Distribution and Genotypic Characterization of Campylobacter jejuni Isolated from Poultry in Split and Dalmatia County,Croatia

Consumption of poultry contaminated with Campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. Therefore, the aim of our study was to investigate the prevalence and genotype diversity of Campylobacter jejuni in poultry meat intended for consumption in Split and Dalmatia County, which is the second biggest County in Croatia. Furthermore, we also wanted to discover possibly stable clones of C. jejuni appearing in different samples and periods of time, which would indicate their ability to persist in or adapt to poultry. In the period from March 2008 until June 2010, 834 samples of poultry from various sources were examined using a surface swab technique. Isolation of C. jejuni was performed by Preston broth and Karmali agar. Identification of the isolates was carried out using biochemical tests. C. jejuni was found in 84 of 574 chicken samples (14.6%) and in nine of 260 samples of turkey (3.5%). Pulse‐field gel electrophoresis (PFGE) was used to analyse 61 obtained isolates using SmaI and KpnI. Of 22 different macrorestriction profiles (MRP) that were found, five were detected in poultry from both different locations and periods of time. Samples from 11 locations were found to be contaminated with more than two different genotypes of C. jejuni. Interestingly, the same MRP were found both in poultry declared to be of domestic origin and in the poultry imported from abroad. The prevalence of C. jejuni in poultry samples was in accordance with previously reported results. Genotypic analysis indicated that the population of C. jejuni in Split and Dalmatia County was diverse and that multiple strains of C. jejuni could be found in the same poultry samples. Furthermore, the same genotypes were identified from the samples obtained from different locations and periods of time, which could support the theory of a global existence of certain MRP that are able to persist in or adapt to poultry.     

Experimental studies on the impact of an increased dose of anionic salts on the metabolism of dairy cows

Summary

This study was initiated to investigate the influence of a daily dose of anionic salts (AS) above the valid upper limit at present on metabolism of dairy cows. Eleven non‐pregnant and non‐lactating Holstein‐Friesian crossbred cows with a permanent rumen cannula were used in a study with a controlled feeding design. The initial daily dose was 2500 meq/day, which resulted in a Dietary Cation Anion Difference (DCAD) of ‐211 meq/kg dry matter. Every seven days, the daily dose was raised by 500 meq. If a cow stopped eating, the application of AS was stopped and these cows were monitored over the next seven days. On day 30 another batch of hay, having the same DCAD but higher concentrations of minerals and energy, was fed. Blood and urine samples were taken to monitor acid‐base balance and calcium concentrations. Acid‐base balance was strongly influenced by AS. Blood pH dropped steadily and reached values around 7.23. Urine pH dropped quickly below 6 and remained at that level regardless of the increased dosage of AS. Net acid base excretion (NABE) fell continuously with the increase of the dosage of AS and reached values below ‐200 mmol/l. Calcium concentrations in the serum were nearly stable, but those in urine increased sharply and remained on an elevated level with increasing doses of AS. A few days before the individual cow's refusal of feed intake, calcium excretion in urine decreased. The majority of cows stopped eating while consuming a diet containing 3500 to 4000 meq AS except two animals who consumed up to 6000 meq/day AS but they received the better hay in the second half of the treatment period. In this time pH in blood increased slowly. NABE remained stable on a low level at ‐100 mmol/l. The results showed that with an increasing amount of AS fed the risk of clinical acidosis increased. The decreasing urine concentrations of calcium indicate a breakdown of the compensation capability of the single cow. Besides the dose of AS fed, the quality of the feed stuff might be another factor concerning the tolerance of cows against AS.     

Effect of injecting amino acids into the egg on embryonic and subsequent growth in the domestic fowl

1. The growing embryo uses about 1% of the total protein available in the egg during the first week of incubation.

2. Injecting the growing embryo with amino acids equivalent to the amount of protein used resulted in higher weight of chicks at all stages up to 56 d.     

Risk ratio as parameter for the genetic characterization of complex binary traits in cattle. A simulation study under various genetic models using halfsib families

The risk ratio (λ_R) is defined as ‘the recurrence risk for a relative of an affected individual divided by the prevalence in the general population’ and is considered as the most important parameter when designing mapping experiments for diseases in humans. In this paper, the risk ratio was introduced as a parameter to genetically characterize complex binary traits such as mastitis in cattle. Simulations were applied to evaluate the properties of λ_R under different genetic models (monogenic, digenic, polygenic and mixed models) and in dependency of their parameters for a design consisting of unbalanced halfsib families typically found in dairy cattle. Population prevalences of the simulated data ranged from 5 to 40% and λ_R was estimated on a phenotypic level. In the discrete loci models complexity of the traits was introduced through different levels of penetrance and the proportions of phenocopies within each genetic background. The risk ratio and the power to reject the null hypothesis of independent halfsibs (λ_R=1) were influenced by the prevalence in all genetic models chosen. Absolute values for λ_R were higher for lower prevalences, for example, λ_R=2.77 and 1.62 for a pure monogenic recessive model with 5 and 20% prevalence, respectively, whereas the power decreases in the case of lower prevalences. For all the prevalences investigated, higher risk ratios were found for discrete loci models compared with the polygenic models, with higher values for the monogenic relative to the digenic models in general. For the mixed models, λ_R was intermediate between the polygenic and discrete loci models. Genes with dominant relative to recessive inheritance for susceptibility caused higher risk ratios in monogenic and mixed models, for example, λ_R=5.16 and 2.77 for a pure monogenic model with 5% prevalence. In the discrete loci models, λ_R decreased with lower penetrance and a higher proportion of phenocopies. Risk ratios increased with the heritability in the polygenic and in addition with the effect of the major gene in mixed models. Consistent patterns of risk ratios were observed under the analysed genetic models and parameters, which indicate that the risk ratio is a parameter well suited to genetically characterize binary traits in unbalanced halfsib families.     

影响牛精子体外获能的因素

本文针对牛胚胎体外生产的关键技术环节体外受精，从温度、卵母细胞、公牛个体、培养液组成、渗透压及pH值等方面阐述了影响牛精子体外获能的因素。     

三种抗球虫药物对家兔CYP1A2 mRNA和蛋白表达的影响

利用Real-time PCR和Western blot分别检测抗球虫药物磺胺喹噁啉、氯苯胍和妥曲珠利对家兔肝脏、肾脏、十二指肠中CYP1A2mRNA和蛋白表达的影响。40只新西兰白兔分为空白对照组、磺胺喹噁啉处理组、氯苯胍处理组和妥曲珠利处理组。空白对照组家兔饲喂正常颗粒饲料,磺胺喹噁啉处理组饲喂含25mg/kg浓度磺胺喹噁啉的颗粒饲料,氯苯胍处理组饲喂含66mg/kg浓度氯苯胍的颗粒饲料,妥曲珠利处理组饲喂含2mg/kg浓度妥曲珠利的颗粒饲料,连续30d。在第31天无菌条件下取出动物的部分肝脏、肾脏以及十二指肠,提取RNA、蛋白分别进行Real-time PCR、Western blot检测。Real-time PCR的结果表明,磺胺喹噁啉、氯苯胍和妥曲珠利都能够诱导家兔肝脏和肾脏CYP1A2mRNA的表达,而妥曲珠利能够显著的抑制十二指肠CYP1A2mRNA的表达。Western blot结果表明,磺胺喹噁啉、氯苯胍和妥曲珠利能够显著的诱导家兔肝脏CYP1A2蛋白水平的表达;磺胺喹噁啉显著的诱导肾脏和十二指肠CYP1A2蛋白表达,而氯苯胍能够显著的抑制肾脏CYP1A2蛋白的表达。结果提示,磺胺喹噁啉、氯苯胍和妥曲珠利对家兔CYP1A2mRNA和蛋白水平的表达均具有组织特异性,在不同的组织器官呈现出显著的诱导和抑制作用。     

苜蓿种带真菌及其致病性测定

对来自我国新疆,内蒙古,吉林,陕西,甘肃,宁夏,山东,黑龙江,山西,河北和江苏等１１个省（自治区）苜蓿产区的３０个审定登记品种的３８个苜蓿种子样品进行了系统的种带真菌检测研究;测定了２１种２５个真菌分离物对苜蓿幼苗根组织的入侵性及１９种真菌对苜蓿种子萌发和幼苗生长的影响。     

两种早熟禾超氧物歧化酶活性测定的适宜条件

以冷地早熟禾 (Poacrymophila)和草地早熟禾 (P .pratensis)为材料 ,探讨光照反应时间、酶浓度以及酶反应体系等对NBT光化还原反应的影响。随着光照反应时间的延长 ,最大光还原值直线上升。 2 5min后光化还原值逐渐趋于稳定。光照反应开始后 ,酶液对NBT光化还原反应速率的抑制百分率迅速升高 ,在 15～ 2 5min时 ,酶液对NBT光化还原反应的抑制百分率保持稳定 ,2 5min后下降速率较大。光照反应时间对SOD酶活性测定的影响较大 ,确定为 2 5min较为适宜。酶浓度在 4 0～ 6 0mg/mL时光化还原值变化速率较一致。测定中反应体系NBT、核黄素的加样量以及光照强度在一定范围内时光化还原值较为稳定     

牛初乳sIgA等电点的测定

采用等电聚焦电泳法测定牛初乳sIgA的等电点。牛初乳sIgA的pI为5．02—5．67,比相关报道更加精确。回归方程：y=0．7543x＋3．1635,相关系数r=0．9989,表明线性关系良好,为牛初乳sIgA的离子交换层析制备条件的选择提供了可靠的理论依据。