Morphological and biochemical changes in the muscle of the marine shrimp Litopenaeus vannamei during the molt cycle

Morphological changes and biochemical composition of abdominal muscles over the molt cycle were investigated in juvenile Litopenaeus vannamei. Eight molt stages were characterized and clear uropod images are presented. Molt frequency was highly correlated with the age of the shrimp. Juvenile shrimps appeared to molt faster at one month of age (4.6 ± 0.5 days/cycle), slow to 11.8 ± 1.7 days/cycle at three months of age, and reach a long molt cycle at six months (17.2 ± 2.7 days/cycle). Myofiber cross-sectional images revealed specific morphological changes in abdominal muscle associated with each molt stage. Expanded fiber size was observed during intermolt stages. Water content and total soluble proteins were balanced throughout the molt cycle. Total DNA concentration increased in intermolt and premolt, while total RNA concentration remained stable except in late-premolt stages. SDS-PAGE analysis showed high levels of actin and myosin in postmolt, reaching a plateau in intermolt and declining in premolt. These results suggest the occurrence of muscle fiber rearrangement in both the premolt and postmolt stages. Abdominal muscle buildup occurs mostly during the intermolt stage.     

中华绒螯蟹蜕皮周期中肝胰腺细胞组成的变化

取一年生未成熟中华绒螯蟹 (Eriocheir sinensis)幼蟹, 甲壳宽度在15~40 mm, 暂养在75 cm×50 cm×45 cm的玻璃缸内, 24 h充气, 自然光照, 每天换水1/3, 每次换水后投喂土豆、杂鱼等食物, 动物适应实验室条件1周后进行实验。为得到蜕皮后的样本, 将处于蜕皮前晚期的个体在相同条件下单独饲养。根据已报道方法, 将中华绒螯蟹的蜕皮周期分为蜕皮间期C期、蜕皮前D₀、D₁和D_3–4期、蜕皮后A-B期等5个时期。采用细胞学和组织学方法观察中华绒螯蟹蜕皮过程中肝胰腺细胞组成的变化, 结果显示, R细胞数量在蜕皮间期和蜕皮前期都占绝对优势, 虽然在蜕皮前早期D₀期数量显著下降(P<0.05), 但从D₁期开始上升, 于蜕皮前晚期D_3–4期达到最高(62.06 ± 3.92)%, 而在蜕皮后A-B期又显著降低(P<0.05)。F细胞的数量除在D₁期显著增加外(P<0.05), 其余时期的变化无统计学差异(P>0.05)。B细胞数量在蜕皮前早期D₀期显著增高(P<0.05), 随后开始下降, 直到蜕皮前晚期D_3–4期恢复到间期水平, 而蜕皮后A-B期数量又显著增高(P<0.05)。E细胞数量除在蜕皮前早期D₀期稍微上升外, 其余时期基本稳定(P>0.05)。饥饿对处于不同蜕皮时期的中华绒螯蟹肝胰腺细胞组成的影响不同, 饥饿48 h后, 处于D₀时期的中华绒螯蟹与正常组相比, 肝胰腺R细胞数量无显著变化, B细胞和E细胞数量显著下降, F细胞数量上升。处于D₁时期的中华绒螯蟹与正常组相比, 肝胰腺R细胞、B细胞、F细胞和E细胞数量均无显著变化。说明中华绒螯蟹蜕皮过程中肝胰腺细胞组成的变化和蜕皮周期密切相关。本研究通过探讨中华绒螯蟹蜕皮过程中肝胰腺组织结构及细胞组成变化, 了解甲壳动物蜕皮过程的基础生物学现象, 并为其健康养殖提供理论基础。

     

中华绒螯蟹RXR基因全长cDNA克隆及表达分析

维甲类X受体(retinoid X receptor,RXR),属于核受体超家族(nuclear receptor super family)成员,是一种重要的调控因子,对甲壳动物生长发育和繁殖具有十分重要的作用。本研究根据陆地蟹 RXR基因保守区序列设计上下游引物,采用逆转录聚合酶链反应(RT-PCR)以及cDNA末端快速扩增(RACE)技术克隆中华绒螯蟹(Eriocheir sinensis)RXR基因并进行各组织间的表达分析。序列分析表明中华绒螯蟹RXR cDNA全长1517bp,编码433个氨基酸。经BLASTn和BLASTx分析表明,中华绒螯蟹RXR氨基酸序列与陆地蟹RXR基因的氨基酸序列相似性最高,为96%。系统进化树分析显示,中华绒螯蟹RXR的氨基酸序列与陆地蟹RXR聚为一支。荧光定量PCR结果显示,RXR在所有检测的组织中均有表达,且在中华绒螯蟹Y器官中表达量最高,肝胰腺、鳃和肌肉中有少量表达,心脏、胃、肠、胸神经节和脑神经节中微量表达。在中华绒螯蟹不同蜕皮时期中,Y器官中RXR表达量在D期最高,与AB期、C期呈显著性差异（P＜0.05）;肌肉中RXR在AB期表达量最高,与其他蜕皮时期呈显著性差异（P＜0.05）;肝胰腺和鳃中RXR在AB期表达量最高,E期表达量最低,但各蜕皮时期表达量之间差异不显著。     

中华绒螯蟹蜕皮激素受体基因（Ers-EcR）的克隆和组织表达分析

蜕皮激素受体(ecdysteroid receptor,EcR)介导调控甲壳动物蜕皮生长、附肢再生等重要生命活动。为了解EcR在人工控制甲壳动物的繁殖和生长中的作用,采用RACE方法结合同源克隆技术,首次从中华绒螯蟹Y-器官中克隆得到蜕皮激素受体基因全长cDNA序列(Ers-EcR,登录号:KF736985),并进行了结构解析和组织表达分析。结果发现,Ers-EcR编码基因全长2 176 bp,开放阅读框为1 638 bp,编码545个氨基酸,具有DNA结合域(DBD)和配体结合域(LBD)等典型的核受体超家族结构域,但不具有信号肽结构。其中,DBD含有8个保守的Cys残基,可以形成2个锌指结构(C156-C159-C173-C176、C192-C198-C208-C211),是典型的DBD特征。多重序列比对分析表明,Ers-EcR氨基酸序列与拳手招潮蟹同源性最高,达到91%。荧光定量PCR结果显示成体中华绒螯蟹ErsEcR基因在Y-器官和肌肉组织中表达量最高,在血液、肠道、卵巢、眼柄、心脏和肝胰腺中有一定表达,在鳃、胸神经节和精巢表达量较低。这表明Ers-EcR基因在中华绒螯蟹各组织器官中的表达不具有典型的特异性,提示Ers-EcR基因可能参与体内多种生命活动的调控。     

中华绒螯蟹Y-器官在蜕皮周期中的超微结构

以光镜和电镜观察处于蜕皮周期不同阶段的中华绒螯蟹(Eriocheirsinensis),结果显示,中华绒螯蟹具Y-器官(YO)1对,卵圆形,直径1 5～2 0mm,苍黄色,位于头胸部鳃腔前端,大颚外侧内收肌腹缘,邻近头胸甲内侧上皮。YO细胞排列成索状,细胞索之间为血窦,YO的细胞索由一种细胞构成,细胞核质比高。YO细胞具有类似脊椎动物合成类固醇激素的典型超微结构特征,蜕皮前期YO细胞的光面内质网、管嵴状线粒体和游离核糖体丰富;蜕皮后期和蜕皮间期的内质网、线粒体和游离核糖体明显减少;青春蜕皮后的YO出现大面积细胞凋亡,YO趋向退化。     

Practical Molt Staging of Penaeus setiferus and Penaeus stylirostris

Molt staging has many applications in penaeid aquaculture as a routine procedure. Using criteria developed for various crustaceans, a method of rapid molt staging was applied to Penaeus stylirostris and Penaeus setiferus adults. The most useful characters were the degree of epidermal retraction from the setal bases and epidermal retraction from the cuticle coupled with the development of new setae. The duration of the molt cycle of P. stylirostris was 11.5 ± 1.0 days ( N = 5) at 27–29 C. The early postmolt, late postmolt, intermolt, early premolt and late premolt stages were readily determined. For P. stylirostris , the duration of postmolt was 27% of the cycle; intermolt, 17%; and premolt, 56%. The length of the molt cycle of P. setiferus at 27–29 C was 13.6 ± 1.0 days ( N = 21) with postmolt occupying 22% of the cycle; intermolt, 19%; and premolt, 59%. Unilaterally eyestalk enucleated P. setiferus had a significantly shorter molt cycle than intact P. setiferus (11.7 ± 1.21 days vs. 13.6 ± 1.02 days, P < 0.05). There were no differences in molt cycle duration between the sexes, whether enucleated or intact ( P > 0.05). A series of illustrations identifies the key characteristics for determining stage of molt in these penaeids.     

Influence de la temperature sur la croissance et la mue de Palaemon serratus en fonction de leur taille

The effects of temperature on growth and on the duration of the molt cycle and inter-molt stages of Palaemon serratus were examined in relation to body size under controlled conditions in the laboratory.Over the size range studied, the best temperature for growth is 22° C, but when the temperature differs from this optimum, young shrimps are less affected than old ones. Growth of the smallest shrimps is depressed especially by extremely low temperatures; on the contrary, high temperatures act negatively on the growth of old animals.The duration of the molt cycle is directly and linearly related to shrimp length; increasing temperature shortens it according to a hyperbolic curve, with a greater effect on the largest animals.Neither body size nor temperature affects significantly the relative durations of the molt stages.     

Spatiotemporal variability in the phenology of the initial intra‐annual molt of American lobster (Homarus americanus Milne Edwards, 1837) and its relationship with bottom temperatures in a changing Gulf of Maine

This study uses the Maine Department of Marine Resources Lobster Sea Sampling data (2000–2016) and logistic models to develop the first time series for the timing and suddenness of onset of the initial intra‐annual molt of American lobster in the Gulf of Maine (GoM), an annual fishery recruitment event crucial to fishermen. Data from three GoM regions (eastern, central, and western coastal Maine) were further divided by sex and estimated maturity of sampled lobsters for analysis. We found differences in the patterns of initial molt timing and suddenness between the regions, sexes, and stages of maturity. Using the Northeast Coastal Ocean Forecasting System hindcast temperatures, seasonal accumulated degrees above 5°C were used to describe the thermal history for each region at ocean depths of about 5 and 110 m. These temperature metrics were used in generalized linear models to investigate the potential effects of seasonal temperatures on the initial molt season. Results showed that initial intra‐annual molting of lobsters was variable from 2000 to 2016, with periods of both earlier and more sudden molts and later and more protracted molts. Warmer temperatures, specifically inshore temperatures, were generally associated with an earlier molt, but without complete uniformity in the direction and magnitude across seasons, regions, and lobster demographics. We also discuss why developing molt time series and quantifying the connection to the bottom temperatures are necessary and emphasized why existing monitoring programs and the applied quantification techniques herein make this relationship difficult to quantify.     

Molting and Mortality of Red Swamp and White River Crawfish Subjected to Eyestalk Ablation: A Preliminary Study for Commercial Soft-Shell Crawfish Production

Eyestalk ablation may reduce the cost of soft-shell crawfish production by reducing the molt interval. In this study, both immature and mature red swamp crawfish Procambarus clarkii and white river crawfish Procambarus zonangulus , formally Procambarus actus actus (Hobbs and Hobbs 1990), were ablated using a pair of modified pliers and placed in a recirculating system. Molting percentages, mortalities and mean molt intervals of the ablated crawfish were analyzed.
Eyestalk ablation resulted in dramatic reduction of molt intervals and mortalities comparable with the current commercial (non-ablation) soft-shell crawfish production systems. The mean molt intervals of the ablated red swamp crawfish ranged from 6.7 to 7.8 days for immature and 9.2 days for mature animals; whereas, the molt interval of ablated white river crawfish was 8.9 and 11.2 days for immature and mature animals, respectively. Mortalities obtained in this study ranged from 20 to 48% and 32 to 66% for immature and mature crawfish, respectively. During the experiments, molting percentages and mortalities were not consistent. Secondary treatments such as air clotting and cauterization did not alter mortality significantly.     

Effect of thermal stress on Hsp70 gene expression and female reproductive performance of giant freshwater prawn,Macrobrachium rosenbergii

Using Hsp70 as a biomarker, thermal stress impinges on reproductive organs, ovary and hepatopancreas were being analyzed by determining the expression of Hsp70 mRNA inside the organs after the adult inter‐molt females were subjected to thermal treatment at 35, 30 and 28°C (Control). Results showed the expression of Hsp70 mRNA under thermal treatment of 35°C after 2 hr recovery in ovary were upregulated at 2, 4, 6, 12, 24 hr and 30 days compared to control whereas in hepatopancreas under similar treatment, the expression of Hsp70 mRNA were significantly higher than control at 6, 24 hr and 30 days. Frequency of reproductive molt at 35°C showed the ovary of females were failed to develop and only entered common molt along three consecutive molt cycles. For 30°C thermal treatment, the expression of Hsp70 mRNA was significantly higher than control after 2 hr recovery but returned to normal afterwards until 30 days’ thermal treatment. Maternal heat shock for 2 hr at 35°C were found to give significantly lower frequency of reproductive molt and longer duration of ovarian development and incubation period whereas maternal heat shock for 2 hr at 30°C gave lower frequency of reproductive molt, slower development of embryo and lower hatching success compared to untreated control. This study suggests that short and long‐term thermal stress at 30 and 35°C were found to affect the induction of Hsp70 mRNA in reproductive organs of Macrobrachium rosenbergii and also influence their reproductive performance.     

Post-molt processes of cuticle formation and calcification in the Japanese mitten crab <Emphasis Type="Italic">Eriocheir japonicus</Emphasis>

This study examines the following in the Japanese mitten crab: (1) the structure of the exoskeleton with special reference to its calcification; (2) the progression of post-molt cuticle formation and calcification. In the crab, the structure and calcification state of the exoskeleton at the molt and during the inter-molt stage were similar to those of other crustaceans. During the inter-molt, the exoskeleton consisted of four cuticle layers; the outermost epicuticle, the exocuticle, the endocuticle and the innermost membrane layer. Intense calcification was observed in the exo- and endocuticle. At the molt, the synthesis of the epi- and exocuticle was already complete, and the addition of the endocuticle began after the molt. Calcification of the exocuticle initiated soon after the molt, but there was a delay between endocuticle matrix synthesis and calcification. Histology showed that the process of calcification was similar to that in other crustaceans. However, calcium concentrations within the exoskeleton continued to increase and never reached the levels of the inter-molt stage at the end of the experiment. This suggests that the Japanese mitten crab is relatively slow to calcify compared to other crustaceans.     

Estimation of terminal molting probability of snow crab <Emphasis Type="Italic">Chionoecetes opilio</Emphasis> using instar- and state-structured model in the waters off the Pacific coast of northern Japan

Terminal molting probability (TMP) of snow crab Chionoecetes opilio was estimated using instar- and state-structured model incorporating the terminal molt in the waters off the Pacific coast of northern Honshu, Japan. TMP is defined as the probability that terminal molt occurs within a year in an instar and is an important parameter for stock assessment. TMP was estimated from the model and the rate of terminal molted crabs (RTM) for instars, which means carapace widths over 30 mm. Carapace widths at RTM of 0.5 were 81 mm for males and 66 mm for females. TMP was estimated as 0.23 and 0.29 for these carapace widths, respectively. Carapace widths at TMP of 0.5 were 94 mm for males and 71 mm for females, and for these carapace widths 57% of male and 50% of female crabs were considered as old-shell which had survived 1 year or more from terminal molt. Changing the fishing or natural mortality by ±0.1, the maximum difference of TMP was less than 0.1 from default estimates.     

Larval molting and growth of the Japanese spiny lobster Panulirus japonicus under laboratory conditions

ABSTRACT:   Ten newly hatched phyllosoma of Panulirus japonicus were cultured individually to monitor body length ( BL ) and intermolt period, and 2000 were cultured in groups to sample specimens for measurement of body weight. Phyllosoma were fed with Artemia and mussel gonad; the culture seawater temperature was 24–26°C. The individually cultured phyllosoma showed an increment in body length by the first molt of approximately 0.5 mm, and the molt increment increased to approximately 1 mm at 5 mm BL ; it was constant to 15 mm BL . Thereafter, the molt increment increased exponentially. The duration of the first instar was 6–7 days. Instar duration increased with development up to approximately 2 weeks at the 20th instar (∼16 mm BL ) and then became constant. Of the 10 larvae reared individually, five metamorphosed to the puerulus stage. The entire phyllosoma life ranged from 245–326 days (mean 289.0 days), and the number of instars ranged from 22–29 (mean 26.2). Body length in the final instar ranged from 28.50–33.10 mm (mean 30.280 mm). For the phyllosoma cultured in groups, relationships between BL and wet/dry body weights ( WW / DW , mg) were expressed as exponential equations: WW  = 0.0686 BL ^2.2023 and DW  = 0.0209 BL ^2.1905.     

日本沼虾蜕皮过程中头胸甲外骨骼超微结构的改变

采用HE染色及石蜡切片的扫描电镜观察方法,描述了日本沼虾(Macrobrachium nipponense)蜕皮间期(C期,intermolt stage)及新旧表皮更替变化剧烈的蜕皮前晚期(D4期,late premolt stage)和蜕皮后期(postmolt stage)A-B期头胸甲外骨骼的形态结构变化特点。结果显示,日本沼虾外骨骼分为上表皮、外表皮、内表皮三层,D4期出现新的上表皮和外表皮,B期出现新内表皮,外表皮强嗜碱性而内表皮弱嗜酸性。扫描电镜观察发现,C期头胸甲内、外表皮均为几丁质-蛋白质纤维构成的平行板层结构且板层内有发达的孔道系统(pore canals,pc),但内外表皮板层间排列紧密程度不同,外表皮板层切面边缘较整齐,结构致密,板层内pc大小均一、近似圆形,内表皮板层切面边缘粗糙,结构较疏松,板层内pc大小不等、多为梭形。在蜕皮前后,新外表皮结构变化显著,与C期相比,D4期新外表皮超微结构与旧内表皮结构相似,而在A期,外表皮超微结构由疏松变为致密,这可能与蜕皮后表皮的钙化相关。本研究旨为阐明沼虾的蜕皮机制提供基础资料。     

中华绒螯蟹蜕壳过程中螯足肌肉组织学及主要蛋白质含量变化

为探讨中华绒鳌蟹(Eriocheir sinensis)蜕皮过程中肌肉生长机制,采用石蜡切片、电镜技术及生物化学方法,以中华绒鳌蟹幼蟹为对象,研究了其蜕壳过程中螯足闭螯肌肌肉的显微结构、超微结构及主要蛋白质含量的变化。结果显示,在蜕皮间期,螯足肌纤维充分伸展,多核,横截面呈不规则圆形或者多边形;肌原纤维排列有序,具有甲壳动物骨骼肌的典型特征,A带、I带及肌质网、二联体、线粒体等细胞器清晰可见。蜕皮前期,肌纤维降解以致其横截面大小不一;肌原纤维内部降解,出现腐蚀性空洞。蜕皮之后,在肌纤维末端的肌节处于超收缩状态,这些肌节长度缩短至正常长度的50%,肌纤维典型结构消失,A带、I带模糊不易区分,但肌质网、二联体、线粒体等结构仍然完整;肌纤维中间部分的肌节结构正常如蜕皮间期。生物化学研究发现,蜕皮前后螯足肌肉中肌原纤维蛋白和可溶性蛋白含量的变化同其结构特征的变化一致。本研究结果表明,中华绒螯蟹蜕皮后螯足肌肉可能的一种生长机制是通过增加新的肌节来伸长,动物刚蜕壳后,这些新形成的肌节以超收缩的形式存在,随着动物吸水身体膨大,外骨骼伸展、硬化,这些较短的肌节由于被拉伸而达到正常肌节的长度,从而完成肌肉的生长。     

The effects of salinity on the survival, growth and haemolymph osmolality of early juvenile blue swimmer crabs, Portunus pelagicus

The blue swimmer crab, Portunus pelagicus, is an emerging aquaculture species in the Indo-Pacific. Two experiments were performed to determine the effects of salinity on survival, growth and haemolymph osmolality of early juvenile P. pelagicus crabs. The salinities tested for the first experiment were 10, 15, 25 and 40 ppt, and for the second experiment 5, 20, 30, 35 and 45 ppt. Each salinity experiment was triplicated, with each replicate consisting of 10 stage 4 juveniles. Each experiment lasted 45 days. Mortalities and incidence of “molt death syndrome” were recorded daily, while the intermolt period, carapace length, carapace width and wet weight were measured at each molt. At the end of the experiments the haemolymph osmolality and dry weights were measured.

Results demonstrate that salinity significantly affects both the survival and growth of early P. pelagicus juveniles. Mortality was significantly higher (p < 0.01) for juveniles cultured at salinities ≤ 15 ppt and at 45 ppt. At a salinity of 5 ppt a complete mortality occurred on day 20. In all salinity treatments, the majority of mortalities were due to “molt death syndrome”. In experiment 1, immediate effects of salinity on growth and development were seen at 10 ppt as the intermolt period was significantly longer (p < 0.01) and the mean carapace size increase was significantly less (p < 0.01) at the first molt compared to the other treatments. Meanwhile, the specific growth rates (carapace length, width and wet weight) were significantly lower (p < 0.05) at high salinities (≥ 40 ppt) due to longer intermolt periods and significantly lower (p < 0.05) carapace size or wet weight increases.

The haemolymph osmolality exhibited a positive linear relationship with the culture medium with an isosmotic point of 1106 mOsm/kg, equal to a salinity of approximately 38 ppt. Based on the osmolality graph, high metabolic cost for osmoregulation due to increased hyper- and hypo-osmotic stress appeared to cause lower survival and specific growth rates of the crabs. The results demonstrate that a salinity range of 20–35 ppt is suitable for the culture of early juvenile P. pelagicus.     

Histological studies on the spent ovaries of aged snow crabs <Emphasis Type="Italic">Chionoecetes opilio</Emphasis> caught in the Sea of Japan

Aged female snow crabs Chionoecetes opilio, 9 or so years old after their final molt, were collected sporadically from artificial reefs established off the coast of Wakasa Bay, Sea of Japan. In order to clarify the degenerative changes that occur in spent ovaries, the ovaries of these crabs were examined histologically. The ovaries were found to be reduced in both size and volume, with most showing significant oosorption by follicular epithelial cells. Reserve fund eggs and unspawned eggs were rarely detected, with the exception of some in atretic follicles. One to dozens of orange to black nodules scattered over the ovary consisted of masses of degenerative ova in various stages of oolysis. Since nodular formation was recognized in 26% of these aged individuals, it seems very likely that such crabs are unable to spawn healthy eggs.     

The effects of holding space on juvenile red king crab,Paralithodes camtschaticus (Tilesius, 1815), growth and survival

Rearing crustaceans communally for aquaculture, stock enhancement or research often results in high rates of cannibalism and low yields. One potential strategy to reduce loss from cannibalism is to rear crustaceans in individual cells. As small holding cell size can result in decreased growth or increased mortality, it is essential to identify the optimal holding cell size, both for mass culturing efforts and for experimental design purposes. In this study, we reared juvenile red king crab, Paralithodes camtschaticus (3.67–8.30 mm carapace length) in 20, 40 and 77 mm diameter holding cells, and monitored growth and survival over a 274‐day experiment. A trend of lower growth per molt in the smallest holding cells resulted in crab 17% smaller than those in the large holding cells at the end of the experiment. In addition, mortality rates were an order of magnitude higher in the small holding cells compared with the large or medium cells. For individual rearing of this size of juvenile red king crab, the medium‐sized cells (40 mm diameter) are the optimal size as there was no increase in mortality and only marginally lower growth rates compared to the large‐sized cells.     

Carbonic anhydrase and Na/K-ATPase activities at different molting stages of the giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis>

ABSTRACT:   In the present study, the role of carbonic anhydrase (CA) and Na/K-ATPase in the gill and epidermal tissues in the giant freshwater prawn Macrobrachium rosenbergii was examined as a function of molting stage. CA activity levels in the front and back gills were low at the intermolt stage C₀, but increased significantly at premolt stage D₃, and then decreased after molting. In the epidermal tissue, activity levels decreased gradually towards premolt to a minimum level at stage D₃, but became elevated at postmolt stages A and B. Na/K-ATPase levels in the front and back gills did not change significantly during the molt cycle. CA in the gill is possibly involved in supplying counter-ions for ion uptake, while CA in the epidermal tissue may play a role in mineralizing the exoskeleton after ecdysis. Na/K-ATPase in the gills may function in ion uptake from the ambient medium; however, since its activity was not influenced by the molt cycle, it probably does not have a major role in osmoregulation in the freshwater environment.     

Setal development and molt prediction in the larvae and adults of the crayfish,Astacus leptodactylus (Nordmann, 1842)

The criteria for determination, given by Drach (1939), have been adapted to divide the molt cycle into different stages for larvae and adults of the crayfish Astacus leptodactylus. The setal formation in the transparent appendages can be used for the definition of stages and substages in the premolt period. In order to evaluate the post-and intermolt stages in more detail, one can combine this method with changes occurring in the cuticle.