Field and laboratory transmission studies of haemic neoplasia in the soft‐shell clam,Mya arenaria,from Atlantic Canada

A two‐year laboratory and field study was initiated in 2001 in response to mass mortalities associated with haemic neoplasia (HN) in 1999 in Prince Edward Island (PEI) soft‐shell clams, Mya arenaria. A laboratory proximity experiment (cohabitation) and an inoculation challenge were conducted with clams and mussels (Mytilus edulis). Three field exposure experiments were also conducted, in which naive clams were held in sediment (in trays) or out of sediment (in mesh bags) at three high HN prevalence sites on PEI. There was a conversion to HN positive in clams in the proximity experiment and in clams injected with whole blood and cell‐free homogenate, but not at statistically significant levels. No mussels or control clams became HN positive. There was a significant conversion to HN positive in as little as 24 and 58 days after transfer with clams held out of sediment and in sediment, respectively. The laboratory and field experiments’ results suggest that HN‐infected clams are spreading the disease through water from infected clams to naïve individuals and via transplantation from affected to unaffected sites. Some environmental conditions (e.g. abnormally high water temperature and hypoxia‐induced sea lettuce [Ulva lacteus] invasion) may make clams susceptible to infections or exacerbate the proliferation of HN.     

Protein‐sparing effects and LPL gene expressions of dietary lipids in the juvenile soft‐shelled turtle,Pelodiscussinensis

The aim of this study was to evaluate the effects of dietary lipids on protein‐sparing and lipoprotein lipase (LPL) mRNA expression in culture using 360 juvenile soft‐shelled turtles (Pelodiscussinensis) (initial weight 4.26 ± 0.14 g). The turtles were allotted to six diets with three duplicates for 60 days. A control diet with 46% protein and 55% fishmeal (CD) and five isonitrogenous diets with 41.3% protein and 45% fishmeal (F, S, L1, L2 and L3) were used, containing the following three lipid types: fish oil, soybean oil and mixed oils (soybean oil: fish oil = 1:1). The results showed that the survival rate was not affected by dietary lipids (P > 0.05). The highest weight gain and lowest feed coefficient ratio were seen in the L3 diets (P < 0.05). Turtles fed with L2 and L3 diets had lower superoxide dismutase activities, higher alanine aminotransferase activities and higher cholesterol concentrations than those exposed to other diets (P < 0.05). Hepatic LPL activity and LPL mRNA expression were higher in the L3 diets than in the other diets (P < 0.05). Overall, there were obvious protein‐sparing effects of dietary lipids and LPL mRNA expression was stimulated by high dietary lipids in soft‐shelled turtles in this study.     

Effects of fishmeal replacement with cottonseed meal protein hydrolysate on growth,digestion and intestinal histology of juvenile Chinese soft‐shelled turtle,Pelodiscus sinensis

A 60‐day feeding experiment was conducted to evaluate the effects of fishmeal (FM) replacement with cottonseed meal protein hydrolysate (CPH) on growth, digestion and intestinal histology of juvenile Chinese soft‐shelled turtle (Pelodiscus sinensis). Five diets were formulated to replace 0, 50, 100, and 150 g/kg fishmeal protein by CPH (CPH0, CPH5, CPH10, CPH15) and CPH15L (CPH15 with micro capsule‐L‐lysine). Weight gain, feed conversion rate and protein efficiency ratio showed no significant differences compared to control group (p > .05). The highest feed intake indicated in CPH15 (p < .05). The composition of whole‐body varied slightly in each groups (p > .05). The trypsin activity significantly elevated when dietary fishmeal protein was replaced by CPH at 30–90 g/kg (p < .05). A significantly higher lipase activities in CPH5 than control group (p < .05). The CPH5‐10 groups showed higher villus height than the other groups (p < .05). The microvillus length in turtles with CPH showed a significant increasing length (p < .05). The results indicated that replacing up to 90 g/kg of dietary fishmeal protein with CPH did not hamper growth or reduce feed intake of turtles. Moreover, CPH replaced 60 g/kg FMP can increase intestine digestive enzymes activities and improve intestinal development.     

Fate and legacy of an invasion: extinct and extant populations of the soft‐shell clam (Mya arenaria) in Grays Harbor (Washington)

1. The soft‐shell clam, Mya arenaria, was intentionally introduced to Washington State during the mid‐1870s. Presently, extensive shell deposits cover parts of the intertidal flats of Grays Harbor, an estuarine ecosystem that has received much attention because of its role as nursery ground for Dungeness crab (Cancer magister), a commercially significant species. 2. Preliminary observations revealed that shells in the deposits belonged to clams that were much larger than extant conspecifics. Shell deposits and extant populations have spatially disjoint distributions; few individuals are found living within the deposits. The purpose of our study was to determine the cause of the different size characteristics between extinct and extant clams, when and how the deposits originated, and why the extant population does not prosper in what appears to be prime habitat. 3. We (i) surveyed extant populations and shell deposits, (ii) followed the extant population through periodic sampling over a 3‐year period, (iii) investigated growth and survival in extant and extinct populations, (iv) experimentally manipulated growth conditions, and (v) searched archival sources for information about the origin of the deposits. 4. We conclude that extinct clams grew faster and lived longer, apparently because they occupied the best habitats available. Archival records suggest that the population expanded rapidly during the period around 20 years after its introduction, but was decimated between 1895 and 1897 by a mass‐mortality episode that resulted in the formation of the deposits. The population since then has never rebounded in its prime habitat, in spite of potential settlers being regularly available. 5. Dungeness crab larvae settle preferentially in the shell deposits, where the abundance of 0+ age juveniles is orders of magnitude higher than in the adjacent flats. We hypothesize that predation by juvenile crabs limits clam recruitment. 6. The introduction of Mya to Grays Harbor during the 1870s has had long‐lasting effects, which persist a century after their once large populations were decimated by a mass‐mortality event. Copyright © 2000 John Wiley & Sons, Ltd.     

Impact of sublethal di‐n‐butyl phthalate on the aquaculture fish species Nile tilapia (Oreochromis niloticus): histopathology and oxidative stress assessment

Phthalates such as di‐n‐butyl phthalate (DBP) and their esters are widely used plasticizers, their ubiquitous presence in daily life, inevitably leads to their restricted use due to important environmental pollution and health impacts and endocrine disruption potential. The aim of this study was to examine the effects of a sublethal concentration of 10 mg L^?1 DBP on haematocrit (HCT) values, gills and liver histology, malondialdehyde (MDA, 2‐thiobarbituric acid‐TBA reactivity) and reduced glutathione (GSH) levels in gills and liver tissue as oxidative stress biomarkers in the aquaculture fish species Nile tilapia (Oreochromis niloticus) after 24 (DBP‐24) and 96 (DBP‐96) h exposure. No differences were found between per cent HCT values in the 24 h exposure groups (P > 0.05). Response of antioxidant defence systems in liver and gill tissues of the fish were dependent on exposure duration and changed to a higher extent during 96 h. MDA levels in liver tissue increased in DBP treated fish in comparison to the control fish. However, the differences between the exposure and control groups were not significant (P > 0.05). A statistically significant decrease (P > 0.05) was recorded in gill MDA levels in the DBP‐96 group when compared to the control and DBP‐24 groups. The liver GSH levels were unchanged in the DBP treated fish. However, GSH levels were significantly lower in the gill tissue of the DBP‐96 group. Exposure to DBP caused several degenerative changes in the histology of gill and liver tissue. Gills displayed hyperaemia, epithelial lifting, oedema, talengiectasia, epithelial hyperplasia and fusion of secondary lamellae, whereas in liver several circulatory anomalies (hyperaemia, blood congestion and sinosoid dilatation) and vacuolization of hepatocytes were observed. Histopathological results demonstrated that the gills were more affected than the liver perhaps due to their direct contact with DBP.     

Acute effects of ammonia exposure on the plasma and haematological parameters and histological structure of the juvenile blunt snout bream,Megalobrama amblycephala,and post‐exposure recovery

In this study, we investigated the toxic effects of ammonia‐N on the plasma and haematological parameters and histological structure of blunt snout bream (Megalobrama amblycephala) juveniles. The fish (initial weight, 14.79 ± 0.01 g) were randomly sorted into six tanks (200 L), and each tank was stocked with 40 fish for culture. The juveniles were exposed to two ammonia‐N levels—0 mg/L (control group) and 25 mg/L (experimental group)—and sampled at 0, 6, 12, 24, 48 and 72 hr, and then they underwent 96 hr of post‐exposure recovery. The results showed that ammonia‐N had significant effects on the plasma and haematological parameters. The treatment group showed increased cortisol, plasma ammonia and haematocrit levels and white blood cell count with increasing exposure time, up to 24 hr, and then the levels and count decreased. A significantly higher plasma glucose level was observed in the treatment group at 12 hr. After 96 hr of post‐exposure recovery, all parameter levels decreased to the control levels. The fish displayed histopathological alterations in the gills, liver and kidney. The results indicate that the severity of the lesions clearly differed among the organs, with the liver showing the most extensive damage, followed by the gills and kidney. Adverse effects to physiological indicators and histological structure increased with increasing exposure time before 24 hr. The fish showed self‐regulation; however, the histological structure could not recover fully, the gill tissue showed irreversible changes and the kidney tissue exhibited the worst recovery ability.     

New Zealand rickettsia‐like organism (NZ‐RLO) and Tenacibaculum maritimum: Distribution and phylogeny in farmed Chinook salmon (Oncorhynchus tshawytscha)

A total of 777 fish from three growing regions of New Zealand Chinook salmon farms comprising of five sites were tested. Quantitative PCR was used to determine the distribution of New Zealand rickettsia‐like organism and Tenacibaculum maritimum. Genetic information from these bacteria were then compared with strains reported worldwide. Using this information, suggested associations of pathogens with clinically affected fish were made. NZ‐RLO was detected in two of the three regions, and T. maritimum was detected in all regions. Three strains of NZ‐RLO were identified during this study. Based on analysis of the ITS rRNA gene, NZ‐RLO1 appears to be part of an Australasian grouping sharing high similarity with the Tasmanian RLO, NZ‐RLO2 was shown to be the same as an Irish strain, and NZ‐RLO3 was shown be closely related to two strains from Chile. Based on multi‐locus sequence typing, the New Zealand T. maritimum was the same as Australian strains. NZ‐RLOs were detected more frequently in fish with skin ulcers than fish without skin ulcers. While additional research is required to investigate the pathogenicity of these organisms, this is the first time that NZ‐RLOs have been associated with the development of clinical infections in farmed Chinook salmon.     

Screening of a high bioflocculant‐producing bacterial strain from an intensive fish pond and comparison of the bioflocculation effects with Rhodococcus erythropolis

A high bioflocculant‐producing bacterial strain BX‐13 isolated from an intensive fish pond by multiple subcultures and measured through primary screening and rescreening using kaolin. The strain BX‐13 was identified to belong to the same branch as Bacillus subtilis by morphological, physiological and biochemical, 16S rDNA gene sequencing, Blast homology search and multiple sequence alignments. The bioflocculation potential of BX‐13 and Rhodococcus erythropolis was compared, using water from the intensive pond where BX‐13 was isolated. The results showed the number of colonies in the BX‐13 treatment was higher than that of the R. erythropolis treatment from day 10 to the end of the experiment (p < 0.05). The numbers of both BX‐13 and R. erythropolis in the water body peaked on day 15, with 2.23 × 10⁷ cfu/L and 9.90 × 10⁶ cfu/L respectively. The bioflocculation volume (BFV) in water increased and then stabilized in both BX‐13 and R. erythropolis treatments. The BFV in BX‐13 peaked on day 25 (52.13 ml/L) and R. erythropolis peaked on day 30 (22.63 ml/L). At the end of the experiment, the BFV of the BX‐13 treatment was higher than the R. erythropolis treatment (p < 0.05). In conclusion, B. subtilis BX‐13 has a very strong flocculation effect, and merits further research into its application for water quality regulation in aquaculture systems.     

Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

Terrestrial prey inputs to streams bordered by deciduous broadleaved forests,conifer plantations and clear‐cut sites in southwestern Japan: effects on the abundance of red‐spotted masu salmon

Terrestrial invertebrates falling from the riparian canopy are a major energy source for fishes in headwater streams. Because quantity and quality of such allochthonous resources can vary depending on riparian conditions, conversion of riparian forests to conifer plantations may affect stream productivity. We compared falling and drifting invertebrate abundances and the diet of masu salmon (Oncorhynchus masou) among stream reaches bordered by deciduous broadleaved forests, conifer plantations (Cryptomeria japonica), and clear‐cut sites in southwestern Japan. We also examined whether among‐reach variation in salmon abundance was related to the riparian vegetation types. The results indicated that, on an annual basis, falling inputs of terrestrial invertebrates at the broadleaved reaches were 2–4 times higher than those at the plantation and clear‐cut reaches. In nonwinter seasons, terrestrial invertebrates made up 40–60% and 30–90% of drift and masu salmon diets, respectively, and drifting invertebrate abundance was higher in the broadleaved reaches than in the plantation reaches. Furthermore, a multivariate analysis of salmon abundance showed that variation in salmon biomass was explained primarily by riparian vegetation type, with broadleaved and clear‐cut reaches having higher biomass than the plantation reaches. These results indicate that terrestrial invertebrates are an important resource for masu salmon, and suggest that streams bordered by conifer plantations receive lower terrestrial prey inputs, which results in lower salmon abundance. In regions where natural forests have been extensively converted to conifer plantations, forest management that allows and facilitates recovery of natural riparian stands is important.     

Co‐Infection with Pseudomonas anguilliseptica and Delftia acidovorans in the European eel,Anguilla anguilla (L.): a case history of an illegally trafficked protected species

Inspections by customs agents at Barcelona airport discovered 420 kg of contraband glass eels prepared for shipment to Hong Kong. After confiscation of these animals by police, they were transported to holding facilities to be maintained until after a judicial hearing. Upon arrival, they were separated into two groups and held under ambient flow‐through conditions in fresh water. During their captivity period, several peaks in mortality occurred and multiple bacterial strains were isolated from moribund animals. Sequencing of 16S rDNA was used to determine specific identity of the isolates. An initial isolation of Pseudomonas anguilliseptica was treated with oxytetracycline. A subsequent isolation of Delftia acidovorans proved resistant to oxytetracycline and was treated with gentamicin in combination with sulphadiazine–trimethoprim. Once the health condition of the animals was stabilized, they were partitioned into groups and subsequently released as part of a restocking effort for the species following the guidelines of Regulation (EC) 1100/2007 (Anon 2007). This represents the first record for both bacterial species in the host Anguilla anguilla in the Spanish Mediterranean.     

Dietary fulvic acid effects on survival and expression of immune‐related genes in Litopenaeus vannamei challenged with Vibrio parahaemolyticus

The effects of fulvic acid (FA) on survival and immune‐related gene expression were investigated in Litopenaeus vannamei challenged with Vibrio parahaemolyticus by immersion. Shrimp were fed with different dietary FA concentrations (1, 2, 4 and 6 g/kg feed) for 20 days (first bioassay) or 8 days (second bioassay, 2 g/kg feed of FA added every 2 days) and then challenged with V. parahaemolyticus. In a third bioassay, the expression of three immune‐related genes (translationally controlled tumour protein [TCTP], superoxide dismutase [SOD] and heat‐shock protein 70 [HSP70]) in haemocytes or hepatopancreas of experimental shrimp was measured by real‐time quantitative PCR at 0, 6, 12, 24, 48, 72 and 96 hr after FA (2 g/kg feed) administration. Fulvic acid increased survival at a concentration of 2 g/kg feed supplied every two days. Interestingly, TCTP gene expression was upregulated, whereas gene expression of SOD and HSP70 was downregulated. In conclusion, dietary fulvic acid improves survival in white shrimp challenged with V. parahaemolyticus and modulates the immune response. Therefore, FA merits further evaluation as prophylactic treatment in commercial shrimp farms.     

Sesbania aculeata leaf meal as replacer of de‐oiled rice bran in aquaculture feed: Growth,IGF‐1 expression,metabolic and biochemical responses in Cyprinus carpio (Linnaeus 1758)

A feeding trial was conducted to evaluate the effect of inclusion of Sesbania leaf meal in the diet of Cyprinus carpio as a replacer of de‐oiled rice bran (DORB). Bacillus subtilis was used to ferment the Sesbania leaf meal (SLM) to produce FSLM (fermented sesbania leaf meal). Fishes were fed with five isonitrogenous (30% CP) and isocaloric diets for 60 days and treatments were viz., C (30% DORB, 0% SLM), R15 (15% DORB, 15% SLM), R30 (0% DORB, 30% SLM), F15 (15% DORB, 15%FSLM) and F30 (0% DORB, 30% FSLM) by replacing DORB with SLM. A similar growth performance with control was observed in 15% FSLM group while there was a growth reduction in all the raw leaf meal fed groups. Similarly, the expression of insulin‐like growth factor‐1 (IGF‐1) was significantly (p < .05) higher in control and 15% FSLM fed groups. Growth and metabolic enzymes such as protease, amylase, ALT and AST activities were significantly higher in the fermented leaf meal fed groups compared to their non‐fermented counterparts. Antioxidant enzymes, serum transaminase enzymes (SGOT, SGPT) and total bilirubin (TBR) were found to be increased by enhanced level of RSLM, however, reduced by inclusion of FSLM. However, 30% inclusion level of FSLM even after fermentation showed detrimental effect on growth and metabolic response at cellular and molecular level. Thus, the results concluded that fermented Sesbania leaf meal (FSLM) using B. subtilis could replace 50% DORB (15% FSLM group) in the diet of C. carpio without affecting the growth and other physiological responses.     

Biotic and abiotic regulation of a low‐density stream‐dwelling brown trout (Salmo trutta L.) population: effects on juvenile survival and growth

The effects of biotic (density‐dependent) and environmental (flow and temperature) factors on the apparent survival, mean length and size variation of a low‐density brown trout population in the juvenile stage, that is, from their first summer (0+) to the end of the second year (1+), were determined. Apparent survival was negatively related to the age class density during the three periods (first summer, first winter and second summer). A significant interaction between the mean flow and 0+ density highlighted a gradient towards strong density dependence acting on fish loss (i.e., mortality or migration) with decreasing summer flow. Conversely, no density dependence was reported at higher mean flows. The mean length was determined by density‐dependent and density‐independent (temperature and flow) factors throughout the study period. The negative relationship between fish length and intracohort density was highly significant during the three periods. The yearling (1+) density was negatively related to 0+ fish length measured after the first summer, suggesting intercohort effects. A positive effect of temperature on fish length was observed. Mean length after the summer seasons (0+ and 1+ fish) was also positively related to mean flow. Fish size variation around the mean measured with the coefficient of variation (CV) increased with increasing 0+ densities, both at the end of the first summer and the first winter. Results suggested that density‐dependent and density‐independent factors acted jointly on apparent survival and growth with a predominance of biotic processes. We discussed the potential implications of density‐dependent regulations on growth and survival for population resilience after catastrophic events.     

Efficacy of Netminder® silicone release coating for biofouling reduction in bay scallop grow‐out and comparative effects on scallop survival,growth and reproduction

Netminder^®, a water‐based silicone barrier release coating, was highly effective in reducing biofouling of lantern nets during grow‐out/overwintering of bay scallops, Argopecten irradians irradians, in a large‐scale restoration programme in New York, USA. Compared with untreated nets, scallops held in Netminder^®‐coated nets experienced reduced survival after 8 months, probably due to higher loads of epibionts on their shells. However, this was countered by higher reproductive and overall condition of scallops in treated vs. untreated nets; there were no consistent differences in shell growth in different net treatments. Although further testing is recommended under a wider range of conditions, we conclude that for the purposes of our scallop restoration work, coating nets with Netminder^® would eliminate the need for a gear change in spring, thus reducing labour costs; the coating process should be refined so that Velcro closures of nets do not become sealed shut.