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.     

European consensus document on mast cell tumours in dogs and cats

In preparing this document the Authors aimed to pool current information on canine and feline mast cell disease. The information was gathered from international studies and a emphasis was placed on material and opinion with a strong evidence base. We intend it to form the basis of our understanding in this disease at the current time and we anticipate that it will be particularly useful for the general practitioner. It should be emphasized that the authors are presenting this work from a European perspective.     

LDSO: a program to simulate pedigrees and molecular information under various evolutionary forces

Simulations are a major tool to evaluate new statistical methods and optimize experimental designs in the genomic era. However, this can only be achieved when the simulations are close enough to reality, as well as diverse enough to be realistic. For mapping studies, it is thus critical to re-create as much as possible the forces generating linkage (mutation, random drift, changes in population sizes, selection and pedigree structure) and the mechanisms producing trait genetic architecture (additivity, dominance, epistasis). We present here a computer program (ldso) simulating these phenomena. Optional outputs provide statistics on the linkage disequilibrium (LD) structure and the identity by descent between chromosomal segments, facilitating further data analyses. Furthermore, ldso enables the simulation of genomic data in known pedigrees, which sticks as precisely as possible to recent population history and structures of the long-range LD, allowing optimization of fine-mapping strategies.     

Antibody prevalence of low-pathogenicity avian influenza and evaluation of management practices in Minnesota backyard poultry flocks

Low‐pathogenicity avian influenza (LPAI) viruses have caused illness in poultry and humans with poultry contact. To determine whether there is evidence of exposure to avian influenza viruses (AIV) among backyard poultry in Minnesota and their human caretakers, 150 flocks of backyard birds were sampled for antibodies to AIV from August 2007 through December 2008. One hundred flocks were tested through routine slaughter surveillance by the Minnesota Board of Animal Health and an additional 50 flocks were contacted and sampled by study investigators. Blood was collected from 10 to 13 birds from each flock and a survey of biosecurity and management practices was administered to the flock owner. Blood samples were tested by agar gel immunodiffusion (AGID) for influenza A antibodies. Tested flocks had a median flock size of 100 birds (range: 12–800 birds), and were most commonly owned for meat for personal use (81% of respondents), fun or hobby (58%) and eggs for personal use (56%). Although 7% of flock owners reported that their birds had shown respiratory signs in the previous 3 months, only 1 of 150 flocks tested positive for influenza by AGID. Antibodies to LPAI H6N1 were detected in the positive flock. The owner of the positive flock did not have antibodies to H6 or other common AIV. Based on the findings of this study, the risk of transmission of LPAI viruses from backyard poultry to owners in Minnesota appears to be low under current conditions and management practices.     

Seroprevalence of Antibodies to Chlamydophila psittaci in Zoo Workers in Brazil

To evaluate the prevalence of antibodies to Chlamydophila psittaci 364 serum samples were collected from veterinarians, biologists, animal scientists, veterinary students, animal keepers and others employees in 20 zoos, and from veterinary practitioners in 10 Brazilian states. Subjects ranged from 15 to 64 years of age, with 268 (74%) males and 96 (26%) females. Chlamydial antibodies were determined by the complement fixation test (CFT) and specific anti‐C. psittaci IgG antibodies were determined by the microimmunoflurescence (MIF) test. Complement fixation test showed 23.9% (87/364) and MIF test showed 4.7% (17/364) positive serum samples. Titres ranged from 16 to 256 in both assays, demonstrating evidence of recent or current infection. Although chlamydial antibodies were detected in workers of seventeen zoos, MIF test only detected specific C. psittaci antibodies in seven of them. Previous psittacosis infection was suspected in eight workers of two zoos, five of whom reported having pneumonia, while employed at the zoos. However, diagnosis was not established in any of these cases in the past. Results indicated the occurrence of infection and previous contact of Brazilian zoo workers with C. psittaci, as well as the zoonotic potential of psittacosis in this risk population. Other studies are necessary to evaluate the risk factors of infection in this population. This seroepidemiological survey confirmed the need to adopt preventive measures to control avian chlamydiosis and protect the health of zoo workers in the country.     

Calcium homeostasis and its relationship to superoxide production in blood and milk neutrophils of lactating goats

Polymorphonuclear neutrophils (PMN), which comprise over 70% of the somatic cells in goat milk, are a major cellular component of innate immunity in the goat mammary gland. However, the function of milk PMNs is modified after diapedesis compared to PMNs in blood. As many aspects of PMN activity depend directly on intracellular Ca²⁺ concentration ((Ca²⁺)_i), the present study aimed to determine the changes in Ca²⁺ homeostasis of milk PMNs from lactating goats compared to autologous blood PMNs, and to examine the significance of these variations to the immuno-competency of milk PMNs. The intracellular Ca²⁺ store of freshly prepared milk cells was estimated from the elevation of (Ca²⁺)_i after ionomycin treatment, which was found to be significantly less than blood PMNs. Replenishment of the intracellular Ca²⁺ store in milk cells after intracellular Ca²⁺ depletion by Bapta-AM followed by spiking with 2.5 mM Ca²⁺ for 20 min was also compared to that of blood PMNs, showing that after depletion/spiking the intracellular Ca²⁺ store in milk cells was much less than blood PMNs. The production of superoxide anion (O₂^?) in vitro in response to (Ca²⁺)_i-dependent or (Ca²⁺)_i-independent modulators was used to evaluate the relevance of altered Ca²⁺ homeostasis on the immuno-competency of milk cells compared to blood PMNs. The results indicated that milk cells produced similarly low levels of O₂^? as blood PMNs when treated with ionomycin. However, the amount of O₂^? produced by milk cells in response to phorbol 12-myristate 13-acetate (PMA) stimulation, although greater than ionomycin treatment, was significantly less than that of blood PMNs. The capacity for O₂^? production by both cell types in response to PMA reverted to the resting state with use of the protein kinase C (PKC) inhibitor, staurosporine. In conclusion, the current study demonstrated an irreversible shortage of intracellular Ca²⁺ in the milk PMNs of lactating goats compared to blood PMNs. It also showed that preliminary O₂^–production, primed by ionomycin treatment, remained unchanged in milk PMNs, despite the shortage in intracellular Ca²⁺, but decreased O₂^? production capacity, mediated via the PKC pathway, in milk PMN. It is suggested that the defects in Ca²⁺ homeostasis in milk PMNs of lactating goats is partially attributable for the post-diapedesis functionality modifications.     

Prednisolone is associated with an increase in serum insulin but not serum fructosamine concentrations in dogs with atopic dermatitis

The aim of this study was to assess the effects of a standard therapeutic protocol of prednisolone (Pred) on glucose homeostasis in atopic dogs and compare it with previously published data for ciclosporin A (CsA). The central aim of the study was to assess and compare the effects of standard therapeutic protocols of prednisolone (Pred) and ciclosporin A (CsA) on glucose homeostasis in dogs with atopic dermatitis (CAD). Both treatments significantly reduced the physical signs of CAD, as determined by the canine atopic dermatitis extent and severity index version 3 (CADESI-03) and the Edinburgh Pruritus Scale (EPS). Post-treatment plasma glucose concentrations were not significantly different in the two groups, but serum insulin concentrations were significantly higher following Pred therapy (P<0.05). Serum fructosamine concentrations were not significantly different pre- and post-treatment with Pred, although previous studies had shown that CsA treatment increased fructosamine concentrations (P<0.005). The two treatment groups were recruited in a similar timeframe, were numerically matched and there were no differences in CADESI-03 and EPS scores between the CsA and Pred groups either before or after treatment. Thus, both CsA and Pred treatment were associated with mild disturbances in glucose metabolism, but only CsA therapy resulted in a significant increase in fructosamine concentrations. This information may be relevant to clinicians when considering therapeutic options for dogs with atopic dermatitis which already have impaired glucose homeostasis.     

Progesterone production in mares and echographic evaluation of the corpora lutea formed after follicular aspiration

Ultrasound‐guided follicular aspiration was performed in 26 Criollo crossbred mares, followed by the evaluation of ultrasonographic images of the Corpus luteum (CL) that was formed after puncture of follicles of different diameters (Group 25–29 mm; Group 30–35 mm and Group >35 mm). Serum progesterone (P₄) concentrations were measured to determine CL function. The size of the CL was measured and the CL was classified based on the following echoscore: 1– anechoic tissue; 2– poorly defined luteal structure with low echogenicity; 3– echogenicity analogous to a luteal structure. The proportion of aspirated follicles that formed a functional CL (based on P₄ concentration) 8 days after aspiration was 57.1% (4/7; CL size 25–29 mm), 75.0% (6/8; CL size 30–35 mm) and 72.7% (8/11; CL size >35 mm), respectively (p > 0.05). The echographic scores of aspirated follicles (indicating the presence or absence of a CL) were consistent with serum P₄ concentrations (p < 0.0001). Of 26 aspirations, 18 resulted in luteal function confirmed by increased progesterone concentrations ([P₄] > 1.0 ng/ml); 17 of these mares (94.4%) had an echoscore (2–3) compatible with luteinization (p = 0.0372). Eight days after aspiration, serum [P₄] > 2.0 ng/ml was associated with high (p = 0.0056) CL echoscore (3) in 15 of 17 mares (88.2%). The echoscore used in this study was valuable as a screening test to detect the presence of a functional CL after aspiration. An echoscore of 3 served as a practical and efficient method to confirm luteinization.     

American trypanosomiasis infection in fattening pigs from the south-east of Mexico

American Trypanosomiasis (AT) is an infectious parasitic disease produced by the protozoa Trypanosoma cruzi (T. cruzi). Infection is acquired by vectorial via but can also be transmitted congenitally, by ingestion of an infected host, by transfusion with contaminated blood or transplant of organs from an infected donor. Currently, AT is widely distributed from the South of the United States to South America. In Mexico, the presence of the parasite has been reported throughout the country where several reservoirs such as dogs, opossums, rats and cats have been identified. Yucatan is in the south-east of Mexico where AT is endemic and has been reported since 1940s. There is little information about the role of pigs as reservoirs of T. cruzi. The frequency of specific antibodies against T. cruzi was determined in fattening pigs from Yucatan, Mexico. After sampling in the 3 main areas of pig production in the state, IgG ELISA and Western blot were performed to identify seropositive cases. Association of farm size, farm area and production system with infected pigs was evaluated. From 273 sampled pigs, 5.4% (n = 15) positive cases were found. No association with evaluated factors and infected pigs was found. Pigs are also reservoirs of T. cruzi in the studied area. These findings are considered important to improve vectorial control in the area in order to avoid the parasite infection in animal populations destined for human consumption and avoid further transmission to humans.