Construction of Eukaryotic Expression Vector of Chicken MHCⅠα and β2m Genes and Its Expression in 293T Cells

To explore the mechanism of MHCⅠ molecule in immune response,chicken MHCⅠα and β₂m genes were cloned by PCR.Then the fragments were inserted into the eukaryotic expression vector with fluorescent protein,and the recombinant plasmids pEGFP-MHCⅠα and pmCherry-MHCⅠβ₂m were constructed.The recombinant plasmids were transfected into 293T cell with lipofectin reagent.The gene products of recombinant plasmids were mainly located to endomembrane system of the cells by fluorescence microscopy,and changed the intracellular localization of the fusion with the fluorescent protein.Moreover,the positive reactions were observed by the method of Western blotting,and the proteins had the molecular weight of 68.3 and 41.3 ku,respectively,in accord with the target proteins.The results showed that the recombinant plasmids were expressed in 293T cells with a good immunological activity,and the proteins had the binding reaction with specific antibodies.     

Effect of supplementation with corn oil on postpartum ovarian activity,pregnancy rate,and serum concentration of progesterone and lipid metabolites in F<Subscript>1</Subscript> (<Emphasis Type="Italic">Bos taurus</Emphasis> × <Emphasis Type="Italic">Bos indicus</Emphasis>) cows

The aim was to evaluate the effect of corn oil supplementation during postpartum anoestrus on ovarian activity, pregnancy rate, progesterone (P₄), and lipid metabolites (cholesterol, CHO; low and high density lipoproteins; LDL and HDL, respectively) concentrations in blood of F₁ (Bos taurus × Bos indicus) grazing cows. Cows were randomly assigned to an experimental group, fed with a supplement containing 4% corn oil on dry matter basis (OG, n = 11), and a control group with the same supplement without corn oil (CG, n = 12). Both supplements contained equivalent amounts of crude protein and metabolizable energy and were fed for 34 days continuously. All cows were induced to estrous 12 days after beginning of supplementation by using a synthetic progestagen and artificially inseminated 56 h after retiring the implants. Pregnancy diagnosis was performed by transrectal palpation 45 days after insemination, evaluating simultaneously ovarian activity. P₄ and lipid metabolites (CHO, HDL, LDL) concentrations were determined in blood samples collected at 3-day intervals, from the beginning of corn oil supplementation and up to 10 days after artificial insemination. Ovarian activity was affected by treatment (p < 0.05), finding ovarian structures in 72.7% of OG cows and in 50% of CG cows. Concentration of P₄ and CHO was higher for OG with respect to CG (2.52 ± 0.65 vs 1.88 ± 0.62 ng/ml and 117.79 ± 11.57 vs 85.71 ± 12.11 mg/dl, respectively), whereas pregnancy rate and blood concentrations of HDL and LDL were not affected by treatment (p > 0.05). Addition of corn oil to the supplement stimulated ovarian activity and increased serum concentrations of progesterone and cholesterol in grazing B. taurus × B. indicus cows with low body condition score showing postpartum anoestrus.     

“Bending” and beyond: Better estimates of quantitative genetic parameters?

Multivariate estimation of genetic parameters involving more than a handful of traits can be afflicted by problems arising through substantial sampling variation. We present a review of underlying causes and proposals to improve estimates, focusing on linear mixed model‐based estimation via restricted maximum likelihood (REML). Both full multivariate analyses and pooling of results from overlapping subsets of traits are considered. It is suggested to impose a penalty on the likelihood designed to reduce sampling variances at the expense of a little additional bias. Simulation results are discussed which demonstrate that this can yield REML estimates that are on average closer to the population values than their unpenalized counterparts. Suitable penalties can be obtained based on assumed prior distributions of selected parameters. Necessary choices of penalty functions and of the stringency of penalization are examined. We argue that scale‐free penalty functions lend themselves to a simple scheme imposing a mild, default penalty which can yield “better” estimates without being likely to incur detrimental effects.     

The Activity and Localization of 3β-hydroxysteroid Dehydrogenase/Δ5-Δ4 Isomerase and Release of Androstenedione and Progesterone by Uterine Tissues During Early Pregnancy and the Estrous Cycle in Pigs

Steroid hormones are produced by the porcine uterus. We hypothesized that the uterus in pigs possesses active 3β-hydroxysteroid dehydrogenase/Δ⁵-Δ⁴ isomerase (3β-HSD) responsible for progesterone and androstenedione production, that uterine steroids may supplement the amount of steroid hormones produced by embryos and corpus luteum and that these steroids are necessary for maintenance of pregnancy. In this study, we examined 1) endometrial and myometrial expression of 3β-HSD mRNA, 2) uterine 3β-HSD protein activity and 3) in vitro production of A₄ and P₄ by uterine slices harvested from pigs on days 10 to 11, 12 to 13 and 15 to 16 of pregnancy and the estrous cycle. The expression of 3β-HSD and the presence and activity of 3β-HSD protein were different in the endometrium and the myometrium during the examined periods of pregnancy and the estrous cycle. Production of A₄ by the endometrium and myometrium was highest on days 12 to 13 of pregnancy and the estrous cycle. Endometrial secretion of P₄ did not differ in the course of early pregnancy and on the respective days of the estrous cycle. The gravid myometrium was the highest source of P₄ in pregnant pigs on days 12 to 13. The release of P₄ by the cyclic myometrium rose during the examined days of the estrous cycle. The steroidogenic activity of the uterus, as described in this study, may support early pregnancy or the luteal phase of the estrous cycle in pigs.     

新&#183;城&#183;疫&#183;诊&#183;断&#183;试&#183;剂&#183;的&#183;改&#183;进

鸡新城疫是危害养鸡业的主要疾病之一。但血凝和血凝抑制（HA-HI）试验中使用的新鲜鸡红细胞保存时间短，洗涤制备需仪器。本试验通过对红细胞醛化使其保存期延长，并对抗原稳定性初步研究，借此建立一种操作简单的新城疫诊断试剂盒。研究结果报告如下。     

Doxycycline treatment for <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> in dogs: impact on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Enterococcus</Emphasis> antimicrobial resistance

Doxycycline is an antibiotic that, in addition to the classic antibacterial use, is also prescribed to fight parasitic diseases, like heartworm disease in dogs. Despite the concern that the overuse of this antibiotic may decrease susceptibility of clinically important bacteria, the consequences of the prolonged doxycycline therapy in heartworm-infected dogs have never been studied before. We have analyzed the impact of this therapy on Staphylococcus aureus and Enterococcus antimicrobial resistance. In this study, 17 heartworm-infected dogs (10 that had completed the doxycycline treatment and 7 dogs that had not yet begun) were included. Twenty-four isolates of Staphylococcus aureus were obtained from two locations of each dog. After treatment, 73.3% of isolates were resistant to at least one antibiotic but only 22.2% of isolates before treatment. Most of doxycycline resistant isolates were obtained from dogs that have received treatment. Erythromycin resistance or intermediate susceptibility was detected in 45.6% of isolates, most of them from dogs after treatment. For Enterococci, 48 isolates were obtained from fecal samples (25 before treatment and 23 after treatment). Before treatment, 32% of isolates were resistant at least to one antibiotic while after, this data increase up to 65%. Comparing isolates before and after treatment, a clear increase in resistance to doxycycline (12% against 21.74%) and erythromycin (20% against 39.13%) was observed. Although the present work is a preliminary research, the results encourages the development of further studies to determinate the effect of prolonged doxycycline therapy on antimicrobial resistance.     

Panton–Valentine leukocidin and some exotoxins of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and antimicrobial susceptibility profiles of staphylococci isolated from milks of small ruminants

The aims of this study were to determine the existence of pvl gene, some toxin genes, and mecA gene in Staphylococcus aureus strains isolated from sheep milk and to examine antimicrobial resistance profiles in staphylococci from sheep and goats' milk. The milk samples were collected from 13 different small ruminant farms in Kirikkale province from February to August 2009. A total of 1,604 half-udder milk samples from 857 ewes and 66 half-udder milk samples from 33 goats were collected. Staphylococcus spp. were isolated and identified from the samples. Toxin genes and mecA gene among S. aureus strains were determined by PCR. Antimicrobial susceptibility of staphylococci was examined by the disk diffusion method on Mueller-Hinton agar, and interpreted according to the Clinical Laboratory Standards Institute (CLSI) guidelines. The prevalence of subclinical intramammary infection in both ewes and goats was 5.2%. The most prevalent subclinical mastitis agents were coagulase-negative staphylococci and S. aureus with prevalences 2.8% (n:46) and 1.3% (n = 21), respectively. The prevalence of resistances in isolated Staphylococcus spp. to penicilin G, tetracycline, erythromycin, gentamicin, and enrofloxacin were found as 26.9% (18), 7.5% (5), 6.0% (4), 3.0% (2), and 1.5% (1), respectively. Only 3 of the 21 S. aureus ewe isolates (13.4%) were shown to harbor enterotoxin genes being either seh, sej or sec. However, fourteen (66.6%) of the 21 S. aureus isolates had pvl gene while none of the isolates harbored mecA gene. In conclusion, Staphylococci were shown to be the most prevalent bacteria isolated from subclinical mastitis of ewes and goats and these isolates were susceptible to most of the antibiotics. In addition, S. aureus strains isolated from ewes were harboring few staphylococcal enterotoxin genes. However, Panton–Valentine leukocidin produced by S. aureus could be an important virulence factor and contribute to subclinical mastitis pathogenicity.