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Biomedical subjects

M J Day

Publications and source records attributed to M J Day.

At least 37 records · Page 2Linked to original sources
PubMed2005

Detection of allelic variants of the canine IGHA gene by fluorescence resonance energy transfer melting temperature examination.

The fluorescence resonance energy transfer (FRET) dual hybridisation probe system has been used for the detection of the accumulation of target DNA during real-time PCR and for the identification of nucleotide polymorphisms through examination of melt curves. This system involves the use of two oligonucleotide probes which are located close to each other and are complementary to an internal segment of a target DNA of interest. Four allelic variants of the gene encoding the hinge region of the immunoglobulin A (IgA) heavy chain (IGHA) have been so far identified in the dog and this variability is due to a combination of single nucleotide polymorphisms and insertion/deletion of nucleic acid motifs. An individual dog may be homozygous or heterozygous for these allelic variants. The purpose of this study was to develop a FRET-based dual probe melting temperature assay to identify the alleles present within an individual dog and to use this assay to determine the frequency of the four allelic variants in different breeds within the canine population. A single pair of oligonucleotide probes were designed that were able to discriminate between the four allelic variants in both homozygous and heterozygous individuals. The genotype of 96 DNA samples obtained from various purebreeds of dogs was determined using this FRET assay. The frequency of each allele differed between the breed groups. The results of this study indicate that it is possible to distinguish relatively complex gene polymorphisms using a single set of oligonucleotide probes. Furthermore, any future comparison of IGHA genotypes between normal and diseased dog populations must take into account the breed variation in allelic frequency.

Alleles↗
PubMed2005

Neutrophilic dermatitis and immune-mediated haematological disorders in a dog: suspected adverse reaction to carprofen.

This report describes the clinical and pathological findings of a suspected idiosyncratic adverse drug reaction in a young dog. The patient presented with sudden onset, severe skin lesions together with episodes of collapse. Investigations revealed a neutrophilic dermatitis with vasculitis, immune-mediated haemolytic anaemia and thrombocytopenia. Similar pathology has been described in human cases of Sweet's syndrome. The chronology of events suggested an adverse drug reaction to carprofen, although two antibiotics had been prescribed within the dog's recent history. Lymphocyte transformation tests were performed and tended to exclude both antibiotics as the cause of the reaction. To the authors' knowledge, lymphocyte transformation tests have not previously been described with regard to drug hypersensitivity assessment in the veterinary literature, and this is the first peer-reviewed case report of neutrophilic dermatitis and vasculitis with immune-mediated haemolytic anaemia and thrombocytopenia occurring as a suspected adverse drug reaction to carprofen in the dog.

Adverse Drug Reaction Reporting Systems↗
PubMed2005

Effect of intestinal inflammation on fecal elastase concentration in dogs.

BACKGROUND: A commercially available ELISA kit for fecal elastase measurement can be used in the diagnosis of exocrine pancreatic insufficiency (EPI) in dogs. However, other causes of diarrhea also may affect fecal elastase concentration. OBJECTIVE: This study was undertaken to determine whether intestinal inflammation alters fecal elastase concentration in dogs. METHODS: Fecal elastase concentration was measured with an ELISA kit in the following groups of dogs: group 1 (n=16), control dogs, without gastrointestinal disease; group 2 (n=14), dogs with diarrhea and no histopathologic evidence of intestinal inflammation; and group 3 (n=12), dogs with diarrhea and histopathologic evidence of intestinal inflammation. Serum trypsin-like immunoreactivity (TLI) was determined in dogs with diarrhea to rule out EPI. RESULTS: All dogs in groups 2 and 3 had serum TLI concentrations >5 microg/L, ruling out EPI. No statistically significant difference was found in fecal elastase concentration among the 3 groups of dogs (P=.969). CONCLUSIONS: The results indicate that intestinal inflammation does not affect fecal elastase concentration, such that test results may be used to exclude a diagnosis of EPI even in animals with inflammatory bowel disease.

Animals↗
PubMed2004

Investigation of nasal disease in the cat--a retrospective study of 77 cases.

A retrospective study was undertaken to determine the prevalence of different diseases in cats referred for investigation of chronic nasal disease, to identify historical, clinical and diagnostic features which may assist in making a diagnosis, and to provide information pertaining to outcome in these cats. Diagnoses included neoplasia (30 cases), chronic rhinitis (27), foreign body (8), nasopharyngeal stenosis (5), Actinomyces infection (2), nasal polyps (2), stenotic nares (2), and rhinitis subsequent to trauma (1). The most common neoplasia was lymphosarcoma (21 cases), with a median survival of 98 days for cats treated with multiagent chemotherapy. Cats with neoplasia were older on average than the other cats, and were more likely to be dyspnoeic and have a haemorrhagic and/or unilateral nasal discharge than cats with chronic rhinitis. Cats with neoplasia were more likely to have radiographic evidence of nasal turbinate destruction, septal changes, or severe increases in soft tissue density than cats with chronic rhinitis. It was unusual for cats with diseases other than neoplasia to be euthanased as a result of their nasal disease.

Animals↗
PubMed2004

Diagnosis of feline haemoplasma infection in Australian cats using a real-time PCR assay.

A total of 147 cats from the Sydney area of Australia that had blood samples submitted to veterinary laboratories were tested using a real-time polymerase chain reaction (PCR) assay able to detect and distinguish the two feline haemoplasma species. This sample number included two cats diagnosed with feline haemoplasma infection by routine blood smear examination. Statistical analysis was performed to evaluate associations between haemoplasma infection, age, sex, breed, haematocrit (HCT) values and anaemia status. One hundred and six cats (72.1%) were negative. Thirty-four cats (23.1%) were positive for 'Candidatus M. haemominutum', six cats (4.1%) were positive for M. haemofelis and one cat (0.7%) was positive for both species. Older, male, non-pedigree cats, with lower HCT values were more likely to be infected with 'Candidatus M. haemominutum'. Significant inverse correlation was found between the amount of M. haemofelis DNA present in the blood and the HCT value. This report documents the existence of, and prevalence of, both haemoplasma species in a sample of cats in Australia and is the first to use quantitative real-time PCR in a prevalence study for haemoplasma infection.

Age Factors↗
PubMed2004

Real-time RT-PCR: considerations for efficient and sensitive assay design.

Real-time RT-PCR has been recognised as an accurate and sensitive method of quantifying mRNA transcripts. Absence of post amplification procedures allows rapid analysis with a greater sample throughput, yet with less risk of amplicon carry-over as reaction tubes are not opened. In order to maximise sensitivity, careful reaction design and optimisation is essential. Several aspects of assay design for real-time RT-PCR are discussed in this paper. We demonstrate the effect of amplicon secondary structure on reaction efficiency and its importance for primer design. Taq-man probes with a deoxyguanosine base at the 5' end fluoresce weakly when labelled with FAM, although weak fluorescence is not a problem when probes are labelled with Texas Red. DNA contamination of RNA samples purified using silica membrane columns is a significant problem but DNase digestion can be used to reduce this, particularly in-solution. MMLV and AMV enzyme systems using a variety of RT priming methods are compared and the problem of primer-dimer formation associated with RT enzymes is described.

Animals↗
PubMed2004

A retrospective study of clinical findings, treatment and outcome in 143 dogs with pericardial effusion.

A retrospective study of 143 dogs with pericardial effusion is presented, including a statistical analysis of survival time. Cases were classified into those in which a mass was seen on echocardiography (echo-positive) and those in which no mass could be identified (echo-negative). Forty-four dogs were echo-positive and 99 were echo-negative. The median survival time (MST) was 1068 days for echo-negative dogs and 26 days for echo-positive dogs. Dogs with a history of collapse were more likely to present with a mass on echocardiography. Those presenting with collapse had an MST of 30 days compared with 605 days for those without collapse. Echo-negative dogs tended to present with ascites and generally had a larger volume of pericardial effusion. The median survival for dogs presenting with ascites was 605 days compared with 45 days for those without ascites. Among echo-negative dogs, 64 per cent had a relapse of their effusion. Subtotal pericardiectomy was performed in 31 echo-negative dogs. The procedure had a perioperative mortality of 13 per cent but provided a favourable long-term prognosis. Dogs undergoing pericardiectomy had a median survival of 1218 days compared with 532 days for those not undergoing surgery.

Animals↗
PubMed2004

Measurement of immunoglobulin concentrations in the feces of healthy dogs.

Selective immunoglobulin A (IgA) deficiency is the most common primary immunodeficiency in humans and may be associated with chronic gastrointestinal disease. This observation has led to the suggestion that the high susceptibility of German shepherd dogs (GSD) to chronic enteropathies is related to a deficiency in mucosal IgA production. Relative deficiencies of IgA has been reported in the serum, saliva, tears, and feces of GSD both with and without alimentary disease; however, the findings of different studies are not consistent. The aim of this study was to confirm whether a relative deficiency of IgA exists in the feces of GSD. Feces were collected from healthy GSD (n = 209), Labrador retrievers (n = 96), beagles (n = 19), and miniature schnauzers (n = 32). Fecal IgA, IgM, and IgG were measured by capture enzyme-linked immunosorbent assays. Fecal IgG concentrations in the four breed groups were not significantly different. IgA concentrations were significantly greater in miniature schnauzers than in GSD (P = 0.0003) and Labradors (P = 0.0004) but not significantly different from those in beagles. IgM concentrations were significantly greater in miniature schnauzers than in GSD (P < 0.0001), Labradors (P < 0.0001), and beagles (P = 0.0098). These findings do not support the hypothesis that GSD have a relative deficiency in fecal IgA. The differences in immunoglobulin concentrations measured from a single defecation, between individuals of the same breed and between breeds, as well as the lack of an internal control molecule, make the determination of a normal reference range for all dogs impossible. Therefore, the usefulness of fecal immunoglobulin quantification for the assessment of intestinal immunoglobulin secretion in dogs is limited.

Animals↗
PubMed2003

Retrospective study of 25 young weimaraners with low serum immunoglobulin concentrations and inflammatory disease.

Twenty-five weimaraners with recurrent infections or inflammatory disease were investigated; their median age was four months (range two to 36 months), and 11 of them were male and 14 female. Twenty of them showed signs of lethargy, anorexia or pyrexia, 13 had been vomiting or had diarrhoea, 12 had shown signs of pain in the joints or bones and been lame, five had had reactions at the site of an injection, five had generalised lymphadenopathy, three had urinary tract infections and two had recurrent or severe pyoderma. They all had a lower concentration of one or more classes of serum immunoglobulin (IgG, IgM and IgA) than the standard control ranges, and their mean concentration of IgG was significantly lower (P<0.005) than the mean concentration of IgG in 15 clinically normal weimaraners. Of 10 cases for which a complete vaccination history was available, nine had developed clinical signs within five days of being vaccinated. Follow-up data were available from 21 of the 25 dogs for a median period of 24.5 months. One dog died during a symptomatic episode, three were euthanased, six were alive at follow-up but had continued to show clinical signs and 11 had made a full recovery.

Animals↗
PubMed2003

Serum IgE and IgG responses to food antigens in normal and atopic dogs, and dogs with gastrointestinal disease.

In human food allergy, with or without concurrent atopy, there may be significant increases in serum allergen-specific IgE. Serological methods have been tried but are not currently recommended for diagnosis of suspected food allergy in dogs. The aim of this study was to investigate humoral immune responses to food antigens in dogs. Serum IgG and IgE antibodies specific for food antigens were measured by enzyme linked immunosorbent assay (ELISA) using polyclonal anti-dog IgG and IgE reagents. Antigens tested were beef, chicken, pork, lamb, chicken, turkey, white fish, whole egg, wheat, soybean, barley, rice, maize corn, potato, yeast and cow's milk. Three groups were examined: normal dogs, dogs with atopic dermatitis (AD); and dogs with one of four types of gastrointestinal (GI) disease: small intestinal bacterial overgrowth (SIBO), inflammatory bowel disease (IBD), food-responsive disease, and infectious diarrhoea. Statistically significant differences in food-specific antibodies were not detected between the GI subgroups. There were statistically significant differences in the IgE concentration between the normal dogs, and dogs with atopic or GI disease, for all of the antigens tested. There were statistically significant differences in the average IgG concentrations between the normal dogs, and dogs with atopic or GI disease, for all of the antigens tested, except egg and yeast. The relationship of antigen responses for pooled data was analysed using principle component analysis and cluster plots. Some clustering of variables was apparent for both IgE and IgG. For example, all dogs (normal and diseased) made a similar IgG antibody response to chicken and turkey. Compared with other groups, atopic dogs had more food allergen-specific IgE and this would be consistent with a Th(2) humoral response to food antigens. Dogs with GI disease had more food allergen-specific IgG compared with the other groups. This may reflect increased antigen exposure due to increased mucosal permeability which is a recognised feature of canine intestinal disease.

Animals↗
PubMed2003

Use of a PCR assay to assess the prevalence and risk factors for Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in cats in the United Kingdom.

Blood samples from 426 healthy and sick cats in the UK were tested in a PCR assay for 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemofelis (basonym Haemobartonella felis). Seventy-two of the cats (16.9 per cent) were positive for 'Candidatus M. haemominutum' alone, six (1.4 per cent) were positive for M. haemofelis alone and one (0.2 per cent) was positive for both. Logistic regression analysis indicated that older male cats were significantly more likely to be infected with 'Candidatus M. haemominutum', but there was no significant association between it and any of the haematological variables measured. M. haemofelis infection was uncommon in the anaemic cats sampled, and there were too few positive cases for multivariable analysis to be performed for M. haemofelis-positive status.

Animals↗
PubMed2003

IgG subclass responses in a longitudinal study of canine visceral leishmaniasis.

Visceral leishmaniasis (VL), caused by Leishmania infantum, is an important disease of domestic dogs. Here, we present data on the IgG subclass antibody response to crude L. infantum antigen in a cohort of naturally infected Brazilian dogs. Specific IgG1-IgG4 responses could be detected in 98, 58, 70 and 82%, respectively of 57 dogs that were seropositive for specific IgG. Levels of all IgG subclasses were strongly inter-correlated. Levels of all IgG subclasses increased at the time of seroconversion, before reaching a plateau after several months. Levels of all IgG subclasses were higher in sick dogs than healthy dogs, and levels of all except IgG2 were higher in parasite-positive (by PCR) than parasite-negative dogs. However, levels of IgG2 relative to IgG1 were lower in sick or parasite-positive dogs compared to healthy or parasite-negative infected dogs. In contrast to previous studies, the results suggest that canine VL is associated with upregulation of specific antibody of all IgG subclasses, particularly IgG1, IgG3 and IgG4.

Animals↗
PubMed2003

Salivary and serum immunoglobulin levels in cats with chronic gingivostomatitis.

The salivary and serum concentrations of immunoglobulins G, M and A (IgG, IgM and IgA), and the salivary concentrations of albumin were measured by ELISA in 30 cats with chronic gingivostomatitis and 32 healthy cats. The cats with chronic gingivostomatitis had significantly higher salivary concentrations of IgG, IgM and albumin, and higher serum concentrations of IgG, IgM and IgA, but significantly lower salivary concentrations of IgA than the healthy cats. The cats with chronic gingivostomatitis were treated with either methylprednisolone, sodium aurothiomalate, metronidazole and spiramycin, or oral hygiene products. After three months of treatment, the cats receiving methylprednisolone had a significant reduction in serum IgG levels compared to the cats treated with sodium aurothiomalate or metronidazole and spiramycin, but after six months of treatment there were no significant differences between the groups. Before the treatments, the levels of oral inflammation were not correlated significantly with any of the serum or salivary immunoglobulin levels. However, the changes in oral inflammation were correlated significantly with the changes in the salivary IgM concentration after three and six months of treatment, and with the change in the salivary IgA concentration after six months of treatment.

Animals↗
PubMed2003

A retrospective study of non-specific rhinitis in 22 cats and the value of nasal cytology and histopathology.

Case records from 40 cats subjected to rhinoscopic examination for investigation of chronic nasal disease were reviewed. Cases in which no specific underlying cause (eg neoplasia) was detected were further selected for detailed retrospective study. In these 22 cats (55% of the initial population), a final diagnosis of non-specific chronic nasal disease was made. The radiographic, rhinoscopic, cytological and histopathological findings were reviewed. Mucosal biopsy specimens were obtained in 20 cases. Despite clinical signs of more than 4 weeks duration, histopathology indicated acute inflammation in four cases. Two cases had chronic lymphoplasmacytic inflammation and 14 had mixed (lymphoplasmacytic and neutrophilic) inflammation. Specimens for cytology were obtained from 17 cases by brush sampling. Three of these samples were not diagnostic due to the poor quality of the slides; one showed normal cytology. Acute inflammation was diagnosed by cytology (n=11) more commonly than chronic (n=1) or mixed inflammation (n=1). Concurrent samples, of quality suitable for both histopathological and cytological interpretation, were collected from 12 cases only. Cytological results were in agreement with the histological results in 25% of these cases, the main discrepancy being the nature of the dominant inflammatory cell type. Therefore cytology does not appear to be a reliable means for detection of chronic inflammation. Further studies are needed in order to investigate the correlation between the nature of mucosal inflammation as defined by both histological and cytological evaluation, and the relationship of these test results to prognosis and therapy.

Animals↗
PubMed2003

Population dynamics and gene transfer in genetically modified bacteria in a model microcosm.

The horizontal transfer and effects on host fitness of a neutral gene cassette inserted into three different genomic loci of a plant-colonizing pseudomonad was assessed in a model ecosystem. The KX reporter cassette (kanamycin resistance, aph, and catechol 2, 3, dioxygenase, xylE) was introduced on the disarmed transposon mini-Tn5 into: (I) the chromosome of a spontaneous rifampicin resistant mutant Pseudomonas fluorescens SBW25R; (II) the chromosome of SBW25R in the presence of a naturally occurring lysogenic-phage (phage Phi101); and (III) a naturally occurring plasmid pQBR11 (330 kbp, tra+, Hgr) introduced into SBW25R. These bacteria were applied to Stellaria media (chickweed) plants as seed dressings [c. 5 x 104 colony-forming units (cfu)/seed] and the seedlings planted in 16 microcosm chambers containing model plant and animal communities. Gene transfer to pseudomonads in the phyllosphere and rhizosphere was found only in the plasmid treatment (III). Bacteria in the phage treatment (II) initially declined in density and free phage was detected, but populations partly recovered as the plants matured. Surprisingly, bacteria in the chromosome insertion treatment (I) consistently achieved higher population densities than the unmanipulated control and other treatments. Plasmids were acquired from indigenous bacterial populations in the control and chromosome insertion treatments. Plasmid acquisition, plasmid transfer from inocula and selection for plasmid carrying inocula coincided with plant maturation.

Bacteriophages↗
PubMed2003

Sterile nodular panniculitis and pansteatitis in three weimaraners.

The clinical and pathological findings in three unrelated weimaraners with pyrexia and multiple subcutaneous nodules are reported. Abdominal pain was an additional feature in two of the dogs and clinical investigations revealed inflammation of subcutaneous, mesenteric and falciform fat. Histopathological findings were consistent with pansteatitis. In the third dog, lesions were apparently limited to the subcutis and, hence, a diagnosis of nodular panniculitis was made. Microbiological examination of tissues was negative in all dogs, and there was no evidence of pancreatic disease. This report thus describes a presumed sterile and idiopathic panniculitis/pansteatitis complex in weimaraner dogs. Although the aetiology is unknown, this may represent an immune-mediated disorder.

Animals↗
PubMed2003

Resolution of chylothorax after resection of rib chondroma in a dog.

A six-year-old, male dobermann was presented with a history of dyspnoea and bouts of coughing. Radiography and computed tomography of the thorax showed pleural effusion and a well-circumscribed, calcified mass of 10 cm in diameter, appearing to originate from the left first rib. Thoracocentesis revealed that the pleural fluid was chylous in nature. An incisional biopsy was performed, which gave a histological diagnosis of chondroma. Resolution of the chylothorax after en-bloc surgical removal of the tumour suggested that the rib tumour was the initiating cause of the chylothorax. Seventeen months later, rib neoplasia recurred without pleural effusion, and was removed successfully. To the authors' knowledge, rib chondroma, which is an unusual tumour in dogs, has not been previously documented as a cause of chylothorax.

Animals↗