novel, rapid, easy-to-use, lateral flow immunoassay offers a practical alternative to traditional laboratory-based immunoassays for measuring serum OS concentra- tions in mares for determining their pregnancy status.