Cloning and sequencing full length of canine Brca2 and Rad51 cDNA.

Mammary tumors are the most common neoplasm in female dogs, Canis canis, and in women. Mutations in human Brca2 confer an increased risk of female breast cancer. Previous studies have shown that the Brca2 tumor suppressor protein interacts with the recombinational repair protein Rad51. We cloned the full-length cDNA of the canine homologues of Brca2 and Rad51 to obtain a basis for studying their relationship with susceptibility to mammary tumors. The canine Brca2 and Rad51 cDNAs are 11 and 1.5 kb long, encoding 3.471 and 339 amino acids, respectively. The amino acid sequence of canine Brca2 showed 68% homology with the human protein, and 58% homology with a murine protein. There were highly conserved regions in the C-terminus of all three proteins, where the Rad51 interacting domain and putative nuclear localization signals are located. Comparing with the partial genomic sequence previously reported, we found possible nuclear polymorphisms in exon 11, some of which result in amino acid substitutions. On the other hand, canine Rad51 protein had extremely high homology (99%) to the human and murine proteins. Expression of both Brca2 and Rad51 was detected in the mammary gland, suggesting that these two genes interact in the canine mammary gland.     

Seroprevalence of Ornithobacterium rhinotracheale infection in commercial laying hens in the north central region of the United States.

This study was the first to examine the seroprevalence of Ornithobacterium rhinotracheale (ORT) within a commercial egg layer population. Serum samples collected from egg production companies were examined by serum plate agglutination test (SPAT) and outer membrane protein-based enzyme-linked immunosorbent assay (ELISA). Results show that 90% of layer flocks were positive by SPAT and 100% by ELISA. Of the pullet flocks examined, 43% and 52% were positive by SPAT and ELISA, respectively. Our study indicates that the prevalence of ORT antibody is high in the commercial layer population, suggesting that this respiratory pathogen can easily spread through multiple-age layer farms from older flocks to newly housed pullet flocks.     

Immunohistochemical demonstration of Leu-7 (HNK-1), Neurone-specific Enolase (NSE) and Protein-Gene Peptide (PGP) 9.5 in the developing camel (Camelus dromedarius) heart

The development of the heart-conducting system has been controversially discussed. The common opinion that these specialized myocytes originate from mesodermal precursors has been challenged when nerve-specific antigens (Leu-7, NF, GIN2) were demonstrated in embryonic hearts of various species, suggesting a neural crest contribution to the embryonic conducting tissue. Anti-Leu-7 (HNK-1) antibodies were reported to reliably mark the conducting system in developing rat, chicken and human hearts. The present investigation was carried out on the hearts of 15 camel fetuses at 35, 45, 60, 75 and 100 cm crown-rump length (three specimens for each stage), in addition to three adult hearts. We investigated the antigenicity of cardiac structures for Leu-7, NSE (Neurone specific Enolase) and PGP (Protein Gene Peptide) 9.5. In all specimens investigated, both NSE and PGP 9.5 were expressed by cardiac nerves and conducting system components. The sinuatrial and atrioventricular nodes, the atrioventricular bundle as well as subendocardial and intramyocardial Purkinje fibers were stained. In contrast, the developing conducting system did not react with anti-Leu-7 antibody, although Leu-7 antigenicity was strongly expressed by the developing cardiac nerves. In adult camel hearts, the same pattern of immunoreactivity for the markers studied was still retained. Our results show that the expression of marker proteins for the developing conducting system is species-specific. Therefore, these markers are of little significance in discussions on the possible neurogenic nature of the heart conducting tissue.     

Possible effect of altered management practices on seroprevalence of bovine leukemia virus in heifers of a dairy herd with history of high prevalence of infection.

A bovine leukemia virus (BLV) control program was instituted at a high-prevalence, purebred Holstein dairy. The specific control program required single use of needles and obstetrical sleeves, disinfection of tattoo equipment before use, dehorning by use of electrical burning devices, replacement of whole milk feeding with high-quality milk replacer, and heat treatment of colostrum before feeding. The effect of the program was estimated by comparing the pre- (1987) and postintervention (1989) prevalences of BLV-agar gel immunodiffusion titers within particular heifer age cohorts (0 to 90, 91 to 180, 181 to 360 days, and 361 days to parturition). Random serologic testing was conducted quarterly within each cohort, but was analyzed on a combined yearly basis. Seroprevalence of BLV decreased in all age cohorts greater than 180 days old, with significant decreases in the 361 days to parturition (P = 0.003) and the combined 181 days to parturition cohorts (P = 0.0004). Prevalence within the latter cohort decreased from 0.44 in 1987 to 0.17 in 1989. These results were obtained without either culling or segregation of BLV-agar gel immunodiffusion-positive cattle.     

Effects of transforming growth factor-α on parathyroid hormone- and parathyroid hormone-related protein- mediated bone resorption and adenylate cyclase stimulation in vitro

The effects of transforming growth factor-alpha (TGF alpha) were determined on the ability of parathyroid hormone (PTH) or parathyroid hormone-related protein (PTHrP) to stimulate bone resorption and adenylate cyclase in vitro. Bovine PTH-(1-34) and human PTHrP-(1-34) were equipotent in their ability to stimulate bone resorption in neonatal mouse calvaria with maximal stimulation (2.9 and 2.8-fold increases in 45Ca release, respectively) at a concentration of 10 nM. Combinations of TGF alpha with bPTH-(1-34) or hPTHrP-(1-34) had additive effects on their ability to stimulate bone resorption when submaximal concentrations of the agonists were used. There was no evidence of synergism between TGF alpha bPTH-(1-34) or hPTHrP-(1-34) in their ability to stimulate bone resorption in vitro, nor was TGF alpha able to increase bone resorption induced by maximal concentrations of bPTH-(1-34) or hPTHrP-(1-34). TGF alpha potentiated the effects of either bPTH-(1-34) or hPTHrP-(1-34) on the stimulation of adenylate cyclase in osteoblast-like ROS 17/2.8 cells. These data indicate that TGF alpha has additive effects with submaximal concentrations of PTH or PTHrP on their ability to stimulate bone resorption which may be important in the pathogenesis of humoral hypercalcemia of malignancy.     

Cerebrospinal elaphostrongylosis in sheep in northern Norway.

Pathologic examination of four sheep from northern Norway exhibiting neurologic signs of paresis, paralysis and vestibular system disease revealed nematodes in the central nervous system (CNS). The worms were identified as Elaphostrongylus rangiferi Mitskevich, 1960, the elaphostrongylid nematode of reindeer (Rangifer tarandus tarandus). Microscopic lesions found in the CNS were focal traumatic encephalomyelomalacia caused by migrating worms, eosinophilic meningitis and choroiditis, lymphohistiocytic and nematode granulomas, and perineural infiltrations. The disease occurred in November and December 1990, after the sheep had been on pasture in areas frequented by considerable numbers of reindeer.     

Small intestinal starch digestion in steers: effect of various levels of abomasal glucose, corn starch and corn dextrin infusion on small intestinal disappearance and net glucose absorption

Eight Holstein steers (four at 300 kg, four at 406 kg) fitted with an elevated carotid artery, hepatic portal and mesenteric venous catheters, and abomasal and ileal cannulas were used in several 4 x 4 Latin square experiments to evaluate small intestinal starch digestion. They were fed alfalfa hay at 1.5% of BW and abomasally infused with water or glucose, corn starch or corn dextrin (one carbohydrate per Latin square) at 20, 40 or 60 g/h, with subsequent determination of small intestinal disappearance and net portal glucose absorption. Increasing the amount of all three carbohydrates infused abomasally increased the amount of carbohydrate disappearing in the small intestine. Increased infusion of glucose caused a continual increase (linear, P less than .01) in net glucose absorption, whereas net glucose absorption for starch and dextrin was maximal at the 20 g/h infusion (quadratic, P less than .05). With the 60 g/h infusion, 94% of the glucose but only 38% of starch and 29% of small intestinal dextrin disappearance could be accounted for as net glucose absorption, leaving a large portion of starch and dextrin disappearance unaccounted for. Of the infused starch and dextrin passing the ileum, 5.8 and 7.3%, respectively, was unpolymerized glucose, indicating that, at least in the distal small intestine, complete starch hydrolysis exceeded the capacity for glucose disappearance. It is concluded that only about 35% of the raw corn starch disappearing in the steer's small intestine resulted in net portal glucose absorption.     

Preanaesthetic use of atropine in small animals

Although it is unwise to recommend any preanaesthetic drug or regimen to be used routinely without consideration for the individual patient, the procedure being undertaken or the other drugs that will be used during the anaesthetic, of all anaesthetic-related drugs that might be under review, atropine is probably one of the least 'toxic' and least likely to cause life-threatening complications when used correctly. In most small veterinary practices there isn't the luxury of a dedicated anaesthetist available to monitor and manage each anaesthetised patient and the best form of cardiac monitor available may be an audible rate monitor. In this situation the advantages combined with the usual indications probably far outweigh the disadvantages of using atropine prophylactically. If a choice must be made, a sinus tachycardia under anaesthesia is probably preferable to a profound bradyarrhythmia. Absolute contraindications for using atropine are rare; the drug, when used alone, has low incidence of toxicity at clinically recommended doses in dogs and cats. Further, the new inhalation agents available to veterinarians (enflurane and isoflurane) do not sensitise the heart to the effect of catecholamines, so that the predominance of sympathetic tone produced when atropine is used is unlikely to precipitate dysarrthythmias during gaseous anaesthesia. Rhythm disturbances that occur under halothane anaesthesia when atropine is used, suggest there is some other cause, or agent, that is arrhythmogenic, such as an alpha 2 agonist, excessively deep level of anaesthesia, hypercapnia or hypoxaemia. Notwithstanding the above arguments, my preference, as a specialist anaesthetist, is that a drug is given only as required (not before) and that when, for example, any unusual heart rhythm occurs, all possible reasons for the occurrence (such as excessively deep anaesthesia or hypoventilation) are eliminated before other drugs like atropine or antiarrhythmics are administered.     

The physical state and plasma biochemical profile of young calves on arrival at a slaughter plant

AIM: To determine the welfare of calves arriving at a slaughter plant by assessing their physical state and plasma biochemical profile. METHODS: Calves (n=7,169) aged 4-10 days, were examined immediately after they were unloaded from trucks (n=20) and categorised on physical grounds as being in an 'unacceptable', 'marginal' or 'acceptable' state. 'Unacceptable' calves were lying down, unable to walk, extremely weak or seriously injured. 'Marginal' calves had a wet umbilicus, were hollow sided, apparently immature, or weak and slow and unsteady on their feet. 'Acceptable' calves were strong, walked freely, round-sided, bright and alert and had a dry umbilicus. After unloading, 27 'unacceptable', 306 'marginal' and 289 'acceptable' calves were weighed, examined clinically and a blood sample was taken from each. A second blood sample was taken from 118 of these calves, 12-15 hours later. The packed cell volume (PCV) and plasma concentrations of glucose, beta-hydroxybutyrate (BHB), total plasma proteins (TPP), gamma-glutamyl transferase (GGT), urea, creatine kinase (CK) and lactate were measured. RESULTS: Only 27 of the 7,169 calves examined (0.4%) were categorised as being in an 'unacceptable' state on arrival and these were euthanased shortly after unloading. Generally, the plasma biochemical profiles of the 'acceptable' calves were similar to those of 'marginal' animals. However the glucose concentration of 'marginal' calves was lower than that of 'acceptable' calves, suggesting that they had lower energy reserves. After 12-15 h in lairage, plasma glucose concentration decreased and the BHB concentration increased in both 'acceptable' and 'marginal' calves compared with concentrations measured on arrival. CONCLUSIONS: The percentage of calves presented for slaughter in an 'unacceptable' state was small (0.4%). However, 4% of calves were classified as 'marginal' and failed to meet all the requirements of the code of recommendations and minimum standards governing the welfare of young calves in New Zealand. The categorisation of calves by a physical assessment was supported by results of plasma biochemistry. As a result, plasma biochemical profiles are considered unnecessary for making decisions regarding the welfare status of calves arriving at a slaughter plant; an assessment of their physical state is sufficient.     

Glutathione, Cysteine, and Ascorbate Concentrations in Clinically Ill Dogs and Cats

Background: Oxidative stress plays a role in the pathogenesis of many systemic diseases. Hospitalized human patients are glutathione, cysteine, and ascorbate deficient, and antioxidant depletion has been correlated with poor clinical outcome. To date little is known about antioxidant concentrations in hospitalized veterinary patients. The purpose of this study was to determine whether ascorbate, cysteine, or glutathione depletion is present in ill dogs and cats compared with healthy controls.
Hypothesis: Clinically ill dogs and cats would be antioxidant depleted, and depletion would correlate with illness severity and clinical outcome.
Animals: Clinically ill client-owned dogs (n = 61) and cats (n = 37), healthy control dogs (n = 37) and cats (n = 33).
Methods: Prospective, observational, case control study. Erythrocyte reduced glutathione, plasma cysteine, and plasma ascorbate were quantified using high-performance liquid chromatography.
Results: Clinically ill dogs had significantly lower erythrocyte glutathione concentrations (1.22 mM, range 0.55–3.61) compared with controls (1.91 mM, range 0.87–3.51; P = .0004), and glutathione depletion correlated with both illness severity ( P = .038) and mortality ( P = .010). Cats had higher ascorbate concentrations when ill (10.65 μM, range 1.13–25.26) compared with controls (3.68 μM, range 0.36–13.57; P = .0009).
Conclusions and Clinical Importance: Clinically ill dogs had decreased erythrocyte glutathione concentrations, which could be a marker of illness severity and prognostic of a poor outcome. Clinically ill cats had an unexpectedly high plasma ascorbate, which could represent a unique species response to oxidative stress.