抗鸡传染性腺胃炎酶标抗体的制备和应用

在前期从患病鸡腺胃内分离到呼肠孤病毒的基础上，以该病毒为免疫原对兔进行免疫制备高免血清，分离纯化IgG，进行辣根过氧化物酶（HRP）标记，并用ELISA双抗体夹心法对山东省青岛、枣庄、泰安等地区采集的传染性腺胃炎的腺胃样品进行检测，结果表明制备的抗鸡传染性腺胃炎呼肠孤病毒的酶标抗体检测传染性腺胃炎特异性强、灵敏度高、较简便。     

血清中IBV特异性IgG及总IgG含量变化规律的研究

本研究采用间接酶联免疫吸附试验对人工感染鸡传染性支气管炎病毒的SPF鸡血清中特异性抗体及总IgG含量进行检测并比较其相关性.结果表明感染IBV-M41第1 d、3 d、5 d、7 d、10d、14d、21 d、28 d的血清中特异性抗体水平随感染天数增加而升高,于第14天达到最高,之后略呈降低趋势,与健康对照比较差异显著(P＜0.01);总IgG含量较健康对照组增幅小,差异显著(P＜0.01).同时,两者的消长趋势基本一致.     

广西猪瘟流行毒株全基因组的克隆与序列分析

参考已发表的猪瘟病毒(CSFV)Shimen株、HCLV株、Paderborn株的核苷酸序列，设计并合成11对引物。应用RT—PcR技术，成功地分11个片段扩增了CSFV广西流行毒株GXWZ02的全基因组，将这11个基因片段克隆，并测定了其核苷酸序列。应用计算机生物软件Vector NT1将11个基因片段进行拼接。确认CSFV GXWZ02株全基因组序列的长度为12296个核苷酸(GenBank收录号AY367767)。将GXXZ02株与国内外已发表的Slhimen、HCLV、39、Brescia、Eystrup、Glentorf、Alfort187、Pader190rn、CS、ALD、GPE、P97、LPC毒株进行全基因组核苷酸序列和推定的氨基酸序列比较，核苷酸同源性分别为85．4％、84．6％、88．7％、85．7％、85．7％、85．3％、85．6％、95．3％、85．7％、85．7％85．4％、83．4％、84．3％，推定的氨基酸同源性分别为92．6％、91．7％、94．2％、93．1％、92．9％、92．3％、93．0％、97．7％、92．6％、93．0％、92．6％、90．5％、90．6％。GXWZ02与Shimen、HCLV的全基因组序列及推定的氨基酸序列有明显差异，表明GXWZ02株在遗传性和抗原性上发生了较明显的变化。系统发育树分析，所比较的14株CSFV被分为2个群：GXWZ02、Paderborn和39这3个毒株被归为群Ⅰ，其余的11个毒株被归为群Ⅱ，GXWZ02与Paderborn的亲缘关系最接近。将GXXZ02与Shimen、HCLV基因组中单个基因分别进行核苷酸及推定的氨基酸序列同源性比较，结果显示，基因E^ms、E2、P7、NS2、NS5A的遗传变异程度大。而NS3、NS4A、NS4B的遗传性则相对保守。     

大豆黄酮对肉仔鸡生长相关激素水平与免疫机能的影响

将640只1日龄艾维茵肉鸡公母分养,分别随机分为4组,分别饲喂添加大豆黄酮0、5、10、15mg/kg的4种饲粮,于试验28、49d采血和屠宰,探讨了日粮中添加不同水平大豆黄酮对肉仔鸡生长相关激素水平及免疫机能的影响。结果显示,日粮中添加大豆黄酮28、49d,均可显著提高公鸡血清生长激素(GH)、胰岛素样生长因子-1(IGF-1)、睾酮(TESTO)的含量(P<0.05或P<0.01),显著降低公鸡血清尿素氮水平(P<0.05或P<0.01),而对母鸡血清GH、IGF-1、尿素氮水平无明显影响;添加大豆黄酮49d,显著提高公鸡T淋巴细胞ANAE阳性率(P<0.05或P<0.01),添加大豆黄酮28d,显著提高公鸡NDV抗体效价(P<0.05),而对母鸡无明显影响(P>0.05);添加大豆黄酮28、49d,对公母鸡免疫器官重量有增加的趋势,但差异不显著(P>0.05)。     

H1N1猪流感广东株血凝素基因的克隆与序列分析

用RT-PCR方法扩增H1N1亚型猪流感病毒广东分离株A／Swine／GuangDong／711／2001HA基因，对其进行了克隆和测序。结果显示，HA基因全长1771bp．共编码579个氨基酸。A／Swine／Guang Dong／711／2001HA基因编码的氨基酸序列中有8个糖基化位点，5个位于HAl基因的第27、28、40、104和287位点，3个位于HA2基因的21、153和213位点。与H1N1亚型猪流感经典毒株比较后发现，血凝素糖基化位点并不是高度保守的。将A／Swine／Guang Dong／711／2001与自Gen Bank读取的1918年人流感毒株A／South Carolina／1／18、1991年人流感毒株A／MD／12／91和1997年猪流感毒株A／Swine／Wisconsin／238／97等进行核苷酸同源性比较分析，A／Swine／Guang Dong／711／2001与A／SouthCarolina／1／18、A／MD／12／91和A／Swine／Wisconsin／238／97等毒株的核苷酸同源性分别为93．9％、94．7％和93．9％。从系统发生树来看，A／Swine／Guang Dong／711／2001与A／South Carolina／1／18和A／Swine／Wisconsin／238／97的亲缘关系相近。     

抗附红细胞体有效药物的筛选

以PCR检测附红细胞体呈现阳性，且镜检染虫率达909／5以上的猪血样为研究对象，选择几种附红细胞体敏感的药物(血虫净，附红净，庆大霉素，博士914，土霉素，三毒清，红弓链914，附红120，红弓链克，乌金土霉素)和对支原体敏感的恩诺沙星，对立克次氏体敏感的红霉素，采用两种方法进行体外药效试验。附红净对附红细胞体的作用效果最明显，而庆大霉素对附红细胞体基本无效，其他药物对附红细胞体也有一定杀灭效果。     

Methyl donors dietary supplementation to gestating sows diet improves the growth rate of offspring and is associating with changes in expression and DNA methylation of insulin‐like growth factor‐1 gene

The study aimed to investigate the effects of maternal dietary methyl donors on the performance of sows and their offspring, and the associated hepatic insulin‐like growth factor‐1 (IGF‐1) expression of the offspring. A total of 24 multiparous sows were randomly fed the control (CON) or the CON diet supplemented with methyl donors (MD) at 3 g/kg betaine, 15 mg/kg folic acid, 400 mg/kg choline and 150 μg/kg VB₁₂, from mating until delivery. After farrowing, sows were fed a common lactation diet through a 28‐days lactation period and six litters per treatment were selected to be fed until at approximately 110 kg BW. Maternal MD supplementation resulted in greater birthweight (p < 0.05) and increased the piglet weights (p < 0.01) and litter weights (p < 0.05) at the age of day 28, compared with that in CON group. The offspring pigs in the MD group had greater ADG (p < 0.05) and tended to lower F:G ratio (p = 0.07) compared with that of CON group from day 28 to 180 of age. The offspring pigs from MD group had greater serum IGF‐1 concentrations and expressions of hepatic IGF‐1 gene and muscular IGF‐1 receptor (IGF‐1r) protein at birth (p < 0.05), and greater hepatic IGF‐1 protein (p = 0.03) and muscular IGF‐1r gene expressions (p < 0.05) at slaughter, than that from the CON group. Moreover, the methylation at the promoter of IGF‐1 gene in the liver of newborn piglets and finishing pigs was greater in the MD group than that of the CON group (p < 0.05). In conclusion, maternal MD supplementation throughout gestation could enhance the birthweight and postnatal growth rate of offspring, associated with an increased expression of the IGF‐1 gene and IGF‐1r, as well as the altered DNA methylation of IGF‐1 gene promotor.     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

紫花苜蓿茎秆组织中木质素的分布与沉积模式

以甘农5号紫花苜蓿(Medicago sativa‘Gannong No.5’)为材料,采用组织化学染色方法研究了茎秆中木质素的发生、分布与沉积规律。结果表明,苜蓿茎秆中木质素的分布与生长部位密切相关,在茎秆顶端初生维管组织中仅木质部有木质素分布,随节间下移,木质素开始在初生韧皮纤维、次生木质部和髓射线中大量沉积;茎秆中存在G、S两种木质素,S木质素的发生迟于G木质素;在维管束之间,两种木质素均以"厚角组织处维管束→厚角组织间维管束"的模式沉积,但在维管束内,木质素沉积表现出异质性,G木质素沉积模式为"初生木质部导管→初生韧皮纤维→次生木质部和髓射线";S木质素沉积模式为"初生韧皮纤维→次生木质部和髓射线→髓薄壁细胞"。分析认为,紫花苜蓿茎秆中木质素特殊的沉积模式可能是其对北温带生长环境的一种适应对策。     

新疆乌鲁木齐市犬布鲁氏菌病流行病学调查

为了解新疆乌鲁木齐市犬布鲁氏菌病流行特征,分析犬感染风险因素,初步掌握养犬人员对布鲁氏菌病的认知程度和行为特点,从而提出合理的预防对策和控制措施,采用流行病学横断面研究方法,通过估计流行率计算抽样数量,然后随机抽样,采用IELISA方法检测光滑型和粗糙型犬布鲁氏菌,同时结合调查问卷,获得乌鲁木齐市犬布鲁氏菌病流行率、感染风险因素和养犬人员对布鲁氏菌病认知情况,并对调查结果采用SPSS和Excel软件进行数据统计分析。结果显示:乌鲁木齐市犬布鲁氏菌病总流行率为25.5%(95%CI:20.9%~30.1%),且犬种和牛羊种并存,公共卫生威胁严重;从不同群体看,牧区流行率最高,为41.5%(95%CI:31.0%~52.0%);外购、散养、常接触牛羊、喂生肉、不处理犬粪便等行为是乌鲁木齐市犬布鲁氏菌感染的潜在风险因素,其中"常接触牛羊"(OR=1.97,95%CI:1.47~15.35)、"喂生肉"(OR=5.06,95%CI:1.69~20.74)是主要风险因素;乌鲁木齐市养犬人员对布鲁氏菌病的认知程度低,个人防护意识差。因此,应加强对犬感染牛羊种布鲁氏菌的公共卫生防控,减少犬与牛羊的直接接触,不喂生肉,定期开展犬布鲁氏菌病专项检测,同时加强宣传,提高养犬人员的布鲁氏菌病认知度。