Analysis of KIT expression and KIT exon 11 mutations in canine oral malignant melanomas

KIT, a transmembrane receptor tyrosine kinase, is one of the specific targets for anti-cancer therapy. In humans, its expression and mutations have been identified in malignant melanomas and therapies using molecular-targeted agents have been promising in these tumours. As human malignant melanoma, canine malignant melanoma is a fatal disease with metastases and the poor response has been observed with all standard protocols. In our study, KIT expression and exon 11 mutations in dogs with histologically confirmed malignant oral melanomas were evaluated. Although 20 of 39 cases were positive for KIT protein, there was no significant difference between KIT expression and overall survival. Moreover, polymerase chain reaction amplification and sequencing of KIT exon 11 in 17 samples did not detect any mutations and proved disappointing. For several reasons, however, KIT expression and mutations of various exons including exon 11 should be investigated in more cases.     

Protein kinase C regulates ezrin-radixin-moesin phosphorylation in canine osteosarcoma cells

The development of metastasis is the most significant cause of death for both canine and human patients with osteosarcoma (OS). Ezrin has been associated with tumour progression and metastasis in human, canine and murine OS. Ezrin activation is dynamically regulated by protein kinase C (PKC) during metastatic progression in human and murine OS. To include the dog in the development of therapeutics that target ezrin biology, we characterized four new canine OS cell lines and confirmed the relationship between PKC and ezrin in these cells. Three of four cell lines formed tumours in mice that were histologically consistent with OS. All cell lines were markedly aneuploid and expressed ezrin and PKC. Finally, both ezrin phosphorylation and cell migration were inhibited using a PKC inhibitor. These data suggest that an association between PKC-mediated activation of ezrin and the metastatic phenotype in canine OS cells.     

Distribution of Campylobacter jejuni isolates from turkey farms and different stages at slaughter using pulsed-field gel electrophoresis and flaA-short variable region sequencing

The aim of this study was to assess the diversity of thermotolerant Campylobacter spp. isolated from turkey flocks at six rearing farms 1-2 weeks prior to slaughter (360 faecal swab samples) and from 11 different stages at the slaughterhouse (636 caecal, environmental, neck skin and meat samples). A total of 121 Campylobacter isolates were identified to species level using a multiplex PCR assay and were typed by pulsed-field gel electrophoresis (PFGE) and flaA-short variable region (SVR) sequencing. All Campylobacter isolates were identified as Campylobacter jejuni. PFGE analysis with KpnI restriction enzyme resulted in 11 PFGE types (I-XI) and flaA SVR typing yielded in nine flaA-SVR alleles. The Campylobacter-positive turkey flocks A, C and E were colonized by a limited number of Campylobacter clones at the farm and slaughter. The present study confirms the traceability of flock-specific strains (PFGE types I, V and IX; flaA types 21, 36 and 161) from the farm along the entire processing line to meat cuts. It seems that stress factors such as high temperature of the defeathering water (54-56 °C), drying of the carcass skin during air chilling (24 h at 2 °C), and oxygen in the air could not eliminate Campylobacter completely. Campylobacter-negative flocks became contaminated during processing by the same subtypes of Campylobacter introduced into the slaughter house by preceeding positive flocks even if they were slaughtered on subsequent days. Proper and efficient cleaning and disinfection of slaughter and processing premises are needed to avoid cross-contamination, especially in countries with a low prevalence of Campylobacter spp. The majority of flaA SVR alleles displayed a distinct association with a specific PFGE type. However, a linear relationship for all strains among both typing methods could not be established. To specify genetic relatedness of strains, a combination of different genotyping methods, is needed.     

Idiopathic osseous hyperplasia of the nasal turbinates in a Welsh terrier

A three-year-old Welsh terrier was presented with a 6-month history of progressive stertor, exercise intolerance and sneezing which was unresponsive to symptomatic treatment. A large calcified mass occupying the entire rostral right nasal cavity was defined with computed tomography. The mass was completely removed via a dorsal rhinotomy. Histopathological examination of the mass revealed osseous hyperplasia of the nasal turbinates. The excision led to complete resolution of clinical signs and there has been no recurrence 18 months after surgery.     

Genetic parameter estimates among scale activity score and farrowing disposition with reproductive traits in swine

The primary objective of this study was to determine if certain behavior traits were genetically correlated with reproduction. If 1 or both of the behavior traits were found to be correlated, a secondary objective was to determine if the behavior traits could be useful in selecting for more productive females. A scale activity score taken at 5 mo of age and a farrowing disposition score taken at farrowing were selected as the behavioral traits. Scale activity score ranged from 1 to 5 and farrowing disposition ranged from 1 to 3. Reproductive traits included age at puberty, number born alive, number born dead, litter birth weight, average piglet birth weight, number weaned, litter weaning weight, average weaning weight, wean-to-estrus interval, ovulation rate including gilts, and postweaning ovulation rate. Genetic correlations between scale activity score and reproduction ranged from -0.79 to 0.61. Three of the correlations, number born alive (P < 0.01), average piglet birth weight (P < 0.001), and wean-to-estrus interval (P = 0.014), were statistically significant but included both favorable and antagonistic correlations. In contrast, all but 1 of the farrowing disposition correlations was favorable and ranged from -0.66 to 0.67. Although only the correlation with litter birth weight was significant (P = 0.018), the consistent favorable direction of all farrowing disposition correlations, except average weaning weight, shows a potential for inclusion of farrowing disposition into a selection program.     

Virological and molecular epidemiological investigations into the role of wild birds in the epidemiology of influenza A/H5N1 in central Thailand

A serological and virological surveillance program to investigate the HPAI H5N1 virus in wild bird populations was undertaken from February 2007 to October 2008. The purpose of the survey was to investigate the infection status in free ranging wild birds in Banglane district, Nakhon Pathom province, central Thailand. Samples from wild birds were collected every two months. Choanal and cloacal swabs, serum and tissue samples were collected from 421 birds comprising 44 species. Sero-prevalence of the virus tested by H5N1 serum neutralization test (using a H5N1 virus clade 1; A/chicken/Thailand/vsmu-3-BKK/2004) was 2.1% (8 out of 385 samples; 95% CI 0.7, 3.5). Species that were antibody positive included rock pigeons (Columba livia), Asian pied starling (Gracupica contra), spotted dove (Streptopelia chinensis), oriental magpie robin (Copsychus saularis), blue-tailed bee-eater (Merops philippinus), myna (Acridotheres spp.), and pond heron (Ardeola spp.). Prevalence by H5N1 virus isolation was 0.5% (2 out of 421 samples; 95% CI 0.0, 1.1); the two H5N1 virus-positive samples were from Asian pied starling (Gracupica contra) and white vented myna (Acridotheres grandis). Positive virological samples were collected in June 2007 while all positive serology samples were collected between May and August except for one sample collected in December 2007. No positive samples were collected in 2008. Molecular studies revealed that the wild bird H5N1 viruses were closely related to poultry viruses isolated in other parts of Thailand. However, there was no poultry H5N1 prevalence study performed in the study site during the time of this wild bird survey. Interpretation of source of virus isolates would include spill-over of H5N1 viruses from contaminated sources due to movement of domestic poultry and/or fomites from other areas; or infection of wild birds within the outbreak locations and then translocation by wild bird movement and interaction with wild birds inhabiting distant locations.     

Associations among albuminuria, C-reactive protein concentrations, survival predictor index scores, and survival in 78 critically ill dogs

Background: Microalbuminuria and C‐reactive protein (CRP) are predictors of morbidity and survival in critically ill human patients. Hypothesis/Objectives: To evaluate results of microalbuminuria assays (untimed single‐sample urine albumin concentration [U‐ALB] and the urine albumin : creatinine ratio [UACR]), serum CRP, and survival predictor index (SPI2) scores as predictors of survival in critically ill dogs. Animals: Seventy‐eight dogs admitted to intensive care units at University of Tennessee (UT) and Colorado State University (CSU). Methods: Prospective observational study. Critically ill dogs were eligible for enrollment, unless euthanized because of financial constraints. Samples were collected within 3 hours of admission. Spearman's rank‐correlation coefficients were determined for U‐ALB, UACR, CRP, and SPI2. U‐ALB, UACR, CRP, and SPI2 were assessed for associations with 7‐ and 30‐day survival by Mann‐Whitney U‐tests and receiver operating characteristic (ROC) curves. P‐values < .0125 were considered significant. Results: UT (n = 49) and CSU (n = 29) patients did not differ significantly. Forty percent (31/78) of dogs died. SPI2 was inversely correlated with U‐ALB (r_s=?0.39, P < .001) and UACR (r_s=?0.41, P < .001). CRP was not correlated with SPI2 (P= .019), U‐ALB (P > .1), or UACR (P > .1). U‐ALB and UACR had very high correlation (r_s= 0.95, P < .001). SPI2, U‐ALB, and UACR differed significantly for survivors and nonsurvivors. SPI2, U‐ALB, and UACR had areas under the ROC curve (AUC) from 0.68 to 0.74 for survival prediction. Conclusions and Clinical Importance: Albuminuria and SPI2, but not CRP, are associated with survival in critically ill dogs. Suboptimal AUCs limit the value of microalbuminuria testing for clinical risk assessment. Additional studies are necessary to determine the usefulness of microalbuminuria testing in patient risk stratification for prospective research.     

Corn distillers dried grains with solubles in diets for growing-finishing pigs: a cooperative study

An experiment involving 560 crossbred pigs (28 replications of 4 to 6 pigs per pen) was conducted at 9 research stations to assess the effects of dietary concentrations of corn distillers dried grains with solubles (DDGS) on pig performance and belly firmness. Fortified corn-soybean meal diets containing 0, 15, 30, or 45% DDGS were fed in 3 phases from 33 to 121 kg of BW. A common source of DDGS containing 90.1% DM, 26.3% CP, 0.96% Lys, 0.18% Trp, 9.4% crude fat, 34.6% NDF, 0.03% Ca, and 0.86% P was used at each station. Diets were formulated to contain 0.83, 0.70, and 0.58% standardized ileal digestible (SID) Lys during the 3 phases with diets changed at 60 and 91 kg of BW, respectively. The DDGS replaced corn and soybean meal, and up to 0.172% Lys and 0.041% Trp were added to maintain constant SID concentrations of Lys and Trp in each phase. At each station, 2 pigs from each pen in 2 replications were killed and a midline backfat core was obtained for fatty acid analysis and iodine value. In most instances, there were differences among stations (P < 0.01), but the station × treatment interactions were few. Body weight gain was linearly reduced in pigs fed the greater amounts of DDGS (0 to 45%) during phase I (950, 964, 921, and 920 g/d; P < 0.01) and over the entire experimental period (944, 953, 924, and 915 g/d; P = 0.03), but ADFI (2.73, 2.76, 2.68, and 2.70 kg) and G:F (347, 347, 345, and 341 g/kg) were not affected (P = 0.15 and P = 0.33, respectively) during the entire test. Backfat depth was reduced (linear, P < 0.02) by increasing amounts of DDGS (22.5, 22.7, 21.4, and 21.6 mm), but LM area (47.4, 47.4, 46.1, and 45.4 cm(2)) was not affected (P = 0.16) by treatments. Estimated carcass fat-free lean was 51.9, 52.2, 52.4, and 52.1% for 0 to 45% DDGS, respectively (linear, P = 0.06). Flex measures obtained at 6 stations indicated less firm bellies as dietary DDGS increased (lateral flex: 11.9, 8.6, 8.4, and 6.6 cm; linear, P < 0.001; vertical flex: 26.1, 27.4, 28.2, and 28.7 cm; linear, P < 0.003). Saturated and monounsaturated fatty acid concentrations in subcutaneous fat decreased linearly (P < 0.001) and PUFA concentrations increased linearly (P < 0.001) with increasing DDGS in the diet. Iodine values in inner (61.1, 68.2, 74.7, and 82.2) and outer (67.9, 73.6, 79.6, and 85.8) backfat increased linearly (P < 0.001) as DDGS in the diet increased. In this study, feeding diets with 30 or 45% DDGS did not have major effects on growth performance, but resulted in softer bellies. Regression analysis indicated that iodine values increased 4.3 units for every 10 percentage unit inclusion of DDGS in the diet.     

Clinical and pathologic responses of American crows (Corvus brachyrhynchos) and fish crows (C ossifragus) to experimental West Nile virus infection

West Nile virus (WNV)-associated disease has a range of clinical manifestations among avian taxa, the reasons for which are not known. Species susceptibility varies within the avian family Corvidae, with estimated mortality rates ranging from 50 to 100%. We examined and compared virologic, immunologic, pathologic, and clinical responses in 2 corvid species, the American crow (Corvus brachyrhynchos) and the fish crow (C ossifragus), following experimental WNV inoculation. Unlike fish crows, which remained clinically normal throughout the study, American crows succumbed to WNV infection subsequent to dehydration, electrolyte and pH imbalances, and delayed or depressed humoral immune responses concurrent with marked, widespread virus replication. Viral titers were approximately 3,000 times greater in blood and 30,000 to 50,000 times greater in other tissues (eg, pancreas and small intestine) in American crows versus fish crows. Histologic lesion patterns and antigen deposition supported the differing clinical outcomes, with greater severity and distribution of lesions and WNV antigen in American crows. Both crow species had multiorgan necrosis and inflammation, although lesions were more frequent, severe, and widespread in American crows, in which the most commonly affected tissues were small intestine, spleen, and liver. American crows also had inflammation of vessels and nerves in multiple tissues, including heart, kidney, and the gastrointestinal tract. WNV antigen was most commonly observed within monocytes, macrophages, and other cells of the reticuloendothelial system of affected tissues. Collectively, the data support that WNV-infected American crows experience uncontrolled systemic infection leading to multiorgan failure and rapid death.