从原始生殖细胞分离昆明小鼠EG细胞及其传代培养

为提高昆明小鼠胚胎生殖细胞(embryonic germ cells, EG cells)建系效率，以怀孕8.5～12.5 d小鼠胎儿为材料，比较了胎儿后1/3部位的组织共培养、生殖嵴共培养、差速贴壁和穿刺生殖嵴4种分离胚胎原始生殖细胞(primordial germ cells，PGCs)的方法以及不同传代方式对EG细胞克隆的影响。结果表明：生殖嵴共培养和差速贴壁方式获得了较好分离EG细胞的效果，与其它两组比较差异显著；手工传代方式与连同成纤维细胞一起消化的传代方式相比获得了较高传代比率。对所分离EG细胞经形态观察、AKP染色和体外分化能力检测，证实其符合小鼠EG细胞的集落状生长、细胞未分化特性及细胞多能性等特征。     

Variation in the immuno-pathological responses of lambs after experimental infection with different strains of Mycobacterium avium subsp. paratuberculosis

Ruminant infection by Mycobacterium avium subsp. paratuberculosis (MAP) causes a granulomatous inflammatory response in the intestine and associated lymph nodes. Differences either in the affected organs or in the inflammatory infiltrate were observed between species and individuals. Such differences are usually attributed to variations in host immune responses or to inconsistent effects among different MAP strains. To evaluate if different MAP strains induce different immuno-pathological responses in lambs, 28 one-month-old individuals were divided into six groups and inoculated with different MAP strains. Groups 1 and 2 were inoculated with two bovine strains isolated in Argentina that showed different genetic patterns after BstEII-IS900-RFLP (hereafter strains E and A respectively). Group 3 was inoculated with a bovine strain isolated in Spain obtained after a previous step of culture (patterns C1). Group 4 was inoculated with a homogenate of intestinal mucosa of a clinical case affected by the same bovine strain as that of group 3. Group 5 was inoculated with an ovine strain that was directly purified from the intestinal mucosa of a clinical case, and group 6 was kept as control (i.e. no inoculation). Peripheral immune responses were assessed until 150 days post-infection (dpi), when lambs were humanely killed. Pathological studies were performed in tissues from the intestine and lymph nodes. Lesion types and inflammatory infiltrates were examined as indicators of pathogenicity. All the lambs infected with bovine MAP strains showed a common lesion pattern regardless of the strain type. Such pattern was characterized by focal lesions mainly in the mesenteric lymph nodes, the presence of fibrous tissue, and, occasionally, necrosis in the granulomas as well as the presence of numerous giant cells. Differences in lesion severity were observed among groups: lambs from groups 1 and 2 had the highest number of granulomas and the largest lymph node area affected. Lesions in animals from group 5 (infected with an ovine strain) were more severe and occurred mostly in the intestinal lymphoid tissue; necrosis, fibrosis or giant cells were never detected in this group. These results indicate that the MAP strain type induces different pathological responses in lambs.     

青岛汇泉湾大叶藻种群遗传多样性的研究

采用聚丙烯酰胺凝胶垂直平板电泳技术对青岛汇泉湾大叶藻种群内部的遗传多样性进行了初步分析,共研究了6种酶,得到该种群多态位点的比例（P％）为56．52％,每个位点平均等位基因数A／L为1．3478,根据其中17个位点,25个等位基因得到平均杂合度的预期值He为0．2122,观测值Ho为0．3251,对其遗传偏离进行D测验表明,大叶藻的杂合度较高,属于遗传多样性较为丰富的物种。     

Notes on hatchery spawning methods for bigmouth sleeper Gobiomorus dormitor

The bigmouth sleeper Gobiomorus dormitor, a popular sport fish within its native range, offers a management alternative to exotic species currently managed for recreational fishing. Hatchery production and stocking are needed to create reservoir sport fisheries because this species usually requires access to marine environments for recruitment. Bigmouth sleepers have not been spawned previously in a hatchery, and hence we used natural, artificial and semi‐natural spawning techniques. No egg deposition or propagation resulted from natural pond spawning. Artificial spawning techniques using Ovaprim^® and Chorulon^® injections followed by hand stripping were more successful. Semi‐natural (hormone injections plus volitional spawning) spawning trials using Ovaprim^® and Chorulon^® displayed the greatest potential, and three consecutive evening injections (1.0 mL kg^?1) of Chorulon^® appeared most conducive. High‐fecundity spawning was achieved by injecting bigmouth sleeper pairs and allowing volitional spawning in aquaria with spawning cavities. Initial success in 2007–2008 was isolated to wild‐caught fish, but captive broodstock were spawned using these techniques in 2009. Eggs hatched in <20 h at 23–24 °C. The resulting larvae, 1.0–1.5 mm in length, displayed large yolk sacs, unpigmented eyes and no apparent mouth. These hatchery propagation efforts suggest that hormone‐induced semi‐natural spawning has the greatest potential for bigmouth sleeper propagation.     

Effect of diet on the lipid composition of the commercial clam Donax trunculus (Mollusca: bivalvia): sex‐related differences

This paper describes the lipid composition of the commercial bivalve Donax trunculus and the differences originated when the animals were fed with two phytoplankton species (Tetraselmis suecica and Chaetoceros sp.) in a hatchery. We also analysed sex‐related differences in lipid classes and fatty acid profile. Total lipids were higher in females than in males. Triglycerides and phospholipids were the major lipid components, and the former dominated in females and the latter in males. The main fatty acids in both sexes were 16:0, 20:5n‐3 (eicosapentaenoic acid) and 22:6n‐3 (docosahexaenoic acid). Females showed higher percentages of saturated and monounsaturated fatty acids and lower levels of polyunsaturated fatty acids than males. Significant differences in total lipid, lipid classes and fatty acid profiles were also found due to diet. Docosahexaenoic acid decreased and total lipids, free fatty acids, arachidonic acid and EPA increased in both sexes. Despite these differences, the condition of the species was maintained and the broodstock even maturated.     

西藏主要农区农田土壤肥力状况研究

西藏由于其特殊的地理位置、气候类型和地质历史以及耕作习惯,使其具有特殊的土壤特征。在2002~2004年间,我们对西藏主要农业区的林芝、拉萨、日喀则、山南等4地(市)的农田土壤进行了采样分析,分析结果显示:4地(市)农田土壤酸碱性由弱酸性至弱碱性(pH6.5~9.0);养分的基本状况是有机质含量在5~50g/kg之间,变幅较大,N素少(全N0.2~2g/kg,速效N20~80mg/kg),P素缺(全P0.22~1.42g/kg,速效P1~133mg/kg),K素(全K1.9~12.2g/kg,速效K80~1290mg/kg)和微量元素比较丰富(Cu10~443mg/kg,Zn47~266mg/kg,Fe1880~7550mg/kg,Mo0.2~6.6mg/kg,Mn196~736mg/kg,B0.5~138mg/kg)。     

Fish invasion in the river systems of Guangdong Province,South China: Possible indicators of their success

Although a large number of fish species have been introduced into Guangdong Province in Southern China, a few species, such as tilapia (Tilapia spp.), North African catfish Clarias gariepinus Burchell, mrigal carp, Cirrhinus mrigala (Bloch) and the sucker mouth catfish (Hypostomus sp.), have established natural populations and can be considered “successful invaders” in large rivers. The specific mechanisms underlying these contrasting results among different introduced fish species remain understudied. The relationship between multiple abiotic–biotic factors and the success of four invasive species was investigated using survey data for the Guangdong Province river ecosystem. In contrast to previous studies that have considered species‐specific traits, the focus was on economic, ecological and anthropogenic factors to predict invasion success. Four main predictive indicators were found: (1) successful invaders were of low or no commercial value; (2) successful invaders tolerated a wide range of environmental conditions, including poor water quality; (3) biodiversity loss accelerated the growth of non‐native populations; (4) human disturbance facilitated population growth and spread of invasive fish species. To lessen the impacts of invasive fish species, the selection of breeding species and breeding areas, maintenance of water quality and reduction in water pollution, protection of the diversity of fish species and reduction of human interference should be addressed.     

Study on an integrated eco‐aquaculture system for rearing the yellow seahorse,Hippocampus kuda Bleeker

Seahorse aquaculture is challenged by insufficient feeding supply and lack of biological and nutritional knowledge, resulting in low survival rate and poor economical profitability. We report here an integrated eco‐aquaculture system in which the yellow seahorse and its natural prey were co‐cultured with fertilized water and seaweed in cement ponds. In the first stage, urea (10 g m^?3) and chicken manure (50 g m^?3) were used to fertilize the cultured water, 5–7 days later, rotifer and micro‐crustaceans were flourishing. Then, seahorse juveniles were stocked at 200 ind m^?3. After 2 weeks, seaweed Gracilaria lichevoides was transplanted into the ponds to regulate water quality, light and to provide holdfast attachment for seahorses. The optimal density of G. lichevoides was controlled between 0.5 and 2 kg m^?3. The introduction of seaweed provides the habitat for natural food of seahorse. Within the cluster of the seaweed, small crustacean density was over 450 individuals per 100 g of fresh weight. Initial size of seahorse juvenile was 1.03 ± 0.091 cm, After the 146‐day rearing period, seahorse survival rates were 70.8%, 57.7% and 42.5%, and body standard lengths were 11.33 cm, 10.84 cm and 10.04 cm in the integrated eco‐aquaculture system (GFA) and in monoculture systems of FA (fertilized + feeding) and BA (only feeding) respectively. This system incorporated traditional Chinese aquaculture technique in which feedings nature food organisms were cultivated by fertilization and ecological regulation. Results from these preliminary experiments suggest that the integrated system could be technically feasible, suitable and exemplary.     

Acrosome reaction of Chinese mitten-handed crab Eriocheir sinensis (Crustacea: Decapoda) spermatozoa: Promoted by long-term cryopreservation

The effects of three media, two temperatures, and fourteen durations of cryopreservation from 0 h to 450 d on in vitro acrosome reaction (AR) of spermatozoa in Chinese mitten-handed crab Eriocheir sinensis (Crustacea: Decapoda) were investigated. The spermatozoa of good quality were obtained from spermatophores by a glass homogenizer in an ice-bath and centrifugation at 4 °C. At 0 h, 2 h, 1 d, 3 d, 15 d, 30 d, 60 d, 90 d, 120 d, 150 d, 180 d, 270 d, 360 d, and 450 d of cryo-storage in Ca²⁺-free artificial seawater, 10% (v/v) glycerol, and 5% (v/v) dimethylsulfoxide at − 80 °C and in liquid nitrogen (− 196 °C), the changes in spermatozoal morphology, the time of beginning AR, and the time of maximum percentage of AR were observed. The relationships of the changes on AR presented with the different media, temperatures, and durations of cryopreservation were speculated. In this study, the cryopreserved spermatozoa all underwent AR in less than 1 h of settlement under room temperature while the percentage of AR in the control was only about 4.9%. Meanwhile, cryopreservation shortened both the time of beginning AR (from 30.11 min of the uncryopreserved spermatozoa to 0 min of cryopreserved spermatozoa on the 30th day) and the time of maximum percentage (from 59.88 min of the uncryopreserved spermatozoa to 0 min of cryopreserved spermatozoa on the 60th day). Whereas the effect of media on sperm cell AR was negligible (P > 0.05), the treatments of spermatozoa with short- and long-term cryopreservation resulted in extremely significant differences in the time of beginning AR as well as in the time of maximum percentage of AR (P < 0.01). The present data indicate that cryopreservation for long or short periods can promote the AR of sperm cells in E. sinensis and physiologically affect the ability to capacitate. It may be that the mechanism of AR in this study is the direct promotion of membrane fusion of the acrosomal cap, or destruction of the proteins inhibiting AR and activation of the proteins promoting AR, by cryopreservation. In addition, the results also show that cryopreservation can protect the spermatozoa because AR can occur in almost all sperm cells cryopreserved for less than 15 d.     

樟子松天然母树林疏伐强度与种子产量的关系

通过对樟子松天然母树林的４种不同疏伐强度标准地的林木开花结实与气象因子观测，进行了疏伐强度对母树林种子产量影响的研究。结果表明：（１）疏伐可有效的促进母树林的开花结实；（２）疏伐强度与母树林单位面积产种量相关密切；（３）疏伐促进林木结实其主要影响因素是光照，地面温度等气象因子；（４）樟子松天然母树林定株疏伐的最佳密度为１９０株ｈｍ＾２。