绿海龟源溶藻弧菌的分离鉴定及药敏分析

【目的】腐皮病是绿海龟(Chelonia mydas)龟苗培育中的常见病,传染性极强,发病率和死亡率高。为确定病因及治疗方法,从病龟鳍状肢的腐皮组织中分离病原菌。【方法】从症状明显的稚龟四肢病灶中分离纯化得到疑似病原菌,编号CMRT91026,开展形态特征、生理生化特性、16SrRNA鉴定和系统发育树构建及相关药物敏感试验。【结果】该菌能在TCBS培养基上生长,菌落呈黄色,为革兰氏阴性短杆菌;在1%NaCl葡萄糖磷酸盐胨水、蔗糖、甘露糖和赖氨酸脱羧酶上的生化特性均显示阳性,并且在3%～10%NaCl胨水均能生长;与NCBI上的溶藻弧菌(Vibrio alginolyticus)同源性达100%。药敏结果显示,该菌对恩诺沙星、萘啶酸、左氧氟沙星、米诺环素、头孢吡肟和头孢曲松敏感;对多西环素、氟苯尼考、妥布霉素、大观霉素、诺氟沙星和氨曲南呈中介;对新霉素、复方新诺明、阿莫西林、庆大霉素、丁胺卡那霉素等15种药物有耐药性。【结论】该菌株鉴定为溶藻弧菌,对喹诺酮类药物(如恩诺沙星)、四环素类药物(如米诺环素)及头孢类(如头孢曲松)等药物较为敏感。试验结果为绿海龟稚龟溶藻弧菌病的诊断及病害防控提供了一定参考。     

Estimation of temporal changes in the growth of green turtles <Emphasis Type="Italic">Chelonia mydas</Emphasis> in waters around the Ogasawara Islands

Biological and life-cycle information is essential to any fisheries management program. We have investigated temporal changes in growth parameters in the von Bertalanffy growth curve for green turtles, Chelonia mydas, in waters around the Ogasawara Islands. The variance model of individual variability of growth used in our study was one derived from the stochastic differential equation with the white noise of constant intensity. Maximum likelihood estimates of the parameters were made using the data of 102 recovery records of tagging experiments from 1974 to 1995. The estimations were carried out for each of two models of linear and sudden changes in the parameters, and the best model was selected using Akaike's information criterion. The selected model was that all of the parameters excepting asymptotic straight carapace length were linear functions of year and that the asymptotic length was constant over the years. The estimated values of the growth coefficient in 1974 and 1995 were 0.0317 and 0.192 per year, respectively, and the estimated value of the asymptotic straight carapace length was 97.2 cm. Implications of the results and future issues to be resolved in near future are discussed.     

Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas

To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6 kDa band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demostration of CD3+ cells in reptiles using specific antibodies.     

广东惠东海龟国家级自然保护区动物多样性调查

了解广东惠东海龟国家级自然保护区动物资源现状,可为保护区建设规划及生态系统维护提供理论参考。采用样线法、红外相机法、市场调查法、悬网捕捞法系统调查了区内的动物种类,分析了多样性指数以及绿海龟(Chelonia mydas)与其他物种的关系。结果表明,调查共记录动物116种,其中无脊椎动物55种,包括浮游动物13种、软体动物25种、节肢动物16种;脊椎动物61种,包括鱼类43种、两栖动物1种、爬行动物2种、鸟类12种、兽类3种。保护区内有国家 级重点保护野生动物1种、 级重点保护野生动物4种、“三有”名录物种11种、《IUCN物种红色名录》濒危物种3种、易危物种3种、近危物种3种。保护区Simpson优势度指数为0.96,Shannon-Wiener多样性指数为3.75,Pielou均匀度指数为0.78;本底动物物种资源丰富,各物种分布较为均匀,群落多样性高,动物资源中既包括了绿海龟的食物来源,也存在绿海龟的天敌;在绿海龟濒危的背景下,制定基于物种多样性的海龟保护管理策略成为当务之急。     

Atypical ocular Chelonoid herpesvirus manifestations in a captive Loggerhead turtle (Caretta caretta)

A captive loggerhead turtle (Caretta caretta) of unknown sex, 3 years of age, presented with bilateral mucoid secretions, severe chemosis, conjunctival hyperemia, and globe retraction. The animal was evaluated ophthalmologically and systemically, and hematological, microbiological, and conjunctival cytological and biopsy samples were collected for complementary diagnosis. The histopathological examination showed amphophilic intranuclear inclusions associated with severe inflammatory infiltrate. The diagnosis of Chelonid alphaherpesvirus 5 (ChAHV 5) was confirmed with end point PCR. Following systemic treatment with L‐lysine, acyclovir and vitamin A, the ocular signs resolved. No amphophilic intranuclear inclusions were seen in a follow‐up biopsy 5 months later, and there has been no recurrence of clinical ophthalmic signs during a 4‐year follow‐up. It is suggested that ChAHV 5 be considered as a differential diagnosis in captive marine turtles that present for conjunctival disease other than fibropapillomatosis.     

Efficacy of praziquantel in the treatment of green sea turtles with spontaneous infection of cardiovascular flukes

Objective To assess the efficacy of praziquantel as a treatment for cardiovascular flukes in turtles.
Procedure Six green sea turtles ( Chelonia mydas ) spontaneously infected with cardiovascular flukes (Digenea: Spirorchiidae) were treated orally with praziquantel, and necropsied 3 or 7 days later to look for flukes in the heart and major blood vessels. Six similar animals were maintained as untreated controls.
Results Absence of flukes in treated, but not control turtles, indicated that a one day course of treatment at a dose rate of 3 times 50 mg/kg body weight is effective.
Conclusion This result should be of value for preventing disease in wild caught green turtles introduced into farms or aquaria.     

Proliferative dermatitis in a loggerhead turtle, Caretta caretta, and a green turtle, Chelonia mydas, associated with novel papillomaviruses

A subadult loggerhead turtle, Caretta caretta, presented with generalized small, white, raised lesions over its neck, shoulders, and all four flippers. A juvenile green turtle, Chelonia mydas, recently treated for fibropapillomatosis, presented with four similar localized lesions on one flipper. To diagnose the conditions, biopsies of the lesions were taken for histopathology, electron microscopy, and molecular diagnostics. Histopathologic findings were similar in the two turtles and skin lesions were characterized by multifocal areas of epidermal hyperplasia accompanied by variation and abnormalities in the nuclear morphology of keratinocytes and a few intranuclear inclusions in some cells. Transmission electron microscopy revealed multiple epithelial cells with large intranuclear aggregates of virions consistent in morphology with papillomavirus. Papillomavirus was detected in samples from both turtles by polymerase chain reaction (PCR). Sequence analysis of the partial sequence of the papillomavirus E1 gene revealed two viruses (CcPV and CmPV) that were distinct from each other and from other species in Papillomaviridae, and likely represent two novel species and perhaps a new genus.