MALDI-TOF-MS用于微生物鉴定的影响因素分析

影响MALDI-TOF-MS微生物鉴定的因素有很多。本研究主要从微生物培养基、培养状态、菌体前处理、上样量等方面进行MALDI-TOF-MS操作技术和方法的探讨。研究结果显示:进行MALDI-TOF-MS细菌鉴定,培养时间在4～24 h之间为佳;尽量选择无色、不含盐的液体培养基;0.1μL菌液是最低鉴定菌量;上样量对结果无影响;对于菌体前处理,除按照标准化前处理程序操作外,也可将菌体加dd H2O制成混悬液进行点靶鉴定,这样更加简便。本研究有助于减少MALDI-TOF-MS操作盲目性,提高鉴定质量,利于操作的规范化和简便化。     

Serine-to-glycine ratios in low-protein diets regulate intramuscular fat by affecting lipid metabolism and myofiber type transition in the skeletal muscle of growing-finishing pigs

Serine and glycine are 2 of the first-affected nonessential amino acids in low crude protein (CP) diets for pigs. Therefore, we explored the effects of different dietary serine-to-glycine ratios on growth performance and lipid metabolism in growing-finishing pigs. A total of 160 crossbred healthy barrows, with a similar body weight of around 59.50 kg, were randomly allotted into 1 of 5 treatments (8 pens per treatment and 4 pigs per pen). The serine-to-glycine ratios of the 5 dietary treatments were as follows: diet A (NORMAL group), 1.18:1 (16% CP); diet B (LOW group), 1.2:1 (12% CP); diet C (S2G1 group), 2:1 (12% CP); diet D (S1G2 group), 1:2 (12% CP); and diet E (S1G1 group), 1:1 (12% CP).We found that the pigs fed a low CP diet (12% CP), when maintaining serine-to-glycine ratio at 1:2 and a total amount of 1.44%, had the same average daily gain as the pigs fed a normal CP diet (16% CP) (P > 0.05), but they had increased intramuscular fat (P < 0.05). Furthermore, they exhibited higher expression of genes involved in lipid oxidation (P < 0.05), which was regulated by modulating methylation levels in the promoters of acyl-CoA oxidase 1 (ACOX1) and acyl-CoA dehydrogenase medium chain (ACADM). When compared with the pigs fed a normal CP diet, these pigs had more oxidative myofibers (P < 0.05), which were regulated by AMPK-PGC-1α and Calcineurin-MEF2/NFAT pathways in a coordinated manner. Our findings suggested that a dietary serine-to-glycine ratio of 1:2 is beneficial for improving meat quality in pigs fed a low CP diet.     

Dietary supplementation with Acanthopanax senticosus extract enhances gut health in weanling piglets

The present study was conducted to investigate the effects of Acanthopanax senticosus extract (ASE) as a dietary additive on gut health in weanling piglets by examining diarrhea frequency, intestinal microbiota and morphology. A total of 96 Duroc× (Landrace × Yorkshire) piglets weaned at 21 days of age with an average initial body weight (BW) of 5.6 ± 0.4 kg were randomly assigned to 3 treatment groups with 4 duplicates of 8 piglets each. The piglets were fed basal diet to which had been added 0 or 1 g/kg of ASE, or 0.7 g/kg antibiotics, respectively. Fecal consistence was monitored twice daily and the frequency of diarrhea was calculated. On day 21 after the initiation of supplementation, 8 piglets were randomly selected from each treatment group (2 piglets per pen) and slaughtered. The jejunum, ileum, colon and cecum were then excised and fixed in 10% neutral formalin solution to determine villus height and crypt depth, after their contents were collected to determine microbiota. The results showed that dietary supplementation with ASE increased (P < 0.05) the density of bacterial populations that co-migrated with Lactobacillus amylovorus, Lactobacillus salivarius, Bacillus subtilis, and Clostridium lituseburens, but decreased (P < 0.05) those co-migrating with Staphylococcus aureus, Salmonella typhimurium, Ruminococcus forques, and E. coli O157:H7 in the PCR-DGGE profiling analysis when compared with the control group. The villus height of the duodenum, jejunum and ileum increased (P < 0.05) by 14.8, 13.7 and 10.0%, while the crypt depth decreased (P < 0.05) by 17.9, 9.1 and 12.1%, respectively, in response to dietary ASE supplementation. Additionally dietary supplementation with ASE or an antibiotic decreased (P < 0.05) the frequency of diarrhea by 55.6 and 52.2%, respectively, compared with the control group. In conclusion, these findings suggest that dietary supplementation with ASE could regulate the microbiota composition and maintain a normal morphology of gut mucosa in weanling piglets, thereby decreasing diarrhea that resulted from weaning stress.     

复方中草药抗乳炎奶牛饲料添加剂的研制

试验结果显示,所研制的3种新制剂对奶牛乳腺炎均有明显的防治效果,同时对提高产奶量,改善牛乳的品质和风味、降低牛乳中的体细胞数也有显著的效果。     

我国部分地区送检样品3种常见猪病血清学和病原学检测

为了解我国部分地区猪瘟、猪繁殖与呼吸综合征和猪伪狂犬病3种常见疫病的流行情况,采用ELISA、聚合酶链式反应（PCR）和反转录PCR（RT-PCR）等方法,对2019年11个省（自治区）172个猪场送检的3 550份猪血清样品进行猪瘟病毒（CSFV）、猪繁殖与呼吸综合征病毒（PRRSV）及伪狂犬病野毒（PRV-gE）抗体检测,对9个省（自治区）72个猪场送检的371份病料进行抗原检测。结果显示：CSFV、PRRSV、PRV-gE抗体阳性率分别为82.75%、84.08%、31.86%,抗原阳性率分别为2.62%、26.15%、1.67%。保育猪群CSFV抗体平均阻断率最低,为44.62%,抗体整齐度差,CV为58.26%,且CSFV抗原主要在哺乳仔猪和保育猪病料中被检出,检出率分别为3.57%、3.00%;除育肥猪群外,其他各阶段猪群的PRRSV抗体S/P平均值为1.0~2.0,抗体整齐度整体较差,且平均抗原阳性率较高（26.15%）,其中保育阶段猪群病原检出率最高（37.68%）;育肥猪、母猪、公猪的PRV-gE抗体阳性率分别为36.82%、35.31%、15.65%,病原主要从死胎/木乃伊胎和哺乳仔猪病料中被检出,检出率分别为8.70%和2.99%。结果表明,PRRSV对我国猪场不同阶段猪群均有较大威胁;保育猪群CSFV抗体水平不理想,暴发疫情风险较高;公猪、母猪及育肥猪群普遍存在PRV感染,尤其是母猪。提议各猪场采取免疫、监测、净化等综合措施,加强这3类常见猪群疫病的防控。     

Optimizing the Conditions for In Vitro Maturation and Artificial Activation of Sika Deer (Cervus nippon hortulorum) Oocytes

With the goal of establishing experimental protocols for cloning sika deer, various conditions for in vitro maturation (IVM) and artificial activation of sika deer oocytes were examined. In vitro maturation was evaluated in seven different culture media. The highest rate of oocyte maturation was 75.4% in 10 μg/ml follicle‐stimulating hormone (FSH), 1 μg/ml LH, 0.2 mm cysteamine and 50 ng/ml epidermal growth factor (EGF) after 24 h of IVM. The efficiency after 24 h of IVM did not differ significantly (p > 0.05) from that observed after 20 h. Cysteamine (0.2 mm ) significantly increased the maturation rates after 20 h (from 59.1% to 67.2%, p < 0.05) and after 24 h (from 63.2% to 71.6%, p < 0.05) of IVM. The IVM rates of oocytes collected during the oestrous season (75.4%) and the anoestrous season (23.3%) were significantly different at 24 h. The 20 μg/ml FSH, 2 μg/ml LH, 0.4 mm cysteamine and 100 ng/ml EGF significantly increased the maturation rates (from 23.3% to 54.2%, p < 0.01) at 24 h during the anoestrous season. For the activation experiments, the most effective method was chemical activation [ionomycin + 6‐dimethylaminopurine (6‐DMAP)], which promoted the development of sika deer oocytes to the blastocyst stage (32.4%). Our results indicate that in vitro matured sika deer oocytes are good candidates for parthenogenetic activation and that chemical treatment is needed for relatively efficient activation of the oocytes. These optimized conditions for IVM and parthenogenetic activation may be useful for efforts to restore populations of the endangered sika deer using the somatic cell nuclear transfer technique.     

山东省野鸟栖息地情况调研

2021年1月,山东省东营市发生天鹅H5N8亚型高致病性禽流感疫情,给山东省禽流感疫情防控与净化带来了极大挑战.为了解山东省野鸟栖息地及野鸟分布信息,采用问卷调查方式,对山东省野鸟栖息地类型、分布、野鸟种类以及迁徙时间等进行调研.同时,采集野鸟粪便,用荧光定量RT-PCR或PCR方法进行H5亚型禽流感、新城疫、 鸭瘟、...     

在种母鸡按标准饲喂条件下雏鸡卵黄囊中钙、磷、锰、锌含量的探讨

从按NRC标准饲养的种母鸡所产种蛋新孵出的AA肉用雏鸡中随机选取80只(公母各半),测定卵黄囊中钙、磷、锰和锌的含量及其与出壳体重和卵黄囊重的相关回归关系。结果表明,雏鸡出壳体重为48.15±0.37g,卵黄囊重为7.50±0.16g,卵黄囊中钙、磷、锰、锌的含量分别为26.86±1 90mg、49.46±1.69mg、0.018±0.002mg、0.222±0.011mg。雏鸡卵黄囊重与出壳体重的相关和回归关系显著(P≤0.05,r=0.60,b=0.25),卵黄囊重对出壳体重的回归方程为:Y=0.25X-3.75,卵黄囊中锌和磷的含量与出壳体重的相关及回归关系显著(P≤0.05,r=0.60,b=0.25),卵苏囊 重对出体重的回归方程为y=0.25x-3.75。卵苏囊中锌和磷的含量与出壳体重的相关及回归关系显著(P≤0.05)r=0.22,0.37:b=0.067,1.69)。因此,可由出壳体重估测卵黄囊重,对通过出壳体重估测卵黄囊中锌和磷的含量,也获得较为满意的结果。     

家畜住肉孢子虫研究进展

住肉孢子虫是一类细胞内的寄生性原虫,广泛发现于爬行类、鸟类和包括人、猴、鲸、各种家畜在内的哺乳动物。根据Ｌｅｖｉｎｅ（１９８６）统计,文献记载的虫种有１２０多种,其中已知生活史的有５６种,寄生在家畜有２０多种,而近１０年来仍不断有新种报道。７０年代生...     

外源病毒检验用抗鸡传染性支气管炎病毒特异性血清的制备与检定

为制备外源病毒检验用抗鸡传染性支气管炎病毒(IBV)特异性血清,对300只3~4周龄的SPF鸡进行基础免疫和加强免疫;最后一次免疫21 d后,采血并分离血清;对血清经混合、分装、冷冻、真空干燥后,进行无菌检验、外源病毒检验、剩余水分测定、中和效价测定和特异性检验。结果表明:本研究制备的抗IBV特异性血清的无菌检验、外源病毒检验和剩余水分测定结果均符合现行《中华人民共和国兽药典》规定;血清中和效价为1:800,与IBV H120种毒株的中和指数为106.3,与鸡传染性法氏囊病毒B87株、鸡新城疫病毒La Sota株、鸡传染性喉气管炎病毒、鸡痘病毒、禽呼肠孤病毒S1133株的中和指数均不大于10;通过AGP、ELISA、HI等试验,确定该血清中不含其他禽源外源病毒抗体。本研究为鸡传染性支气管炎活疫苗或毒种的外源病毒检验提供了具有良好特异性和高效价的中和用血清。