Bonamia in Ostrea angasi: Diagnostic performance,field prevalence and intensity

Bonamia spp. parasites threaten flat oyster (Ostrea spp.) farming worldwide. Understanding test performance is important for designing surveillance and interpreting diagnostic results. Following a pilot survey which found low Bonamia sp. intensity in farmed Ostrea angasi, we tested further oysters (n = 100–150) from each of three farms for Bonamia sp. using heart smear, histology and qPCR. We used a Bayesian Latent Class Model to assess diagnostic sensitivity (DSe) and specificity (DSp) of these tests individually or in combination, and to assess prevalence. Histology was the best individual test (DSe 0.76, DSp 0.93) compared to quantitative polymerase chain reaction (qPCR) (DSe 0.69, DSp 0.93) and heart smear (DSe 0.61, DSp 0.60). Histology combined with qPCR and defining a positive from either test as an infected case maximized test performance (DSe 0.91, DSp 0.88). Prevalence was higher at two farms in a high‐density oyster growing region than at a farm cultivating oysters at lower density. Parasite intensities were lower than in New Zealand and European studies, and this is probably contributed to differences in the performance of test when compared to other studies. Understanding diagnostic test performance in different populations can support the development of improved Bonamia surveillance programs.     

Two epizootic Perkinsus spp. events in commercial oyster farms at Santa Catarina,Brazil

Perkinsus spp. have been detected in various bivalve species from north‐east Brazil. Santa Catarina is a South Brasil state with the highest national oyster production. Considering the pathogenicity of some Perkinsus spp., a study was carried out to survey perkinsosis in two oyster species cultured in this State, the mangrove oyster Crassostrea gasar and the Pacific oyster Crassostrea gigas. Sampling involved eight sites along the state coast, and oyster sampling was collected during the period between January 2013 and December 2014. For the detection of Perkinsus, Ray's fluid thioglycollate medium (RFTM) and histology were used, and for the identification of the species, PCR and DNA sequencing were used. Perkinsus spp. was found by RFTM in C. gigas and C. gasar from São Francisco do Sul. This pathology was also detected in C. gasar from Balneário Barra do Sul both, by RFTM and histology. Perkinsus marinus was identified in C. gigas and C. gasar from São Francisco do Sul and Perkinsus beihaiensis in C. gasar from Balneário Barra do Sul. This is the first report of P. marinus in C. gigas from South America. Results of this preliminary study suggest that both oyster species tolerate the species of Perkinsus identified, without suffering heavy lesions.     

First report of Shewanella sp. and Listonella sp. infection in freshwater cultured loach,Misgurnus anguillicaudatus

A novel disease outbreak occurred among cultured loach, Misgurnus anguillicaudatus in China. The diseased fish displayed lethargy, loss of appetite, black discolouration, anasarca and ventral skin hyperaemia. Clinical signs were observed in most of the fish examined, which consisted of discoloured liver, swollen spleen and petechial haemorrhage in the intestine and peritoneal membranes. The causative agent was found to be two species of bacteria by experimental infection, identified as Shewanella sp. and Listonella sp. The result showed that these were likely to be Shewanella putrefaciens and Listonella anguillarum respectively. Bacterial identification consisted of physiological and biochemical tests as well as 16S rRNA sequence analysis. In addition, antibiotic susceptibility testing was performed, revealing that Shewanella putrefaciens and Listonella anguillarum were susceptible to enrofloxacin, ciprofloxacin, levofloxacin, pipemidicacid and norfloxacin. This is the first report of S. putrefaciens and L. anguillarum from cultured loach.     

Nutrient removal efficiency of Hydropuntia cornea in an integrated closed recirculation system with pink shrimp Farfantepenaeus brasiliensis

In this study, we have tested the effect of seaweed stocking density in an experimental seaweed biofilter using the economically important red seaweed Hydropuntia cornea integrated with the cultivation of the pink shrimp Farfantepenaeus brasiliensis. Nutrient removal efficiency was evaluated in relation to seaweed stocking density (2.5, 4, 6 and 8 g fw L^?1). Total ammonia nitrogen (TAN) was the main nitrogen source excreted by F. brasiliensis, with concentrations ranging from 41.6 to 65 μM of NH₄⁺‐N. H. cornea specific growth rates ranged from 0.8 ± 0.2 to 1.4 ± 0.5% day^?1 with lowest growth rates at higher seaweed stocking density (8 g fw L^?1). Nutrient removal was positively correlated with the cultivation densities in the system. TAN removal efficiency increased from 61 to 88.5% with increasing seaweed stocking density. Changes in the chemical composition of the seaweed were analysed and correlated with nutrient enrichment from shrimp effluent. The red seaweed H. cornea can be cultured and used to remove nutrients from shrimp effluents in an integrated multi‐trophic aquaculture system applied to a closed recirculation system. Recirculation through seaweed biofilters in land‐based intensive aquaculture farms can also be a tool to increase recirculation practices and establish full recirculation aquaculture systems (RAS) with all their known associated benefits.     

Functional identification of ToLAAO genes and polymorphism association analysis of Cryptocaryon irritans resistance in Trachinotus ovatus

‘Marine white spot disease’ is caused by Cryptocaryon irritans infection and can lead to high mortality in Trachinotus ovatus. L-Amino acid oxides (LAAOs) play a key role in antibacterial activity and parasitic activity. To investigate the function of the LAAO (ToLAAO) and LAAO-like (ToLAAO-like) genes of T. ovatus, this study explored the sequence characteristics and relationship between polymorphisms and traits of anti-C. irritans. The ToLAAO and ToLAAO-like ORF sequences obtained from the whole genome of T. ovatus were 1563 and 1584 bp, which encoded 520 and 527 amino acids respectively. Both sequences contained a highly conserved flavin adenine dinucleotide-binding domain and a similar amino oxidase domain. Sequence multiple alignment analysis showed that ToLAAO and ToLAAO-like had the highest homology to the LAAO sequence of Larimichthys crocea. Quantitative real-time polymerase chain reaction (qRT-PCR) results showed that ToLAAO and ToLAAO-like mRNA were generally expressed in 10 tissues. ToLAAO mRNA was highly expressed in the testis, while ToLAAO-like mRNA was highly expressed in muscle tissue. After C. irritans infection, ToLAAO and ToLAAO-like mRNA were significantly upregulated in the skin and spleen, while only ToLAAO mRNA was significantly upregulated in the liver and head kidney, and only ToLAAO-like mRNA was significantly upregulated in the gills. Five SNP sites were identified from the ToLAAO and ToLAAO-like genomic sequence fragments, and two sites (6200C/T and 6237G/A) of LAAO were significantly associated with resistance to C. irritans. These results suggest that ToLAAO and ToLAAO-like genes play crucial roles in defending against the immune response to C. irritans.     

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.     

Recent advances of genome mapping and marker‐assisted selection in aquaculture

Aquaculture is the fastest growing sector in agriculture. Substantial genetic gains have been achieved in a few cultured species using conventional selective breeding approaches. However, the majority of fish and shellfish species remain in their wild state. Due to the recognition of the enormous potential of marker‐assisted selection (MAS) to speed up genetic gain through early selection, aquaculture scientists have constructed linkage maps in over 40 species and mapped quantitative trait loci (QTL) for important traits in over 20 species since the 1990s. Although MAS and genomic selection (GS) have not been widely used in aquaculture, their application in breeding programmes is expected to be a fertile area of research. In this paper, I summarized the recent advances of linkage and QTL mapping, as well as MAS in aquaculture species. I also discussed the potentials of genome‐wide association studies (GWAS) and GS in aquaculture species.     

Time kinetics of fatty acid changes in phospholipids following enrichment and starvation of Artemia franciscana with a main focus on docosahexaenoic acid

The main objective was to study time kinetics of change in important highly unsaturated fatty acids (HUFAs) in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) of Artemia franciscana nauplii and juveniles following enrichment and subsequent starvation. Samples of Artemia nauplii were taken at variable times (0.5–24 h) following enrichment and starvation. Samples of Artemia juveniles were taken after 2, 3 and 4 days of cultivation. No docosahexaenoic acid (DHA) was found in PC and PE of Artemia nauplii during the first hour of enrichment, while a significant (P < 0.05) increase was found in total lipids (TLs). The content of DHA in PC and PE increased thereafter steadily from 1 to 8 h of enrichment. DHA in PC and PE during enrichment (1–8 h) and following starvation (8–24 h), respectively, increased and decreased significantly (P < 0.05), but at a lower rate than that in TL. Moreover, juvenile Artemia (2–4 days) contained a relatively low level of DHA in TL compared with enriched Artemia nauplii, but the content of DHA in PC and PE was similar. The results open perspectives for both industry and science. For scientific studies, the lag phase in HUFA enrichment makes it possible to produce Artemia nauplii with variable relative HUFA enrichments in phospholipids and TL.     

Multiscalar land suitability assessment for aquaculture production in Uruguay

This article presents a highly detailed territorial assessment for aquaculture production in Uruguay and a methodology to identify the most suitable areas in order to develop the activity. Furthermore, it proposes the basis to generate a dynamic modelling tool of high spatial resolution to support the decision‐making process regarding national aquaculture activities. The modelling structure of aquaculture suitability was developed by the construction of a hierarchical model, combining a multicriteria assessment and a geographic information system. Forty‐one attributes were integrated in six aptitude models: one model for each one of the five common production systems intended to be the most viable in Uruguay and a sixth aptitude indicator that summarizes those five systems, representing the maximum suitability for aquaculture activities. In addition, the hierarchical model allows measurement and identification of relative weights of each model, according to the planning scale, ranging from basin to cell.     

Development of an in vitro model to assess protein bioavailability in diets for common octopus (Octopus vulgaris)

An in vitro model designed to assess protein bioavailability in diets for growing Octopus was developed. The model required a previous assessment of some functional features of protein digestion in this species like the main producing organs, optimum pH for activity and total production per g tissue. The main producing organs identified were the salivary glands and the hepatopancreas (HP), being optimum pH for protease activity quite different in both organs (mostly alkaline in the posterior salivary glands and acid in the HP). In spite of the high‐specific protease activity measured in the salivary glands, a major role of the HP in protein hydrolysis is suggested due to the much bigger size of this viscera. All this information was used as a basis to develop an in vitro two‐step hydrolysis process, which simulated protein hydrolysis performed by these two organs using the Octopus enzymes. The assay was used to evaluate differences in amino acid bioavailability from several protein sources (casein, gelatin, fish meal, squid meal and krill meal) that could be used as feed ingredients for this species. As significant differences were detected both in total amount and in rate of release of the amino acids from such proteins, the model is proposed as a complementary tool in the selection and nutritional evaluation of protein ingredients to be used in diets for Octopus.     

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.     

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.     

Utilization of Caridina nilotica (Roux) meal as a protein ingredient in feeds for Nile tilapia (Oreochromis niloticus)

The effects of replacing fish meal with Caridina nilotica as a protein ingredient on growth performance, nutrient utilization, carcass, proximate composition and economic benefits in Nile tilapia (Oreochromis niloticus) culture was evaluated. Replacement of the FM with C. nilotica was done at 25%, 50%, 75% and 100% (D25, D50, D75 and D100) and the substitution effects was compared with the control diet (D0, 0% C. nilotica). After 140 days of culture, the best growth performance, nutrient utilization and economic benefits occurred in fish groups fed diets with 25% C. nilotica inclusion. However, growth performance in fish fed diets D50 and D75 were comparable with the control (P > 0.05). At 100% substitution level of FM with C. nilotica, the growth performance and fish survival was lower than control. Protein and lipid contents in the fish and their digestibilities were highest in diet D25 and decreased with increasing levels of substitution of FM with C. nilotica. This study demonstrate that utilization of local protein sources (C. nilotica) can be effectively used to replace up to 75% of FM in the diets without compromising growth performance, survival, nutrient utilization and economic benefits in O. niloticus culture.     

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.     

Inbreeding and disease in tropical shrimp aquaculture: a reappraisal and caution

The disease crisis facing shrimp aquaculture may be propelled, in part, by an interaction between management practices that cause inbreeding, and the amplification by inbreeding of susceptibility to disease and environmental stresses. The study describes and numerically simulates gene flow from Penaeus (Litopenaeus) vannamei hatcheries that employ a ‘Breeder Lock’ to discourage use of their PL as breeders, through ‘copy hatcheries’ that breed the locked PL, to inbred shrimp in farm ponds. Re‐analysis of published data shows that inbreeding depression under stress is exceptionally strong in shrimp. Inbreeding is currently overlooked as a problem because: (1) procedures recommended for well‐managed hatcheries do not consider their implications for the copy hatcheries that supply most farmed shrimp (estimated 70%), (2) inbreeding in hatcheries is often reported as zero even though zero is the mathematical expectation of the usual estimator (Fis, fixation index) whatever the true genealogy of the broodstock. Simulation shows, however, that inbreeding can be estimated with Wang's trioML estimator, that Fis can differentiate Breeder Locked from copy PL and that simple tests can verify the lock status of PL. The importance of inbreeding should be re‐evaluated in the context of disease and environmental stress. Unrecognized inbreeding may increase the incidence, prevalence and lethality of WSSV, IHHNV, EMS (AHPND) and other diseases.     

clpV is a key virulence gene during in vivo Pseudomonas plecoglossicida infection

Interaction between bacterial pathogen and aquatic animal host is exceedingly complex, which involves large dynamic changes in gene expression during different stages of the disease. However, research on identifying key virulence genes based on the dynamics of gene expression changes of a one‐sided bacterial pathogen in tissue has not been reported so far across different stages of infectious disease. The clpV for the T6SS of Pseudomonas plecoglossicida was identified for a candidate for key virulence gene based on dynamic changes of gene expression. For the Epinephelus coioides infected using clpV‐RNAi strain, no deaths were observed up to 20 dpi. The spleens, kidneys and livers of all the E. coioides that received clpV‐RNAi strain failed to develop visible nodules at 5–8 dpi, with the swelling gradually disappearing. The burdens of clpV‐RNAi strain in the spleen and blood were greatly reduced at most of the time points after injection, and the burdens of clpV‐RNAi strain in the head kidneys and trunk kidneys also had a sharp reduction from 72 to 120 hpi. This paper provides a new insight into the discovery of key virulence genes of pathogens in infected tissue systems.     

Current knowledge of nocardiosis in teleost fish

Nocardia sp. is the causative agent of nocardiosis, a lethal granulomatous disease of the skin, muscle, and various inner tissues affecting various teleost and shellfish. Four species of Nocardia have been isolated from diseased fish and shellfish, namely Nocardia asteroides, Nocardia seriolae, Nocardia salmonicida and Nocardia crassostreae. Therefore, in fish aquaculture, nocardiosis has caused severe economic losses, especially in the Asian region. Considerable research has been performed, since the first report of identified Nocardia sp. in fish, to characterize Nocardia sp. and identify rapid detection techniques, immune response against infection and prophylactic approaches. In this review, the current state of knowledge about nocardiosis in fish has been presented, including the pathogenesis, diagnosis, host immune response and vaccine development.     

The efficacy of MS‐222 as anaesthetic agent in four freshwater aquarium fish species

The efficacy of anaesthetic tricaine methanesulfonate (MS‐222) was evaluated in four freshwater aquarium fish species, Zebrafish (Danio rerio), Guppy (Poecilia reticulata), Discu (Symphysodon discus) and Green swordtail (Xiphophorus helleri). The correct dose of anaesthetic should induce the plane 4 of anaesthesia in less than 180 s, recovery in less than 300 s and must survive when exposed during 30 min to anaesthetic. Fishes were exposed to six concentrations of anaesthetic (75, 100, 125, 150, 200 and 250 mg L^?1) and the time of fish reaching plane 4 of anaesthesia, post exposure recovery, and the percentage of survival when fish were subject to 30 min in the anaesthetic were recorded. The optimal doses varied according to the species: D. rerio – 75, 100 and 125 mg L^?1, P. reticulata – 125, 150 and 200 mg L^?1, S. discus – 75 and 100 mg L^?1 and X. helleri – 125 and 150 mg L^?1. The induction time generally decreased significantly with increasing concentration of MS‐222 for all of the species evaluated. The recovery time had a tendency to increase with the increase of the MS‐222 concentration for D. rerio, P. reticulata and S. discus. On the other hand, X. helleri recovery time decreased with the increase of MS‐222 concentration. MS‐222 proved to be effective in anaesthesia for all the freshwater ornamental species studied. The main results clearly show that the optimal dose to anesthetize is fish species dependent and it is completely wrong to extrapolate optimal anaesthetic concentrations between different species.