Digestive enzyme spectra in crustacean decapods (Paleomonidae, Portunidae and Penaeidae) feeding in the natural habitat

This work describes the profile of five proteases and four carbohydrases from the crustacean decapods Macrobrachium australiense (Holthuis), Scylla serrata (Forskal), Portunus pelagicus (Linnaeus), Penaeus esculentus, Penaeus plebejus (Hess) and Metapenaeus bennettae (Racck & Dall), feeding in the natural habitat, in order to provide an indication of their digestive capabilities. The results raised the following points. First, species from each family showed a particular suite of digestive enzymes. Second, the activity of cellulase from M. australiensis and S. serrata , using AZCL-HE cellulose as the substrate, was around 90% higher than that observed with AZO-CM cellulose. However, for P. pelagicus and P. esculentus , the enzyme activity was better with AZO-CM cellulose. Third, M. australiense displayed the highest ratio of amylase to protease activity. In contrast, Portunidae specie s, P. pelagicus and S. serrata showed the lowest ratios. Fourth, comparison of the laminarinase activity of M. bennettae and P. esculentus in October (Spring) and December (early Summer) showed a significant decrease in December. Finally, the wide distribution of digestive enzymes in these crustaceans may reflect different feeding habits and habitats.     

Susceptibility,behaviour, and retention of the parasitic salmon louse (Lepeophtheirus salmonis) differ with Atlantic salmon population origin

Atlantic salmon populations across the world have diverse ecological and evolutionary histories, from wild anadromous or landlocked, to domestication and genetic modification. The natural host behaviours confer protection from infestation by ectoparasitic salmon lice Lepeophtheirus salmonis, yet whether genetic origin results in different behaviours and thus susceptibility to infestation is unknown. In common garden experiments, we tested antiparasite behaviours, susceptibility and retention of salmon lice in wild anadromous, wild landlocked, domesticated and genetically modified domesticated strains. Within domesticated strains, we tested two infestation histories (previously infested and naïve) and a new phenotype (albino colouring). Farmed stocks initially acquired 24%–44% higher levels of parasite density than the wild and landlocked strains. Burst swimming and displacement behaviours were higher in the domesticated groups, and jumping was more prevalent in the domesticated strains. At 34 days post‐infestation, domesticated strains and the wild anadromous strain did not differ significantly from each other; however, landlocked salmon had increased infestation levels considerably. Domesticated strains lost ~20% (±9.9%–16.5%; 95% CI) of their initial parasite load, while parasite load increased by 5.5% (±30.1%) for wild salmon and 20.1% (±28.5%) in landlocked salmon. This study provides early evidence for diverged host–parasite interactions associated with domestication in this system.     

Parasites of cultured and wild brown‐marbled grouper Epinephelus fuscoguttatus (Forsskål, 1775) in Lampung Bay,Indonesia

A total of 210 Epinephelus fuscoguttatus (brown‐marbled grouper) was examined for parasites. During three consecutive seasons (two rainy and one dry season from 2002 to 2004), 35 specimens each taken from floating net cages of the National Sea Farming Development Centre (Balai Budidaya Laut) and from wild catches in Lampung Bay, South Sumatra, Indonesia were studied. Twenty‐five (cultured grouper) and 30 (wild grouper) parasite species/taxa were identified, with an infracommunity ranging from one to nine (cultured) and three to 14 parasite species (wild), demonstrating a species‐rich parasite fauna even in the cultured fish. Protozoans (1 species), microsporeans (1), myxozoans (1), digeneans (8), monogeneans (5), cestodes (3), nematodes (8), acanthocephalans (2) and crustaceans (6) were found. The most abundant parasites were the monogeneans Pseudorhabdosynochus epinepheli and Pseudorhabdosynochus lantauensis for both, cultured and wild grouper during all seasons. For the cultured fish, the prevalence of monoxenous ectoparasites (e.g. P. epinepheli, P. lantauensis, Capsalidae gen. et sp. indet., Benedenia epinepheli) was in most cases higher than that of heteroxenous endoparasites. This contrasts the wild grouper, where heteroxenous parasites such as Allopodocotyle epinepheli and Raphidascaris sp. occurred at a similar prevalence compared with the fairly abundant Pseudorhabdosynochus spp. No seasonality of infestation was observed for both cultured and wild fish. The high levels of infestation of potentially pathogenic monogeneans throughout the year could result in significant parasite outbreaks at the locality studied.     

Epizootiology of Parvicapsulaminibicornis in Fraser River sockeye salmon, Oncorhynchusnerka (Walbaum)

Late-spawning Fraser River sockeye salmon, Oncorhynchus nerka , stocks have suffered significant prespawn mortality associated with an unusually early freshwater migration pattern and the myxosporean parasite Parvicapsula minibicornis . Surveys of migrating adult salmon from several spawning populations were conducted in 1999 and 2000 to determine the extent of infection with P. minibicornis , when and where the parasite first becomes detectable during migration, and whether early migrating stocks might be used as sentinels to assess risk of infection in late-spawning stocks. Posterior kidney, preserved in 95% ethanol, was examined for P. minibicornis in stained histological sections and using a polymerase chain reaction (PCR) test. The prevalence of this parasite in all Fraser River sockeye salmon stocks examined was high (range 47–100% infected). In contrast, P. minibicornis was not detected in the fish tested from the two sockeye salmon stocks outside the Fraser River drainage in either 1999 or 2000. The parasite was also not detected histologically or by PCR in the kidney tissue of the fish from the Fraser River that were sampled in salt water or early during their freshwater migration up the river. These findings and the progression in the prevalence and intensity of infection as the fish from three stocks (early Stuart, Weaver Creek and Cultus Lake) were monitored over time, suggest salmon acquired the parasite either in the lower Strait of Georgia or in the lower Fraser River before the confluence of the Harrison River. In both 1999 and 2000 the parasite was present in all Fraser River sockeye salmon stocks sampled, which suggests that early Stuart salmon may be valuable as a sentinel stock for the presence of the parasite in later-spawning stocks.     

锯缘青蟹幼体饵料的营养强化

用酵母、水球藻、鱼油强化和豆油强化四种不同方式培养轮虫，再分别投喂锯缘青蟹幼体，分析测定轮虫和体的生化组成，结果显示，（1）不同方式培养的轮虫之间以及摄食这些轮虫的锯缘青蟹幼体之间的蛋白质含量都没有显著差异；（2）轮虫的脂类含量和脂肪酸组成与培养方式密切相关，小球藻轮虫的脂类含量最高，20：5n-3(EPA)占总脂肪酸的比例也最高 ，为18.05%，鱼油轮虫则含有最多的22：6n-3(DHA)，占总脂肪酸3.16%，脂类含量仅次于小球藻轮虫；（3）锯缘青蟹幼体的脂类含量和脂肪酸组成受相应饵料营养成分的影响。另外，幼体培育实验也发现，饵料营养成分影响幼体的存活率，结果表明，提高轮虫的EPA和DHA含量，尤其晨DHA含量，将有利于锯缘青蟹幼体的存活和发育。     

Spawning Characteristics of the South African Mudcrab Scylla serrata (Forskål) in Captivity

Scylla serrata is a potential aquaculture species in Southern Africa. Information about its reproductive biology is required as a prerequisite to establishing hatchery technology. Adult female S. serrata were caught in the Umlalazi estuary on the subtropical east coast of South Africa and kept in captivity to observe and record spawning characteristics. Data collected included crab size and mass, time in captivity prior to spawning, fecundity per batch, relative fecundity, individual egg mass and size, size of zoea 1 larvae, incubation time, and hatch success rate. Of the 119 crabs kept in captivity 83% spawned in the maturation system– most within 40 d of capture. The crabs were highly fecund (mean relative fecundity per batch = 10,655 ± 4,069 eggs/g female) and the majority of the batches hatched within 288 h (12 d) at 27 C. Spawning in captivity occurred throughout the year, with a peak in late winter/early spring. This differs slightly from records of ovarian maturity stages of the crabs in the wild. A pattern of synchronous spawning was recorded where the females were observed to extrude their eggs in groups, commonly within 3 d of one another, separated by long periods of inactivity, suggesting an exogenous spawning cue. A total of 1,374,488 zoea larvae were obtained per kg of female per month. This means that if sufficient mature females can be caught from the wild, these could be used for stocking hatchery operations. The crabs were easy to maintain, mature, and spawn in captivity. This will facilitate future domestication which will eventually reduce the need for wild caught broodstock. The spawning characteristics of South African 5. serrata At in well with those observed for the genus throughout its distribution implying that ecological and fisheries management could be similar.     

The parasitic barnacle, Sacculina granifera Boschma, affecting the commercial sand crab, Portunus pelagicus (L.), in populations from two different environments in Queensland

Abstract. The parasitic barnacle, Sacculina granifera , changed the abdominal morphology of the host crab, Portunus pelagicus (L.). Three different shapes were recognizable with increase of the host size. Crabs carrying externae could be found throughout the year with higher percentages in the dry season (May-October) in the Gulf of Carpentaria and in warm months (October-March) in Moreton Bay. The parasite infested more males than females in the Gulf but both sexes of crabs equally in the Bay. Generally, the parasite matured when the host (always immature) reached the size of mature crabs. The smallest crab carrying an externa was 70 mm carapace width from the Gulf and 90 mm CW from the Bay. Overall, 6.6% of P, pelagicus were found with externae in the Bay, a subtropical area where crabs are heavily exploited by amateur and commercial fishermen. In comparison, only 1.2% of crabs were found with externae in the Gulf, a tropical area where no commercial fishery is targeted at this species.     

Immunization of grouper, Epinephelus coioides, confers protection against a protozoan parasite, Cryptocaryon irritans

The present study aimed to determine whether protection is conferred by immunization of grouper, Epinephelus coioides, against a protozoan parasite, Cryptocaryon irritans. The immunization of E. coioides was carried out by a low level exposure of fish to live C. irritans theronts from predetermined number of tomonts and by an intraperitoneal injection of a vaccine consisting of formalin-killed C. irritans theronts.

Mucus titers detected by ELISA were significantly higher in fingerling and adult grouper subjected to the low level of exposure to C. irritans theronts at 3-week post-exposure compared to fish that had no previous exposure. In addition, significantly smaller tomonts were produced from adult grouper after three successive exposures than the tomonts produced after a single exposure to the parasite.

In the vaccine-immunization experiment, no mortality was monitored in fish that received high dose vaccine (100 μg/fish), while 40% cumulative mortality and 100% cumulative mortality were recorded in low dose group (10 μg/fish) and control group (PBS-injected), respectively. In the succeeding replicate, the vaccine-immunized group (high dose) had 37.5% cumulative mortality and 100% cumulative mortality for the control. In addition, a total of 1830 tomonts were collected at 5-day post-challenge from the control group while none from the vaccine-immunized group. Significantly fewer trophonts and tomonts were enumerated at 5-day and 7-day post-challenge, respectively, in the vaccine-immunized group than the control.

Results suggest that a protective immunity has been conferred on the immunized grouper as indicated by high antibody titers in the mucus of C. irritans-exposed fish and higher survival and fewer parasites in vaccine-immunized fish than the control groups. The conferred immunity played a major role in preventing or limiting the adhesion, invasion, and development of C. irritans theronts on the skin of the immunized grouper.     

Encephalomyelitis associated with microsporidian infection in farmed greater amberjack, Seriola dumerili (Risso)

An outbreak of a disease characterized by a peculiar spiral movement in farmed greater amberjack, Seriola dumerili (Risso), occurred in Kagoshima Prefecture, Japan, in May 2008, immediately after importing the fish from China. Although neither bacteria nor viruses were detected in routine diagnostic tests, histopathological observations of the affected fish revealed severe inflammation in the tegmentum of the brain including the medulla oblongata and the anterior part of the spinal cord. In addition, a microsporidian parasite was observed in the nerve cell bodies or axons in the inflamed tissues. We identified a microsporidian small subunit rRNA gene (SSU rDNA) from the lesion, and the sequence showed 96.1% identity with that of Spraguea lophii. Subsequent in situ hybridization using probes presumably specific to the SSU rRNA confirmed that the parasite observed in histopathology harboured the identified SSU rRNA. Apparently degenerated microsporidian cells or spores were also frequently observed in tissue sections. Thus, the disease was most probably caused by the infection of a hitherto unknown microsporidian parasite that has a genetic affinity to the genus Spraguea, in the central nervous system of the amberjack.     

Immunization of channel catfish, Ictalurus punctatus Rafinesque, against Ichthyophthirius multifiliis (Fouquet): killed versus live vaccines

Abstract. Fish surviving infection with the pathogenic ciliated protozoan, Ichthyophthirius multifiliis (Fouquet, 1876), become resistant to subsequent infection by the parasite. The acquired immunity suggests that development of a vaccine against the parasite may be possible. Because of the advantages of immunoprophylaxis for treatment of the disease, an effort has been made to determine whether fish exposed to killed parasite preparations can resist subsequent lethal challenge. Both the route of administration and the effects of stage specific antigens have been examined. Channel catfish vaccinated by intraperitoneal (i.p.) injection or bath immersion with killed I multifiliis tomites show 100% mortality following a standard challenge protocol. Similarly, 100% mortality was observed in test groups injected with tomite cilia. In both cases, a consistent difference in days to death between control and test group animals was observed. Although complete mortality was seen with fish injected with tomite preparations, fish vaccinated with killed trophonts (the feeding stage of the parasite) had a much greater degree of protection with approximately 50% of fish surviving an otherwise lethal challenge. Finally, animals injected intraperitoneally with live tomites showed nearly complete immunity and were identical in their response to fish which survive natural infection. The response of fish vaccinated with live parasites indicates that animals injected intraperitoneally can develop surface immunity and that i.p. injection is a suitable route of administration for potential I. multifiliis vaccines.     

利用AFLP技术筛选锯缘青蟹性别差异DNA片段

采用高盐和酚氯仿异戊醇 (PCI)结合法提取DNA ,利用AFLP技术 ,应用 5 2个引物组合 ,检测了锯缘青蟹 (Scyllaser rata)雌雄基因组DNA的多态性 ,筛选与锯缘青蟹性别相关的分子标记。实验中共扩增出 4 312条带 ,筛选出候选差异DNA片段 74 8条。这些差异DNA片段的获得 ,为研究锯缘青蟹性别的分子标记奠定了基础     

Humoral immune response of turbot, Scophthalmus maximus (L.), to antigens from Tetramicra brevifilum Matthews & Matthews, 1980 (Microspora)

Abstract. The humoral immune response of turbot, Scophthalmus maximus (L.), to antigens from the microsporean parasite Tetramicra brevifilum Matthews & Matthews, 1980, was studied. Thirty days after intraperitoneal immunization with whole T. brevifilum spores in Freund's complete adjuvant, double indirect ELISA indicated that initial production of antibodies to parasite surface antigens was considerably higher than production of antibodies to the antigens contained in a crude extract (CE) of spores. Following re-immunization without adjuvant on day 30, levels of antibodies to surface antigens gradually declined, whilst levels of antibodies to CE antigens increased. The antibody response of intraperitoneally immunized fish was characterized by Western blotting of total soluble antigens obtained by heating and reduction of T. brevifilum spores at 95–100°C in Tris-HCl buffer containing SDS and dithiothreitol: a series of bands with molecular weights between 20 and 53 kDa was recognized by immunized turbot sera. Four additional bands (with molecular weights between 15 and 18kdA) were recognized by serum from re-immunized fish. ELISA studies of sera from naturally infected fish revealed a surprisingly low incidence of strong T. brevifilum seropositivity (61% individuals); antibodies to surface antigens predominated in seropositive individuals. The low background response levels and high sensitivity of the ELISA used in this study indicate that the assay is of value for the monitoring of serum antibody levels in turbot. However, given the relatively low seropositivities observed in naturally infected turbot, particularly to CE antigens, the use of anti- T. brevifilum serum antibody levels for the diagnosis of infection by this parasite may lead to false negative results.     

应用PCR方法检测患“黄水病”锯缘青蟹中的血卵涡鞭虫

"黄水病"是目前锯缘青蟹养殖过程中的主要疾病之一,病蟹主要症状表现为肌肉白浊,体液呈土黄色或浊白色牛奶状。因该病流行范围广、发病率和死亡率高,给青蟹养殖业主造成巨大经济损失,严重地影响了青蟹养殖的健康发展。本研究从病蟹体液中发现大量疑似血卵涡鞭虫的寄生原虫,应用已建立的梭子蟹血卵涡鞭虫病的PCR检测方法对患"黄水病"青蟹进行检测。结果从患病青蟹组织的DNA中扩增出产物大小为585bp的特异性目的片段,经序列分析比较,与三疣梭子蟹上发现的血卵涡鞭虫的序列同源,同源性达99.7%。综合病原流行病学调查、组织病理学、电镜观察等分析结果,初步确定血卵涡鞭虫是引起养殖青蟹"黄水病"的重要病原。     

Cardiac myxosporiosis of pearl-spot, Etroplus suratensis (Bloch), due to Myxobolus etropli sp. nov

A new myxosporean, Myxobolus etropli sp. nov., was found to infect the bulbus arteriosus of Etroplus suratensis (Bloch) from brackishwater lagoons of Muttukkadu, Chennai coast, India. A survey from May 1993 to October 1994 revealed a prevalence rate of 33.7% of this parasite. Macroscopic discoloured foci/cysts were seen in the bulbus arteriosus of the fish. The parasite showed strict host and site specificity. Histopathology showed that the infection was restricted to the bulbus. This is the first report of a myxosporean from E. suratensis.     

锯缘青蟹亲蟹选择及培育技术

亲蟹质量及亲蟹培育是锯缘青蟹人工育苗的基础。本文就亲蟹的选择、亲蟹暂养期的环境、亲蟹的饵料及饲养条件进行了探讨。在相同的理化因子条件下,海捕亲蟹的抱卵率(62.13％)大约为养殖亲蟹抱卵率(26.67％)的2.31倍。     

Dietary modulation of humoral immune response and anaemia in rainbow trout, Oncorhynchus mykiss (Walbaum), infected with Cryptobia salmositica Katz, 1951

Abstract. The humoral immune response in Cryptobia-salmositica -infected rainbow trout on pantothenic-acid-deficient/low protein (19%) diets was depressed. There was no significant difference in parasitaemia between fish on the pantothenic-acid-deficient diet and those on the supplemented diet; however, the parasitaemia in fish on 19% protein diet was significantly lower than that in fish on 38% protein diet. The low humoral responders, in contrast to the high responders (on 38 and 29% protein diets), had less severe anaemia and the packed cell volume at the end of the experiment was not significantly different from that at 2 weeks post-infection. Red cells from the low responders were Coombs' negative while those from the high responders were Coombs' positive from 4 to 6 weeks post-infection. The authors suggest that, in the low responders, red cell destruction is essentially due to the'lytic component' of the parasite antigen in contrast to the involvement of both the ^llytic' and the 'immune complex-forming' components in the high responders. The present study indicates that certain deficient diets are useful in modulating the parasitaemia and also in decreasing red cell destruction. Therefore, dietary modification may help in altering the course of such infections and the progression of the disease.     

Cutaneous myxidiosis in European eel,Anguilla anguilla (Linnaeus, 1758): histopathology,histochemistry and laminin immunohistochemistry

Histopathology, histochemistry and immunohistochemistry of the integument of European eel, Anguilla anguilla (Linnaeus, 1758), infected by Myxidium sp. are reported. Skin samples from affected and unaffected eels were dissected, formalin fixed, paraffin embedded, sectioned and stained with H&E, Periodic acid–Schiff's staining method, Alcian Blue 8 GX pH 2.5/Periodic acid–Schiff's and McCallum‐Goodpasture's Gram stain. Moreover, immunohistochemistry was performed using a primary polyclonal laminin antibody. Histologically, cysts (diameter 2–3 mm) were observed mainly under the scale pockets, encircled by a thin collagen layer, lined by elongated, flattened fibroblasts and containing bipolar, PAS‐ and Gram‐positive spores with opposite polar capsules. The epidermis stretched by the underlying cyst appeared dysplastic, thinned with a significant reduction in mucous cells number. Only inconsistent and aspecific inflammatory reaction was noted around the cysts at the dermis/epidermis interface. Intense laminin‐like protein immunolabel was documented in the plasmodial ectoplasm and related to host anergia. This was the first report of laminin immunolabel in a member of the Myxozoa. Epidermal dysplasia represents likely an aspecific response against the underlying tensile force exerted by the developing parasite cyst, while fibroblast and collagen encapsulation denote a parasite‐driven host response protecting, rather than harming, the encircled parasite.     

Dietary protein requirement of giant mud crab Scylla serrata juveniles fed iso-energetic formulated diets having graded protein levels

The protein requirement of juvenile mud crab Scylla serrata (body weight=0.25±0.051 g, carapace width=9.3±0.04 mm) fed with different iso-energetic, iso-lipidic diets with graded protein levels (15–55% crude protein at 5% intervals) was determined. The feeding trial was conducted for a period of 63 days to determine the minimum and optimum protein requirement of juvenile S. serrata . The crabs fed with 15% and 20% dietary protein levels showed 100% and 12.5% of mortalities respectively. The mortalities observed in the above treatments were associated with the prolonged intermoult duration (46 and 32 days respectively). All other treatments recorded 100% survival. The best growth performance as well as the nutrient turn-over was recorded in crabs fed with 45% crude protein in the diet. Second-order polynomial regression of specific growth rate (SGR) as well as body protein gain vs. dietary protein levels suggested that 46.9–47.03% dietary protein is required for the best growth response and protein deposition in juvenile S. serrata . An extrapolation of 'SGR' and 'daily protein gain' upon the 'dietary protein level' axis ( Y =0) showed that 14.7–16.2% dietary protein is necessary for the minimum maintenance metabolism.     

Eimeria scardinii Pellérdy and Molnár, 1968 in the kidneys of Rutilus rutilus (L,) and Scardinius erythrophthalmus (L.) from northern Greece

Abstract. The occurrence of Eimeria scardinii in roach, Rutilus rutilus (L.), and rudd, Scardinius erythrophthalmus (L.), from a lake in northern Greece is reported. The coccidium was found in the renal haemopoiettc tissue, mainly in the form of mature sporulated oocysts. The morphology and dimensions of the parasite, as well as the host reaction, were very similar in both species of fish examined. The overall prevalence of infection in roach and rudd was 6·67% and 6·3%, respectively. The prevalence of infection of roach was higher during summer (22·22%) but less than 5% in other seasons.