DNA methylation status of bovine blastocyst embryos obtained from various procedures

DNA methylation is an important factor for the regulation of gene expression in early embryos. It is well known that the satellite I sequence is more heavily methylated in bovine somatic cell nuclear transfer (NT-SC) embryos than in embryos derived from in vitro fertilization (IVF). However, the methylation status of bovine embryos obtained by other procedures is not well known. To clarify DNA methylation levels of bovine embryos obtained from various procedures, we examined satellite I sequences in bovine blastocyst (BC) embryos derived from NT-SC, NT using embryonic blastomeres (NT-EM), in vivo (Vivo), IVF and parthenogenetic treatment (PA). Furthermore, in order to evaluate the efficacy of DNA demethylation by the NT procedure, we determined the DNA methylation levels in bovine embryos in which NT was recapitulated (Re-NT). Although the DNA methylation levels in the NT-SC embryos were higher than those in the other embryos, the NT-EM embryos exhibited lower DNA methylation levels. The satellite I sequence in the NT-SC embryos was more demethylated than that in the donor cells. Although the DNA methylation level in the individual NT-SC embryos showed variation, the full-term developmental efficacy of these embryos were not different. These findings suggest that the methylation level of the satellite I sequence at the BC stage is not related to the abnormalities of bovine embryos produced by NT-SC. There was no difference in methylation levels between Re-NT and NT-SC embryos. Our results indicated that the DNA methylation status differed among embryos produced by various methods and that at least some of the demethylation of the donor cell genome occurred in the recipient cytoplast after NT-SC, but the demethylation ability of the NT procedure was noted in the first NT but not in the second NT.     

Implications of global and regional patterns of highly pathogenic avian influenza virus H5N1 clades for risk management

This paper analyses the publicly available data on the distribution and evolution of highly pathogenic avian influenza virus (HPAIV) H5N1 clades, whilst acknowledging the biases resulting from the non-random selection of isolates for gene sequencing. The data indicate molecular heterogeneity in the global distribution of HPAIV H5N1, in particular in different parts of East and Southeast Asia. Analysis of the temporal pattern of haemagglutinin clade data shows a progression from clade 0 (the ‘dominant’ clade between 1996 and 2002) to clade 1 (2003–2005) and then to clade 2.3.4 (2005 onwards). This process continuously produces variants, depending on the frequency of virus multiplication in the host population, which is influenced by geographical variation in poultry density, poultry production systems and also HPAI risk management measures such as vaccination. Increased multilateral collaboration needs to focus on developing enhanced disease surveillance and control targeted at evolutionary ‘hotspots’.     

ECOLOGY OF MYCOPLASMAS IN CLINICALLY HEALTHY CATS

Mycoplasmas were isolated from various sites and organs of a series of 319 clinically healthy cats. They include M. felis, M. gateae, M. Arginini, A. laidlawii, feline ureaplasmas, M. pulmonis, M. arthritidis, and M. gallisepticum. In addition, there were 10 strains of mycoplasmas which could not be identified with the specific antiserums by growth inhibition tests. Antibody against M. felis was demonstrated by haemagglutination-inhibition and complement-fixation tests in cats which were over 6 months of age. However, no such antibody against M. felis was detected in animals which were less than 6 months old. No antibody against A. laidlawii, M. gateae, M. arginini and feline ureaplasmas was demonstrated by the same serological methods. The significance of these mycoplasmas in cats is described.     

Effects of beta-glucanase-producing Lactobacillus strains on growth, dry matter and crude protein digestibilities and apparent metabolisable energy in broiler chickens

The effects of beta-glucanase expressed by transformed Lactobacillus strains on growth performance, apparent digestibilities of dry matter and crude protein, and apparent metabolisable energy were studied. Two hundred and forty 1-d-old chicks (Avian-43) were randomly divided into three dietary treatment groups and fed with the following diets: (i) basal diet (control) (BD); (ii) basal diet with parental Lactobacillus strains (BDP) and (iii) basal diet with transformed Lactobacillus strains (BDT). At 21 d of age, the body weight, body weight gain and feed conversion ratio of the BDT-fed chickens were significantly improved. At 14 and 21 d of age, the proportions of dry matter in the duodenum, jejunum, ileum, caeca and excreta of chickens given the BDT diet were significantly higher than those of chickens given the BD and BDP diets. Apparent metabolisable energy, digestibilities of crude protein and dry matter were also significantly improved (by 3.5, 5.6 and 3.5%, respectively) by the BDT diet. These results showed that the transformed Lactobacillus strains improved digestibility as well as enhanced the growth performance of chickens.     

富马酸二甲酯的合成和应用

添加化学防腐剂是防止饲料腐败变质的一种常用方法。目前,我国经常使用的化学防腐剂有苯甲酸(盐)、山梨酸(盐)、尼泊金酯、丙酸钙和双乙酸钠等。这些防腐剂有的应用pH值和抗菌谱窄,有的价格较高。富马酸二甲酯是一种高效低毒的化学防腐剂,对霉菌有良好的抑制作用。     

Isolation of chicken taste buds for real‐time Ca2+ imaging

We isolated chicken taste buds and used a real‐time Ca²⁺ imaging technique to investigate the functions of the taste cells. With RT‐PCR, we found that isolated chicken taste bud‐like cell subsets express chicken gustducin messenger RNA. Immunocytochemical techniques revealed that the cell subsets were also immunopositive for chicken gustducin. These results provided strong evidence that the isolated cell subsets contain chicken taste buds. The isolated cell subsets were spindle‐shaped and approximately 61–75 μm wide and 88–98 μm long, and these characteristics are similar to those of sectional chicken taste buds. Using Ca²⁺ imaging, we observed the buds' response to 2 mmol/L quinine hydrochloride (a bitter substance) and their response to a mixture of 25 mmol/L L‐glutamic acid monopotassium salt monohydrate and 1 mmol/L inosine 5′‐monophosphate disodium salt, umami substances. The present study is the first morphological demonstration of isolated chicken taste buds, and our results indicate that the isolated taste buds were intact and functional approaches for examining the taste senses of the chicken using Ca²⁺ imaging can be informative.     

The α2A‐adrenoceptor subtype plays a key role in the analgesic and sedative effects of xylazine

Xylazine, the classical α₂‐adrenoceptor (α₂‐AR) agonist, is still used as an analgesic and sedative in veterinary medicine, despite its low potency and affinity for α₂‐ARs. Previous pharmacological studies suggested that the α_2A‐AR subtype plays a role in mediating the clinical effects of xylazine; however, these studies were hampered by the poor subtype‐selectivity of the antagonists used and a lack of knowledge of their bioavailability in vivo. Here, we attempted to elucidate the role of the α_2A‐AR subtype in mediating the clinical effects of xylazine by comparing the analgesic and sedative effects of this drug in wild‐type mice with those in α_2A‐AR functional knockout mice using the hot‐plate and open field tests, respectively. Hippocampal noradrenaline turnover in both mice was also measured to evaluate the contribution of α_2A‐AR subtype to the inhibitory effect of xylazine on presynaptic noradrenaline release. In wild‐type mice, xylazine (10 or 30 mg/kg) increased the hot‐plate latency. Furthermore, xylazine (3 or 10 mg/kg) inhibited the open field locomotor activity and decreased hippocampal noradrenaline turnover. By contrast, all of these effects were abolished in α_2A‐AR functional knockout mice. These results indicate that the α_2A‐AR subtype is mainly responsible for the clinical effects of xylazine.     

Sensitivities of three county health department surveillance systems for child-related dog bites: 261 cases (2000)

OBJECTIVES: To determine incidence of child-related dog bites and sensitivities of 3 county health department dog-bite surveillance systems. DESIGN: Retrospective study. STUDY POPULATION: Child-related dog-bite data obtained from surveillance systems of 3 counties in Georgia in the year 2000. PROCEDURE: To characterize the sensitivity of health department dog-bite surveillance systems, 9 other potential sources of dog-bite records that matched records by victim name, age, gender, and incident date were evaluated. The number of reported bites and the most productive sources for identifying additional cases were determined. The Chandra Sekar-Deming capture-recapture method was used to estimate the number of unreported bites, and estimates of dog-bite incidence rates were made. RESULTS: 40, 36, and 185 dog bites were reported in the 3 counties, respectively. Capture-recapture calculations estimated an additional 9, 5, and 128 dog bites in these counties, respectively. Local health departments recorded 45.5% to 82.5% of dog bites. Local hospital emergency departments, police departments, and a rabies-testing laboratory received additional reports. Among these data sources, local hospital emergency department records were the best source for identifying additional cases. CONCLUSIONS AND CLINICAL RELEVANCE: Dog bites are a preventable cause of childhood injuries, and surveillance is a critical tool for tracking childhood dog bites in the community. Counties should use combined data from local health departments, local hospital emergency departments, and police departments to implement or revise dog-bite prevention programs.     

Effects of extracerebral dopamine on salsolinol‐ or thyrotropin‐releasing hormone‐induced prolactin (PRL) secretion in goats

The aim of the present study was to clarify the effect of extracerebral dopamine (DA) on salsolinol (SAL)‐induced prolactin (PRL) secretion in goats. An intravenous injection of SAL or thyrotropin‐releasing hormone (TRH) was given to female goats before and after treatment with an extracerebral DA receptor antagonist, domperidone (DOM), and the PRL‐releasing response to SAL was compared with that to TRH. DOM alone increased plasma PRL concentrations and the PRL‐releasing response to DOM alone was greater than that to either SAL alone or TRH alone. The PRL‐releasing response to DOM plus SAL was similar to that to DOM alone, and no additive effect of DOM and SAL on the secretion of PRL was observed. In contrast, the PRL‐releasing response to DOM plus TRH was greater than that to either TRH alone or DOM alone and DOM synergistically increased TRH‐induced PRL secretion. The present results demonstrate that the mechanism involved in PRL secretion by SAL differs from that by TRH, and suggest that the extracerebral DA might be associated in part with the modulation of SAL‐induced PRL secretion in goats.     

Efficacy of extended pirlimycin therapy for treatment of experimentally induced Streptococcus uberis intramammary infections in lactating dairy cattle

Streptococcus uberis is an important cause of mastitis in dairy cows throughout the world, particularly during the dry period, around the time of calving, and during early lactation. Strategies for controlling S. uberis mastitis have not received adequate research attention and are therefore poorly defined and inadequate. Objectives of the present study were to evaluate the efficacy of extended therapy regimens with pirlimycin for treatment of experimentally induced S. uberis intramammary infections in lactating dairy cows during early lactation and to evaluate the usefulness of the S. uberis experimental infection model for evaluating antimicrobial efficacy in dairy cows. The efficacy of extended pirlimycin intramammary therapy regimens was investigated in 103 mammary glands of 68 dairy cows that became infected following experimental challenge with S. uberis during early lactation. Cows infected with S. uberis in one or both experimentally challenged mammary glands were randomly allocated to three groups, representing three different treatment regimens with pirlimycin, including 2-day (n = 21 cows, 31 mammary quarters), 5-day (n = 21 cows, 32 quarters), and 8-day (n = 26 cows, 40 quarters). For all groups, pirlimycin was administered at a rate of 50 mg of pirlimycin hydrochloride via intramammary infusion. A cure was defined as an experimentally infected mammary gland that was treated with pirlimycin and was bacteriologically negative for the presence of S. uberis at 7, 14, 21, and 28 days after treatment. Experimental S. uberis intramammary infections were eliminated in 58.1% of the infected quarters treated with the pirlimycin 2-day regimen, 68.8% for the 5-day regimen, and 80.0% for the 8-day regimen. Significant differences (P <.05) in efficacy were observed between the 2-day and 8-day treatment regimens. The number of somatic cells in milk decreased significantly following therapy in quarters for which treatment was successful in eliminating S. uberis. However, there was no evidence to suggest that extended therapy with pirlimycin resulted in a greater reduction in somatic cell counts in milk than the 2-day treatment. The S. uberis experimental infection model was a rapid and effective means of evaluating antimicrobial efficacy during early lactation at a time when mammary glands are highly susceptible to S. uberis intramammary infection.