Immunohistochemical detection of tumor suppressor gene p53 protein in feline injection site-associated sarcomas

Sarcomas associated with injection sites are a rare but important problem in cats. Immunohistochemical detection of p53 protein may correlate to mutation of the p53 tumor suppressor gene, a gene known to be important in oncogenesis. The expression of nuclear p53 protein in 40 feline injection site-assocated sarcomas was examined by immunohistochemical staining. In 42.5% (17/40), tumor cell nuclei were stained darkly; in 20% (8/40), tumor cell nuclei were stained palely; and in 37.5% (15/40), tumor cell nuclei were unstained. Immunohistochemical detection of p53 protein in a proportion of injection site-associated sarcomas suggests that mutation of the p53 gene may play a role in the pathogenesis of these tumors.     

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.     

Serum antibody responses to vaccinal antigens in lean and obese geriatric dogs

The immune responses in control dogs [1 to 4 years of age, body condition score (BCS): 4 to 5 out of 9] were compared to those of aging dogs (based on breed and body size) either categorized as lean (BCS: 4 to 5 out of 9) or obese (BCS: 8 to 9 out of 9). Of interest were the serum titers to the following common agents found in vaccines, canine parainfluenza virus (CPIV), canine parvovirus (CPV), canine distemper virus (CDV), canine respiratory coronavirus (CRCoV), and Bordetella bronchiseptica. There were no statistical differences in the antibodies to CPIV, B. bronchispetica, and CRCoV, among the age/weight categories, nor among the age/weight categories and the time, in days, between the date of sample collection and the date of the last recorded vaccination for CPIV, B. bronchiseptica, CPV, and CDV. For CPV, the control dogs had significantly (P < 0.002) higher serum neutralization (SN) titers than the lean geriatric dogs and the obese geriatric dogs. For CDV SN titers, the only statistically significant (P = 0.01) difference was that the control dogs had higher SN titers than the lean geriatric dogs.     

Cutaneous periocular Habronema infection in a dromedary camel (Camelus dromedarius)

A 6‐year‐old castrated dromedary camel (Camelus dromedarius) presented with a non‐healing, severely pruritic, ulcerative fibrotic plaque located at the medial canthus. Histological examination of surgical biopsies identified degenerating nematode larvae within eosinophilic granulomas. Treatment involved repeated debridement of the lesion, injectable ivermectin and anti‐inflammatory therapies, and injectable and topical antibiotics . A specially constructed mask with goggles to prevent the camel from continuing to self‐traumatize the eye and lesion was also placed. Full recovery occurred approximately 1 month after diagnosis. Because of the location of the lesion, time of year, the gross and microscopic characteristics of the lesion, the presence of a likely nematode larva and the response to treatment, a diagnosis of cutaneous habronemiasis was made.     

Isolation of Leptospira interrogans serovar bratislava from sows in Iowa

Leptospira interrogans serovar bratislava was recovered from 2 of 10 sows examined from an Iowa slaughterhouse. Isolations were made from the kidney and genital tract of each sow. Serovar bratislava is not included in vaccines because it has not been previously isolated in the United States.     

Evaluation of formalin-fixed paraffin-embedded tissues from feline vaccine site-associated sarcomas for feline foamy virus DNA

OBJECTIVE: To evaluate a group of vaccine site-associated sarcomas (VSS) for the presence of feline foamy virus (FeFV) DNA, using polymerase chain reaction (PCR) methods. SAMPLE POPULATION: 50 formalin-fixed paraffin-embedded (FFPE) tissue blocks from VSS of cats. PROCEDURE: DNA was extracted from FFPE sections of each tumor, and regions of the gag and pol genes of FeFV were amplified by use of PCR methods, using 1 primer set for each region. Sensitivity of the method was compared between fresh and FFPE cells, using mouse kidney tissue that was injected with FeFV-infected cultured cells and using agarose-cell pellets. Results-Feline foamy virus DNA was not detected in VSS tissues. Sensitivity of the method was 10 times greater in fresh versus FFPE mouse tissues. Sensitivity of the method in fresh FeFV-infected cultured cells versus FFPE agarose-cell pellets was equal when fixation was 24 or 48 hours and 10 times greater when fixation was 72 hours or 1 week. CONCLUSIONS AND CLINICAL RELEVANCE: A PCR-based method can be successfully applied to FFPE tissues for FeFV DNA detection. Results suggest there is no direct FeFV involvement in the pathogenesis of VSS in cats.     

The influence of rider:horse bodyweight ratio and rider-horse-saddle fit on equine gait and behaviour: A pilot study

The effect of rider weight on equine welfare and performance requires further investigation. The objective of this prospective, cross-over, randomised trial was to assess gait and behavioural responses of horses to riders of similar ability, but different bodyweights. Six nonlame horses in regular work were ridden by each of four riders: Light (L), Moderate (M), Heavy (H) and Very Heavy (VH). Saddle fit was assessed subjectively throughout the study. Each horse was ridden twice by riders L and M, and once by rider H. Rider VH rode five horses once and one twice. Each horse-rider combination undertook a standardised, 30-min ‘dressage-test' which was abandoned if we observed lameness grade ≥ 3/8 in one limb, grade ≥ 2/8 in ≥ 2 limbs, or ≥ 10/24 behavioural markers of pain. Horses were reassessed in hand 45–60 min after any abandonment. Mean rider bodyweights, body mass index (BMI) values and rider:horse bodyweight percentages for the L, M, H and VH riders were respectively: 60.8, 77.8, 91.0, 142.1 kg; 23.2, 28.0, 26.3, 46.9 kg/m²; 10.0–11.7%, 12.8–15.0%, 15.3–17.9%, 23.6–27.5%. All 13 H and VH rider tests (lameness, n = 12; behaviour, n = 1) and one of 12 M rider tests (lameness) were abandoned. Lameness was confirmed using inertial measurement unit data. All horses trotted sound after test abandonment and completed the study moving well when ridden. Limitations of the study were saddle fit was not ideal in all horse-rider combinations and abandonment criteria were subjective. The conclusions and clinical relevance of the study were that large riders can induce temporary lameness and behaviours consistent with musculoskeletal pain. This may relate to rider bodyweight and/or weight distribution. Riders M and H had similar BMI but markedly different test abandonment rates, therefore bodyweight is likely to be more relevant than BMI. Further work is required to determine if horse fitness, adaptation to heavier weights and better saddle fit for heavier/taller riders will increase horses' weight-carrying capacity.     

Population-based study of fecal shedding of Clostridium perfringens in broodmares and foals

OBJECTIVE: To determine the percentage of broodmares and foals that shed Clostridium perfringens in their feces and classify the genotypes of those isolates. DESIGN: Prospective cross-sectional study. ANIMALS: 128 broodmares and their foals on 6 equine premises. PROCEDURES: Anaerobic and aerobic bacteriologic cultures were performed on feces collected 3 times from broodmares and foals. All isolates of C. perfringens were genotyped. RESULTS: Clostridium perfringens was isolated from the feces of 90% of 3-day-old foals and 64% of foals at 8 to 12 hours of age. A lower percentage of broodmares and 1- to 2-month-old foals shed C. perfringens in their feces, compared with neonatal foals. Among samples with positive results, C. perfringens type A was the most common genotype identified (85%); C. perfringens type A with the beta2 toxin gene was identified in 12% of samples, C. perfringens type A with the enterotoxin gene was identified in 2.1% of samples, and C. perfringens type C was identified in < 1% of samples. CONCLUSIONS AND CLINICAL RELEVANCE: Clostridium perfringens was identified from the feces of all but 6 foals by 3 days of age and is likely part of the normal microflora of neonatal foals. Most isolates from broodmares and foals are C. perfringens type A; thus, the clinical relevance of culture results alone is questionable. Clostridium perfringens type C, which has been associated with neonatal enterocolitis, is rarely found in the feces of horses.     

Bovine mononuclear phagocytic cells: identification by monoclonal antibodies and analysis of functional properties

Monoclonal antibodies (mAb) which react with bovine monocytes have been produced. These include three mAb (P8, IL-A22 and IL-A24) that recognize the majority of monocytes and granulocytes in peripheral blood; two of these mAb were also shown to react with 30-40% of cells in bone marrow, including both monocytic and granulocytic cells, and with variable percentages of tissue macrophages. Thus these mAb can act as markers for myeloid cells in haemopoietic tissues and for monocytes in cell populations devoid of granulocytes. A further two mAb (IL-A23 and IL-A25) recognize monocytes and/or macrophages. The reactivity of one of these mAb (IL-A25) appears to be mainly restricted to pulmonary macrophages. The other mAb reacts with a variable proportion of blood monocytes and generally with a higher percentage of tissue macrophages, suggesting that its expression may relate to activation or maturation of monocytes. In order to study the functional properties of peripheral blood monocytes, techniques were developed for obtaining populations of peripheral blood mononuclear cells (PBM) depleted of monocytes to less than 0.2% and monocyte populations of greater than 97% purity. Removal of monocytes from PBM abrogated the capacity of the cells to proliferate in response to Con A and PBS, although addition of 2-mercaptoethanol to the cultures restored proliferation. In both allogeneic and autologous mixed leukocyte cultures (MLC), monocytes were required in the stimulator cell populations for induction of the proliferative responses, and both responses could be elicited with purified monocytes. However, proliferation in the autologous MLC occurred only with responder cell populations that were depleted of monocytes. Moreover, it was shown that addition of more than 5% unirradiated monocytes to the autologous MLC suppressed proliferation. These findings indicate that monocytes play an important role in the induction and regulation of cellular immune responses in cattle. Two of the mAb that react with monocytes and granulocytes were tested for their capacity to inhibit proliferative responses of PBM to mitogens, alloantigens or the soluble antigen, KLH.(ABSTRACT TRUNCATED AT 400 WORDS)     

The effect of the Halothane and Rendement Napole genes on carcass and meat quality characteristics of pigs

The objective of this study was to determine the effects of the Halothane (N) and Rendement Napole (RN) genes on carcass and meat quality characteristics in pigs. Halothane and RN carrier (Nn/RN- rn+) Hampshire boars (n = 4) were mated to dams that were homozygous for the normal allele of both genes (NN/rn+ rn+) to produce progeny of four genotypes: 1, NN/rn+ rn+ (n = 31); 2, Nn/rn+ rn+ (n = 27); 3, NN/RN- rn+) (n = 30); and 4, Nn/RN- rn+ (n = 23). A DNA test was used to determine Halothane genotype, and longissimus glycolytic potential was used to predict the RN genotype. Pigs were reared under standard conditions to approximately 120 kg live weight and slaughtered at a commercial plant, and carcass characteristics and meat quality were evaluated. Halothane carriers (Nn/ _ _), in comparison to Halothane normal (NN/_ _) pigs, had shorter carcasses, lower longissimus ultimate pH, higher Minolta L* and b* values, and greater drip loss. Rendement Napole gene carriers (_ _/RN- rn+) had higher L* and b* values and drip and cooking loss and lower longissimus ultimate pH than homozygous recessive animals (_ _/rn+ rn+). There were Halothane x RN genotype interactions (P < 0.05) for subjective color, firmness, and marbling scores, and for shear force. Animals that were normal for both genes (NN/rn+ rn+) had the highest subjective scores for color (2.60, 1.88, 1.85, and 1.95, SE = 0.181, P < 0.05), firmness (2.53, 2.03, 2.10, and 1.89, SE = 0.182, P < 0 .05), and marbling (2.11, 1.44, 1.53, and 1.55, SE = 0.153, P < 0 .05) for genotypes 1, 2, 3, and 4, respectively, suggesting darker, firmer muscle with a higher level of marbling for this genotype. Shear force was highest for Nn/rn+ rn+ animals (3.83, 4.41, 3.79, and 3.70, respectively, SE = 0.172, P < 0.05). Gilts had less s.c. backfat thickness, greater longissimus muscle area, and lower subjective marbling scores than barrows. There was no effect (P > 0.05) of gender on other meat quality traits. This study illustrates the negative effects of the Halothane and RN genes on fresh pork quality and suggests that in combination the detrimental effects of the two genes are additive for ultimate pH, objective color, and water-holding capacity.