Heterozygosity testing and multiplex DNA panel screening as a potential tool to monitor health and inbreeding in a small, closed dog population.

BACKGROUND: Selective breeding in populations with a limited effective population size may result in a loss of genetic diversity, which can cause an increased concentration of specific disease liability genes. The Dutch Shepherd Dog (DSD) in the Netherlands is an example of such a breed with a small effective population. OBJECTIVE: To evaluate the measurement of genetic diversity and multiplex DNA panel screening for implementation in a breeding strategy for the Dutch Shepherd Dog (DSD) and to investigate the clinical relevance of potentially identified mutations in the multiplex DNA panel screening. RESULTS: Genome-wide SNP testing showed genetic isolation and reduced genetic diversity within coat variety subgroups of the DSD. Panel screening identified a Von Willebrand's Disease type I mutation. Although decreased Von Willebrand's Factor proteins were significantly lower in DSDs carrying the VWD-I allele compared to the wildtype, clinical follow-up did not show a significant association between the clinical phenotype and VWD-I genotype. CONCLUSIONS: Genetic relationship measurement within a breed population may be a useful tool to enable breeding strategies to conserve genetic diversity. Results from a disease panel screening need to be evaluated for clinical relevance before breed selection restrictions can be considered.

Hemostatic Findings in Ascitic Fluid: A Cross-Sectional Study in 70 Dogs.

BACKGROUND: Ascitic fluids of horses and humans have fibrinolytic activity, independent of the underlying mechanism of fluid formation. OBJECTIVE: To determine whether coagulation and fibrinogenolytic/fibrinolytic activity (ie, low fibrinogen and increased fibrin-fibrinogen degradation products [FDPs], D-dimer, or both) occur in all types of ascitic fluid in dogs. ANIMALS: A total of 70 client-owned dogs with ascites. METHODS: In this cross-sectional study, dogs were categorized based on the pathophysiology of fluid formation into 4 groups: transudates due to decreased osmotic pressure, transudates due to increased hydrostatic pressure, exudates, and hemorrhagic ascites. Fibrinogen, FDPs, and D-dimer concentrations were measured and then compared in both ascitic fluid and plasma. RESULTS: Ten dogs had transudates due to decreased colloid osmotic pressure, 18 had transudates due to increased hydrostatic pressure, 13 had exudates, and 29 had hemorrhagic ascites. Ascitic fibrinogen concentrations (n = 70) were significantly lower (median = 59 mg/dL; range: 59-122 mg/dL) than those in the plasma (median = 168 mg/dL, range: 59-879 mg/dL; P < .0001). Ascitic FDPs concentrations (n = 70) were significantly higher (<5 µg/mL: 3/70 dogs, ≥5 to <20 µg/mL: 11/70 dogs, ≥20 µg/mL: 56/70 dogs) than those in the plasma (<5 µg/mL: 17/70 dogs, ≥5 to <20 µg/mL: 28/70 dogs, ≥20 µg/mL: 25/70 dogs; P < .0001). Ascitic D-dimer concentrations (n = 70) were significantly higher (median = 3.98 µg/mL, range: 0.02-9.19) than those in the plasma (median = 0.11 µg/mL, range: 0.01-4.08; P < .0001). Analysis of the data for each of the 4 different types of ascites showed similar results to those of all the data analyzed together. CONCLUSIONS AND CLINICAL IMPORTANCE: Ascitic fluid of dogs has evidence of coagulation activation and fibrinogenolytic/fibrinolytic activity and that this phenomenon occurs independent of the underlying mechanism that leads to the formation of ascites.

High intravascular tissue factor expression in dogs with idiopathic immune-mediated haemolytic anaemia.

A high mortality occurs in dogs with idiopathic immune-mediated haemolytic anaemia (IMHA) during the first 2 weeks after the diagnosis. The aim of this study was to investigate the inflammatory response and coagulation abnormalities in dogs with IMHA in relation to the prognosis and to establish the contribution of whole blood tissue factor (TF) and IL-8 gene expressions. Gene expressions in dogs with IMHA were compared to healthy dogs, dogs with DIC, dogs with sepsis, and in two groups of dogs that underwent intensive care treatment but had no evidence for either DIC or sepsis. The whole blood TF and IL-8 expressions were up regulated in all non-IMHA groups. Similarly, the TF expression in IMHA dogs was high, but the intravascular IL-8 expression was not increased. The dogs with IMHA had a pronounced inflammatory response that included a high WBC, left shift and monocytosis in comparison to the other disease groups. Coagulation factor activities in IMHA dogs were decreased fitting consumptive coagulopathy and the acute phase proteins FVIII and fibrinogen were increased. The platelet parameters suggested platelet activation and high platelet turnover in IMHA dogs. The model that best explained mortality contained monocytosis, increased activated partial thromboplastin time and elevated creatinine. Whole blood TF gene expression is up regulated and may contribute to consumptive coagulopathy in dogs with IMHA. Increased TF expression by activated platelets is an alternative explanation and should be investigated.

Idiopathic immune-mediated hemolytic anemia: treatment outcome and prognostic factors in 149 dogs.

BACKGROUND: Canine idiopathic immune-mediated hemolytic anemia (IMHA) is associated with a high mortality, especially in the 1st 2 weeks after diagnosis despite treatment. OBJECTIVES: To determine treatment outcome and identify prognostic variables in order to define areas of future research. ANIMALS: One hundred forty-nine dogs with hematocrit <30% and either a positive Coombs' test or spherocytosis and with no evidence of disease that can trigger IMHA were included. METHODS: Retrospective cohort study. All dogs were treated with prednisolone and azathioprine according to a standard protocol. Survival analysis was performed by the Kaplan-Meier method. Variables recorded at the time of diagnosis were tested as possible prognostic variables in a univariate and multivariate Cox proportional hazard model. RESULTS: The main predictors for mortality in dogs with idiopathic IMHA are the presence of increased plasma urea concentration, bands, thrombocytopenia, and petechiae at the time of diagnosis. The estimated Kaplan-Meier half-year survival was 72.6% (95% confidence interval [CI]: 64.9-81.3%). Mortality occurred mostly within the 1st 2 weeks. Cox proportional hazards analysis indicated that increased plasma urea concentration, icterus, and petechiae were the major independent predictors of mortality in the 1st 2 weeks. In most dogs that survived IMHA, a 3-month protocol of azathioprine with prednisolone maintained clinical remission. The estimated half-year survival for dogs that survived the 1st 2 weeks was 92.5% (95% CI: 86-99.3%). CONCLUSIONS AND CLINICAL IMPORTANCE: If the dogs survived IMHA, a 3-month protocol of prednisolone and azathioprine was effective with regard to survival and clinical outcome. Future research should be directed at identifying whether thrombotic tendency in dogs with IMHA is the main contributor to the development of increased plasma urea concentration, icterus, thrombocytopenia, and petechiae.

[Ehrlichia and Babesia infections in dogs in The Netherlands]. / Ehrlichia- en Babesia-infecties bij de hond in Nederland.

A retrospective study was performed at the Department of Clinical Sciences of Companion Animals at Utrecht University amongst 75 dogs diagnosed with a Babesia canis and/or an Ehrlichia canis infection. The majority of the dogs had visited an endemic area (most often the Mediterranean area or the Dutch Antilles), but two dogs became infected with Babesia in the Netherlands. Babesia infections were associated with a stay in an endemic area and an incubation period that are both significantly shorter (less than 3 months) than those for Ehrlichia and co-infections (more than 3 months). Reasons for the owner to seek veterinary attention (lethargy, anorexia, fever), findings from the physical examination (pale mucous membranes, hepato-/splenomegaly) and laboratory results (anemia, thrombocytopenia, hypo-albuminemia) were highly aspecific, making serology or PCR mandatory for diagnosing infections. Antigenic stimulation by the parasite sometimes resulted in immune-mediated diseases such as immune-mediated hemolytic anemia, thrombocytopenia, glomerulonefritis, and polyarthritis and in the case of ehrlichiosis in hypergammaglobulinemia. Specific therapy (imidocarb-diproprionate and/or doxycycline) was necessary, and because combined infections were common, it was considered appropriate to administer both drugs while the definitive diagnosis was being established. The prognosis was reasonably good, with almost half of all patients showing no clinical signs after treatment, although Babesia and co-infections were associated with a significantly longer survival sometimes resulted than Ehrlichia infections.

Pyothorax in nine dogs.

The results of treatment of pyothorax using systemic antibiotics, drainage, and lavage of the pleural space, are reported for 9 dogs. All 9 dogs recovered completely. In 8 of the 9 dogs the follow-up period was at least 6 months and in none was there a relapse. The results obtained with this treatment are excellent in comparison with the results that have been reported for treatment with systemic antibiotics and drainage of the pleural space but without lavage. Apart from the addition of pleural lavage to the treatment protocol, the better result might be because migrating plant related foreign bodies did not seem to play an important role in the pathogenesis of pyothorax in this group of dogs.

[Pyothorax in nine dogs]. / Pyothorax bij negen honden.

Nine dogs with pyothorax were treated with systemic antibiotics, and drainage and lavage of the pleural space. All nine dogs recovered completely. Eight dogs were followed up for 6 months or longer and none showed signs of relapse. The results of this treatment are excellent compared with those for treatment with systemic antibiotics and drainage of the pleural space but without lavage. Apart from the addition of pleural lavage to the treatment protocol, this difference in efficacy may be due to the fact that migrating grass seed awns do not seem to play an important role in the pathogenesis of pyothorax in dogs in the Netherlands.

Evaluation of the results of a L-asparaginase-based continuous chemotherapy protocol versus a short doxorubicin-based induction chemotherapy protocol in dogs with malignant lymphoma.

The results of an L-asparaginase-based continuous chemotherapy protocol (n = 52) versus a short doxorubicin-based induction chemotherapy protocol (n = 65) were evaluated in 117 dogs with malignant lymphoma. There were no differences between the two groups in patient characteristics or incidence of protocol-related toxicity. Complete remission was induced in 71.2% of the dogs treated with the L-asparaginase protocol and in 67.7% of the dogs treated with the doxorubicin-plus protocol. The calculated Kaplan-Meier one- and two-year survival fractions in the L-asparaginase group were 48% and 26%, and in the doxorubicin-plus group 35%, and 22%, respectively. Differences in remission and survival between the two treatment groups were not significant. A multivariate Cox proportional hazards survival analysis revealed that elevated pretreatment plasma creatinine concentration and prior treatment with prednisolone were associated with shorter survival times. An elevated pretreatment plasma creatinine concentration and total leucocyte count were associated with a decrease in the disease-free period. Differences in efficacy and toxicity between the two protocols were not significant. There is no apparent advantage in using the continuous L-asparaginase protocol, and the shorter doxorubicin-plus protocol is less expensive and less time consuming.

[Tumor lysis syndrome in a dog]. / Tumor lysis syndroom bij een hond.

The acute tumour lysis syndrome in humans as well as in dogs is caused by the acute lysis of tumour cells following chemotherapy. The release of intracellular products and their metabolites result in hyperkalemia, hyperphosphatemia, and hyperuricemia. The precipitation of uric acid and calciumphosphate in the kidneys can lead to hypocalcemia and acute renal failure which leads to fatal azotemia and arrhythmia's due to electrolyte disturbances. In this article the occurrence of the acute tumour lysis syndrome in a dog with malignant lymphoma is described. Suggestions to prevent the tumour lysis syndrome are made.

Long-term follow-up of avascular necrosis of the femoral head in the dog.

In a clinical study of 35 dogs with avascular necrosis of the femoral head, 60 per cent were Yorkshire terriers; the mean age of 33 of the dogs at onset of clinical signs was seven months. The dogs had the following signs: muscle atrophy (n = 25), shortening of the affected leg (n = 14), pain on passive movement of the hip joint (n = 28), and crepitation of the hip joint (n = 8). Radiographic findings were irregular density and flattening of the femoral head in combination with degenerative joint disease. Conservative treatment consisted of exercise therapy, and surgical treatment of a standard femoral head and neck excision. In 17 of the dogs the results of therapy were evaluated with the help of a questionnaire. It is concluded that femoral head and neck excision is indicated when conservative treatment fails to lead to clinical improvement within four weeks. Femoral head and neck excision has a good long term prognosis; however, slight intermittent lameness may remain.

Isolation of a new human herpesvirus producing a lytic infection of helper (CD4) T-lymphocytes in peripheral blood lymphocyte cultures--another cause of acquired immunodeficiency?

A new human helper (CD4) T-lymphotropic herpesvirus (HTLHV) was first isolated in February 1985 from the cultured peripheral blood lymphocytes (PBL) of a patient with the acquired immunodeficiency syndrome, and subsequently from the PBL of 1 patient with hairy cell leukaemia and 2 patients with lymphoproliferative disease associated with human T-lymphotropic virus type I infection. The viruses could be serially subcultured in umbilical cord PBL cultures in which they infected helper (CD4) T-lymphocytes producing multinucleate giant cells with intranuclear inclusions followed by cell lysis. Electron microscopy of infected cultures revealed that the isolates were herpesviruses. Specific DNA probing showed that the 4 isolates were related to one another but were distinct from cytomegalovirus, Epstein-Barr virus, Herpesvirus hominis types 1 and 2, and varicella-zoster virus. HTLHV lyses the same target cell as human immunodeficiency virus in PBL cultures suggesting that it may have a similar potential to cause acquired immune deficiency. The development of an unequivocally diagnostic serological test is a priority, so that the epidemiology and pathogenesis of HTLHV infection can be studied.

Postnatal and antenatal laboratory diagnosis of glutaric aciduria II in a South African family.

Glutaric aciduria type II (GA II) was proved in a neonate who presented shortly after birth with respiratory distress, metabolic acidosis, non-ketotic hypoglycaemia and a sweaty-feet-like odour. The diagnosis was based on elevated levels of glutaric and other acids in the urine and on studies on cultured skin fibroblasts where defective metabolism of fatty acids of varying chain length was demonstrated. Antenatal diagnosis was performed on a subsequent pregnancy in this family where an abnormal amniotic fluid organic acid profile together with defective fatty acid oxidation in cultured amnion cells was indicative of GA II in the fetus. This is the first report of this genetic disorder in a South African family and it should be considered in suspected organic acidaemia in the neonatal period.

[Diagnosis of intrathoracic lipoma by computer assisted tomography: a case report]. / Rekenaartomografiese diagnose van intratorakale lipoom.

A case of intrathoracic lipoma in a child is presented. The diagnosis was made before operation by means of computer-assisted tomography, a non-interventional procedure.