Avoidance response of juvenile Pacific treefrogs to chemical cues of introduced predatory bullfrogs

Bullfrogs (Rana catesbeiana), native to eastern North America, were introduced into Oregon in the 1930's. Bullfrogs are highly efficient predators that are known to eat a variety of prey including other amphibians. In laboratory experiments, we investigated whether juvenile Pacific treefrogs (Hyla regilla) recognize adult bullfrogs as a predatory threat. The ability of prey animals to acquire recognition of an introduced predator has important implications for survival of the prey. We found that treefrogs from a population that co-occurred with bullfrogs showed a strong avoidance of chemical cues of bullfrogs. In contrast, treefrogs from a population that did not co-occur with bullfrogs, did not respond to the bullfrog cues. Additional experiments showed that both populations of treefrogs use chemical cues to mediate predation risk. Treefrogs from both populations avoided chemical alarm cues from injured conspecifics.     

冰乙酸和胃蛋白酶依次提取新鲜林蛙皮胶原蛋白

采用冰乙酸和胃蛋白酶依次对新鲜林蛙皮胶原蛋白进行提取.在冰乙酸提取过程中,首先通过控制单一因素变量对料液比、pH值、冰乙酸浓度、酸解时间和温度等5个因素对新鲜林蛙皮胶原蛋白提取率的影响进行研究,并采用正交实验考察林蛙皮胶原蛋白酸解提取的最佳工艺;其次,在冰乙酸提取的最优条件下,利用胃蛋白酶将酸解后的残余物进行再次酶解提取.实验结果表明,最佳酸解的工艺条件是:料液比为1∶30,pH为7.5,酸解时间为48 h,酸解温度为4℃,冰乙酸浓度为0.5 mol/L.在此工艺条件下,胶原蛋白的提取率为11.85％,胃蛋白酶对蛙皮残渣的提取率为5.25％,所得胶原蛋白的纯度为64.70％.     

中国林蛙皮中胶原蛋白的提取研究

采用酶解法从中国林蛙皮中提取胶原蛋白。考查了提取次数、液料比、酶解时间、酶解温度以及酶用量对胶原蛋白提取率的影响。单因素考察结合正交试验设计得到胶原蛋白的优化提取条件为:提取次数为2次,液料比为60,提取时间为72 h,提取温度为37℃,酶用量为1.5%。此条件下中国林蛙皮胶原蛋白的产率可达26.2%。     

超声波-双酶法协同提取林蛙皮多糖的工艺优化

以野生中国林蛙皮为原料,优化超声波-双酶法协同提取林蛙皮多糖的工艺参数。结果表明:超声波提取优化工艺条件为料液比1∶40(g/mL),超声功率600W,超声处理时间25min,提取多糖得率为1.18%。在超声波优化结果基础上,进行双酶法处理,得出最佳酶解条件是pH9.0,酶解温度45℃,碱性蛋白酶添加量2.3%、胰蛋白酶添加量2.8%,酶解时间4h,多糖得率为2.286%。由此可见,超声波和酶法协同处理是提取林蛙皮多糖的一种有效的提取方法。     

利用线粒体DNA上的cytB鉴定林蛙及林蛙油

目的：以线粒体DNA上cytB为研究对象,采用聚合酶链式反应(PCR)手段,建立林蛙及林蛙油鉴定方法。方法：采用市售成年雌蛙的新鲜组织和林蛙油为实验材料,通过试剂盒提取总DNA;以已报道的东北林蛙、中国林蛙、黑斑蛙和蟾蜍的cytB为模板,设计四对引物;PCR扩增产物回收测序。结果：仅东北林蛙引物的PCR体系获得1100 bp左右大小的产物,产物序列与东北林蛙cytB序列一致性为99%,说明实验对象林蛙为东北林蛙,林蛙油来源为东北林蛙。讨论：利用mtDNA独特的优势,应用PCR扩增cytB,并测序确定了林蛙品种,建立了林蛙种属以及林蛙油基源的鉴定方法,为林蛙油类药材的鉴定提供科学理论依据。     

Fatty acid composition, extraction, fractionation, and stabilization of bullfrog (Rana catesbeiana) oil

The oil extracted from the fat-storage organ (fat body) of the bullfrog (Rana catesbeiana) was characterized for its fatty acid composition. The main fatty acids were palmitic (18.1%), stearic (4.1%), myristic (2.7%), oleic (31.7%), and linoleic (12.9%) acids. Long-chain polyunsaturated fatty acids were also present in significant amounts, i.e., eicosapentaenoic (1.5%) and docosahexaenoic (4.7%), and were probably derived from the fish meal content of the diet. A partially fractionated oil was extracted from the homogenized and frozen fat body with an oleic acid content of 43.2%. The natural alkaloid boldine, added at 0.5 mg/g oil level, improved the oxidative stability by a factor ranging from 1.7 to 2.4, as assessed by the Oil Stability Index method between 90 and 110°C. The stabilization effect of boldine was higher than that of naringenin, morin, and quercitin and for the synthetic antioxidant butylated hydroxytoluene at the same concentration level.     

响应曲面法优化碱性蛋白酶提取林蛙残体胶原蛋白

以提取林蛙油后的剩余林蛙干残体为原料,通过碱性蛋白酶法提取胶原蛋白。以胶原蛋白提取率为指标,对影响胶原蛋白提取pH 值、加酶量、酶解温度和酶解时间4 因素进行研究,在单因素试验基础上,采用响应曲面分析法,得出最优酶提取工艺条件,并根据实际生产优化为：pH10、加酶量1.5%、酶解温度50℃、酶解时间2.5h,在此工艺条件下,林蛙残体胶原蛋白提取率为55.38%,胶原蛋白成品的纯度64.15%。     

石蛙的酶解液化及风味改良

在酶适宜的环境条件下,利用木瓜蛋白酶与中性蛋白酶酶解石蛙肉及利用乳酸菌、黄酒酵母对酶解液进行风味改良。结果表明,中性蛋白酶对石蛙的水解效果较好,乳酸菌发酵液中氨态氮含量较高,乳酸菌发酵液风味改良效果优于黄酒酵母。     

高压脉冲电场提取中国林蛙多糖的研究

通过单因素考察及正交试验优化了用高压脉冲电场提取中国林蛙多糖的试验条件，并与碱提取法、酶提取法以及复合酶提取法进行了比较。结果显示用0．5％KOH提取液，在电场强度为20kv／cm和脉冲数为6μs的条件下用高压脉冲电场提取林蛙多糖的提取率最大为55．59％。比较高压脉冲电场提取法与碱法、酶法以及复合酶法在林蛙多糖提取率、总糖含量方面的差异，高压脉冲电场提取的林蛙多糖提取率和总糖含量均高于其他三种方法，其提取率是复合酶法的1．77倍，总糖含量高于复合酶法6．34％，且提取物中杂质少。因此，高压脉冲电场提取多糖是一种很有希望的多糖提取新方法。     

林蛙油对冷应激小鼠抗氧化能力的影响

研究林蛙油对冷应激小鼠抗氧化能力的影响,为林蛙油的合理开发和利用提供依据。本实验将40 只雄性昆明品系小鼠随机分为4 组：对照组、冷应激组、林蛙油5 倍剂量组和25 倍剂量组,每组10 只。对照组和冷应激组灌胃生理盐水,5 倍剂量组和25 倍剂量组灌不同剂量的林蛙油,每组连续灌胃28d。灌胃到第15 天,每天除对照组灌胃后置室温外,其他各组灌胃30min 后,置－ 10℃冰箱30min,连续14d,杀鼠,测血清和脑组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果表明,林蛙油可以增加小鼠血和脑组织中SOD 活性(p ＜ 0.01),降低MDA 含量(p ＜ 0.01),在低温条件下林蛙油能增强小鼠抗氧化能力。