Biocontrol method in aquaculture for rearing the swimming crab larvae Portunus trituberculatus

The aim of this work is to control the waterenvironment for culturing larvae of the swimmingcrab, Portunus trituberculatus, using microorganisms.The bacterial strain PM-4 (Thalassobacter utilis)improved the survival rate of crab larvae andrepressed the growth of Vibrio anguillarum (bacterium)and Haliphthoros sp.(fungus) in seawater. PM-4 wascultured and added daily to seawater during the firstto third zoean growth stage of the crab with diatomsand rotifers. Numbers of PM-4 decreased in culturewater during the first 3 days, because of feeding bythe first zoean stage of larvae. The finalconcentration of PM-4 was 10⁵ to 10⁶ cellsml^–1according to the plate count method in larval rearingwater.During 1989 to 1993, we tried seed productions of aswimming crab in 200 m³ containers at TamanoStation, Japan Sea-Farming Association. In 33trials of the biocontrol methods, average survivalrate of crab larvae was 28.3% when the bacterialstrain PM-4 was added. In 42 trials in which the strainPM-4 was not added, average survival rate of crab larvae was15.6%. We conclude that thebacterial strain PM-4 is effective as a biocontrolagent.     

三疣梭子蟹精子顶体反应过程中的形态和结构变化

用离子载体A2 3187和卵水人工诱导三疣梭子蟹精子的顶体反应 ,分别获得 75 33%和 84 83%的顶体反应率。应用光镜和电镜技术观察了顶体反应前后精子形态和结构的变化。未处理精子呈陀螺形 ,由顶体、核杯和 5 - 10条核辐射臂组成。顶体包括顶体囊和顶体管。顶体囊的伞形头帽拥有约 70条辐射肋。连续发生的精子顶体反应过程被人为地分为四个阶段 :(1)头帽鼓起 ;(2 )顶体囊外翻 ;(3)穿孔器前伸 ,顶体囊膜翻转 ;(4 )顶体囊膜脱落 ,顶体丝形成。直到第四阶段才观察到钉状精子的辐射臂开始收缩。探讨了辐射臂和穿孔器前冲在精子入卵中的功能     

三疣梭子蟹卵附着机制及相关形态学特征

Using histological methods and scanning electron microscopy, the structure of cement gland and formation of outer egg-membrane and egg-stalk of the swimming crab Portunus trituberculatus were studied to explain the mechanism of egg-attachment and function of the cement gland. Eggs adhered to the setae of pleopodal basipodite and endopodite for hatching followed by spawning but no egg attached on the setae of exopodite. The setae of pleopodal basipodite and endopodite were smooth for egg attachment while the setae of exopodite were branched and suitable for collecting and protecting eggs. Cement gland distributed along longitudinal axis of the pleopodal basipodite and endopodite under epithelia cells. Cells of the cement gland were elliptical and its size was about 50 times of the epithelium. Some glue secreted to the surface of the pleopod along the seta, while the other glue secreted to the surface of the pleopod by the channel which across the integument of the pleopod. The first spawned egg was transferred to the setae that were far from the pore, not attached to the setae of the pleopodal basipodite and endopodite near the genital pore. And the egg attachment became nearer and nearer to the pore with further spawning. Since the eggs to glide rather than roll, there was almost no glue on the contrary surface of the egg at first: and with further secretion, the whole surface was surrounded by glue. The thick part of glue formed one short bud during the glide of the egg and gradually developed into a full egg-stalk. On the other hand, the else glue developed into an outer egg-membrane, which was “trichromatic egg membrane” originally. Each egg attached to the setae of pleopodal basipodite and endopodite with its own egg-stalk. The egg-stalk was a plane strap at the formation stage and later it rolled into a rope shape, but it could be reverted to the former by outside force. Four types of egg attachment : an egg to a seta, an egg to several setae, several eggs to a seta and several eggs to several setae were discovered. In summary, the cement gland existed in the pleopod of the swimming crab and formed outer egg membrane and egg-stalk during the egg attachment [Acta Zoologica Sinica 50 (5) : 873 - 879, 2004].     

三疣梭子蟹两种养殖模式浮游动物调查

于2012年6-12月,在宁波市象山县东盛水产养殖有限公司对主养三疣梭子蟹池塘两种混养模式（模式Ⅰ：三疣梭子蟹与日本对虾、黑鲷、菲律宾蛤仔混养;模式Ⅱ：三疣梭子蟹与日本对虾、黑鲷混养）浮游动物进行了调查与比较。结果显示：共检出浮游动物13属19种,其中,原生动物3属3种;轮虫2属5种;枝角类2属3种;桡足类6属8种。浮游动物平均密度模式Ⅰ（370 ind./L）〉模式Ⅱ（209 ind./L）;平均生物量模式Ⅰ（17.974 mg/L）〈模式Ⅱ（18.102 mg/L）。Margalef指数为0.22~0.81;Shannon-Wiener指数为1.28~3.03;Pielou均匀度指数为0.54~0.96,均值都是模式Ⅰ〉模式Ⅱ。     

三疣梭子蟹精子顶体反应前后胞内Ca~(2+)的变化

应用激光扫描共聚焦显微镜(LSCM)和Fluo-3/AM荧染技术对三疣梭子蟹精子顶体反应前后的胞内Ca2 变化进行了观察和检测.结果显示,在精子顶体反应过程中,胞内Ca2 主要分布在细胞核、穿孔器和胞质膜残存处,胞内Ca2 浓度([Ca2 ]I)总体上呈现先上升后下降的趋势.顶体反应前精子的平均荧光强度为35.95±5.71;穿孔器前伸、顶体囊膜翻转阶段精子的平均荧光强度为66.80±7.35;顶体囊膜脱落、顶体丝形成阶段精子的平均荧光强度为3.87±2.82;上述各阶段间精子荧光强度有极显著差异(P<0.01).顶体反应穿孔器前伸、顶体囊膜翻转阶段的精子相比顶体反应前精子,[Ca2 ]I显著提高;而在顶体囊膜脱落、顶体丝形成阶段,[Ca2 ]I则急剧下降,只在顶体丝基部胞质膜残存处有微量Ca2 存在.初步探讨了三疣梭子蟹精子顶体反应前后胞内Ca2 变化的功能.     

Asymmetric swimming pattern of Vibrio alginolyticus cells with single polar flagella

The swimming pattern of bacteria with single polar flagella has usually been described as "run and reverse". We observed the swimming traces of monotrichously flagellated Vibrio alginolyticus cells and examined the relationship between the swimming pattern and the sense of progress. Swimming in regions other than a solid surface was confirmed to be linear run and reverse. Near a solid surface, the traces consisted of "run and arc"; the cells were found to curve sharply during backward swimming, while they progressed linearly during forward swimming. The "run and arc" swimming pattern may play an important role in the chemotaxis strategy of marine bacteria at solid surfaces.     

Major histocompatibility complex class IIB allele polymorphism and its association with resistance/susceptibility to Vibrio anguillarum in Japanese flounder (Paralichthys olivaceus)

The full length of major histocompatibility complex (MHC) class IIB cDNA was cloned from a Chinese population of Paralichthys olivaceus by homology cloning and rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). The MHC IIB genomic sequence is 1,864 bp long and consists of 34-bp 5′UTR, 741-bp open reading frame, 407-bp 3′UTR, 96-bp intron1, 392-bp intron2, 85-bp intron3, and 109-bp intron4. Phylogenetic analysis showed that the putative MHC class IIB amino acid of the Chinese P. olivaceus shared 28.3% to 85.4% identity with that of the reported MHC class IIB in other species. A significant association between MHC IIB polymorphism and disease resistance/susceptibility was found in Chinese P. olivaceus. Thirteen different MHC IIB alleles were identified among 411 clones from 84 individuals. Among the 280 (268) nucleotides, 32 (11.4%) nucleotide positions were variable. Most alleles such as alleles a, b, c, d, e, f, j, k, i, m were commonly found in both resistant and susceptible stock. Via χ² test, allele d was significantly more prevalent in individuals from susceptible stock than from resistant stock, and their percentages were 23.80% and 7.14%, respectively. In addition, allele g occurred in 9 and allele h in 4 of 42 resistant individuals that were not present in the susceptible stock; their percentages were 21.4% and 9.52%, respectively. Although allele l was found only in 8 individuals from the susceptible stock, its percentage is 19.05%.     

Characterization of a fatty acid binding protein from the swimming crab Portunus trituberculatus and its effects on the composition of fatty acids in different tissues

Fatty acid binding proteins (FABPs) belonging to the lipocalin protein family, have an affinity for fatty acids (FAs) and participate in both the transport of FAs and lipid metabolism. In this study, a 909-bp full-length pt-FABP cDNA containing a 411-bp open reading frame (ORF) was obtained from the swimming crab Portunus trituberculatus. Quantitative real-time PCR revealed that during larval development, pt-FABP expression increased gradually from the zoea to the megalopa, and decreased in the juvenile crab. pt-FABP mRNA was detected at the highest level in the testis, followed by the muscles, with the lowest level in the hepatopancreas. The composition of fatty acids in three different tissues from crabs of different sex was detected by gas chromatography–mass spectrometry (GC-MS). The results indicated that the FA profile was in accordance with the expression of pt-FABP, which suggested that pt-FABP participates in the unsaturated fatty acid transportation and storage system of P. trituberculatus.     

Variants of the CD40 ligand gene are not associated with increased susceptibility to tuberculosis in West Africa

Evidence for linkage between tuberculosis and human chromosomal region Xq26 has previously been described. The costimulatory molecule CD40 ligand, encoded by TNFSF5 and located at Xq26.3, is a promising positional candidate. Interactions between CD40 ligand and CD40 are involved in the development of humoral- and cell-mediated immunity, as well as the activation of macrophages, which are the primary host and effector cells for Mycobacterium tuberculosis. We hypothesised that common variation within TNFSF5 might affect susceptibility to tuberculosis disease and, thus, might be responsible for the observed linkage to Xq26. Sequencing 32 chromosomes from a Gambian population identified nine common polymorphisms within the coding, 3 and 5 regulatory sequences of the gene. Six single nucleotide polymorphisms (SNPs) and a 3 microsatellite were genotyped in 121 tuberculosis patients and their available parents. No association with tuberculosis was detected for these variants using a transmission disequilibrium test, although one SNP at –726 showed some evidence of association in males. This finding, however, did not replicate in a separate case control study of over 1,200 West African individuals. We conclude that common genetic variation in TNFSF5 is not likely to affect tuberculosis susceptibility in West Africa and the linkage observed in this region is not due to variation in TNFSF5.Sadly, Professor Steve Bennett passed away in March 2003