Development and characterization of monoclonal antibodies to subgroup A avian leukosis virus

Avian leukosis virus subgroup A (ALV‐A) is a retrovirus which infects egg‐type chickens and is the main pathogen of lymphoid leukosis (LL) and myeloid leukosis (ML). In order to greatly enhance the diagnosis and treatment of clinical avian leukemia, two monoclonal antibodies (MAbs) to ALV‐A were developed by fusion between SP2/0 and spleen cells from mice immunized with expressed ALV‐A env‐gp85 protein. Using immunofluorescence assay (IFA), two MAbs reacted with ALV‐A, but not with subgroups B and J of ALV. Western blot tests showed that molecular weight of ALV‐A envelope glycoprotein recognized by MAbs was about 53 kD. Isotyping test revealed that two MAbs (A5C1 and A4C8) were IgG1 isotypes. These MAbs can be used for diagnosis and epidemiology of ALV‐A.     

《畜产品加工》课程教学内容的研究与设计

通过介绍《畜产品加工》课程教学内容设计的全过程,重点对畜产品加工相关岗位典型工作任务和职业能力进行分析,围绕课程设计的理念和思路,紧密结合教学规律和学生的认知规律,对课程教学内容的设计进行了论述,旨在为探索中等职业学校《畜产品加工》课程教学内容改革提供参考。     

饲粮半胱氨酸水平对獭兔生产性能和SLC1A4基因表达的影响

试验旨在研究饲粮中半胱氨酸含量对獭兔生长性能、毛皮品质和SLC1A4基因表达的影响。选取90日龄健康獭兔144只,随机分为4组(每组36个重复,每个重复1只)。对照组饲喂基础饲粮(半胱氨酸含量为0.25%),试验组饲喂半胱氨酸含量分别为0.60%、0.70%和0.80%的试验饲粮(在基础饲粮中分别添加0.35%、0.45%和0.55%半胱氨酸)。预饲期7d,正饲期40d。试验期间每隔10d,每组随机屠宰4只獭兔,取样测定皮毛品质和SLC1A4基因表达。结果表明:(1)饲粮半胱氨酸水平为0.70%的试验组末重和平均日增重显著提高(P0.05);料重比显著降低(P0.05);四个组间平均日采食量差异不显著(P0.05)。(2)饲粮半胱氨酸水平为0.60%和0.70%的试验组饲喂到130日龄时皮张面积显著增大(P0.05);三个试验组獭兔在100日龄时被毛密度均显著大于对照组(P0.05);半胱氨酸水平为0.70%的试验组被毛密度在130日龄时显著大于其他三个组(P0.05);半胱氨酸对獭兔皮张厚度无显著性影响(P0.05)。(3)饲粮半胱氨酸含量对獭兔皮肤、肝脏和十二指肠组织中SLC1A4基因的表达量影响极显著(P0.01)。SLC1A4基因的表达量均呈现先增高后降低的趋势,饲粮中半胱氨酸水平为0.70%的试验组达到最高值。这一趋势在皮肤组织中、肝脏和十二指肠组织中分别于110日龄和100日龄时表现得较为显著。由此可见,獭兔饲粮中半胱氨酸的最优水平为0.70%,且在110日龄前使用效果最佳。     

复方抗菌剂对鲫鱼细胞色素P4503A活性的影响

试验研究了五倍子、复方抗菌剂对鲫鱼CYP3A酶活性的影响。设计复方抗菌剂浓度分别为30mg/kg、10 mg/kg和五倍子浓度为30 mg/kg,采用灌肠给药对鲫鱼进行处理;在试验的第3、7、10天取样测定其肝和肾组织中CYP3A的酶活性变化。结果表明:复方抗菌剂30 mg/kg处理组和五倍子30 mg/kg处理组的肝CYP3A活性较对照组显著降低(P<0.05),且在处理第10天,肝CYP3A酶活性较前2次的降低极显著(P<0.01);而复方抗菌剂10 mg/kg组的肝CYP3A酶活性无显著变化。对于肾组织,各药物组的CYP3A酶活性均无明显变化。     

Effects of aromatase inhibitors on reproduction in male three-spined sticklebacks, Gasterosteus aculeatus, exposed to long and short photoperiods

Three-spined stickleback Gasterosteus aculeatus, males were implanted with Silastic capsules filled with different aromatase inhibitors; 1,4,6-androstatriene-3,17-dione or the non-steroidal CGS16949 A, 4-(5,6,7,8-tetrahydrimidazol [1,5-a]pyridin-5-yl) benzonitrile monohydrochloride or empty capsules. The fish were then exposed to long or short photoperiod. Under the long photoperiod most fish in all treatments displayed a hypertrophied kidney (a secondary sexual character in sticklebacks) and completed, quiescent spermatogenesis, similar as in the natural spawning period. Under the short photoperiod the controls had unstimulated kidneys and an active spermatogenesis, whereas the males implanted with both aromatase inhibitors had stimulated kidneys, though not to the extent as in the long photoperiod, and completed, quiescent spermatogenesis. These findings suggest that aromatization is of importance for the inhibitory effects of short photoperiod on reproduction in the stickleback. This revised version was published online in July 2006 with corrections to the Cover Date.     

Preparation and comparison of biological properties of recombinant carp (Cyprinus carpio) growth hormone and its Cys-123 to Ala mutant

Carp growth hormone (cGH) cDNA, in which Cys-123 was mutated to Ala, was prepared, transferred to the expression vector, expressed in Escherichia coli and the mutant was purified to homogeneity. The mutation only slightly improved yield of the monomeric fraction, indicating that Cys-123 is not involved in improper refolding. As compared to cGH, the mutant (cGH-C123A) exhibited lower binding affinity toward homologous liver receptors and lower bioactivity in a 3T3-F442A preadipocyte bioassay despite the fact that both hormones exhibited almost identical cross-reactivity with anti-cGH antibodies. These results, along with those of a structural comparison to hGH, suggest that Cys-123 is located in the hydrophobic core of the hormone, and is most likely affecting the conformation of the binding site. Dimeric forms of the hormone and its mutant were less active than their respective monomers. Homologous binding experiments using a carp liver microsomal fraction revealed a single receptor population with Kd = 0.77 nM and B_max = 241 fmol/mg microsomal protein.

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Résumé Un ADN complémentaire (cDNA) de l'hormone de croissance de carpe (cGH), dans lequel l'acide aminé Cys-123 a été muté en Ala, a été préparé, inséré dans un vecteur d'expression, et exprimé dans Escherichia coli. Le mutant a ensuite été purifié jusqu'à homogénéité. La mutation améliore seulement faiblement la production de la fraction monomérique, indiquant que le Cys-123 n'est pas impliqué dans un repliement erroné. Comparé à la cGH, sa forme mutée (cGH-C123A) montre une plus faible affinité de liaison vis à vis de récepteurs hépatiques homologues, et une plus faible activité biologique dans un test réalisé sur des préadipocytes 3T3-F442A; cela en dépit du fait que les deux hormones présentent des réactions croisées presque identiques avec un anticorps anti-cGH polyclonale. Ces résultats, associés à une comparaison à la structure de l'hGH, suggèrent que le Cys-123 est localisé dans la partie hydrophobique de l'hormone, et affecte, le plus vraisemblablement, la conformation du site de liaison. Les formes dimériques de l'hormone et de sa forme mutée sont moins actives que leurs monomères respectifs. Les études de liaison homologue, réalisées avec des fractions microsomales de foie, révèlent une population unique de récepteurs de Kd = 0,77 nM et de B_max = 241 fmol/mg de proteine microsomale.

     

A comparison of retinol, retinal and retinyl ester concentrations in larvae of Atlantic halibut (Hippoglossus hippoglossus L.) fed Artemia or zooplankton

Atlantic halibut (Hippoglossus hippoglossus L.) larvae were fed enriched Artemia or zooplankton in duplicate tanks from 0 to 60 days after first‐feeding. Both diets and the larvae were analysed for vitamin A (VA) in order to confirm earlier findings, in which Artemia fed larvae had lower levels of VA compared with larvae fed zooplankton. Furthermore, we wanted to investigate the composition of the retinoids in the larvae. The results showed that Artemia and zooplankton contains low levels of VA, probably too low to sustain the assumed requirement. Nevertheless, larvae fed Artemia had the same level of retinal and retinol as larvae fed zooplankton. We found a significant lower level of retinyl esters in larvae fed Artemia. The total VA level was lower in larvae fed Artemia only at the end of the feeding trial after the onset of metamorphosis. Our conclusion is that feeding Artemia to Atlantic halibut larvae is not likely to cause VA deficiency.     

应用间接荧光抗体技术研究水中嗜水气单胞菌的数量与鱼类出血性败血病的关系

应用间接荧光抗体技术（ＩＦＡＴ），并结合嗜水气单胞菌（Ａｈ）地区优势血清型，对鱼类出血性败血病的致病菌Ａｈ进行水中数量的测定。ＭＭ感染试验和对生产鱼塘调查的结果表明，水中Ａｈ菌量与疾病呈正相关，致病浓度又随鱼体健康低下、水中氨氮偏高、溶氧偏低等多种因子的影响而下降。本试验方法快速简便，检测对象为水体，非常适用于预测疾病的发展趋势。     

山姜对益生芽孢杆菌体外抑菌活性试验研究

为研究中草药山姜粗提液对益生芽孢杆菌体外抑菌效果,特采用试管二倍稀释法和管碟法检测不同浓度的山姜煎煮液和乙醇提取液在体外对蜡样芽孢杆菌PAS38菌株和枯草芽孢杆菌Pab04菌株的最小抑菌浓度(MIC)和抑菌直径,并以肠毒性大肠杆菌(ETEC)、4μg/mL硫酸庆大霉素作阳性对照,生理盐水作阴性对照。结果表明,山姜煎煮液和乙醇提取液对两株益生芽孢杆菌和ETEC无抑菌作用,硫酸庆大霉素对蜡样芽孢杆菌PAS38和枯草芽孢杆菌Pab04菌株MIC均为1.25μg/mL,对ETEC的最低抑菌浓度MIC为0.625μg/mL。山姜煎煮液和乙醇提取液对受试菌株抑菌直径与生理盐水相比差异不显著(P>0.05),硫酸庆大霉素与其相比差异极显著(P<0.01)。山姜粗提液对两株益生芽孢杆菌无明显抑菌作用。     

Distribution and induction of cytochrome P450 1A1 in the rainbow trout brain

Cytochrome P450 (CYP) 1A1 participates in the activation as well as detoxification of environmental pollutants such as aromatic hydrocarbons. This CYP form is also efficiently induced by aromatic hydrocarbons. The presence of CYP 1A1 in the brain might thus be of physiological and toxicological importance. In the present investigation on rainbow trout, the distribution of 7-ethoxyresorufin-O-deethylase (EROD) activity, a cytochrome CYP 1A1 catalyzed reaction, was measured in whole tissue homogenates from brain parts. In control fish, a relatively high activity was found in the rainbow trout olfactory bulb compared to the other brain parts. Although an EROD induction (3 to 7-fold) by β-naphthoflavone (BNF) was recorded in all brain parts from the rainbow trout, the highest induced activity was measured in the olfactory bulbs. To ascertain the distribution of EROD activity in cells, whole brain tissue was subfractionated by differential centrifugation. The fractionation scheme separated mitochondria (P2 fraction) and microsomes (P3 fraction) as determined by marker enzymes and electron microscopy. In control rainbow trout, a low EROD activity could be measured in the P2 fraction. BNF induced the EROD activity in both P2 and P3 fractions. Western blotting showed the induction by BNF of a protein band in the P2 and P3 fractions with a molecular mass around 58,000 when highly specific anti-cod CYP 1A1 antibodies were used. ELISA measurements confirmed the induction of CYP 1A1 protein in the rainbow trout brain subcellular fractions.