Regulation of bovine pyruvate carboxylase mRNA and promoter expression by thermal stress

Pyruvate carboxylase (PC) catalyzes the rate-limiting step in gluconeogenesis from lactate and is a determinant of tricarboxylic acid cycle carbon flux. Bovine PC 5' untranslated region (UTR) mRNA variants are the products of a single PC gene containing 3 promoter regions (P3, P2, and P1, 5′ to 3′) that are responsive to physiological and nutritional stressors. The objective of this study was to determine the direct effects of thermal stress on PC mRNA and gene expression in bovine hepatocyte monolayer cultures, rat hepatoma (H4IIE) cells, and Madin-Darby bovine kidney epithelial (MDBK) cells. Hepatocytes were isolated from 3 Holstein bull calves and used to prepare monolayer cultures. Rat hepatoma cells and MDBK cells were obtained from American Type Culture Collection, Manassas, VA. Beginning 24 h after initial seeding, cells were subjected to either 37°C (control) or 42°C (thermal stress) for 24 h. Treatments were applied in triplicate in a minimum of 3 independent cell preparations. For bovine primary hepatocytes, endogenous expression of bovine PC mRNA increased (P < 0.1) with 24 h of thermal stress (1.31 vs. 2.79 ± 0.49, arbitrary units, control vs. thermal stress, respectively), but there was no change (P ≥ 0.1) in cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) mRNA expression. Similarly, exposure of MDBK cells to thermal stress increased (P < 0.1) expression of bovine PC mRNA without altering (P ≥ 0.1) PEPCK-C mRNA expression. Conversely, there was no effect (P ≥ 0.1) of thermal stress on endogenous rat PC (0.47 vs. 0.30 ± 0.08, control vs. thermal stress) or PEPCK-C (1.61 vs. 1.20 ± 0.48, arbitrary units, control vs. thermal stress, respectively) mRNA expressions in H4IIE cells. To further investigate the regulation of PC, H4IIE cells were transiently transfected with bovine promoter-luciferase constructs containing either P1, P2, or P3, and exposed to thermal stress for 23 h. Activity of P1 was suppressed (P < 0.1) 5-fold, activity of P2 was unchanged (P ≥ 0.1), and activity of P3 was increased (P < 0.1) by 5.4-fold. These data indicate that response of bovine PC gene to thermal stress is through promoter regulation and suggest that there are unique characteristics of bovine PC promoters that may contribute to the physiological response to thermal stress.     

Comparison of the Value of Pulsed-field Gel Electrophoresis,Random Amplified Polymorphic DNA and Amplified rDNA Restriction Analysis for Subtyping Taylorella equigenitalis

Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzymes, including AluI and MboI, of the 1,500 bp fragments of rDNA amplified by polymerase chain reaction did not discriminate the genomic variations among the eight strains of T equigenitalis. Thus, pulsed-field gel electrophoresis was shown to discriminate these eight organisms better than random amplified polymorphic DNA analysis, while amplified rDNA restriction analysis was found to be unsuitable for subtyping T equigenitalis.     

Cysteamine Supplementation of In vitro Maturation Media: A Review

Under in vitro culture conditions, oxidative modifications of cell components via increased reactive oxygen species (ROS) represent a major culture induced stress. Anti‐oxidant systems such as glutathione (GSH) can attenuate the deleterious effects of oxidative stress by scavenging ROS. It has been suggested that GSH content in oocytes may serve as a reservoir protecting the zygote and the early embryos from oxidative damage before genomic activation and de novo GSH synthesis occur. Addition of low molecular weight compounds to culture media, such as cysteamine, can increase GSH levels by increasing cysteine uptake. Quite naturally, effects of supplementation of in vitro maturation (IVM) media with low molecular weight thiols have been studied in various species. This article reviews the use of cysteamine supplementation for IVM, its effects on maturation rates and further embryo development.     

Emesis in dogs: a review

Emesis is a common presenting sign in small animal practice. It requires a rational approach to management that is based upon a sound understanding of pathophysiology combined with logical decision making. This review, which assesses the weight of available evidence, outlines the physiology of the vomiting reflex, causes of emesis, the consequences of emesis and the approach to clinical management of the vomiting dog. The applicability of diagnostic testing modalities and the merit of traditional approaches to management, such as dietary changes, are discussed. The role and usefulness of both traditional and novel anti-emetic drugs is examined, including in specific circumstances such as following cytotoxic drug treatment. The review also examines areas in which common clinical practice is not necessarily supported by objective evidence and, as such, highlights questions worthy of further clinical research.     

Comparison of inhibition of polymerisation of mammalian tubulin and helminth ovicidal activity by benzimidazole carbamates

The correlation between the inhibition of hatching of Haemonchus contortus eggs and inhibition of mammalian tubulin polymerisation by benzimidazole carbamates has been investigated. The hatching process was observed to be independent of the biomass (eggs plus debris) over a 6-fold range and the early (E1-E3) stages of egg development, but was dependent on the concentration of co-solvent (DMSO) and time of incubation. Benzimidazole carbamates with strong inhibitory activity against mammalian tubulin were potent inhibitors of egg hatch, while non-inhibitors failed to prevent hatching. It is postulated that the primary mode of action of these drugs on nematode eggs is the inhibition of microtubule-dependent processes within the developing egg. The implications and limitations of this correlation are discussed.     

Trimethoprim-sulfadiazine in the horse: serum, synovial, peritoneal, and urine concentrations after single-dose intravenous administration

Six healthy adult mares were given a single IV injection of trimethoprim (TMP)-sulfadiazine (SDZ) at a dosage rate of 2.5 mg of TMP/kg of body weight and 12.5 mg of SDZ/kg. Serum, synovial, peritoneal, and urine TMP-SDZ concentrations were measured serially over a 48-hour period. The highest measured mean concentrations of TMP and SDZ were found in the first (0.5 hour) sample of serum, synovial fluid, and peritoneal fluid. The mean peak concentrations of TMP and SDZ averaged 4.37 micrograms/ml and 21.81 micrograms/ml for serum, 2.95 micrograms/ml and 15.31 micrograms/ml for synovial fluid, and 3.88 micrograms/ml and 19.52 micrograms/ml for peritoneal fluid, respectively. Urine concentrations of the drugs were relatively high and peaked early. The elimination rate for TMP and SDZ averaged 0.41 and 0.26 hour-1, while the elimination half-life was 1.91 and 2.71 hours, respectively, and the volume of distribution averaged 0.59 and 0.52 L/kg, respectively.     

Risk Analysis of Antibiotic Resistance, Berlin, BfR, 10-12 November 2003: activities of the Codex Alimentarius Commission related to antibiotic resistance

The FAO/WHO Codex Alimentarius Commission develops recommendations concerning risk management related to regulatory issues, on the basis of scientific advice provided by FAO and WHO. The complex question of anti-microbial resistance that requires a multidisciplinary approach, has been considered from different point of views in the framework of Codex, especially food hygiene and veterinary drugs and to a lesser extent, plant protection. Antibiotic resistance was considered from the perspective of the food chain and a risk profile was developed. Risk management options were also considered and may be grouped in two main categories: prevention of contamination through the food chain; and prudent use of anti-microbials. It is recognized that further scientific advice is necessary before developing specific risk management options in the framework of Codex. It is expected that the results of the FAO/OIE/WHO Expert Consultations on anti-microbial resistance will provide the necessary guidance for further action.     

Housing- and management-specific effects on the health of swine

Important diseases with regard to inadequate housing and management conditions in pig production are given. Typical examples like claw lesions, cannibalism and infectious factorial diseases are described. Beside negative factors of pig housing and management which are of particular pathogenic importance some aspects of disease control and herd health are listed finally.     

Growth patterns of Angus, Hereford and shorthorn cattle. I. Comparison of inbred and noninbred lines, changes in patterns over time and effects of level of inbreeding and reproductive performance

Mature weight (A) and rate of maturing (K) were estimated for 283 Angus, 140 Hereford and 280 Shorthorn cows utilizing the asymptotic regression equation Yt=A(1-Be-Kt). The Yt was weight of the individual at age t; and B was an estimate related to early life weight changes and provided for a Y-intercept (A-B). Each breed consisted of four inbred and two noninbred lines. Regression of estimated growth curve parameters on levels of inbreeding of the individuals and of their dams and effects of early reproductive performance (EREPRO) were studied as sources of variation in addition to line differences and trends in line values over years. Considered jointly with inbreeding of the dam and with EREPRO, inbreeding of the individual was negatively related (P less than .01) to estimates of A in the three breeds and unrelated (P greater than .10) to estimates of K. Each 1% increase in inbreeding of the individual was associated with about 2 kg decrease in estimated mature weight. An increase in inbreeding of the dam of an individual was negatively related to estimates of K in Angus (P less than .01) and Shorthorns (P less than .05). Inbreeding of dams was positively related to estimates of A in the three breeds, but only in Shorthorns could the relationship be declared significant. Estimates of A were about 46 kg heavier and estimates of K about .010 less for each year an individual failed to produce a calf during her first three opportunities. In general, A values declined in all three breeds during the study. Significant changes were observed in all Hereford analyses and after inbreeding was included in the Angus analysis, while inclusion of inbreeding in the Shorthorn analysis caused the change to become unimportant. The K values increased in all three breeds during the study; however, the change could be declared significant only in Herefords.     

Brown recluse spider (Loxosceles reclusa) envenomation in small animals

Objective – To provide a comprehensive review of relevant literature regarding the brown recluse spider (BRS) and to define those criteria that must be satisfied before making a diagnosis of brown recluse envenomation.
Etiology – The complex venom of the BRS contains sphingomyelinase D, which is capable of producing all the clinical signs in the human and some animal models.
Diagnosis – There is no current commercially available test. In humans there are many proposed guidelines to achieve a definitive diagnosis; however, there are no established guidelines for veterinary patients.
Therapy – Currently, no consensus exists for treatment of BRS envenomation other than supportive care, which includes rest, thorough cleaning of the site, ice, compression, and elevation.
Prognosis – Prognosis varies based on severity of clinical signs and response to supportive care.