Procambarus clarkii Girard as a vector for the crayfish plague fungus, Aphanomyces astaci Schikora

Abstract. Procambarus clarkii Girard, a native freshwater crayfish species of Lousiana, USA, was found to harbour the crayfish plague fungus, Aphanomyces astaci Schikora, in its cuticle as a benign infection. Under certain conditions, P. clarkii dies as a result of this parasite, and the A. astaci infection then becomes acute and can be transmitted to Astacus astacus (L.).
Therefore, it is now shown that at least three different species of North American crayfish ( Pacifastacus leniusculus Dana, Orconectes limosus Raff, and Procambarus clarkii Girard) can carry the crayfish plague fungus, A. astaci , can transfer the disease to other crayfish species, and under certain circumstances can die of its own infection.     

Preliminary Results on the Fatty Acid Composition of Freshwater Crayfish, Astacus astacus and Pacifastacus leniusculus, Held in Captivity

The fatty acid compositions of the tail muscle and hepatopancreas of captive noble crayfish Astacus astactus and of signal crayfish pacfastacus leniusculus were investigated. The major fatty acids in the tail muscle of A. astacus were 16:0. 18:ln-7 + 9, 20:4n–6 and 20:5n–3 and the fatty acid composition of hepatopancreas was principally 16:0, 18:In-7 + 9 and 20: ln-9+ 11. These animals had been fed marine fish (sprat and herring) for 2 mo. They were kept in a flowthrough system with gravity-fed fresh water from a nearby lake with a high concentration of calcium and pH above 8. The major fatty acids in the tail muscle of P. leniusculus were 16:0. 18:ln-7+9, 18:2n–6 and 20:5n–3, and in the hepatopancreas 16:0, 18:1n–7 + 9 and 18:2n–6. These animals had been fed mainly plant material (green peas, maize and alder leaves) in a different laboratory for a period of about 2 yr and kept in a flow-through system with ambient water of pH around 7. In A. astacus the total fatty acid content was nearly 13 times higher in the hepatopancreas than in the tail muscle, however, three fatty acids 18:0, 20:4n–6 and 20:Sn-3 were, respectively, 289, 403, and 320% more concentrated in the fatty acids of tail muscle than in the hepatopancreas. Total fatty acids of the hepatopancreas of male P. Ieniusculus was 42 times higher than in the tail muscle and was three times greater than in the hepatopancreas of A. astacus .     

Lethal limits of high temperature for two crayfishes, the native species Cambaroides japonicus and the alien species Pacifastacus leniusculus in Japan

ABSTRACT: Lethal limits of high temperature were studied to clarify the effects on the survival of the endangered Japanese crayfish species Cambaroides japonicus and the alien species Pacifastacus leniusculus . After the acclimation period for 2 weeks at 16°C, the temperature was raised at a rate of 1°C per week. As a result, the ultimate upper lethal temperatures of C.   japonicus and P.   leniusculus were 27.0 and 31.1°C, respectively, and the lethal temperature for P.   leniusculus was significantly higher than for C.   japonicus . The natural distributions of these two species are discussed in terms of the temperature tolerance.     

Mating and spawning of freshwater crayfish, Austropotamobius pallipes Lereboullet, under laboratory conditions

Abstract. The study includes mating and spawning data of freshwater crayfish, Austropotamobius pallipes Lereboullet, obtained in four consecutive years (1987–1990). A total number of 586 wild-caught female A. pallipes in the 21·65 mm to 45·60mm carapace length (CL) size were held under laboratory conditions at different dates in September–October. Initial densities at mating time were 21 and 24 crayfish/m² with a 1:2 male:female ratio. High percentages of mating were obtained (97·4–100%) except in 1989 (57·5%). In all years, most of the matings (90%) were concentrated in a period of 8–12 days with mean water temperatures between 12°C and 13·5°C. Spawning took place a few days after mating (minimum 2 days, maximum 14 days) at mean water temperatures between 8·2°C and 10·8°C. In the first 3 years, almost 100% of mated females spawned. However, in the fourth year (1990), when the mean size of crayfish was smaller, only 72·4% of mated females spawned. The mean number of pleopodal eggs was 64 (maximum 220, minimum 18). Egg diameter ranged between 2·30 and 3·25 mm (mean value 2·78 mm). Pleopodal egg number was positively correlated with carapace length ( r = 0·72).     

Aphanomyces astaci pathogenicity under laboratory and field conditions

Abstract. The high pathogenicity of Aphanomyces astaci to susceptible crayfish has often been mentioned but not well investigated in publications on crayfish plague. This paper presents a study of that pathogenicity and considers the relationships between size of zoospore challenge and challenge temperature, which results in the extremely complex series of pathologies which comprise crayfish plague.     

Occurrence of the Enigmatic, Unicellular Psorospermium Organism in Several Cultured, Sympatric Populations of the Freshwater Crayfishes Procambarus clarkii and Procambarus zonangulus

The parasitic Psorospermium organism was studied in cultured sympatric populations of mature Procambarus clarkii and Procambarus zonangulus during four consecutive annual crayfish seasons from 1991–1992 through 1994–1995. Most P. clarkii were infected while fewer than half of the P. zonangulus were infected throughout the study period. The numbers of Psorospermium were four to eight times greater in P. clarkii and showed annual changes not seen in P. zonangulus . Both crayfishes demonstrate 1yr life cycles in Louisiana and this seemed to explain the variations seen in Psorospermium dynamics in the two hosts. In general, there was no consistent association between crayfish size and the number of parasites found in them for either crayfish species. There was some indication of an association between the presence of infection and the number of parasites found in P. zonangulus .     

The immunomodulatory effect of laminaran [β(1,3)-D-glucan] on Atlantic salmon, Salmo salar L., anterior kidney leucocytes after intraperitoneal, peroral and peranal administration

Anterior kidney leucocytes obtained from Atlantic salmon, Salmo salar L., 2 days after administration of laminaran, were assayed for their capacity to reduce nitroblue tetrazolium to formazan, and for their activity of lysosomal acid phosphatase after intraperitoneal (15 mgkg^?1), peroral (150 mg kg^?1) or peranal (150 mg kg^?1) administration. Leucocytes obtained from salmon treated by an intraperitoneal injection of laminaran produced significantly more superoxide anion than cells obtained from fish treated with dextran or sodium chloride immediately after cell isolation. Immediately after extraction, the activity of acid phosphatase in anterior kidney leucocytes obtained from salmon injected with laminaran was significantly higher than in cells harvested from fish treated with dextran or sodium chloride. Furthermore, cells obtained from salmon treated by peroral instillation of laminaran showed significantly enhanced production of superoxide anion compared with leucocytes from fish treated with either sodium chloride or dextran. The acid phosphatase activity in anterior kidney leucocytes from salmon treated by peroral and peranal instillation of laminaran was significantly higher than in cells from fish treated either with sodium chloride or dextran. Finally, fluorescence microscopic examination of tissue sections from fish treated peranally by intubation with fluorescein labelled laminaran revealed fluorescent vesicles in intestinal epithelial cells and in anterior kidney macrophages.     

饲料铜含量对亚硝酸盐胁迫下斑节对虾血细胞的影响

研究了饲料铜离子含量对亚硝酸盐胁迫下斑节对虾（Penaeus monodon）血细胞参数的影响。向斑节对虾投喂不同铜离子添加量（0,10,25,40,55和110 mg/kg）的饲料8周后,进行24 h 的20 mg/L亚硝酸氮胁迫,取样测定对虾的血细胞总数（THC）、血细胞活性氧（ROS）含量和血细胞凋亡率。结果显示,经亚硝酸盐胁迫后,25-55 mg/kg饲料组对虾能保持较高的THC,以及较低血细胞ROS含量和凋亡率;110 mg/kg饲料组对虾的THC最低,血细胞ROS含量和凋亡率均最高。这些结果表明饲料中添加25-55 mg/kg的铜离子能有效降低亚硝酸盐胁迫对血细胞造成的氧化损害,但过量添加（110 mg/kg）反而对血细胞造成更为严重的损害。     

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.     

Histopathology of oxygen intoxication in the juvenile red abalone, Haliotis rufescens Swainson

Abstract. Juvenile abalone were exposed to supersaturated oxygen conditions (about 150–200% of saturation) in order to reproduce similar conditions encountered in intensive husbandry systems and to then study the resultant lesions. Depigmentation, lethargy and swelling of tissues were observed clinically. Histopathology showed that the cytoplasmic vacuoles of large haemocytes were enlarged. Oxygen emboli were observed throughout the muscular tissue and connective tissue but separation of fibrous neural sheath from nerve cell bodies and surrounding tissue was the predominant lesion. Chromatin of the nuclei of nerve cells became marginated. Gaseous emboli were observed at various other locations in the vascular system. At termination of the experiment all animals sampled bacteriologically showed systemic infection with Vibrio alginolyticus . The clinical signs appeared to result from mechanical interference caused by the accumulation of oxygen and from dysfunction of neural structures. The predominant histopathological changes occurred after only 3 h or less exposure time. Normal histology of the haemocytes and nerve structures is described.     

流式细胞术在克氏原螯虾血细胞的分类、活性和免疫功能研究中的应用

应用流式细胞术(FCM)对克氏原螯虾(Procambarus clarkii)血细胞的分类、活性和免疫功能进行了研究。结果显示:血细胞可分为透明细胞、小颗粒细胞和大颗粒细胞三个亚群,组成比例分别为(26.25±5.29)%、(51.44±7.02)%和(11.20±1.82)%;螯虾血细胞的平均总凋亡率约为3.12%;血细胞对荧光大肠杆菌的吞噬活力显著(P<0.05)高于荧光微球,吞噬率分别为17.04%和14.57%;血细胞在自然生理状态下含有一定量的活性氧,其在两类颗粒细胞的含量显著(P<0.05)高于透明细胞,在大颗粒细胞中最高。结果表明,FCM能较好地应用于虾类的血细胞分类和功能研究。     

应用流式细胞术检测虾类血细胞总数方法的建立

通过对流式细胞术(FCM)检测虾类血细胞总数(THC)的条件和方法进行优化,为虾类血细胞学研究提供快捷、准确的测定方法。2012年7-9月,应用SYBR Green I作为荧光染料标记完整血细胞,设置3个不同染料浓度(1×、10×和100×),测定不同孵育时间下染色细胞比例的变化。结果显示,染料终浓度为10×时染色效果最佳,其最佳孵育时间为60 min;应用建立的FCM方法和显微计数方法测定10尾凡纳滨对虾(Litopenaeus vannamei)的THC,平均值分别为(16.68±1.57)×106个/m L和(15.09±1.76)×106个/m L,2种方法测定结果的相关性极显著(R2=0.8064,P<0.01)。凡纳滨对虾经不同浓度(0.5 mg/L和5.0 mg/L)的Cd2+胁迫,利用建立的FCM方法测定Cd2+胁迫下对虾THC的变化。结果显示,0.5 mg/L和5.0 mg/L Cd2+胁迫下,对虾THC随着胁迫时间的延长不断下降,胁迫48 h时分别下降至对照组的78.7%和64.7%,可见Cd2+胁迫对虾类血细胞产生毒性,抑制了血细胞活性,表明该方法适用于虾类的血细胞学研究。     

Pathogenicity and culture studies comparing the Aphanomyces involved in epizootic ulcerative syndrome (EUS) with other similar fungi

Pathogenicity and cultural experiments described here provide futher evidence that a distinct species of Aphanomyces is responsible for much of the characteristic pathology of epizootic ulceration syndrome (EUS). Zoospores from 58 fungal isolates were injected intramuscularly in snakehead fish, Channa striata (Bloch). These fungi comprised: Aphanomyces strains isolated from EUS-affected fish; saprophytic Aphanomyces , Achlya and Saprolegnia spp. from infected waters; and further saprolegniaceous fungi involved in other diseases of aquatic animals. Only the Aphanomyces strains isolated from fish affected by EUS, Australian red spot disease (already considered synonymous with EUS) or mycotic granulomatosis described from Japan were able to grow invasively through the fish muscle and produce the distinctive EUS lesions. In contrast to Aphanomyces astaci Schikora, the EUS- Aphanomyces was shown to be unable to infect noble crayfish, Astacus astacus L. The snakehead-pathogenic strains were further distinguished from all the other fungi under comparison by their characteristic temperature-growth profile and inability to grow on certain selective fungal media.     

饥饿胁迫对虾夷扇贝几种免疫因子的影响

在(17±1)℃条件下,研究了壳长(8.83±0.52)cm的虾夷扇贝在不同饥饿时间(5、10、15、20、30 d)处理后,血液中血细胞总数、血细胞吞噬水平、酸性磷酸酶、过氧化物歧化酶的变化。结果表明,扇贝的血细胞总数随着饥饿时间的延长逐渐降低,饥饿20 d以上的扇贝血细胞数明显降低,与投喂组差异极显著(P<0.01),扇贝血细胞吞噬率和酸性磷酸酶活性呈现出先小幅上升后下降的趋势,饥饿253、0 d与投喂组差异显著(P<0.05),饥饿253、0 d的扇贝过氧化物歧化酶的活性有所升高,饥饿30d,与投喂组差异显著(P<0.05)。试验表明,饥饿超过20 d,扇贝的免疫系统受到较为严重的破坏。     

Intestinal absorption of immunomodulatory laminaran and derivatives in Atlantic salmon, Salmo salar L.

Abstract. Radiolabelled (¹²⁵I, ³H) immunomodulatory laminaran (isolated from Laminaria hyperborea ), a β(1,6)-branched β(1,3)-D-glucan and different radiolabelled sulphated analogues of laminaran were administered peranally to Atlantic salmon, Salmo salar L. Intestinal absorption and tissue distribution were examined by means of radioactive tracer techniques. The intestinal uptake was highest with native laminaran and laminaran with a low degree of sulphatation, while highly sulphated laminarans were poorly absorbed. The tissue distribution analysis revealed high amounts of radiolabelled compound in the liver, and anterior and posterior kidney, whereas the spleen contained low amounts. Peak serum and organ concentrations were reached about 30 min after administration. The results were confirmed by autoradiography of tissue sections from salmon after peranal administration of radiolabelled laminaran. Finally, the peranal administration of fluorescence labelled laminaran revealed epithelial supranuclear fluorescent vacuoles containing laminaran. It is concluded that native laminaran and slightly sulphated laminaran are absorbed from the posterior intestine and that they are distributed to tissues rich in immunocompetent cells. Thus, these compounds may have potential as immunomodulatory feed additives.     

枯草芽孢杆菌基因组DNA对中华绒螯蟹非特异性免疫因子的影响

将枯草芽孢杆菌(Bacillus subtzlis)基因组DNA按5、20和80μg/kg剂量体腔注射中华绒螯蟹(Eriocheir sinensis)(约0.1 mL/只),设注射等体积TE缓冲液为对照组,分别在第1、3、5、7天采样,测定该细菌DNA对中华绒螯蟹的免疫刺激作用。结果显示:注射上述3种剂量的枯草芽孢杆菌基因组DNA均能不同程度地提高中华绒螯蟹的THC、血细胞呼吸爆发活性和血清PO、ACP活性,试验组蟹的上述免疫指标的活性显著高于对照组(P<0.05),并以20μg/kg体重剂量的刺激效果最好;但试验剂量的细菌DNA对血清ALP活性无显著影响(P>0.05)。     

The immunomodulatory effect of LPS, laminaran and sulphated laminaran [β(l,3)-D-glucan] on Atlantic salmon, Salmo salar L., macrophages in vitro

Abstract. The stimulatory effect of LPS ( Vibrio anguillarum ), laminaran and sulphated laminaran, aqueous soluble β(1,6)-branched β(l,3)-D-glucan obtained from Laminaria hyperborea , on head kidney macrophages of Atlantic salmon, Salmo salar L., is reported. The macrophages, after stimulation with LPS or laminaran, showed pronounced spreading, membrane ruffling and increased organellc content when examined by light microscopy. LPS stimulation induced enhanced phagocytic and pinocytic activity, higher intraccllular production of superoxide anion, and higher activity of acid phosphatase compared to control cells. Native laminaran stimulated the cells to pinocytose more fluid phase, increase the intracellular production of superoxide anion and to elevate the activity of acid phosphatase. Sulphated laminaran induced higher production of superoxide anion and higher activity of acid phosphatase by macrophages than in control cells.     

Freshwater Crayfish Growth Under Culture Conditions: Proposition for a Standard Reporting Approach

A review of approaches currently used to measure growth in freshwater crayfish under experimental and pond grow-out conditions is presented. The review examined 206 papers on growth and production of freshwater crayfish of which 34 were selected for detailed investigation. Selection was based on the amount of information provided in the published report and on the species studied. The review was restricted to six crayfish species: Cherax tenuimanus (marron), C. destructor (yabbie), C. quadricarinatus (red claw), Procambarus clarkii (red swamp crayfish), Astacus astacus (noble crayfish), and Pacifastacus leniusculus (signal crayfish). Growth indices reported included instantaneous growth rate (IGR), absolute growth rate (AGR), slope of log weight against time (GRE) and specific growth rate (SGR, IGR100). Studies were performed for varying time intervals under a wide range of stocking densities. Most reports did not express growth rate as an index but simply reported growth in terms of initial and final weights. The range of approaches used to express growth performance in freshwater crayfish growth studies hinders comparisons of growth rate and production findings. It is suggested that the reports of studies of growth performance in cultured crayfish include a growth rate parameter as well as initial and final weights. SGR is recommended as the parameter of choice since it is based on an exponential growth trend which is typically observed in measurements of weight increase with time in cultured juvenile and early adult crayfish.