Hosts and transmission of the crayfish plague pathogen Aphanomyces astaci: a review

The crayfish plague pathogen, Aphanomyces astaci Schikora, has become one of the most well‐studied pathogens of invertebrates. Since its introduction to Europe in the mid‐19th century, it has caused mass crayfish mortalities, resulting in drastic declines of local populations. In contrast, North American crayfish usually serve as latent carriers, although they may also be negatively affected by A. astaci infections under some circumstances. Recent research benefiting from molecular tools has improved our knowledge about various aspects of A. astaci biology. In this review, we summarize these advances, particularly with respect to the host range and transmission. We highlight several aspects that have recently received particular attention, in particular newly confirmed or suspected A. astaci hosts, latent A. astaci infections in populations of European crayfish, and the relationship between A. astaci genotype groups and host taxa.     

Immunostimulatory effects of dietary fructooligosaccharides on red swamp crayfish,Procambarus clarkii (Girard)

Fructooligosaccharides (FOS) are widely researched as prebiotics and immunostimulants in humans and terrestrial animals. However, limited researches on immune functions of FOS were reported in crustacean. In the present study, red swamp crayfish Procambarus clarkii were fed with different concentrations of FOS (2.0, 5.0, 8.0, 10.0 g kg^?1 diet), and then the immunostimulatory effects of FOS were evaluated by measuring immune parameters and examining immune‐related gene expression. PO and SOD activities were significantly increased in groups fed with 8.0 or 10.0 g kg^?1 of FOS compared with those in control group. In addition, significant elevations of SOD and phagocytic activities were also observed in group fed with 5.0 g kg^?1 of FOS. Among examined immune‐related genes, expressions of crustin1, lysozyme, SOD and pro‐PO were significantly up‐regulated by feeding with 8.0 or 10.0 g kg^?1 of dietary FOS. Survival rates of crayfish challenged with Aeromonas hydrophila were significantly improved by feeding with 5.0, 8.0 or 10.0 g kg^?1 of dietary FOS for 30 days. Phagocytic activities and bacterial clearance efficiencies were also significantly increased in groups fed with 8.0 or 10.0 g kg^?1 of FOS. The results indicated that dietary FOS effectively induced innate immune responses of red swamp crayfish in a dose‐dependent way and could be used as potential immunostimulant in crustacean.     

Variation in the hyphal growth rate and the virulence of two genotypes of the crayfish plague organism Aphanomyces astaci

Crayfish plague, a devastating disease of freshwater crayfish, is caused by an oomycete organism, Aphanomyces astaci. Currently five genotypes of A. astaci are known, but variable features between the strains or genotypes have not been studied extensively. This study analysed 28 isolates of the As genotype and 25 isolates of the Ps1 genotype and reveals that the radial growth rate is significantly (P < 0.001) different between these two genotypes, although highly variable inside the genotype As. Two Ps1 genotype isolates and two As genotype isolates with different radial growth rates were tested in an infection trial. Clear differences were detected in the development of mortality in the test groups. The representatives of the Ps1 genotype caused total mortality within a short time span. The As genotype isolates were much less virulent. The slow‐growing As isolate showed higher virulence than the As isolate with a high growth capacity. Although slow growth could be one survival strategy of the pathogen, several other mechanisms are involved in the pathogenicity and warrant further studies.     

Growth responses of juvenile red swamp crayfish, Procambarus clarkii Girard, to several diets under controlled conditions

The growth responses of juvenile crayfish, Procambarus clarkii Girard, to diets based on common fresh feedstuffs available from agriculture and freshwater ecosystems were studied under controlled laboratory conditions. The dietary treatments were: fresh leaves and shoots of watercress, fresh potato tubers, fresh fish meat from mosquitofish, and all the possible combinations of those ingredients on alternate feeding days. After 22 days, the results showed a significantly higher growth rate (P= 0.05) in crayfish fed with mosquitofish meat. The differences were not significant between the other treatments, but those which included fish meat induced growth rates consistently higher than those based only on plant ingredients. This trend was maintained until the end of the experiment. These results suggest that a dietary supplement based on animal ingredients might significantly improve growth rate and yield, provided the economic feasibility of such a production process be confirmed in field trials. However, it was not clear whether this was because of protein, or lipid concentration and composition of mosquitofish meat. Further research must also clarify the role of vegetable ingredients in dietary composition, as well as the effect of microbial populations occurring during the decomposition of plant material on its nutritional properties for crayfish.     

维生素E对克氏原螯虾生殖的影响

实验用不同的维生素E(VE)含量的饲料投喂克氏原螯虾亲虾,研究对其繁殖性能的影响,目的是为研制亲虾的人工饲料提供指导.结果显示,在添加VE量分别为基础饲料干重的0%、0.004%、0.02%、0.1%和0.5%的5个饲料组中,0.02%VE组不仅可以增加产卵雌虾的数量比例,而且也能提高克氏原螯虾雌虾的产卵量.随着VE含量的升高,产卵雌虾的比例呈下降趋势,单个雌虾的产卵量与0.02%VE组差不多,这表明饲料中适当添加VE可促进克氏原螯虾产卵,但饲料中VE含量过高对克氏原螯虾的繁殖产生负面影响.在饲料中添加适量的VE能够提高亲虾繁殖能力,方法简单,效果明显,适合规模化生产的需要,可以推广.     

维生素E对克氏原螫虾生殖的影响

实验用不同的维生素E（VE）含量的饲料投喂克氏原螯虾亲虾，研究对其繁殖性能的影响，目的是为研制亲虾的人工饲料提供指导。结果显示，在添加VE量分别为基础饲料干重的0％、0．004％、0．02％、0．1％和0．5％的5个饲料组中，0．02％VE组不仅可以增加产卵雌虾的数量比例，而且也能提高克氏原螯虾雌虾的产卵量。随着VE含量的升高，产卵雌虾的比例呈下降趋势，单个雌虾的产卵量与0．02％VE组差不多，这表明饲料中适当添加VE可促进克氏原螯虾产卵，但饲料中VE含量过高对克氏原螯虾的繁殖产生负面影响。在饲料节添加适量的VE能够提高亲虾繁殖能力，方法简单，效果明显，适合规模化生产的需要，可以推广。     

Effects of simulated precipitation extremes on crayfish (Procambarus clarkii Girard) oviposition in artificial burrows

Mature female Procambarus clarkii were housed individually in artificial burrows simulating conditions in natural burrows. Crayfish were exposed to conditions of simulated temporary flooding dislocation (Experiment 1) or simulated drought (Experiment 2). In Experiment 1, females were dislocated from burrows to simulate 0, 1, 3 or 6 temporary flooding dislocations, and in Experiment 2, females were exposed to the following simulated drought conditions: wet control (no drought), dry control (no free water), month‐long periods without free water during July, August, September and October (1 month drought), two consecutive months without free water (August–September), and three consecutive months without free water (August–October). In Experiment 1, no significant differences in survival or oviposition rate were observed following transfer of individuals from ‘settled/sealed’ burrows to ‘new burrow’ containers, even after six consecutive dislocations. In Experiment 2, simulated drought of 2 or 3 month's duration had a significant negative impact on oviposition. Survival was significantly reduced only among females exposed to no free water during the 6‐month study. No spawning occurred without the presence of free water in the burrows, and when free water was absent during the peak spawning months of September and/or October, oviposition was delayed until free water was returned. These results indicate that brood female P. clarkii can withstand short‐term conditions without free water in the burrow with little effect, but reproduction and survival are significantly reduced by prolonged drought.     

The influence of haemocyte number on the resistance of the freshwater crayfish, Pacifastacus leniusculus Dana, to the parasitic fungus Aphanomyces astaci

Abstract. The number of circulating haemocytes in crayfish, Pacifastacus leniusculus Dana, was reduced after injection of yeast cell walls (Zymosan) into the animals. Crayfish harbouring the parasitic fungus A. astaci as a 'latent' infection in melanized areas in the cuticle died due to unrestricted growth of unmelanized hyphae when they were injected with Zymosan. The β–1,3–gIucan laminaran, introduced into the haemocoel of P. leniusculus harbouring A. astaci hyphae in the cuticle, also caused a decrease in the total haemocyte counts (THC), but these animals survived the treatment and no unmelanized hyphae of A, astaci were found. The recovery of THC was found to be more rapid after laminaran injection than after Zymosan injection, which could explain why laminaran treated crayfish retained their resistance and could combat the A. astaci hyphae in the cuticle. The importance of the number of haemocytes in defence and well-being of crayfish is discussed.     

稻田养殖克氏原螯虾关键技术探讨

克氏原螯虾俗称小龙虾,是淡水经济虾类,因可食部分较多、肉质细嫩、味道鲜美、营养价值高、蛋白质含量高的优点深受人们的欢迎.其养殖业也逐步引起人们的重视,由于稻田养殖克氏原螯虾可以把种植业与养殖业有机结合起来,充分利用人工建设的生态系统,使其发挥各自的作用.正因为有这样的优点,很多地方都开展了稻田养殖克氏原螯虾的试验[1-3].     

克氏原螯虾繁殖生物学的研究

结合实验室研究和室外调查等方法,就比较有争议的有关克氏原螯虾繁殖生物学的一些问题进行了较为详细的研究。认为克氏原螯虾雌雄鉴别最可行的方法是根据二者之间腹肢的不同。雄虾的第1、2对腹肢的基部和中段呈红色,末端变白,而雌虾没有这样的腹肢。克氏原螯虾没有生殖蜕皮的习性,交配之前不一定需要蜕皮。武汉地区克氏原螯虾群体一般每年有2个产卵期,一个为5月左右,一个为9月下旬到10月中旬。雌虾的抱卵量在200~300粒。水温22~25℃时,克氏原螯虾胚胎发育的时间为25~33d。     

Direct visualization of the novel pathogen,Spiroplasma eriocheiris,in the freshwater crayfish Procambarus clarkii (Girard) using fluorescence in situ hybridization

Spiroplasma eriocheiris is the first spiroplasma strain known to be pathogenic to freshwater crustaceans. It has caused considerable economic losses both in the freshwater crayfish Procambarus clarkii (Girard) and in some other crustaceans. The monitoring of the pathogen in crustacean populations and study of its behaviour in the laboratory require the development of reliable diagnostic tools. In this article, we improved microscopic identification of S. eriocheiris by combining in situ hybridization with specific fluorescently labelled oligonucleotide probes. The established fluorescence in situ hybridization (FISH) allowed simultaneous visualization, identification and localization of S. eriocheiris in the tissues of diseased crayfish P. clarkii and exhibited low background autofluorescence and ideal signal‐to‐noise ratio. With the advantages of better tissue penetration, potentially more specific and stable, we designed three species‐specific oligonucleotide probes utilizing the sequences of 16S‐23S rRNA intergenic spacer regions (ISRs) of S. eriocheiris. Positive hybridization signals were visualized in haemocytes and connective tissues of hepatopancreas, cardiac muscle and gill from diseased crayfish. This unique distribution pattern matched the pathological changes when diagnosed by H&E staining and indicated that S. eriocheiris probably spread throughout the tissues in P. clarkii by hemokinesis. This assay will facilitate our understanding of the pathogenesis of S. eriocheiris and enhance the early diagnosis of the novel pathogen.     

小龙虾池塘养殖试验

小龙虾又名克氏原螯虾(Procambarus clarkii),主要分布于长江中下游地区。小龙虾病害少,耐低氧,能在池塘、河沟、湖泊、稻田、沼泽地等水体中生长繁殖,对不良环境的耐受力很强,因此逐渐形成养殖热潮,小龙虾产业得到了快速发展[1]。2018年石家庄市水产技术推广站承担了河北省水产重点技术示范与推广项目-克氏原螯虾养殖技术示范项目,开展小龙虾养殖技术示范与推广,发展本地的小龙虾产业,探索调整池塘养殖品种结构,发展生态、高效养殖。     

Growth performance and muscle composition response to reduced feeding levels in juvenile red swamp crayfish Procambarus clarkii (Girard, 1852)

Overfeeding in aquaculture can lead to suboptimal growth and increased production costs. Red swamp crayfish, Procambarus clarkii, is one of the most noteworthy species cultured in China, but little information is available on the optimal feeding levels of this species, especially in ponds cultured with macrophytes. In this study, we tested the effects of five different feeding levels (20%, 40%, 60%, 80% and 100% satiation) of an artificial diet on growth performance and muscle composition of juvenile P. clarkii in 15 concrete ponds cultured with the macrophyte Hydrilla verticillata (three replicates for each treatment). The results showed that growth performance decreased only when feeding levels were below 60% satiation. Muscle composition analysis revealed that the moisture and ash contents of muscle did not vary significantly with feeding levels but that the lipid and protein contents of muscle significantly decreased when P. clarkii was fed to 40% satiation. Stable isotope analysis suggested a shift in crayfish diets to easily available H. verticillata when feeding levels decreased. With this study, we hope to encourage crayfish farmers to reduce feeding levels and increase natural food items such as macrophytes in cultured ponds to maximize crayfish yields and reduce production costs.     

Estimates of genetic parameters for growth-related traits of the red swamp crayfish,Procambarus clarkii

In this study, technologies of family construction and grow-out culture in the same pond were studied for red swamp crayfish. A total of 77, 110 and 78 families were constructed in 2011, 2012 and 2013, respectively. Heritability, genetic and phenotypic correlations were estimated for body weight, body length and carapace length through performance testing in progeny of 2012 at 5 and 10 months of age, resulting from a nested mating design. At 5 months of age, none of the estimated heritabilities were significant (0) based on sire components (h _s ² ) for these traits, the genotypic correlations, either; only the phenotypic correlations were significant (0.8561–0.9420). At 10 months of age, the estimated heritabilities (h _s ² ) were low for the three traits (0.1429–0.1632); the phenotypic and genetic correlations were significant, 0.8148–0.9052 and 0.9981–0.9993, respectively. It was concluded that genetic improvement through selective breeding conducted on red swamp crayfish might be feasible and would be an efficient method to obtain a positive response to selection for the 10-month-old red swamp crayfish.     

Response of crayfish, Procambarus clarkii, haemocytes infected by white spot syndrome virus

White spot syndrome virus (WSSV) is a serious pathogen of aquatic crustaceans. Little is known about its transmission in vivo and the immune reaction of its hosts. In this study, the circulating haemocytes of crayfish, Procambarus clarkii, infected by WSSV, and primary haemocyte cultures inoculated with WSSV, were collected and observed by transmission electron microscopy and light microscopy following in situ hybridization. In ultra-thin sections of infected haemocytes, the enveloped virions were seen to be phagocytosed in the cytoplasm and no viral particles were observed in the nuclei. In situ hybridization with WSSV-specific probes also demonstrated that there were no specific positive signals present in the haemocytes. Conversely, strong specific positive signals showed that WSSV replicated in the nuclei of gill cells. As a control, the lymphoid organ of shrimp, Penaeus monodon, infected by WSSV was examined by in situ hybridization which showed that WSSV did not replicate within the tubules of the lymphoid organ. In contrast to previous studies, it is concluded that neither shrimp nor crayfish haemocytes support WSSV replication.     

Invasive dynamics of the crayfish Procambarus clarkii (Girard, 1852) in the international section of the River Minho (NW of the Iberian Peninsula)

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重金属对克氏原螯虾的影响及其对策

介绍了我国在重金属对克氏原螯虾影响方面的研究和应用现状,提出了如何规避重金属对克氏原螯虾影响的加工及食用的办法和措施。     

An unidentified gland-like tissue near the androgenic gland of red swamp crayfish, Procambarus clarkii

ABSTRACT:   Among all of the decapods that have been studied, only one kind of gland, the androgenic gland, has been found attached to the subterminal ejaculatory region of the vas deferens. In the present study of the red swamp crayfish, it was found that two kinds of gland-like tissues were attached to the subterminal ejaculatory region of the vas deferens. One of the tissues, which is referred to as TIB, is located inside the body cavity, whereas the other tissue, which is referred to as TIC, is inside the coxa. The characteristics of TIB resemble those of the androgenic gland of other malacostracans. The TIC cells, however, are smaller and more basophilic due to the small size and high density of their cytoplasm and nuclei. Moreover, black fibers, similar to the nerve fibers of the ganglion, were detected among the TIC cells after Bodian's stain. The conclusion is that TIC may be a newly found male sexual characteristic and further research is needed to confirm its functions.