云南腾冲北海湿地自然保护区两栖动物多样性现状

21世纪以来,全球近半的两栖动物在物种和种群水平正受到威胁,我国云南尤为突出。云南腾冲北海湿地具有独特的漂浮状苔草沼泽植被,两栖类是评价生态系统健康的指示类群,进行两栖类资源调查有助于推进湿地保护工作。2019年5月、7月和8月以及2020年5月、6月和9月采用了以样线法为主、访问为辅的方法进行了野外调查,共记录两栖动物21种,包括入侵种1种,其中20种土著种属于2目7科17属。与上一次调查名录（2008年）相比,此次调查增加北海湿地两栖动物新记录5种,包括腾冲掌突蟾（Leptobrachella tengchongensis）、红蹼树蛙（Rhacophorus rhodopus）、白颌大树蛙（Zhangixalus smaragdinus）、背条螳臂树蛙（Chiromantis doriae）以及入侵种牛蛙（Lithobates catesbeianus = Rana catesbeiana）;新修订2科8种。用G-F指数进行数据分析,与周边县域进行平均动物区系相似性比较。结果显示,北海湿地两栖类G-F指数为0.432;由平均动物区系相似性比较可知,北海湿地与腾冲（高黎贡山国家级自然保护区）、泸水（高黎贡山国家级自然保护区）、龙陵小黑山省级自然保护区和铜壁关自然保护区盈江片区为周缘关系;外来入侵种牛蛙数量远多于土著种,亟需采取针对性保护措施;需建立北海湿地两栖动物监测体系,作为湿地生态系统健康的指示标准之一。     

牛蛙食性的研究

牛蛙(Rana catesbeiana Shaw)属于脊椎动物门(Vertebrata)、两栖纲(Amphibia)、无尾目(Anura)、蛙科(Ranidae)、蛙亚科(Raniinae)、蛙属(Rana)。牛蛙原产北美洲洛矶山一带沼泽地区,因其鸣叫声远闻似牛叫而得名“牛蛙”。牛蛙的种类较多,如美国牛蛙、春蛙、沼泽牛蛙、非洲牛蛙、非洲大牛蛙等。由于牛蛙生长发育快,体重可达0.5-1.5公斤,肉质鲜美可口、商品性好、食用价值高而先后被加拿大、古巴、墨西哥、南美洲以及日本等国引进饲养。我国八十年代引养的牛蛙属美国牛蛙,又叫“喧蛙”,有的称“食用蛙”。     

五肢牛蛙

牛蛙(Rana catesbeiana)原产北美洲,后来逐渐传至菲律宾、日本及我国台湾,继而马来西亚、新加坡,随后牛蛙移植到我国云南。 今年5月底由楚雄二中张燕翔等同学在楚雄尹家咀水库放养的牛蛙池塘里捕获到一个牛蛙     

同一生境牛蛙与昭觉林蛙蝌蚪的食性比较

对采自昆明东北郊同一水塘的牛蛙和昭觉林蛙蝌蚪进行食性分析。结果表明 ,牛蛙蝌蚪Ⅰ、Ⅱ期的食物种类分别为 8大类 5 1种和 37种 ,而昭觉林蛙蝌蚪Ⅱ期的食物种类有 6大类 30种 ;两者的食物组成均以硅藻门、绿藻门、蓝藻门为主 ,但昭觉林蛙蝌蚪Ⅱ期与牛蛙蝌蚪Ⅰ期的摄食种类差异极显著 ,与牛蛙Ⅱ期的不显著。昭觉林蛙蝌蚪Ⅱ期与牛蛙蝌蚪Ⅰ、Ⅱ期的食物生态位重叠度分别为 0 6 95 2和 0 6 887,单位体重摄食的生物量分别是后两者的 10 96倍和 15 4 8倍 ,但百分率相似性指数并不高 (分别为 5 4 95 %和 5 8 11% )。与等重量牛蛙蝌蚪相比 ,昭觉林蛙蝌蚪食物量较大 ,即昭觉林蛙蝌蚪为生存所需要的生物量远高于牛蛙蝌蚪。本文结果及野外观察表明 ,昭觉林蛙与牛蛙蝌蚪存在较激烈的食物竞争 ,且前者处于不利地位。     

舟山群岛变态后牛蛙的食性研究

牛蛙(Ranacatesbeiana)是极其危险的外来入侵种,对当地生物类群的捕食是其主要的危害机制,但目前有关其食性的研究均只局限于一个研究地点。为了全面评估牛蛙对当地生物类群的捕食影响,2005年6月30日至8月11日期间,作者采用洗胃法对舟山群岛8个岛屿(岱山、六横、秀山、佛渡、桃花、虾峙、册子和普陀山)变态后牛蛙的食性进行了研究。共采集到变态后的牛蛙391只,包括牛蛙成体113只和亚成体278只。食性分析结果表明:(1)按食物体积计算,成年牛蛙以十足目、鞘翅目、蜻蜓目、中腹足目、蛙形目和鲤形目为主要食物,亚成体牛蛙以十足目、鞘翅目、鲤形目、蜻蜓目、直翅目、膜翅目、鳞翅目幼虫、中腹足目和蛙形目为主要食物;成体和亚成体牛蛙胃中食物的平均大小均随其体长增加而增大;(2)不同岛屿上的牛蛙在主要食物类型构成上存在差异。这表明牛蛙在不同地方对不同生物类群造成的捕食危害是不一样的,充分反映了其生态危害的广泛性。     

浙江东部牛蛙的自然种群及潜在危害

牛蛙(Rana catesbeiana)的入侵已成为一些本地两栖类种群数量下降或灭绝的主要因素之一。1959年牛蛙被引入中国大陆,但直到1990年才开始在国内大范围饲养。牛蛙的自然种群在国内一直未见报道。我们于2003年9—11月调查了浙江省东部5区3县的牛蛙种群,发现在岱山、秀山、六横及佛渡岛上有牛蛙分布,共捕获23只牛蛙,包括4只成年个体,18个亚成体和1只蝌蚪。于2004年1月和2月在岱山捕获28只蝌蚪。结果显示：(1)牛蛙的蝌蚪能在当地越冬成活,说明牛蛙在上述4岛成功地建立了自然种群;(2)牛蛙主要分布于水库、池塘和溪流,牛蛙的生境与6种当地蛙有部分重叠;(3)养殖时管理不善造成的牛蛙逃逸,以及牛蛙价格过低而导致养殖户将牛蛙弃置野外可能是牛蛙入侵的主要原因。牛蛙对当地蛙的潜在威胁很大,开展牛蛙的分布、食性和种群动态研究是当务之急。     

第三次全国牛蛙驯化会议简讯

1963年11月18日到21日,中华人民共和国水产部在广州召开了第三次全国牛蛙驯化会议。北京、上海、天津、南京、广州、福建、宁波、湛江、中山等地代表汇报了全国第二次牛蛙驯化会议后的情况,并着重交流了有关牛蛙驯化的技术经验。国家科委、水产部、广东省水产厅等领导同志对会议做了重要指示。     

一株牛蛙源金黄杆菌的分离鉴定及其致病特性

【背景】美国牛蛙养殖过程中病害问题非常突出,尤其是细菌性病害,其病原种类多、病原菌复杂多样、蔓延速度快、发病死亡率高,一直是牛蛙养殖过程中防控的难点。【目的】确定从患病牛蛙体内分离到的一株细菌NW1203的分类地位和致病性。【方法】无菌操作从牛蛙体内取样划线分离细菌,通过形态观察、生理生化试验、16S rRNA基因序列比对进行种属鉴定,通过人工感染、溶血性试验和病理切片观察分析其致病特性。【结果】经形态和生理生化鉴定及16S rRNA基因序列比对,菌株NW1203为金黄杆菌属细菌,与Chryseobacterium sp. F30的相似性达100%,进化树也显示该菌与金黄杆菌属细菌聚类;溶血性试验表明,菌株NW1203对绵羊、小鼠和牛蛙的血细胞都呈完全溶血;人工感染试验及感染病蛙的组织切片观察显示,菌株NW1203对牛蛙具有较强致病性,可引起牛蛙肝、肾、脾等主要组织严重病变,LD50为4.753×103 CFU/g。【结论】明确了菌株NW1203为牛蛙新病原,为牛蛙疾病的防控提供了理论依据。     

饲料中添加胆汁酸对牛蛙生长性能、体组成和营养物质表观消化率的影响

试验研究了饲料中添加胆汁酸对牛蛙Rana (Lithobates) catesbeiana生长性能、体成分和营养物质表观消化率的影响。基础饲料以鱼粉、豆粕为主要蛋白源, 棕榈油为主要脂肪源, 在基础饲料中分别添加0、100、200和300 mg/kg的胆汁酸(含量99.1%)制作4种等氮等能的试验饲料。试验将144只牛蛙初始体重(75.014.23) g随机分为4组, 每组3个重复, 每个重复12只牛蛙, 进行为期8周的生长试验。结果显示: 各试验组牛蛙成活率和摄食率差异不显著(P0.05)。200 mg/kg胆汁酸添加组牛蛙的增重率、特定生长率、饲料效率、蛋白质效率和氮保留率显著高于对照组(P0.05)。在饲料中添加胆汁酸显著降低了牛蛙脏体指数和全体的脂肪含量(P0.05), 但不影响肌肉粗脂肪以及全体和肌肉水分、粗蛋白、粗灰分含量(P0.05)。各处理组间血清总胆固醇、甘油三酯、高密度脂蛋白胆固醇和低密度脂蛋白胆固醇浓度均无显著变化(P0.05), 血清尿素氮含量随着饲料中胆汁酸添加量的增加而降低, 添加300 mg/kg胆汁酸组牛蛙血清尿素氮含量显著低于对照组(P0.05)。血糖水平则随着胆汁酸添加量的增加而先降低后升高, 其中100 mg/kg胆汁酸添加组牛蛙血糖水平最低(P0.05)。100 mg/kg胆汁酸添加组牛蛙肠道脂肪酶活力显著高于对照组和300 mg/kg胆汁酸添加组(P0.05), 与200 mg/kg胆汁酸添加组之间差异不显著(P0.05), 蛋白酶活力与脂肪酶活力有相似的变化趋势, 而淀粉酶活力未受胆汁酸添加水平影响。各处理组饲料营养物质的表观消化率随饲料中胆汁酸添加量的增加而先升高后降低, 其中饲料添加200 mg/kg胆汁酸显著增加了牛蛙对饲料营养物质的表观消化率(P0.05)。结果表明: 在饲料中添加适量的胆汁酸能提高牛蛙对饲料的消化率, 提高饲料效率, 从而促进牛蛙的生长, 同时可促进蛙体脂肪代谢, 降低机体脂肪沉积, 提高牛蛙可食部分比例。根据本试验结果, 建议牛蛙饲料中胆汁酸添加水平为200 mg/kg。       

牛蛙肥大细胞的组织化学与形态学

目的鉴定牛蛙组织中肥大细胞的存在。方法用于肥大细胞研究的一些常规组织化学技术与形态学方法。结果牛蛙的舌、肠、肠系膜和脾中肥大细胞数量较多,少量也见于神经、心、肾、肝和皮肤等多种组织中。肥大细胞有沿血管周和神经分布的倾向。脾脏中的肥大细胞形状比较一致,呈圆形或卵圆形,而在其它部位的肥大细胞则形态多样。Bouin氏液及Carnoy氏液是牛蛙肥大细胞优良的固定液。然而,与哺乳动物的黏膜肥大细胞相似的是,中性缓冲福尔马林(NBF)固定显著的阻断了牛蛙肠黏膜肥大细胞(MMC)的染色。有趣的是,甲苯胺蓝是牛蛙肥大细胞的最佳染料,它比阿尔新蓝能很好地显示牛蛙的肥大细胞。透射电镜下证实,牛蛙肥大细胞中含有大量特征性的胞浆颗粒。肥大细胞靠近雪旺氏细胞,并可见于神经束膜间,甚至以其突起与神经束膜相连。结论通过组织化学与形态学研究证实了牛蛙组织中肥大细胞的存在,再次证实肥大细胞与外周神经之间存在密切的解剖学关系。     

The emerging amphibian pathogen Batrachochytrium dendrobatidis globally infects introduced populations of the North American bullfrog, Rana catesbeiana

Batrachochytrium dendrobatidis is the chytridiomycete fungus which has been implicated in global amphibian declines and numerous species extinctions. Here, we show that introduced North American bullfrogs (Rana catesbeiana) consistently carry this emerging pathogenic fungus. We detected infections by this fungus on introduced bullfrogs from seven of eight countries using both PCR and microscopic techniques. Only native bullfrogs from eastern Canada and introduced bullfrogs from Japan showed no sign of infection. The bullfrog is the most commonly farmed amphibian, and escapes and subsequent establishment of feral populations regularly occur. These factors taken together with our study suggest that the global threat of B. dendrobatidis disease transmission posed by bullfrogs is significant.     

Diet composition of post-metamorphic bullfrogs (Rana catesbeiana) in the Zhoushan archipelago, Zhejiang Province, China

Bullfrogs (Runa catesbeiana) are listed as one of the 100 worst invasive alien species in the world.They are generalist predators and thus may affect native species through predation.In previous studies,the food contents of bullfrogs were mostly examined at a single site.In the present study,the diet composition of post-metamorphic bullfrogs on eight islands (Daishan,Liuheng,Xiushan,Fodu,Taohua,Xiashi,Cezi,and Putuoshan) in the Zhoushan Archipelago,Zhejiang Province of China,were examined by using the stomach flushing method from June 30 to August 11 in 2005.A total 0f391 individual frogs were measured,including 113 adults and 278 juveniles.The analysis of the stomach contents shows that,for adult bullfrogs,the most important prey (by diet volume) overall were Decapoda,Coleoptera,Odonata,Mesogastropoda,Raniformes,and Cypriniformes.For juvenile bullfrogs,these were Decapoda,Coleoptera,Cypriniformes,Odonata,Orthoptera,Hymenoptera,Lepidoptera larvae,Mesogastropoda,and Raniformes.Moreover,the prey size and diet volume increased with the body size of both adult and juvenile bullfrogs.The diet composition of primary preys of bullfrogs was significantly different among the islands.The results indicate that bullfrogs exert different predatory influences on native fauna at different sites and that bullfrogs are generalist predators with extensive ecological impacts on native fauna.     

Population genetics of introduced bullfrogs, Rana (Lithobates) catesbeianus, in the Willamette Valley, Oregon, USA

The American bullfrog, Rana (Lithobates) catesbeianus, is endemic to eastern North America, but has been introduced to approximately 40 countries on four continents and is considered one of the hundred worst invasive alien species in the world. Here, we investigated the genetics of invasive bullfrogs in the Willamette Valley, Oregon, USA, where bullfrogs are widespread and abundant to determine: (1) the minimum number of bullfrog introductions; (2) the native source population(s); and (3) whether genetic variation is reduced compared to source populations. To answer these questions, we analyzed partial sequences of the mitochondrial cytochrome b gene for 251 bullfrogs from the Willamette Valley and the native range. We found that bullfrogs from the Mississippi River basin and Great Lakes region were introduced at least once to the Willamette Valley. Genetic variation measured as haplotype diversity (h) and nucleotide diversity (?? _n ) was not significantly different between Willamette Valley and source populations. Our results were in contrast to a recent genetic analysis of invasive bullfrog populations in Europe, which found that genetic variation in European bullfrog populations was much lower than in source populations. European bullfrogs also originated from different source populations than Willamette Valley bullfrogs. The difference in genetic composition between Willamette Valley and European bullfrogs is likely due to differences in their invasion histories and may have implications for the potential of bullfrogs in these different regions to adapt and expand.     

Attenuated cardiovascular effects of prostaglandin I2 and prostaglandin F2 alpha in cold acclimated American bullfrogs, Rana catesbeiana

American bullfrogs, Rana catesbeiana respond to prostaglandins with changes in heart rate and blood pressure. These studies compare responses of warm (22 degrees C) and cold acclimated (5 degrees C) bullfrogs to prostaglandins. Gas chromatographic analysis determined equivalent fatty acid profiles in total lipids of heart and artery tissue from warm and cold acclimated animals. Arachidonic acid was the fatty acid precursor found in greatest abundance in both groups. For cardiovascular experiments, bullfrogs were cannulated by using a T-cannula implanted in the right sciatic artery. In warm acclimated bullfrogs, preinfusion systemic arterial pressure (SAP) was 14.7 +/- 0.5 mm Hg, and heart rate was 33.0 +/- 1.7 beats/min. Cold acclimated bullfrogs had SAP values of 8.0 +/- 0.8 mm Hg, and heart rate was 6.9 +/- 0.3 beats/min. Arachidonic and eicosapentaenoic acid infusions (2,000 micrograms/kg body weight [bw]) were hypertensive in cold acclimated and hypotensive in warm acclimated animals. These effects were blocked by indomethacin (4 mg/kg bw). In both warm and cold acclimated bullfrogs, prostaglandin F2 alpha (3-100 micrograms/kg bw) was hypertensive, while prostaglandin I2 (0.03-3 micrograms/kg bw) was hypotensive, with both prostaglandins stimulating a greater absolute response in warm acclimated animals. In addition, both prostaglandins increased heart rate in warm but not in cold acclimated bullfrogs. The results suggest diminished cardiovascular sensitivity to prostaglandins at low environmental temperatures.     

An exotic species is the favorite prey of a native enemy

Although native enemies in an exotic species'' new range are considered to affect its ability to invade, few studies have evaluated predation pressures from native enemies on exotic species in their new range. The exotic prey naiveté hypothesis (EPNH) states that exotic species may be at a disadvantage because of its naïveté towards native enemies and, therefore, may suffer higher predation pressures from the enemy than native prey species. Corollaries of this hypothesis include the native enemy preferring exotic species over native species and the diet of the enemy being influenced by the abundance of the exotic species. We comprehensively tested this hypothesis using introduced North American bullfrogs (Lithobates catesbeianus, referred to as bullfrog), a native red-banded snake (Dinodon rufozonatum, the enemy) and four native anuran species in permanent still water bodies as a model system in Daishan, China. We investigated reciprocal recognition between snakes and anuran species (bullfrogs and three common native species) and the diet preference of the snakes for bullfrogs and the three species in laboratory experiments, and the diet preference and bullfrog density in the wild. Bullfrogs are naive to the snakes, but the native anurans are not. However, the snakes can identify bullfrogs as prey, and in fact, prefer bullfrogs over the native anurans in manipulative experiments with and without a control for body size and in the wild, indicating that bullfrogs are subjected to higher predation pressures from the snakes than the native species. The proportion of bullfrogs in the snakes'' diet is positively correlated with the abundance of bullfrogs in the wild. Our results provide strong evidence for the EPNH. The results highlight the biological resistance of native enemies to naïve exotic species.     

Diet composition of post-metamorphic bullfrogs (Rana catesbeiana) in the Zhoushan archipelago, Zhejiang Province, China

Bullfrogs (Rana catesbeiana) are listed as one of the 100 worst invasive alien species in the world. They are generalist predators and thus may affect native species through predation. In previous studies, the food contents of bullfrogs were mostly examined at a single site. In the present study, the diet composition of post-metamorphic bullfrogs on eight islands (Daishan, Liuheng, Xiushan, Fodu, Taohua, Xiashi, Cezi, and Putuoshan) in the Zhoushan Archipelago, Zhejiang Province of China, were examined by using the stomach flushing method from June 30 to August 11 in 2005. A total of 391 individual frogs were measured, including 113 adults and 278 juveniles. The analysis of the stomach contents shows that, for adult bullfrogs, the most important prey (by diet volume) overall were Decapoda, Coleoptera, Odonata, Mesogastropoda, Raniformes, and Cypriniformes. For juvenile bullfrogs, these were Decapoda, Coleoptera, Cypriniformes, Odonata, Orthoptera, Hymenoptera, Lepidoptera larvae, Mesogastropoda, and Raniformes. Moreover, the prey size and diet volume increased with the body size of both adult and juvenile bullfrogs. The diet composition of primary preys of bullfrogs was significantly different among the islands. The results indicate that bullfrogs exert different predatory influences on native fauna at different sites and that bullfrogs are generalist predators with extensive ecological impacts on native fauna. __________ Translated from Biodiversity Science, 2006, 14 (5): 363–371 [译自: 生 物多样性]     

First Evidence of <Emphasis Type="Italic">Batrachochytrium dendrobatidis</Emphasis> in China: Discovery of Chytridiomycosis in Introduced American Bullfrogs and Native Amphibians in the Yunnan Province,China

Although the chytrid fungus Batrachochytrium dendrobatidis (Bd), the etiological agent of amphibian chytridiomycosis, has been implicated in mass mortality and population declines on several continents around the world, there have been no reports on the presence of Bd infections in amphibians in China. We employed quantitative PCR and histological techniques to investigate the presence of Bd in introduced North American bullfrogs (Rana catesbeiana) (referred to hereafter as bullfrog) and native amphibians in bullfrog-invaded areas of the Yunnan Province, China. A total of 259 samples at five wild sites were collected between June and September in 2007 and 2008, including bullfrogs and four native amphibian species (Rana pleuraden, Rana chaochiaoensis, Odorrana andersonii, and Bombina maxima). In addition, 37 samples of adult bullfrogs were obtained from a food market. Bd infections were discovered in bullfrogs and three native amphibian species from all of the surveyed sites. Of the 39 Bd-positive samples, 35 were from wild-caught bullfrog tadpoles, postmetamorphic bullfrogs, R. pleuraden, R. chaochiaoensis, and O. andersonii, and four were from adult bullfrogs from the market. Our results provide the first evidence of the presence of Bd in Chinese amphibians, suggesting that native amphibian diversity in China is at risk from Bd. There is an urgent need to monitor the distribution of Bd in amphibians in China and understand the susceptibility of native amphibian species to chytridiomycosis. Strict regulations on the transportation of bullfrogs and the breeding of bullfrogs in markets and farms should be drafted in order to stop the spread of Bd by bullfrogs.     

Development of a novel enzyme immunoassay for the measurement of in vitro GnRH release from rat and bullfrog hypothalamic explants

Gonadotropin-releasing hormone (GnRH) is critical for the initiation and maintenance of reproduction in vertebrates. Information regarding GnRH release is abundant in mammals, but absent in poikilothermic tetrapods. In this study, we established a novel GnRH enzyme immunoassay (EIA) to measure GnRH release over time from hypothalamic explants isolated from mature field-caught and commercially-acquired male bullfrogs, Rana catesbeiana. Hypothalamic explants from rats were used as a positive control to test the sensitivity and accuracy of our EIA and to ensure our in vitro system could detect GnRH pulses. Prominent GnRH pulses were present in the majority (9/10) of rat hypothalamic explants, but absent in all (17/17) of the commercial bullfrogs and the majority (5/8) of field-caught bullfrogs. In three cases where GnRH pulses were observed in field-caught bullfrogs, there was only one pulse during the 2-h incubation period; high-frequency pulses similar to those observed in rats were not observed. Veratridine, which opens voltage-gated sodium channels, stimulated GnRH release in all explants cultured in the presence of Ca(2+), demonstrating explant viability. The levels of both spontaneous and veratridine-induced GnRH release were significantly higher in field-caught than commercial bullfrogs. This study demonstrated, for the first time, the temporal pattern of GnRH release in a poikilothermic tetrapod. Further, our results suggest the levels and patterns of GnRH output in bullfrogs are subject to the dynamic regulation by physiological and environmental cues.