温度和盐度突变对菲律宾蛤仔斑马蛤耗氧率和排氨率的影响

为研究温度和盐度对蛤仔新品种斑马蛤耗氧排氨的影响,以野生蛤仔为对照,实验设置15、20、25、30和35℃五个温度梯度和20、25、30、35和40五个盐度梯度,结果表明:温度和盐度对斑马蛤的耗氧率和排氨率影响显著（P < 0.05）。在温度15-35℃内,随着温度的增加,耗氧率和排氨率整体上呈增加的趋势。在20-40盐度内,耗氧率随着盐度的升高先减少后增加,排氨率随着盐度的升高先增加后减少,在盐度为30时达到最高值。在水温为15℃,盐度20-40内,斑马蛤的O:N为9.534-62.008;在盐度为35,水温在15-35℃内,斑马蛤的O:N是20.700-74.138。与野生蛤仔比较,斑马蛤的耐高温能力要强于野生蛤仔,从Q₁₀的变化反映出斑马蛤对温度的敏感性相对较弱,适应温度变化的能力比较强;斑马蛤的耐低盐和耐高盐能力强于野生蛤仔。研究结果为进一步完善蛤仔斑马蛤的人工养殖技术提供参考。     

海洋污染物对菲律宾蛤仔的免疫毒性

环境污染能够影响养殖贝类的免疫能力,是导致贝类大规模死亡的重要原因之一.探讨了大连周边4个海区污染物对采集的菲律宾蛤仔(Ruditapes philippinarum)免疫毒性影响.结果发现:污染物浓度和种类对蛤仔的免疫和生理指标具有重要影响,在重金属和石油污染物浓度较低的皮口海区,血细胞总数、亮氨酸氨基肽酶活性和血淋巴溶菌酶活性均显著高于其它3个海区(P＜0.05),而蛤仔的脂质氧化水平则较低;在重金属和石油污染物浓度较高的黑石礁海区,蛤仔血淋巴谷胱甘肽含量显著高于其它3个海区(P＜0.05);在重金属浓度较高的庄河海区,蛤仔表现出较高的超氧化物歧化酶活性(P＜0.05).     

温度和盐度对皱肋文蛤幼贝存活与生长的影响

在室内控制条件下,研究了不同海水温度和盐度对皱肋文蛤(Meretrix lyrata)幼贝存活与生长的影响。结果表明:皱肋文蛤幼贝适宜生存温度为12.2-35.6℃,最适生存温度为24-30℃;适宜生长温度为23.5-33.0℃,最适生长温度为27-30℃,属典型的南方滩涂贝类。皱肋文蛤幼贝适宜生存盐度为4.3-40.5,最适生存盐度为11-31;适宜生长盐度为17.1-33.4,最适生长盐度为19-23,属广盐性滩涂贝类。该贝低温和高温敏感起始点分别为21℃和33℃;低盐和高盐敏感起始点分别为9和33。皱肋文蛤幼贝对极端温、盐度具有一定的耐受力:在37℃下仍可保持6d,100%不死亡,在39、41℃下分别在3d和5d内全部死亡;在4、6、8、10、12℃下则可分别100%存活3、4、6、9、11d;在盐度为0、5、7、9时保持100%存活的时间分别为5、8、10、10d;在盐度为33、35、37、39时保持100%存活的时间分别为7、5、3、3d,盐度41时当天即出现死亡。     

铜藻苗种繁育研究

2011—2013年, 以浙江省南麂岛马祖岙的野生铜藻作为亲本, 详细观察了铜藻繁殖特性、胚胎发育和幼孢子体的早期发育过程, 开展了铜藻幼孢子体早期发育条件(温度、盐度、光照强度、光照周期等)的优化研究。结果发现, 南麂岛铜藻的自然繁殖盛期为每年3月下旬至5月上旬, 室内海水温度为13—18℃, 较适宜于雌雄生殖托的形成。在试验条件下, 铜藻受精卵发育的适宜温度为10—20℃, 盐度是20—30, 在该培育条件下, 经过48h的孵化, 受精卵可以大批量同步化发育成具有假根的幼孢子体。人工育苗结果表明, 适宜的环境因子(温度20℃, 盐度20—30, 光照强度4500 lx, 光照时间12—18h/d)较利于幼孢子体的生长发育。铜藻幼孢子体的前期培育连续流水和充气为宜, 经18—20d的室内培育, 幼孢子体可发育成具有2—3叶片的幼苗。质地坚硬、表面粗糙的水泥砖较适合铜藻幼孢子体的附着和生长, 可满足后期藻场修复的要求。     

定点突变Ca~(2+)平衡位点对粘质沙雷氏菌脂肪的影响

为了研究粘质沙雷氏菌(Serratia marcescens)脂肪酶lip A中的Ca~(2+)对酶活力的影响,采用定点突变的方法对平衡Ca~(2+)的位点D388进行定点突变,对突变体进行诱导表达后,纯化目的蛋白并进行酶学性质分析。与野生酶相比,突变酶的最适温度由40℃降低到25℃,最适p H值由8.5降到8.0,T_(1/2)值比野生酶提高了2℃。由此可知,该Ca~(2+)对脂肪酶lip A酶学性质有着至关重要的作用。     

温度和盐度对青蛤孵化及幼虫、稚贝存活与生长变态的影响

在9个温度梯度(10-34℃)和10个盐度梯度(盐度3‰-50‰)条件下,研究了温度和盐度对青蛤孵化及幼 虫、稚贝生存与生长变态的影响。结果表明,青蛤孵化和浮游幼虫生长的适温范围为24-32℃,最适温度均为26- 30℃,稚贝生长的适温范围为22-32℃,最适温度为24-30℃。在最适温度下,D形幼虫变态率达80．7％-88．2％, 浮游幼虫和稚贝的存活率分别为86．2％-88．7％和81．5％-84．0％;孵化及浮游幼虫的生长适宜盐度为15‰- 30‰,稚贝为10‰-35‰,最适盐度均为20‰-25‰。在最适盐度下,D形幼虫的成活率、变态率、生长速度皆最高, 分别达到86．9％、77．5％和9．38×11．0μm/d,匍匐幼虫经14-14．5d发育至双管期稚贝,至双管期稚贝的成活率 82．5％-85．0％,日平均生长达13．1μm以上。与大多数滩涂贝类一样,青蛤属于广温广盐性贝类,且稚贝对低盐的 适应能力强于对高盐的适应能力。     

辽东湾北部近海沙蜇的动态分布

通过分析2008-2011年5月下旬至7月下旬辽东湾北部近海的大型水母锚流网监测数据并结合8月份的渔业资源拖网调查和渔港访问的数据,探讨辽东湾北部近海沙蜇的动态分布移动规律并着重分析了沙蜇幼水母的生态类型,为渔业资源生产和灾害性预警预报提供科学依据.结果显示:6月份,辽东湾北部近海发现大量沙蜇幼水母,并且在双台子河河口近海5 m等深线两侧水域形成密集区,6月上中旬沙蜇海区平均渔获密度52.8条·网-1·h-1,6月下旬海区平均渔获密度46条·网-1·h-1,最高渔获密度可达667条·网-1·h-1,出现在双台子河口5 m等深线内;7月份,沙蜇个体增大,逐渐进入成体阶段,调查海区沙蜇渔获数量比6月份大幅度减少,并且渔获密度的中心区向5-10 m等深线内偏深水域移动,7月上旬海区平均渔获密度5.1条·网-1,h-1,7月中下旬海区平均渔获密度1.8条·网-1·h-1;8月中旬辽东湾北部近海5 m等深线内沙蜇数量极少,在10-20 m等深线内海域往往出现成体沙蜇的密集区并形成渔汛.沙蜇幼水母生存的海水表层温度为17.7-27.3 ℃,表层盐度为24.3-31.9;渔获密度300条·网-1·h-1以上的沙蜇幼水母密集分布区海水表层温度为20.4-24.4℃,表层盐度为27.1-31.6.辽东湾北部河口区是沙蜇的可能发生地之一,相对于沙蜇成体阶段表现出来的低温高盐的生态属性而言,河口近海相对高温低盐水域往往出现沙蜇幼水母渔获的中心,沙蜇幼水母阶段表现出相对高温低盐的生态特性.沙蜇自幼水母至成体阶段,海区分布密集区自浅水位移至深水.     

菲律宾蛤仔莆田群体与大连群体生物学比较

对菲律宾蛤仔莆田和大连群体形态及生物学进行了比较。菲律宾蛤仔大连群体和莆田群体的生物学零度(BZP)分别为6.77℃和10.99℃;有效积温(EAT)分别为315.21和172.79℃.d。通过人工促熟,大连群体繁殖期比自然海区提早1个月,莆田群体比自然海区提早5个月。莆田的蛤仔第1次最高产卵量可达104万粒,大连的蛤仔最高产卵量可达65万粒,大连的蛤仔卵径(66.13±2.40)μm(n=40),莆田的蛤仔卵径(71.88±3.14)μm(n=40),二者差异显著(p<0.01)。大连的蛤仔D形幼虫大小平均96.13(±5.83)μm×76.75(±4.46)μm(n=40);莆田的蛤仔D形幼虫大小平均107.63(±5.55)μm×82.88(±4.22)μm,二者差异显著(p<0.01)。莆田的蛤仔幼虫整个浮游期平均日增长9.63μm×9.79μm,大连的蛤仔幼虫整个浮游期平均日增长12.20μm×12.47μm,后者快于前者(p<0.01)。大连的蛤仔在水温25℃以上条件下,浮游期为10d左右,附着大小为179.9μm×174.4μm,附着至变态需8~9d,变态大小230.0μm×220.0μm,相同条件下,莆田的蛤仔浮游期仅5~6d,附着至变态只需4~5d,附着大小为174.9μm×171.4μm,变态大小仅为196.7μm×190.8μm。     

盐度对三疣梭子蟹摄食行为及生长特性的影响

从行为角度出发, 设置了 5、 10、 20、 30 和 40 五个盐度梯度 , 探讨不同盐度对梭子蟹幼蟹和成蟹的摄食行为及特性的影响。结果表明 : 在盐度 5 时, 除摄食活动外 , 幼蟹不外出游动且深潜底沙(约 0.5 cm); 在盐度 40 时, 幼蟹浅潜沙层 , 仅盖住背甲 (约 0.1 cm); 盐度为 10、 20 和 30 时, 除觅食活动外 , 幼蟹均外出游动。日摄食次数由幼蟹的 1 -2次增加到成蟹的 2-4 次。盐度为 10-40 时, 幼蟹平均日摄食量均维持在 0.0999 g·g−1 以上(每天每只梭子蟹单位体重所摄取食物的重量, 下同 ), 其中盐度为 20 时的日摄食量为最大(0.1974 g·g−1), 幼蟹生长的适宜盐度范围为 10-40, 最适为 20。此外 , 各盐度对成蟹的行为压迫不大, 对应的日平均摄食量亦无显著差别。     

海南豇豆枯萎病病原鉴定及生物学特性初步研究

从海南乐东豇豆地采集到的枯萎病病样,经形态学观察、致病性测定和真菌ITS序列分析,将其病原鉴定为尖孢镰刀菌菜豆专化型(Fusarium oxysporum f.sp.phaseolus)。生物学特性分析表明:该病原菌菌丝生长适宜温度为20~30℃,最适温度为25℃,50℃时不能生长;适宜p H为6~10,最适p H值为8.0;全黑暗和全光照有利于菌丝的生长;可利用多种碳、氮源,最适碳源为果糖和D-甘露醇,氮源为蛋白胨和硝酸钠。     

The clam sipho as indicator for growth indices in the soft-shell clam <Emphasis Type="Italic">Mya arenaria</Emphasis>

Due to the species’ deep burrowing behaviour, growth parameters of the soft-shell clam Mya arenaria L. 1758 are difficult to estimate, especially in deeper habitats which are not directly accessible. In this study, we analysed 192 specimens of M. arenaria and found, contrary to results of most other studies that the sigmoid Gompertz growth model (GGM) is appropriate to describe the growth. Predictions from this model confirm the finding of a life span up to 8 years and as a consequence of this, a maximum shell-length of 60 mm. Individual growth rates calculated from the first deviation indicate a nearly exponential growth in young individuals. The relationship between the clam sipho-width and age, shell-length and biomass conform to the GGM. This is a new approach to assess growth of M. arenaria and to solve practical problems that arise in field studies.     

Reproduction and development of the freshwater clam Corbicula australis in southeast Australia

The freshwater clam Corbicula australis is an important component of the macrobiota of the river systems of southeast Australia. Reproduction of two populations of this clam in the Nepean River at Douglas Park and Menangle was investigated to document the gametogenic cycle, larval morphology and to determine when they incubate embryos. C. australis is a simultaneous hermaphrodite and broods its young in the inner demibranchs. The gonads are ovotestes with oogenic and spermatogenic regions in each ascinus. The sperm are biflagellate, a condition unique in the Bivalvia to clonal corbiculids. Gametogenesis was continuous and did not exhibit a seasonal pattern. In contrast, spawning and incubation of embryos was limited to the warmer months of the year. Embryos were present in the gills from October to May. C. australis develops through a modified veliger larva with a vestigially ciliated velum which is not used for swimming or particle capture. The velum is covered by microvilli and it is suggested that the velar epithelium may be specialised for nutrient uptake in the marsupial environment. C. australis produces several clutches each year and the young are released as advanced juveniles with a well-developed foot. Reproductive output differed between the two populations. This was in part due to the larger size of the clams from Menangle and may also reflect the enhanced productivity at this site. The suite of life history traits exhibited by C. australis: hermaphroditism, potential for self-fertilization/androgenesis, brooding progeny to the crawl-away juvenile stage and a high reproductive output, provide for rapid colonization and population growth in this clam which typically inhabits disturbance prone sandy lotic habitats.     

Gymnophallid digenean Parvatrema duboisi uses Manila clam as the first and second intermediate host

To identify the metacercariae of a gymnophallid trematode in the Manila clam Ruditapes philippinarum from the Ariake Sea, experimental infection and molecular analysis were conducted. Based on the morphology of adult worms obtained from experimentally infected mice, the parasite was identified as Parvatrema duboisi. Comparison of internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) sequences for metacercariae and sporocysts from Manila clams and adult worms collected from wild wigeon Anas penelope showed minor differences ranging from 0 to 0.8%. These data strongly suggest that in the Ariake Sea, the parasite has a lifecycle using the Manila clam as the first and second intermediate hosts and wigeon as the definitive host.     

Valve closure responses of the Asiatic clam Corbicula fluminea exposed to cadmium and zinc

The valve movement patterns of immobilized Asiatic clams (Corbicula fluminea) were monitored during exposure to constant concentrations of cadmium (0.0, 0.1, 0.2, 0.3, and 0.4 mg 1^–1) or zinc (0.0, 0.1, 0.3, 0.5, and 0.9 mg 1^–1) for 24 h following a 24-h acclimation period. Data indicate that the duration of response was concentration dependent and toxicity related. Durations of periods with valves parted declined as the concentration of heavy metal increased. Behavior was consistent for both mean time to first closure following the initial exposure and mean time per valve parting episode over a 24-h exposure period. Mean time per valve parting episode during the 24-h exposure period ranged from 600 minutes for control trials to 36 and 69 minutes for the highest concentrations of cadmium and zinc tested, respectively. There was no association between durations of periods with valves sealed and solutions of cadmium; however, lengthening durations of periods with valves sealed coincided with exposure of clams to progressively more concentrated zinc solutions. In addition, Asiatic clams demonstrated a greater rate of response (decline in the duration of periods with valves parted) to progressively more concentrated solutions of cadmium than to comparable increases in the concentrations of zinc solutions.     

d-Alanine metabolism in the lucinid clam Lucinoma aequizonata

The chemoautotrophic symbiont-bearing clam Lucinoma aequizonata contains very high levels of free d-alanine in all tissues. The possible sources for this amino acid and its involvement in the clams' metabolism were investigated. Very low levels of d-alanine (generally below 1 mol·l^-1) were measured in the sediment porewaters from the habitat of the clams. Experiments with ¹⁴C-labeled tracers demonstrate an active metabolism of d-alanine in the clams rather than a role as inert waste product. d-alanine is metabolized at about 0.12 mol·g fw^-1·h^-1. Label from aspartate, but not glucose and CO₂, is incorporated into d-alanine. Incubation with labeled d-alanine did not result in formation of radioactive l-alanine. Tests for alanine racemase (EC 5.1.1.1) and d-amino acid oxidase (EC 1.4.3.3.) did not show activity in either gill, i.e. symbiont and host, or foot tissue. d-Alanine amino transferase (EC 2.6.1.b.) was demonstrated in gill and foot tissues. Two sources for d-alanine are proposed: a degradation of cell walls of symbiotic bacteria and production by the host using a d-specific alanine transaminase.Abbreviations aa amino acid(s) - fw fresh weight - HPLC high-performance liquid chromatography - MBH methyl benzethonium hydroxyde - NAC N-acetyl-l-cysteine - OPA ortho-phthaldialdehyde - TCA tricarbonic acid     

Heat shock protein response to thermal stress in the Asiatic clam,Corbicula fluminea

A recent approach to evaluate environmenta induced damages has been damages has been suggested, based on the stress response. The approach involves the detection of stress protein induction in organisms to infer about environmental conditions in their surroundings. However, to be an indicator of adverse biological effects in the environment, the elevation of stress proteins should be compared to a response pattern for the experimental species. JuvenileCorbicula fluminea, collection from a control site, were submited to heat-shock stress in the laboratory to obtain the stress response pattern under normal and extreme conditions. Acclimated to 26°C, the specimens were submited to 29, 32, 35 and 38°C, for 96 h. After 1, 2, 4, 8, 24, 48, 72 and 96 h of exposure, clams were removed from each vial and prepared for stress protein analysis. Animals from the control site were frozen in liquid nitrogen at the time of collection, and prepared for stress protein analysis. Hsp60 and 70 were detected by immunoreactivity after separation on 12.5% polyacrylamide gels and transference to nitrocellulose by western blotting, to determine the stress protein concentrations. The result showed that hsp70 increased at 4h from the beginning of the experiment and progressed over the 96 h experimental period in animals exposed to 35°C. However hsp70 levels decreased between 4 h and 24 h for the clams stressed at their lethal temperature of 38°C. Immunoblotting with hsp60 showed similar reactivity. At 38°C there was an increase in the amount of hsp60 at 4h, reaching a maximum eight-fold level at 8h. By 96h, the amount decreased to levels lower than those observed at 4h. At 38°C the level of hsp60 began to decrease at 8 h and continue to decline to 24 h when the clams died. The data support the hypothesis of increasing concentrations of stress protein until the heat shock approaches the thermal limits for the species. The results of this research suggest the usefulness of using the stress response as a diagnostic in environmental toxicology. They confirm that the sps response may serve as a valid biomonitoring tool under chronic, sublethal exposures when it is still possible to prevent effects at organismal or higher organizational levels.     

A genetic comparison of two species of clam shrimp in the genus Eulimnadia: An electrophoretic approach

Herein we report the first genetic comparison among species in the genus Eulimnadia. Multilocus genotypic patterns (using six allozyme loci) were compared for a total of 2277 clam shrimp from nine populations from Arizona and New Mexico. Seven of these populations were morphologically typed as Eulimnadia texana Packard and two were typed as Eulimnadia diversa Mattox. All populations were hermaphrodite-biased, and highly inbred (inbreeding coefficients ranging from 0.33 to 0.98). Genetic distances showed the two species to be within the range described for other arthropods. One of the two E. diversa populations appeared to be a hybrid between E. texana and E. diversa, showing electrophoretic patterns similar to both species, although morphologically, they were typed as E. diversa. A phenogram (generated using coancestry distances and a neighbor joining algorithm) placed this hybrid population half-way between these two species, and a breakdown of individuals within this hybrid population (based on allozyme scores) indicated individuals very similar to the second E. diversa population, and two groups of apparent hybrid individuals. Therefore, the distinction between these two species is questionable due to their apparent hybridization in this area of Arizona. Genetic population structuring was noted among the seven E. texana pools. Estimated migration rates were less than one migrant per generation. Even in the geographically close pools in New Mexico, which were separated by only hundreds of meters, significant sub-structuring was noted, and estimates of migration rate were less than two migrants per generation.     

The larval development of an Australian limnadiid clam shrimp (Crustacea, Branchiopoda, Spinicaudata), and a comparison with other Limnadiidae

The adult morphology of the Australian Limnadopsis shows some remarkable differences to that of other Limnadiidae. These differences are not reflected in its larval development. In Limnadopsis parvispinus, larval development comprises six stages. In stages I and II only the three naupliar appendages are present: the antennule as a small bud, the biramous antenna as the main swimming organ, and the mandible. The antennal protopod bears two endites, the proximal naupliar process and a more distal endite. In stage III a bifid naupliar process (in earlier stages not bifid) and the first signs of the carapace and trunk limb anlagen (undifferentiated rudiments) appear. In stage IV the carapace anlagen become more pronounced. The number of trunk limb anlagens increases to five, and differentiation has commenced. In stage V the first five pairs of trunk limbs are differentiated to varying degrees. The anterior-most four pairs of trunk limbs are subdivided into five endites, a small endopod, an exopod and an epipod. The bivalved carapace covers the anterior-most limbs. In larval stage VI the carapace is larger and the trunk limbs are further differentiated. A general pattern in the sequence of larval stages is the increasing number of sensilla on the antennules. From the last larval to the first postlarval stage, a significant change in morphology takes place. The trunk limbs are now used for swimming. Typical larval organs are much smaller than in the last larval stage. A comparison with other representatives of the Limnadiidae shows a high degree of correspondence, with most differences explained by the heterochronous appearance of characters during development. Five to seven stages are described for all studied Limnadiidae, including one particular stage in which four fully developed setae, a bifid naupliar process and the first signs of carapace anlagen are present. These characters are found in stage III in L. parvispinus, Limnadia stanleyana, Eulimnadia texana, and Imnadia yeyetta but in stage IV in E. braueriana and L. lenticularis. Based on a comparison of the larval stages of six limnadiid and one cyzicid species, we conclude that at least six naupliar stages belong to the limnadiid ground pattern.     

Within-season reproductive and somatic energy allocation in a freshwater clam,Anodonta piscinalis

The effect of a change in the environment on reproductive and somatic energy allocation in an iteroparous freshwater clam Anodonta piscinalis was studied at different time of the seasonal reproductive cycle. Environmental change was produced by reciprocal transplant experiments among sites of varying productivity. In addition, clams were caged at high density to reduce the availability of resources. Transplanting females before fertilization (from May to June), or during the early development of the brood (from July to August) had no detectable effect on reproductive output. Early-season environment, however, affected body mass and percent fat content of females from two populations. This suggests that maintaining the level of reproductive allocation when resources are reduced early in the season leads to lower allocation to somatic growth and biochemical storage. Transplanting females late in the season (from September to November) had a substantial effect on reproductive output and body mass, but not on fat content, suggesting that late in the season allocation to biochemical energy storage is important. Hence, late in the season reproductive allocation may be adjusted to prevailing conditions in preparation for winter. Indeed, over-wintering site had a significant effect on percent fat content, body mass and shell growth when females were kept in a new environment from September to March. Variation among transplant sites in female body mass matched the estimated productivity of the sites, suggesting that it occurred in response to differences in the productivity of the habitats. The results emphasize the importance of taking seasonal changes in the priorities of energy allocation and the seasonality of reproductive processes into account when developing or testing models of optimal energy allocation.     

菲律宾蛤仔EST_SSR标记与生长性状的相关分析

研究利用20个微卫星标记对菲律宾蛤仔斑马蛤F2代家系107个个体进行遗传多样性分析,并对标记位点与生长相关性状进行分析。在20个微卫星位点共检测到41个等位基因,各位点等位基因数为2—3个,等位基因片段大小为109—430 bp,平均等位基因数为2.05个。平均有效等位基因数为1.71个,观测杂合度平均值为0.504,期望杂合度的平均值为0.431,平均多态信息含量为0.324。经卡方检验,3个位点SSR11,SSR164和SSR213的基因型分布显著偏离了孟德尔定律(P0.01)。运用SPSS 20.0对20个微卫星位点与菲律宾蛤仔斑马蛤家系生长性状的相关性(壳长、壳宽、壳高和体重)进行连锁显著性检验。结果表明,SSR9位点与壳高存在显著的相关关系(P0.05),SSR135和SSR164位点与壳宽呈显著相关(P0.05),SSR142位点与体重呈显著性相关(P0.05)。研究结果可为菲律宾蛤仔的分子标记辅助选育提供参考。