Pathogenic threats and probiotic use in larviculture of the scallop,Pecten maximus

During a series of experiments, bacteriological elements in scallop larval rearing were investigated: larvae susceptibility to pathogens as a function of their age, and the use of probiotic bacteria during larviculture. Younger larvae (d5 PF) were highly more susceptible to pathogenic‐challenge than their older siblings, which were challenged at an older age (d15 PF). A challenge with 10⁴ CFU mL^?1 of V. pectenicida killed 100% of d5 PF larvae 7 days following challenge, yet killed only 9% of d15 PF larvae 9 days following challenge. Use of the probiotics Phaeobacter gallaeciensis, Alteromonas macleodii 0444 and Neptunomonas sp. 0536, provided for larger larvae, a high yield of competent larvae and, perhaps more importantly, protection against pathogen‐challenge similar to levels achieved from antibiotic use. When challenged with V. pectenicida, d29 survivals were 20.3%, 85.1% and 75.0% respectively for control (no probiotic), antibiotic treated, and ‘probiotic mix’ administered larvae. Use of potential probiotic Pseudoalteromonas sp. D41 appeared to hinder scallop larvae. Future use of probiotics in scallop larval rearing would benefit from combined use of P. gallaeciensis, A. macleodii 0444 and Neptunomonas sp. 0536.     

Increased gonad growth of the purple sea urchin (Strongylocentrotus purpuratus) fed the giant kelp (Macrocystis pyrifera) and the sea lettuce (Ulva lactuca) enriched with nutrients

Adult (Strongylocentrotus purpuratus) purple sea urchins were fed giant kelp (Macrocystis pyrifera) and sea lettuce (Ulva lactuca) macroalgae that were unenriched and enriched with nutrients. Urchins were fed over 10 weeks (5% body weight per day) under laboratory conditions to determine the effect of macroalgae species and treatment (enriched and unenriched) on growth, gonad index and gonad quality. Significant differences were observed when urchins were fed enriched Macrocystis pyrifera (MPE). Gonad weight increased from 0.7 ± 0.1 g (mean ± SD) to 5.9 ± 1 g, and gonad index increase from 3.5 ± 1.5% to 14.9 ± 2.3% over 10 weeks. Significant differences were also noted in gonad quality, and gonad colour and firmness were better in urchins that were fed unenriched M. pyrifera (MPUE) and MPE – whereas texture did not differ between M. pyrifera treatments and unenriched U. lactuca (ULUE). With regard to taste, gonads were sweeter in MPE‐fed individuals. Enriched U. lactuca (ULE) failed to grow gonads significantly; as a result, their quality was unacceptable for the market. These results suggest that S. purpuratus experience increased gonad production and quality in short periods when fed nutrient‐enriched M. pyrifera.     

Evaluation of dried macrophytes as an alternative diet for the rearing of the sea urchin Paracentrotus lividus (Lamarck, 1816)

The aim of this work was to assess the potential use of different dried macroalgae as food in the rearing of Paracentrotus lividus. Growth, consumption and food conversions were compared in adult sea urchins fed with fresh or dried thalli of four macroalgae species. Six experimental diets were tested: (a) fresh Palmaria palmata; (b) fresh Saccharina latissima; (c) dry P. palmata; (d) dry S. latissima; (e) dry Laminaria digitata and (f) dry Grateloupia turuturu. Linear growth rates were similar for all treatments. Specific growth rate was higher in sea urchins fed with fresh P. palmata, but no difference was found between animals fed with fresh S. latissima and those fed with dried diets. Regarding daily food consumption (DFC), sea urchins consumed the same amount of dried macroalgae as fresh but exhibited a higher food conversion efficiency (FCE) when fed with fresh P. palmata. However, this FCE was only significantly higher when compared to sea urchins fed with dry L. digitata. Dried G. turuturu is not a suitable diet due to its rapid degradation after rehydration. The results suggest that P. lividus adults can be reared on dried macroalgae thalli without detriment to their somatic growth, especially over short periods. The low cost of feeding sea urchins with this diet could help small shellfish farmers to diversify their production into echinoculture.     

The role of rpoS in the regulation of Vibrio alginolyticus virulence and the response to diverse stresses

Vibrio alginolyticus is a leading aquatic pathogen, causing huge losses to aquaculture. rpoS has been proven to play a variety of important roles in stress response and virulence in several bacteria. In our previous study, upon treatment with Cu²⁺, Pb²⁺, Hg²⁺ and low pH, the expression levels of rpoS were downregulated as assessed by RNA‐seq, while impaired adhesion ability was observed, indicating that rpoS might play roles in the regulation of adhesion. In the present study, the RNAi technology was used to knockdown rpoS in V. alginolyticus. In comparison with wild‐type V. alginolyticus, RNAi‐treated bacteria showed significantly impaired abilities of adhesion, growth, haemolytic, biofilm production, movement and virulence. Meanwhile, alterations of temperature, salinity, pH and starvation starkly affected rpoS expression. The present data suggested that rpoS is a critical regulator of virulence in V. alginolyticus; in addition, rpoS regulates bacterial adhesion in response to temperature, pH and nutrient content changes. These are helpful to explore its pathogenic mechanism and provide reference for disease control.     

Recent update on the prevalence of Vibrio species among cultured grouper in Peninsular Malaysia

Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene.     

Investigations on the role of the salmon louse,Lepeophtheirus salmonis (Caligidae), as a vector in the transmission of Aeromonas salmonicida subsp. salmonicida

A bacteria–parasite challenge model was used to study the role of sea lice, Lepeophtheirus salmonis (Copepoda), as a vector of Aeromonas salmonicida subsp. salmonicida. Three hypotheses were tested: (i) L. salmonis can acquire A. salmonicida subsp. salmonicida via water bath exposure; (ii) L. salmonis can acquire the bacteria via parasitizing infected Atlantic salmon, Salmo salar; and (iii) L. salmonis can transmit the bacteria to naïve Atlantic salmon via parasitism. Adult L. salmonis exposed to varying A. salmonicida subsp. salmonicida suspensions (10¹–10⁷ cells mL^?1) for 1.0, 3.0 or 6.0 h acquired the bacteria externally (12.5–100%) and internally (10.0–100%), with higher prevalences associated with the highest concentrations and exposures. After exposure to 10⁷ cells mL^?1, viable A. salmonicida subsp. salmonicida could be isolated from the external carapace of L. salmonis for 120 h. Lepeophtheirus salmonis also acquired the bacteria externally and internally from parasitizing infected fish. Bacterial transmission was observed only when L. salmonis had acquired the pathogen internally via feeding on ‘donor fish’ and then by parasitizing smaller (<50 g) ‘naive’ fish. Under specific experimental conditions, L. salmonis can transfer A. salmonicida subsp. salmonicida via parasitism; however, its role as a mechanical or biological vector was not defined.     

Extreme ocean acidification reduces the susceptibility of eastern oyster shells to a polydorid parasite

Ocean acidification poses a threat to marine organisms. While the physiological and behavioural effects of ocean acidification have received much attention, the effects of acidification on the susceptibility of farmed shellfish to parasitic infections are poorly understood. Here we describe the effects of moderate (pH 7.5) and extreme (pH 7.0) ocean acidification on the susceptibility of Crassostrea virginica shells to infection by a parasitic polydorid, Polydora websteri. Under laboratory conditions, shells were exposed to three pH treatments (7.0, 7.5 and 8.0) for 3‐ and 5‐week periods. Treated shells were subsequently transferred to an oyster aquaculture site (which had recently reported an outbreak of P. websteri) for 50 days to test for effects of pH and exposure time on P. websteri recruitment to oyster shells. Results indicated that pH and exposure time did not affect the length, width or weight of the shells. Interestingly, P. websteri counts were significantly lower under extreme (pH 7.0; ~50% reduction), but not moderate (pH 7.5; ~20% reduction) acidification levels; exposure time had no effect. This study suggests that extreme levels – but not current and projected near‐future levels – of acidification (?pH ~1 unit) can reduce the susceptibility of eastern oyster shells to P. websteri infections.     

Spat availability of commercial bivalve species recruited on artificial collectors from the northern Gulf of California. Seasonal changes in species composition

This study reports a year‐round recruitment of spat of four commercial bivalve species; Pteria sterna, Euvola vogdesi, Pinctada mazatlanica and Pinna rugosa collected in the region of Puerto Peñasco, north‐eastern coast of the Gulf of California. Bimonthly recruitment of commercial bivalve spat on netlon^® collectors was evaluated for six sites from June 2007 to August 2008. To describe spat recruitment abundances with environmental parameters, sea surface temperature (°C) and surface chlorophyll a concentration (mg m^?3) were characterized by means of monthly Aqua/MODIS satellite data. For each species a repeated measures anova was used to evaluate differences in the number of spat between months, sites and depths. Maximum sea surface temperature was recorded in August–September (~31.5°C) and the minimum in January–February (~15°C), while the minimum surface chlorophyll a was observed in June–September (mean range = 1.5–2 mg m^?3) and the maximum in January–March (mean range = 2–5 mg m^?3). Spat recruitment showed distinct patterns; P. sterna can be characterized as having a Winter–Spring pattern, E. vogdesi a winter pattern, while P. mazatlanica and P. rugosa a summer spat recruitment pattern. This information constitutes part of the fundamental data needed for the development of aquaculture and conservation initiatives in the region based on wild spat supply.     

SNP discovery and gene annotation in the surf clam Mesodesma donacium

The main objective of this research was to identify single‐nucleotide polymorphisms (SNPs) from an Expressed Sequences Tags (EST) data set generated by 454 pyrosequencing in the soft clam Mesodesma donacium. A total of 180 159 ESTs were yielded from a M. donacium cDNA library. De novo assembly was performed using stringent calling parameters, producing 10 178 contigs and 41 765 singletons. Here, a total of 2594 SNPs were discovered related to 613 consensus sequences, achieving a frequency of 1 SNPs per 260 bp. SNP variants showed that A/G, A/T and C/T were the most abundant among the identified polymorphisms. We validated a total of 12 SNPs loci by HRMA for annotated genes such as heat shock protein‐70 and the translation elongation factor 1‐alpha. The Gene Ontology analysis regarding molecular function level revealed that sequences with SNPs were mainly classified to protein and nucleotide binding, as well hydrolase activity, ion binding and oxidoreductase activity. Further, biological processes like cellular and metabolic process, biogenesis, localization and biological regulation were highly annotated. The most expressed genes were related to the mitochondrial electron transport chain, senescence‐associated protein, ubiquitin and actin. Interestingly, some relevant genes related to immune response and biomineralization showed a high abundance, such as tumor necrosis factor (TNF)‐alpha‐receptor‐like protein, serine protease inhibitor, heat shock protein, aragonite‐binding protein and ferritin. This study contributes to relevant genes associated with functional polymorphisms and gives an overview for future genetic investigations.     

Effect of Ocimum sanctum Linn. (Tulsi) extract on the immunity and survival of Labeo rohita (Hamilton) infected with Aeromonas hydrophila

A 60 days study was conducted to evaluate the efficacy of water extract of Ocimum sanctum Linn. leaf on the immune response and disease resistance of Labeo rohita fingerlings against the Aeromonas hydrophila infection. Ocimum sanctum extract was incorporated in the diets (at 0.0%, 0.05%, 0.1%, 0.2%, 0.5% and 1%) of Labeo rohita, rohu fingerlings (6.6 ± 0.013 g). After 42 days blood, plasma and serum were sampled to determine super oxide anion production, lysozyme activity, total immunoglobulin in plasma, blood glucose, serum total protein, albumin, globulin, albumin:globulin ratio, WBC, RBC, haemoglobin content. Fish were challenged with A. hydrophila after 42 days and mortalities were recorded over 18 days post infection. The results demonstrate enhanced super oxide anion production, lysozyme activity, total immunoglobulin in plasma, serum total protein, globulin, total RBC counts, total WBC counts and haemoglobin content (P < 0.05) in treatments group compared with control group. Dietary O. sanctum extracts of 0.2% showed significantly (P < 0.05) higher protection relative percentage survival (RPS 40.00 ± 5.773%) against A. hydrophila infection than control. These results indicate that O. sanctum leaf extract stimulates the immunity and makes L. rohita more resistant to bacterial infection (A. hydrophila).     

Use of planted biofilters in integrated recirculating aquaculture‐hydroponics systems in the Mekong Delta,Vietnam

The feasibility of using planted biofilters for purification of recirculated aquaculture water in the Mekong Delta of Vietnam was assessed. The plant trenches were able to clean tilapia aquaculture water and to maintain good water quality in the fish tanks without renewal of the water. NH₄‐N was removed efficiently in the plant trenches, particularly in the trenches with Canna glauca L., probably because of plant uptake and nitrification–denitrification. Plant uptake constituted 6% of N and 7% of P in the input feed. Approximately 1.0 m³ of water was needed per kg of fish produced, and 370, 97 and 2842 g fresh aboveground biomass of Ipomoea aquatica Forssk., Lactuca sativa L. and C. glauca, respectively, were produced. The leafy vegetables provide some extra income besides fish products, whereas C. glauca provides nice flowers and contributes to a significant nutrient removal with annual uptake rates of 725 kg N and 234 kg P ha^?1 year^?1. This research demonstrates that integrated recirculating aquaculture‐hydroponics (aquaponics) systems provide significant water savings and nutrient recycling as compared with traditional fish ponds.     

Histopathological changes on the gills of asp (Aspius aspius) and European catfish (Silurus glanis) caused by Lamproglena pulchella and a Lamproglena sp. (Copepoda: Lernaeidae), respectively

In a parasitology survey of Hungarian fishes, heavy infections of parasitic copepods Lamproglena pulchella and a Lamproglena sp. were found in the gills of the asp and the European catfish, respectively. Individuals of both fish species were emaciated and infected with hundreds of Lamproglena. Copepods located close to the tip of gill filaments and formed a depression at the attachment sites. In histological sections, cell degenerations and local haemorrhages were present adjacent to the maxillipeds and where the maxillary claws pierced the gill tissue. Around maxillae and in the midgut of the Lamproglena, damaged piscine blood cells and remains of the gill tissue were observed. Host reaction was expressed by proliferation of epithelioid cells, increase in both number and size of goblet and mast cells and formation of giant cells.     

Ecology and conservation of the American eel in the Caribbean region

The majority of American eel, Anguilla rostrata LeSueur, knowledge is derived from temperate regions in the United States and Canada, with little known from its tropical Caribbean distribution. Findings of original research on American eel distribution, abundance, population biology, habitat ecology and threats from the Caribbean island of Puerto Rico were synthesised. American eel were captured from 48 of 116 sites (41.4%) in 26 of 49 river basins (53.1%) during 2005–2016, and it was extirpated upstream of dams and migration barriers >3.0 m high (38.9% of habitat). Mean density and biomass were 438.9 fish/ha and 23.44 kg/ha, respectively. Upstream habitats favoured larger individuals, and females were larger than males. The swim‐bladder parasite Anguillicoloides crassus Kuwahara, Niimi & Hagaki was not found in 120 eels examined. Realised threats include dams and other migratory barriers, habitat loss and alteration and pollution; exotic species and commercial fishing are impending threats; and the least understood is climate change.     

Multiple dietary administrating strategies of water hyacinth (Eichhornia crassipes) on enhancing the immune responses and disease resistance of giant freshwater prawn,Macrobrachium rosenbergii

The different products of Eichhornia crassipes leaves including dried E. crassipes powder (DEP), hot‐water treated E. crassipe (HTE), hot‐water extract of E. crassipe (ECE) and dreg of hot‐water extract of E. crassipe extract (dECE) were produced and incorporated into the diet of the prawn, Macrobrachium rosenbergii, as an immunostimulant. Results showed that prawn fed the HTE‐, ECE‐ and dECE‐containing diets for 4 months had increased total haemocyte count, different haemocyte count, phenoloxidase activity, respiratory bursts, superoxide dismutase activity, glutathione peroxidase activity especially in HTE and ECE treatments. The phagocytic activity and clearance efficiency against Lactococcus garvieae of prawn fed the HTE‐ and ECE‐containing diets were significantly higher than those of prawn fed the control diet at 2–4 months. The relative percentage survival of prawn fed the DEP‐, HTE‐, ECE‐ and dECE‐containing diets for 4 months following 144 h challenging with L. garvieae were 19.0%, 38.1%, 38.1% and 33.3%. It was concluded that E. crassipes leaves containing an active component which was easily extracted by hot water can enhance innate immunity and resistance against pathogen of M. rosenbergii by dietary long‐term administration, and the administration of HTE in the diet was the best strategy due to the availability and convenience.     

Declines in the energy content of yearling non‐native alewife associated with lower food web changes in Lake Michigan

Juveniles of non‐native alewife, Alosa pseudoharengus (Wilson), were collected in Lake Michigan in 1998, 1999, 2010, 2011 and 2013 to evaluate changes in energy content during a period of major ecosystem changes. Consistent with historical data, energy content of yearling alewife declined from late winter into late spring and was at its lowest point in June. Energy density and length‐adjusted, entire‐body energy were lower in 2010, 2011 and 2013 than in 1998 and 1999. Energy losses over the first winter in the lake were more severe for the 2010 year class (56% decrease) than for the 1998 year class (28% decrease). Alewife diets in late spring of 2010–2013 reflected the loss of major prey such as Diporeia spp. and a shift towards lower energy prey. The recent decline in energy content of yearling alewife can be linked to recent changes in productivity and abundance of key components of the lower food web of Lake Michigan following the dreissenid invasion.     

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.     

Bacillus subtilis CK3 used as an aquatic additive probiotics enhanced the immune response of crayfish Procambarus clarkii against newly identified Aeromonas veronii pathogen

To identify the aetiology of the disease outbreak in cultured Procambarus clarkii, a dominant bacteria strain from the diseased crayfish hepatopancreas was isolated. It was identified as Aeromonas veronii based on biochemical identification and 16S rDNA sequencing. The symptoms of artificially infected crayfish were similar to those of naturally infected P. clarkii. The A. veronii was sensitive to norfloxacin, ciprofloxacin, ceftazidime, cefuroxime and ceftriaxone antibiotics, while resistant to penicillin, ampicillin, oxacillin and piperacillin. Virulence genes such as the enterotoxin genes (act, alt and ast) and haemolytic toxin genes (hlyA and aerA) were detected. After A. veronii infection, the connections between some liver tubules disappeared, the vacuoles appeared in the brush border and the mitochondria were enlarged. The antagonistic action of previous identified B. subtilis CK3 against A. veronii was also detected. The supernatant of B. subtilis CK3 exhibits a significant bactericidal effect on A. veronii. After B. subtilis CK3 immersion, the antioxidant enzymes and immune-related enzyme activities in hepatopancreas were significantly higher than control. The accumulative mortality caused by infection of A. veronii can be significantly reduced by adding B. subtilis CK3 into the aquatic water. These results demonstrated that B. subtilis CK3 could act as a water additive to improve the immune response of P. clarkii against newly identified A. veronii. The present study will provide a new way for the prevention and control of crayfish bacterial diseases and also provide technical support for the healthy cultivation of P. clarkii.     

Effects of native medicinal herbs (Alternanthera sessilis,Eclipta alba and Cissus quadrangularis) on growth performance,digestive enzymes and biochemical constituents of the monsoon river prawn Macrobrachium malcolmsonii

The medicinal herbs such as Alternanthera sessilis, Eclipta alba and Cissus quadrangularis were used to assess the growth promoting ability, enhancement of muscle biochemical constituents and profiles of amino acid in the freshwater prawn, Macrobrachium malcolmsonii. The herbal powders were separately incorporated with basal diet, and these diets were fed to M. malcolmsonii postlarvae for a period of 90 days. At the end of experiments, the survival of prawn significantly improved in herbal incorporated feed. In addition, the growth parameters, such as weight gain, specific growth rate and protein efficiency rate were significantly (P < 0.05) higher in herbal diets fed groups when compared with control. The activities of digestive enzyme, biochemical constituents, profile of amino acid, energy utilization parameters, concentrations of vitamin C and E were significantly (P < 0.05) elevated in experimental diets fed groups when compared with control. The profiles of essential amino acids were also found elevated in herbal incorporated diets fed PL. Among the three herbs tested, C. quadrangularis incorporated diet fed group showed better performance followed by the E. alba and A. sessilis. The present study suggests that these herbs can be incorporated in artificial feed formulations for Macrobrachium culture.