Effects of temperature on cellular and biochemical parameters in the crab Carcinus aestuarii (Crustacea, Decapoda)

The effects of temperature on cellular and biochemical parameters of the crab Carcinus aestuarii were evaluated. Crabs were kept for 7 days at 4, 17 (reference value) and 30 °C (salinity of 35 psu), and total haemocyte count (THC), haemocyte volume, haemocyte proliferation, phenoloxidase (PO) activity in both haemocyte lysate (HL) and cell-free haemolymph (CFH), CFH total protein and glucose levels, superoxide dismutase (SOD) and catalase (CAT) activities in both gills and digestive gland were evaluated. The lowest and the highest temperature significantly decreased THC, whereas haemocyte volume and haemolymph glucose concentration did not differ significantly among experimental conditions. Haemolymph protein concentration significantly reduced in crabs maintained at 30 °C, when compared with that of animals kept at 4 and 17 °C. Haemocyte proliferation increased significantly in crabs kept at 4 and 30 °C, when compared with that of crabs held at 17 °C. Likewise, a significantly higher PO activity was recorded in CFH from crabs kept at 4 and 30 °C, than in control crabs. Conversely, PO activity did not vary significantly in HL. With regard to antioxidant enzyme activities, a significant decrease in CAT activity was observed in gills from crabs kept at 4 °C, when compared to that of crabs kept at 17 °C and 30 °C. Results obtained demonstrated that the highest and lowest temperature tested influenced crab biological responses, and indicated that C. aestuarii modulated its cellular and biochemical parameters (mainly haemocyte proliferation, CFH protein concentrations and CFH PO activity) in order to cope with temperature.     

Can starvation influence cellular and biochemical parameters in the crab Carcinus aestuarii?

Crustacea experience periods of starvation during moulting or when limited food availability occurs. The effects of starvation on Crustacea physiological responses have been demonstrated, whereas the effects of starvation on Crustacea immune parameters remain to be more fully studied. In the present study the effects of starvation on immune parameters and antioxidant enzyme activities of the crab Carcinus aestuarii were evaluated for the first time. Treated crabs were starved for 7 days, whereas control crabs were fed daily with mussels. Total haemocyte count (THC), haemocyte diameter and volume, haemocyte proliferation, cell-free haemolymph (CFH) glucose and total protein levels, and phenoloxidase (PO) activity in both haemocyte lysate (HL) and CFH were measured in crabs. In addition, superoxide dismutase (SOD) and catalase (CAT) activities were evaluated in both gills and digestive gland from crabs, in order to evaluate whether starvation induced oxidative stress in C. aestuarii. THC increased significantly in starved crabs, with respect to controls, whereas no significant variations were observed in haemocyte diameter, volume and proliferation. In CFH of starved animals glucose concentration significantly increased, whereas total protein concentration significantly reduced. A significantly higher PO activity was recorded in HL from starved crabs, than in control crabs. Conversely, PO activity did not vary significantly in CFH. Starvation did not cause significant alterations in antioxidant enzyme activities in both gills and digestive gland. Results obtained demonstrated that starvation influenced crab immune parameters, but did not induce oxidative stress. Results also indicated that C. aestuarii can modulate its cellular and biochemical parameters in order to cope with starvation.     

The nonsteroidal anti-inflammatory drug,ibuprofen, affects the immune parameters in the clam Ruditapes philippinarum

The effects of ibuprofen (IBU) on the immune parameters of the clam Ruditapes philippinarum were investigated after a 7-day exposure to sublethal IBU concentrations (0, 0 + ethanol, 100, 500, and 1000 μg/L). Total haemocyte count (THC), haemocyte diameter and volume, haemocyte proliferation and uptake of the vital dye Neutral Red (NR) were measured. The cytotoxicity (assessed by the lactate dehydrogenase assay, LDH) and the capability of IBU to induce DNA fragmentation (indicative of apoptosis) were also investigated. The exposure of clams to the highest IBU concentration significantly reduced their THC, whereas no significant changes were observed in either the diameter or volume of haemocytes. Significant increases in haemocyte proliferation were recorded in clams that were exposed to the two highest tested concentrations of IBU. Exposure of clams to 1000 μg IBU/L significantly reduced NR uptake and increased haemolymph LDH activity. Conversely, IBU did not induce DNA fragmentation in haemocytes. Although the IBU concentrations tested in this study were higher than those generally recorded in aquatic environments, results obtained indicate that exposure of clams to IBU induces significant alterations in the immune parameters and suggest potential immunosuppression in treated clams.     

低溶解氧对凡纳滨对虾酚氧化酶原激活系统和相关免疫因子的影响

本文研究了凡纳滨对虾在低溶解氧条件下酚氧化酶原系统的激活机制。结果表明:低溶氧(1.5mg/L,3.0mg/L)胁迫对凡纳滨对虾血淋巴中多巴胺含量、血细胞数量、酚氧化酶原激活系统相关酶活力、血淋巴中类α2-巨球蛋白活力影响显著(P<0.05),且各指标表现出明显的时间效应性。在实验时间内,DO为3.0mg/L处理组,总血细胞、小颗粒细胞和透明细胞数量均在12h内降低而后趋于稳定,大颗粒细胞数量在24h内降低后保持稳定,DO为1.5mg/L处理组,总血细胞数量和三类血细胞数量均在24h内降低而后趋于稳定;DO为3.0mg/L和1.5mg/L处理组凡纳滨对虾血淋巴中多巴胺含量分别在24h、12h内呈峰值变化,至12h、6h时达到最大值,在24h、12h后恢复至对照组水平并趋于稳定;血细胞中酚氧化酶原活力分别在12h、6h时显著降低,且均在24h时趋于稳定;丝氨酸蛋白酶活力在6h时均显著降低,在24h时达到最低值,而后趋于稳定,蛋白酶抑制剂活力显著降低,但类α2-巨球蛋白活力48h内呈峰值变化,与对照组比较均显著升高,48h后趋于稳定。实验期内对照组DO为5.5mg/L各指标变化不显著。     

Physiological responses of juvenile and adult shore crabs Carcinus maenas (Crustacea: Decapoda) to pyrene exposure

This study measured aspects of the physiology of juvenile [(<35mm carapace width (CW)] and adult (>60mm CW) Carcinus maenas to test the hypothesis that these different life-history stages exhibit different sensitivities to environmental contamination. Newly-collected juveniles had significantly (P<0.05) lower immunocompetence (phagocytosis and cellular integrity), lower metabolic energy (haemolymph glucose) and increased scope for growth compared with adults. Seven day exposure to a sub-lethal concentration (200microgL(-1)) of pyrene significantly (P<0.01) reduced immunocompetence, elevated basal heart rate and decreased respiration (at rest) for juveniles but had no overall impact on adult crab physiology. Results confirm that physiological differences exist between juvenile and adult shore crabs, and cause juveniles to be more susceptible to the effects of pyrene exposure than adults. Such differences in sensitivity to contamination between life-cycle stages of the same species have to be taken into account during the risk assessment process.     

Changes in the tissue concentrations and contents of calcium, copper and zinc in the shore crab Carcinus maenas (L.) (Crustacea: Decapoda) during the moult cycle and following copper exposure during ecdysis

The tissue concentrations and contents of calcium, copper and zinc were determined at different stages in the moult cycle of the shore crab Carcinus maenas. The crabs were dissected into the following tissues: the midgut gland, the gill, the ‘rest tissue’ (the latter defined as the left over tissue, i.e. muscles and exoskeleton), and the haemolymph, and analysed as such. One group of crabs was maintained in clean seawater. In these crabs, calcium was lost from haemolymph during premoult and apparently stored in the midgut gland. At ecdysis approximately 90% of the whole body calcium content was lost with the exuviae while the haemolymph calcium content doubled. During early postmoult calcium stored in the midgut gland was used in combination with calcium absorbed from the surrounding seawater for a rapid calcification of the new exoskeleton. Both the copper and zinc concentrations declined in the ‘rest tissue’ compartment during premoult. At ecdysis the haemolymph copper and zinc contents increased 35 and 86%, respectively, from premoult to the newly moult stage. At the same time, however, the copper and zinc haemolymph concentrations decreased. During postmoult, copper (25%) and zinc (45%) were lost from haemolymph and mainly excreted but a minor fraction (5–10%) was stored in the midgut gland.A second group of crabs was maintained in copper-contaminated seawater (0.1 mgl⁻¹) during ecdysis and postmoult. Following exposure of the animals to raised ambient levels of copper, the calcium and zinc contents of the midgut gland were reduced while the copper contents of the midgut gland and most other tissues were increased in comparison to crabs held in clean seawater.     

Shell use by sympatric hermit crab species in a seasonally open coastal lagoon in Northeastern Brazil

There are gaps in what is known about the patterns of gastropod shell use by hermit crabs in Brazilian lagoon areas, especially in Northeastern Brazil. However, this is important because the understanding of selection patterns provides information on life history and eco‐evolutionary conceptions of paguroids. The present study investigated the use of gastropod shells occupied by Pagurus criniticornis and Clibanarius sclopetarius in a coastal lagoon seasonally connected to an estuary and to the sea, correlating eco‐evolutionary aspects and hydrodynamic characters. The study was carried out between February 2013 and January 2014. Hermit crabs and their shells were identified and measured (hermit crabs’ cephalothorax shield length and width, major propodus length and height, weight, shell total length and width, shell aperture length and width). Partial least squares regression was used to analyse the morphometric data. Additionally, a multinomial proportions test was performed to infer patterns (inter‐/intra‐specific) of shells’ occupation. Hermit crab species occupied a total of 13 types of shells, predominantly those of Neritina virginea (67.83%). The shell weight was the most important determinant of shell occupancy in the morphometric model (variable importance in projection >1). The proportions of N. virginea shells used were similar in both species of hermit crabs, except between the sexes of P. criniticornis. Presumably, the high utilization rate of N. virginea shells is related to its abundance and dispersal, and to the shells’ suitability for hermit crabs, which is reflected by the morphometric model, as well as by the hydrodynamics of the lagoon‐estuarine environment. The relative occupation of different types of shells for each species of hermit crabs studied appears to be associated with regulation by inter‐/intra‐specific competition, which fosters the co‐occurrence of those populations. These results endorse coastal lagoons as a refuge and recruitment area for aquatic fauna. In addition, it highlights a challenge in the management and conservation of paguroid species whose population dynamics depend on the resources coming from the Gastropoda community.     

A non-destructive assessment of the exposure of crabs to PAH using ELISA analyses of their urine and haemolymph

Urine and haemolymph can be repeatedly sampled from crabs with no (or limited) damage to the organism. Their analysis offers a measure of the animals' exposure to biologically available contaminants. Shore crabs (Carcinus maenas) were exposed to the PAHs phenanthrene and pyrene at concentrations ranging from 20 to 200 microg l(-1). After 48 h, urine and haemolymph samples were taken and analysed using ELISA and UV-fluorescence spectrophotometry. High correlations were recorded between the two sets of results from the urine analyses (r2 = 0.83 for phenanthrene and r2 = 0.88 for pyrene). Contaminant concentrations were much lower in haemolymph than in the urine. Analyses of urine taken from crabs collected from clean and contaminated sites confirm the suitability of these analyses for environmentally exposed organisms. Again, a good correlation was recorded between the ELISA and spectrofluorimetric analysis (r2 = 0.83). In this instance, difficulties were experienced with haemolymph analyses owing to a lack of sensitivity.     

Seasonal differences in the physiology of Carcinus maenas (Crustacea: Decapoda) from estuaries with varying levels of anthropogenic contamination

This study reports the seasonal variability in aspects of the physiology of the shore crab Carcinus maenas from three estuaries in South-west England, each with varying anthropogenic inputs: Avon Estuary (‘relatively low’ impact), Yealm Estuary (‘intermediate’ impact) and Plym Estuary (‘relatively high’ impact). Crabs collected over 12 months from the Avon had a significantly ‘lower’ physiological condition in winter and spring compared to summer and autumn; in particular, haemocyte phagocytic capability (a general indicator of immune function) was significantly higher in winter and spring compared to summer and autumn, and total haemolymph antioxidant status (an indicator of oxidative stress) was significantly lower in winter compared to the remainder of the year. Potentially, shore crabs may be more susceptible to the effects of contaminant exposure, such as increased immunotoxicity (thus, reduction of immune function) and/or oxyradicals (or reactive oxygen species) exposure) especially in seasons of increased susceptibility i.e. summer/autumn (lower phagocytic capability) and winter (lowest antioxidant function). As the Avon was taken to represent the ‘reference’ site, this pattern is considered to reflect the ‘normal’ seasonal variability in shore crab physiology. Shore crab physiological condition from the ‘relatively high’ impact estuary (Plym) revealed increased cellular viability and antioxidant status in autumn and winter compared with that of the ‘standard’ pattern (Avon) However, crabs from the intermediate impact estuary (Yealm) only demonstrated significant physiological differences in summer as shown by a lower cellular viability. All crabs had been exposed to PAHs (confirmed by the presence of PAH metabolites in their urine) which may account for the observed differences in shore crab physiology. In conclusion, to aid understanding of the potential contaminant impacts on biota it is imperative that the ’normal’ seasonal variability of physiological condition be established. Biological effects-based monitoring studies should therefore be employed seasonally to potentially highlight ‘windows of sensitivity’ to contaminant impact.     

注射生物胺对凡纳滨对虾免疫指标的影响

研究了注射生物胺(多巴胺、5-羟色胺)对凡纳滨对虾Litopenaeus vanna mei免疫指标的影响。结果表明,注射生物胺(10-9mol.尾-1)对凡纳滨对虾血细胞数量、血淋巴酚氧化酶活力、溶菌和抗菌活力影响显著(p<0.05),而对照组各免疫指标无明显变化;在实验时间内,注射生物胺处理组各免疫指标呈明显峰值变化,血细胞数量均在3h达到最小值,血淋巴酚氧化酶活力和溶菌、抗菌活力均分别于6h时达到最大值和最小值,且多巴胺处理组各免疫指标比5-羟色胺处理组变化更明显,其中大、小颗粒细胞和血淋巴酚氧化酶活力在12h后保持稳定,总血细胞和透明细胞数量18h后趋于稳定,血淋巴溶菌和抗菌活力在24h后稳定,而且稳定后各免疫指标与对照组均无显著差异。     

Implementing geostatistical analysis to study spatio-temporal distribution patterns of swimming crabs(Portunus trituberculatus)

This study attempted to compare the performance of local polynomial interpolation,inverse distance weighted interpolation,and ordinary kriging in studying distribution patterns of swimming crabs.Cross-validation was used to select the optimum method to get distribution results,and kriging was used for making spatial variability analysis.Data were collected from 87 sampling stations in November of 2015(autumn) and February(winter),May(spring) and August(summer) of 2016.Results indicate that swimming crabs widely distributed in autumn and summer:in the summer,they were more spatially independent,and resources in each sampling station varied a lot;in the winter and spring,the abundance of crabs was much lower,but the individual crab size was bigger,and they showed the patchy and more concentrative distribution pattern,which means they were more spatially dependent.Distribution patterns were in accordance with ecological migration features of swimming crabs,which were affected by the changing marine environment.This study could infer that it is applicable to study crab fishery or even other crustacean species using geostatistical analysis.It not only helps practitioners have a better understanding of how swimming crabs migrate from season to season,but also assists researchers in carrying out a more comprehensive assessment of the fishery.Therefore,it may facilitate advancing the implementation in the pilot quota management program of swimming crabs in northern Zhejiang fishing grounds.     

中国对虾血淋巴抗菌活力的测定

采用溶藻胶弧菌（Ｖｉｂｒｉｏ ａｌｇｉｎｏｌｙｔｉｃｕｓ）、副溶血弧菌（Ｖ． ｐａｒａｈａｅｍ ｏｌｙｔｉｃｕｓ）、哈维氏弧菌（Ｖ．ｈａｒｖｅｙｉ）、嗜水气单胞菌（Ａｅｒｏｍ ｏｎａｓｈｙｄｒｏｐｈｉｌａ）对中国对虾（Ｐｅｎａｅｕｓ ｃｈｉｎｅｎｓｉｓ）血浆样品、血细胞溶解物（ＨＬＳ）样品的抗菌活力进行了测定。表明中国对虾血浆样品对四种细菌均表现出一定的抗菌活力。其中,三种弧菌活细胞数在孵育１ｈ 时呈现最低;嗜水气单胞菌活细胞数在孵育４ｈ 时最少。在三种弧菌中,血浆样品对哈维氏弧菌表现出的抗菌活力最强,而对副溶血弧菌表现出的抗菌活力最弱。血细胞溶解物样品对三种弧菌几乎没有表现出抗菌活力;嗜水气单胞菌在与血细胞溶解物样品孵育２ｈ 时,其活细胞数呈现最低,说明血细胞溶解物样品对该菌表现出了一定的抗菌活力。由此显示,抗菌活力在中国对虾血浆样品中表现得较强,而在血细胞溶解物样品中表现得较弱,血淋巴抗菌因子可以在短时间内对细菌表现出其最强的活力,但对不同种细菌其抗菌作用强弱不同。     

Patterns of genetic and morphometric diversity in the marbled crab (Pachygrapsus marmoratus, Brachyura, Grapsidae) populations across the Tunisian coast

The present study reports on population structure analysis of the marbled crabPachygrapsus marmoratus (Fabricius, 1787) from the Tunisian coast, an appropriate location to study biogeographical process...     

Elucidating cellular and behavioural effects of contaminant impact (polycyclic aromatic hydrocarbons,PAHs) in both laboratory-exposed and field-collected shore crabs,Carcinus maenas (Crustacea: Decapoda)

Monitoring effects of contaminants at lower levels of biological organisation (e.g. biochemical and cellular) allows for mechanistic evaluation of effects of contaminant exposure through laboratory exposures. However, higher level organism effects (e.g. physiological and behavioural) are deemed more ecologically-relevant. In the present study, cellular (cell viability and immune function), physiological (cardiac activity) and behavioural (foraging behaviour) responses were evaluated in field-collected shore crabs Carcinus maenas from three estuaries [a ‘relatively’ (polycyclic aromatic hydrocarbon) PAH-contaminated site (Plym Estuary) and two ‘comparatively clean’ field sites (Avon and Yealm Estuaries)] and compared with responses of crabs exposed in the laboratory to a model organic contaminant (PAH) pyrene (200 μg l⁻¹ for 28 days). The hypothesis that behavioural indicators may be more sensitive than other more traditional methods was tested. No significant impacts were observed at the cellular or physiological level in Plym-collected crabs (the ‘relatively’ contaminated site), but foraging behaviour was significantly altered (increased prey handling time) compared to individuals collected from the ‘relatively’ uncontaminated sites (Avon and Yealm). When given a cockle as a prey item, both Plym-collected and laboratory-exposed crabs took longer to handle and break into cockle shells. Therefore, ecologically-relevant behavioural observations may serve as valuableindicators of environmental quality.     

三疣梭子蟹乳化病血液生化指标诊断技术的研究

对患乳化病的三疣梭子蟹进行了血液生化指标检测．结果表明：与健康三疣梭子蟹相比，病蟹肌酐、总胆固醇、Na^＋、Cl^-、Mg^2＋、Ca^2＋等含量明显上升，而血糖、总蛋白、白蛋白、谷丙转氨酶、谷草转氨酶、乳酸脱氢酶、碱性磷酸酶等含量却显著下降．通过感染试验的追踪检测，并与对照组相比，感染后24h含量发生明显变化的血液生化指标为血糖、总胆固醇、总蛋白、谷草转氨酶和Mg^2＋，此5种指标可作为三疣梭子蟹乳化病早期诊断的参考指标．     

逐步线性回归法实现中华虎头蟹(Orithyia sinica) 形态指标与体重的通径分析

为探讨中华虎头蟹(Orithyia sinica)形态指标与体重的关系, 为良种选育提供最佳的测量指 标, 实验以中华虎头蟹为材料, 测量甲宽、甲长、眼间距、侧齿间距、大螯不动指长、大螯宽、步足 长节长、体高、体重等26 个指标, 借助逐步线性回归、相关分析和通径分析等方法, 分析形态指标 与体重的关系。结果表明, 除第Ⅳ侧齿间距(A₈)外, 实验所测形态指标与体重的相关系数均达到了显 著水平(P<0.05); 采用逐步线性回归的方法建立了多元回归方程Y=-273.841+10.4X₁+2.867X₂, 其中 Y 为体重(g), X₁ 为右大螯宽(mm), X₂ 为第Ⅱ侧齿间距(mm), 定量分析了形态指标对体重的影响结果。     

口服免疫药物后中国对虾某些血淋巴因子的测定及方法研究

于１９９１年４－９月，以海捕中国对虾亲虾及养殖中国对虾为材料，运用单向免疫扩散（ＳＲＩＤ）方法测定对虾血淋巴免疫因子，并采用经改进后的Ｈｏｒｏｗｉｔｚ等人的方法测定对虾体内的酚氧化酶（ＰＯ）活性，目的在于运用该两种方法对口服不同免疫药物的养殖对虾的血淋巴免疫因子及ＰＯ活力进行测定。对亲虾和养殖对虾口服不同免疫药物前后的测定结果表明，几种情况下的对虾血淋巴中均存在类似ＩｇＭ的因子，血细胞中存在酚氧化     

大治河河口无齿螳臂相手蟹生长和成熟的研究(英文)

无齿螳臂相手蟹是长江口潮间带和潮上带的优势种之一,具有重要的生态功能。为探求无齿螳臂相手蟹不同阶段的异速生长和成熟状况,于2009年10月至2010年9月期间采集样本,通过聚类和分段线性回归等分析方法对雌雄蟹头胸甲、螯、腹部等形态数据进行分析,建立区别不同生长阶段的判别函数并用逻辑斯蒂回归估计其50%形态成熟点。结果如下:雌雄蟹50%形态成熟点分别为头胸甲宽18.22mm和16.36mm。雄蟹的螯宽和雌蟹的腹宽均表现出明显的异速生长;根据相对生长率的不同,雄蟹异速生长可分为三个阶段,雌蟹分为两个阶段;不成熟的雄蟹异速生长拐点发生在壳宽11.78mm,不成熟和成熟雌蟹腹部在壳宽13.04～18.64mm范围内重叠。同时研究了50%的生理成熟大小,雌雄成熟大小分别为17.50mm,17.20mm。雄性形态成熟大小小于生理成熟大小,而雌性则相反。     

Impacts of Habitat Complexity on Physiology: Purple Shore Crabs Tolerate Osmotic Stress for Shelter

Purple shore crabs, Hemigrapsus nudus (Crustacea: Decapoda), can survive indefinitely in salinities of 8 (25% SW), but also tolerate short-term exposure to salinities as low as 2. In the laboratory their salinity preference range, determined from choice chamber experiments, is 22–32 and they can discriminate between pairs of salinities separated by a difference of 2. These crabs show a strong positive thigmotaxis and a weak negative phototaxis and tend to choose environments with available shelter. The presence of shelter significantly alters the behaviour of this species. When shelter is available the salinity preference range is 10–32. Even in salinities below this preference range, the presence of shelter prolongs the time spent in the lower test salinities. This change in behaviour has implications on the crab's physiology: the haemolymph osmolality falls to lower levels when crabs remain in low salinity under shelters. In the field, H. nudus is found in creeks with salinities close to freshwater and they may remain in this salinity for up to 11 h, if there are rocks under which to shelter. An increase in habitat complexity increases the number of crabs that are found within the creek. These crabs in the low salinity environment have a lower haemolymph osmolality than crabs on the nearby open shore. In H. nudus the behavioural selection of a shelter appears to outweigh the physiological costs associated with osmotic regulation of the body fluids. Therefore, the distribution of H. nudus in estuaries may depend more on the availability of suitable habitats rather than the salinity tolerance of this species.     

有机污染提高对虾对病原菌易感性研究

1995年5～11月,以人工合成饵料作为有机污染源,取体长8、10和12cm以上中国对虾,在0.9m3水族箱中,分批进行有机污染提高对虾对病原体易感性试验.对虾在上述环境中培育2～3周后,测定其对付溶血性孤菌易感性及体内与抗病有关一些酶的活性.结果:与对照组相比,试验组对虾对病原菌易感性提高1～3倍;体内SOD、PO及溶菌酶活性分别下降约22%、50%和28%～86%.血细胞数量减少50%左右.这与人工合成饵料在水环境中经分解后,产生一些对对虾有害物质,其中以氨氮产生和DO减少尤为显着有关.