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1.
Enzyme Immunoassays for the Determination of Ovine LH and FSH   总被引:2,自引:0,他引:2  
The development of competitive enzyme immunoassays for ovine plasma LH (oLH) and FSH (oFSH) is described. Standards and plasma samples were preincubated with diluted antiserum to oLH or oFSH and the reacted solution (100 μl per well) was transferred to plates previously coated with oLH or oFSH, respectively. The second antibody used was anti‐rabbit IgG horseradish peroxidase. The measuring range was 0.39–50 ng/ml for each hormone and the 50% relative binding sensitivity was 9 ng/ml for oLH. The respective value for oFSH was 3.5 or 34 ng/ml with different hormone and antibody preparations used for the assay. The enzyme immunoassays were used to determine oLH and oFSH levels in plasma from ewes of two breeds during the oestrous cycle. The assays detected the first FSH surge coincident with the LH surge, the second FSH surge about 24 h later and the periodic fluctuations of FSH concentrations during the luteal phase of the oestrous cycle. These enzyme immunoassays are an efficient and economic alternative to the established radioimmunoassays (RIA) for oLH and oFSH.  相似文献   
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The aim of this study was to examine the effect of varying intracellular reactive oxygen species (ROS) levels during oocyte in vitro maturation with enzymatic ROS production systems (xanthine + xanthine oxidase or xanthine + xanthine oxidase + catalase), scavenger systems (catalase or superoxide dismutase + catalase) or cysteine on porcine oocyte maturation. Oocyte ROS levels showed an increase when H2O2 or O2? production systems were added to the culture medium (p < 0.05). On the other hand, the presence of ROS scavengers in the maturation medium did not modify oocyte ROS levels compared with the control after 48 h of maturation, but the addition of cysteine induced a decrease in oocyte ROS levels (p < 0.05). The ROS production systems used in this work did not modified the percentage of oocyte nuclear maturation, but increased the decondensation of sperm head (p < 0.05) and decreased the pronuclear formation (p < 0.05). In turn, the addition of O2? and H2O2 scavenging systems during in vitro maturation did not modify the percentage of oocytes reaching metaphase II nor the oocytes with decondensed sperm head or pronuclei after fertilization. However, both parameters increased in the presence of cysteine (p < 0.05). The exogenous generation of O2? and H2O2 during oocyte in vitro maturation would not affect nuclear maturation or later sperm penetration, but most of the spermatozoa cannot progress to form the pronuclei after fusion with the oocyte. The decrease in endogenous ROS levels by the addition of cysteine would improve pronuclear formation after sperm penetration.  相似文献   
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Oocyte maturation depends on the metabolic activity of cumulus–oocyte complex (COC) that performs nutritive and regulatory functions during this process. In this work, the enzymes [phosphofructokinase (PFK) and malate dehydrogenase (MDH)] were tested to elucidate the metabolic profile of porcine COCs during the in vitro maturation (IVM). Enzymatic activity was expressed in U/COC and U/mg protein (specific activity) as mean ± SEM. In vitro maturation was performed with 2‐oxoglutarate (5, 10 and 20 mm ) or hydroxymalonate (30, 60 and 100 mm ) inhibitors of PFK and MDH, respectively. The PFK and MDH activities (U) remained constant during maturation. For PFK, the U were (2.48 ± 0.23) 10?5 and (2.54 ± 0.32) 10?5, and for MDH, the U were (4.72 ± 0.42) 10?5 and (4.38 ± 0.25) 10?5 for immature and in vitro matured COCs, respectively. The specific activities were significantly lower after IVM, for PFK (4.29 ± 0.48) 10?3 and (0.94 ± 0.12) 10?3, and for MDH (9.08 ± 0.93) 10?3 and (1.89 ± 0.10) 10?3 for immature and in vitro matured COCs, respectively. In vitro maturation percentages and enzymatic activity diminished with 20 mm 2‐oxoglutarate or 60 mm hydroxymalonate (p < 0.05). Viability was not affected by any concentration of the inhibitors evaluated. The U remained unchanged during IVM; however, the increase in the total protein content per COC provoked a decrease in the specific activity of both enzymes. Phosphofructokinase and MDH necessary for oocyte IVM would be already present in the immature oocyte. The presence of inhibitors of these enzymes impairs the meiotic maturation. Therefore, the participation of these enzymes in the energy metabolism of the porcine oocyte during IVM is confirmed in this study.  相似文献   
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The efficacy of trout oil (TO), extracted from trout offal from the aquaculture industry, was evaluated in juvenile Murray cod Maccullochella peelii peelii (25.4±0.81 g) diets in an experiment conducted over 60 days at 23.7±0.8 °C. Five isonitrogenous (48% protein), isolipidic (16%) and isoenergetic (21.8 kJ g?1) diets, in which the fish oil fraction was replaced in increments of 25% (0–100%), were used. The best growth and feed efficiency was observed in fish fed diets containing 50–75% TO. The relationship of specific growth rate (SGR), food conversion ratio (FCR) and protein efficiency ratio (PER) to the amount of TO in the diets was described in each case by second‐order polynomial equations (P<0.05), which were: SGR=–0.44TO2+0.52TO+1.23 (r2=0.90, P<0.05); FCR=0.53TO2–0.64TO+1.21 (r2=0.95, P<0.05); and PER=–0.73TO2+0.90TO+1.54 (r2=0.90, P<0.05). Significant differences in carcass and muscle proximate compositions were noted among the different dietary treatments. Less lipid was found in muscle than in carcass. The fatty acids found in highest amounts in Murray cod, irrespective of the dietary treatment, were palmitic acid (16:0), oleic acid (18:1n‐9), linoleic acid (18:2n‐6) and eicosapentaenoic acid (20:5n‐3). The fatty acid composition of the muscle reflected that of the diets. Both the n‐6 fatty acid content and the n‐3 to n‐6 ratio were significantly (P<0.05) related to growth parameters, the relationships being as follows. Percentage of n‐6 in diet (X) to SGR and FCR: SGR=–0.12X2+3.96X–32.51 (r2=0.96) and FCR=0.13X2–4.47X+39.39 (r2=0.98); and n‐3:n‐6 ratio (Z) to SGR, FCR, PER: SGR=–2.02Z2+5.01Z–1.74 (r2=0.88), FCR=2.31Z2–5.70Z+4.54 (r2=0.93) and PER=–3.12Z2–7.56Z+2.80 (r2=0.88) respectively. It is evident from this study that TO could be used effectively in Murray cod diets, and that an n‐3:n‐6 ratio of 1.2 results in the best growth performance in Murray cod.  相似文献   
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The effect of finishing extensive farming period, to reduce fat content and manipulate the fatty acid profile of fish muscle, was evaluated in rainbow trout. Fish were stocked in an artificial lake, in which fish were fed only on naturally available nutrients with no supply of artificial feed, for different lengths of time from 0 to 120 days. No weight loss was noted during the whole finishing period while total length increased from 228±7 to 269±3 mm and the condition factor decreased from 1.41±0.04 to 0.89±0.02. The total fat content of the fillets decreased considerably from 4.7±0.6% at the beginning to 2.4±0.4% and 0.7±0.2% after 45 and 120 days respectively. Fillet fatty acid composition was affected by the time of stocking in the extensive farm. In contrast to the reduction in C18:1n‐9, C18:2n‐6, total monounsaturated fatty acid and total n‐6 percent values, an increase in the C20:5n‐3, C22:6n‐3, total polyunsaturated fatty acid and total n‐3 percent values was observed. It was shown that while other finishing strategies for salmonids have some disadvantages, the extensive culture system seems to be a potentially useful tool for increasing the general quality of the end product.  相似文献   
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Meat chickens housed on a commercial broiler farm in Australia were vaccinated once at 10 to 11 days-of-age by aerosol with live V4 Newcastle disease virus (NDV) vaccine. Groups of vaccinated and unvaccinated birds were flown to Malaysia, where they were challenged with a virulent strain of NDV. Survival rates in vaccinated chickens challenged 7, 14, 21 or 31 d after vaccination were 0.47, 0.77, 0.97 and 0.92, respectively. All unvaccinated chickens died due to Newcastle disease (ND) following challenge. Chickens in Australia and Malaysia were bled and the serums tested for haemagglutination-inhibiting (HI) antibody to NDV. Many vaccinated birds with no detectable antibody, and all birds with a log2 titre of 2 or greater, survived challenge. The results showed that this V4 vaccine induced protective immunity in a significant proportion of chickens within 7 d of mass aerosol vaccination. This early immunity occurred in the absence of detectable circulating HI antibody. Non-HI antibody mediated immunity continued to provide protection up to 31 d after vaccination. Almost all vaccinated birds were protected within 3 w of vaccination. It is concluded that the V4 vaccine is efficacious and could be useful during an outbreak of virulent ND in Australia.  相似文献   
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SUMMARY Big liver and spleen disease (BLS) was reproduced experimentally by intravenous (IV) and oral (PO) administration of BLS inocula to susceptible broiler breeder hens 34 to 36 weeks of age. Serological and pathological signs of BLS similar to those seen in the natural disease occurred in inoculated and in-contact birds. Splenomegaly was the earliest and often the only necropsy finding, with hepatomegaly and kidney enlargement occurring in some birds later in the course of the disease. After IV administration, serum antigen was detected between 2 and 4 weeks, and antibody between 3 and 5 weeks. After PO administration, antigen was detected between 2 and 4 weeks, and antibody between 3 and 6 weeks. Antibody persisted in all birds to the end of the experiment (6 weeks), and horizontal transmission probably occurred since in-contact birds developed BLS. Liver probably contained the highest concentration of BLS agent because it had the highest infectivity.  相似文献   
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