Three‐dimensional visualization of green fluorescence protein‐labelled Edwardsiella tarda in whole Medaka larvae

The invasive fish pathogen Edwardsiella tarda is common in aquatic environments and causes the environmentally and economically destructive emphysematous putrefactive disease called edwardsiellosis. In order to understand the organism's infection pathway, medaka larvae (Oryzias latipes) were immersion‐infected with E. tarda labelled with green fluorescence protein (GFP) and then visualized in three dimensions under confocal laser microscopy and light‐sheet fluorescence microscopy. Confocal microscopy revealed GFP‐labelled E. tarda in the mouth, head, gill bridges, gill cover, skin, membrane fin, gastrointestinal tract and air bladder, and in the caudal vein, somite veins, caudal artery and caudal capillaries. Light‐sheet microscopy additionally showed GFP‐labelled E. tarda in the pharyngeal cavity, muscle of the pectoral fin and cardiac atrium and ventricle. These findings suggest that during its infection of fish, E. tarda initially adheres to, and invades, the epithelial cells of the skin, gills and gastrointestinal tract (through the pharyngeal cavity); E. tarda then enters the blood vessels to access organs, including the air bladder and heart.     

Forecasting the Genetic Impacts of Net Pen Failures on Gulf of Mexico Cobia Populations Using Individual‐based Model Simulations

Offshore net pen fish farming provides a cost‐efficient means for production of marine finfish, and there is great interest in development of net pen operations in domestic waters. However, there are concerns over the possible genetic and ecological impacts that escaped fish may have on wild populations. We used individual‐based simulations, with parameter values informed by life history and genetic data, to investigate the short‐term (50 yr) impacts of net pen failures on the genetic composition of cobia, Rachycentron canadum, stocks in the Gulf of Mexico. Higher net pen failure rates resulted in greater genetic impacts on the wild population. Additionally, the use of more genetically differentiated source populations led to larger influxes of non‐native alleles and greater temporal genetic change in the population as a result of net pen failure. Our results highlight the importance of considering the appropriate source population for broodstock collection in net pen aquaculture systems and help to provide a general set of best management practices for broodstock selection and maintenance in net pen aquaculture operations. A thorough understanding of the genetic diversity, stock structure, and population demography of target species is important to determine the impact escapees can have on wild populations.     

Retracted: Study of testis histology and sperm morphology in the Bunnei,Barbus sharpeyi (Gunther, 1874) induced by luteinizing hormone‐releasing hormone analogue and carp pituitary extract

This study was conducted to determine the effects of hormone treatment on testis structure in Barbus sharpeyi, as well as the morphology of sperm examined by scanning electronic microscopy (SEM). Male B. sharpeyi were divided into three groups (three fish per group) and injected with luteinizing hormone – releasing hormone analogue (LHRH–A₂) or carp pituitary extract (CPE). The first and second groups were treated with 10 μg kg^?1 LHRH–A₂ and metoclopramide (MET), and their testis were sampled pre‐ and Poststripping respectively. The third group received 2 mg kg^?1 CPE and were killed pre‐stripping. Based on the histological results obtained, the testicular connective tissue of the lumen was thicker, and seminal vesicles were of a lower volume, in fish injected with CPE in comparison to the other groups. After treatment with LHRH–A₂ and MET, not all spermatozoa within the testis were ejaculated, and only a small amount of sperm was obtained by abdominal stripping. The highest and lowest diameters of connective tissue within lobules were observed in fish receiving CPE and LHRH–A₂ treatments respectively. There was a significant difference (P < 0.05) in lobule space between the fish injected with the CPE and the fish injected with the LHRH–A₂ and MET. The SEM results revealed that the spermatozoa of B. sharpeyi were composed of a spherical to elliptical head, a cylindrical midpiece, and a lengthy flagellum. In conclusion, it was found that injection with LHRH–A₂ and MET improved the spermatogenic process in comparison to injection with CPE.     

Potential application of baited remote underwater video to survey abundance of west coast rock lobster Jasus lalandii

The first assessment of the baited remote underwater video system (BRUVs) for monitoring rock lobster Jasus lalandii (Milne‐Edwards) in South Africa was compared to annual ring net and trap survey data. Count data from 58 BRUVs, 36 ring net and 95 trap samples were fitted with generalised additive models using depth, habitat, time and spatial gradient as independent variables. A power analysis was used to determine the number of samples needed per annum to detect a 5 and 10% increase and decrease in population size. The BRUVs outperformed the nets and traps in terms of sampling effort and were better at detecting changes in abundance of lobster than abundance of three fish species. Considering factors such as cost, analysis time, additional data and environmental impact, traps are the most feasible method in deeper waters but BRUVs could replace nets and traps in depths less than 60 m, low‐density areas and marine reserves.     

Nano-calcium carbonate reinforced polypropylene and propylene-ethylene copolymer nanocomposites: Tensile vs. impact behavior

Improvement of both the tensile and impact strength of the same polymeric material has always been a great challenge for the plastic industry. The study focuses on the effect of incorporation of calcium carbonate nanoparticles (0.3 wt% to 15 wt%) into three polypropylene (PP) based matrices viz. PP homopolymer, propylene-ethylene (PP-PE) copolymer and the blend of PP:PP-PE (30:70) to improve their impact behavior without hampering the tensile strength much. A loss in both the tensile and impact properties was observed in PP based nanocomposite. However, PP-PE based nanocomposites showed a significant improvement in impact strength (47 %) at 10 wt% loading with a loss of tensile strength by 22 %. To minimize this loss a blend of PP:PP-PE (30:70) was explored as a matrix. At 10 wt% loading, this matrix showed an improvement of 30 % in impact strength whereas the tensile loss was minimized to 10 %. Further, silane coupling agent which promoted good interfacial adhesion was used for best compositions. The variation of crystalline morphology of the nanocomposites with various formulations was analyzed using differential scanning calorimetry and X-ray diffraction.     

Effect of Growth Regulators and Ethanol on Termination of Dormancy in Potato Tubers

The main objective of this study was to find the best practice of inducing the sprouting of dormant potato tubers. We compared two protocols of breakage of dormancy, which are based on dipping excised potato eyes in an aqueous solution of gibberellic acid (GA₃) and kinetin (standard 1) or in the aqueous solution of GA₃, thiourea, and daminozide (standard 2), with a newly reported approach based on ethanol. We tested the effect of ethanol alone or in combination with GA₃ and/or kinetin on dormancy release and sprouting of the potato tubers. As a model, we used two potato genotypes (cultivars Pasat and Dorota), with long dormancy of 5 and 10 weeks respectively. We showed that the standard 2 was the most effective treatment both for dormancy breaking and in promoting sprout growth, especially for cv. Dorota, for which the treatment induced 82.3% of tuber eye-plugs to sprout 28 days after treatment and to produce 93.2% of emerged plants after subsequent 28 days of cultivation in the greenhouse. For this cultivar, similar efficacy was observed for the combination of 4% ethanol with GA₃ and kinetin. The same concentration of ethanol combined with GA₃ but without kinetin was the most efficient treatment for breaking dormancy of cultivar Pasat. However, the difference between the various treatment combinations was statistically insignificant. Ethanol alone or in combination with kinetin poorly induced breakage of dormancy, confirming the main role of GA₃ in artificial dormancy breaking. Thus our study showed that the standard 2 is the most effective approach for breakage of dormancy at least with long term-dormancy cultivars.     

Local adaptation studies and conservation: Parasite–host interactions between the endangered freshwater mussel Unio crassus and its host fish

1. Parasite–host interactions can involve strong reciprocal selection pressure, and may lead to locally adapted specializations. The highly threatened unionoid mussels are temporary parasites on fish, but local adaptation has not yet been investigated for many species.
2. Patterns of local adaptation of one of Europe's most threatened unionoids, the thick‐shelled river mussel (Unio crassus) were investigated. Eurasian minnows (Phoxinus phoxinus) from two rivers (separate drainage areas) were cross‐infested in the laboratory with sympatric and allopatric mussel larvae, while bullheads (Cottus gobio), inhabiting only one of the rivers, were infested with sympatric or allopatric mussel larvae. Larval encystment, juvenile mussel excystment and survival were measured.
3. For one river, but not the other, juvenile excystment from P. phoxinus was highest when infested with sympatric mussels. The opposite pattern was found for C. gobio in this river, where juvenile excystment and post‐parasitic juvenile survival from allopatric C. gobio were highest. The results thus cannot confirm local adaptation of U. crassus to P. phoxinus in the study rivers, as excystment was not consistently higher in all sympatric mussel–host combinations, whereas there were potential maladaptive signs of U. crassus in relation to C. gobio. There was no loss of encysted larvae 3 days after infestation until juvenile excystment. Most juveniles were excysted between 17 and 29 days after infestation, and the numbers of excysted juveniles increased with fish size.
4. The results have implications for parasite–host ecology and conservation management with regard to unionoid propagation and re‐introduction. This includes the need to (1) test suitability and adaptation patterns between U. crassus and multiple host fish species, (2) evaluate the suitability of certain unionoids and host fish strains after more than 3 days, and (3) determine whether large fish produce more juvenile mussels than smaller fish.

     

A bacterium with close genetic identity to Pseudomonas mandelii associated with spring fish kills in wild bluegill Lepomis macrochirus Rafinesque and pumpkinseed sunfish Lepomis gibbosus (Linnaeus)

Pseudomonas fluorescens are known bacterial pathogens in fish. The P. fluorescens group contains at least nine different bacterial species, although species from fish have rarely been differentiated. Two isolated fish kills affecting wild bluegills, Lepomis macrochirus Rafinesque, and pumpkinseed sunfish, Lepomis gibbosus (Linnaeus), occurred in the spring of 2015 during cool water temperatures (12.5°C–15.5°C). Disease signs included severe bacteraemia with rare gross external signs. Pure bacterial cultures isolated from kidneys of all affected fish were identified as P. fluorescens using the API 20NE system, while no bacteria were isolated from asymptomatic fish. To further identify the species of bacterium within the P. fluorescens complex, genetic analysis of the 16S rRNA, rpoD and gyrB genes was conducted. DNA sequences of bacterial isolates from both mortality events were identical and had close identity (≥99.7%) to Pseudomonas mandelii. Although likely widespread in the aquatic environment, this is the first report of a bacterium closely resembling P. mandelii infecting and causing disease in fish. The bacterium grew at temperatures between 5°C and 30°C, but not at 37°C. It is possible that infections in fish were a result of immunosuppression associated with spring conditions combined with the psychrotrophic nature of the bacterium.     

Growth performance and whole‐body composition of yellow catfish (Pelteobagrus fulvidraco Richardson) under feeding restriction

Fish growth is an important index in aquaculture practice. However, the effect of growth under restricted feeding on quality of fish grown to the same size remains poorly unknown. In this study, yellow catfish were reared by feeding them at three different feeding rates such as low (FR‐L group), medium (FR‐M group) and high (apparent satiation) ration levels for 82, 58 and 34 days to reach the same size, respectively. After this period, the growth performance, whole‐body amino acid composition and fatty acid profile of the fish were evaluated. Feeding restriction led to significant decreases in specific growth rate and feed conversion efficiency. Significant increases in dry matter, protein and energy contents were observed in fish under both restricted feeding treatments. FR‐L‐treated fish had significantly lower whole‐body essential amino acid contents, and significantly higher Ala and non‐essential amino acid contents compared to fish fed to satiation. Fish in restricted feeding treatments had significantly higher n‐3 PUFA and n‐3/n‐6 ratio compared to the fish fed to satiation. The present results indicate that from a quality perspective, there are certain advantages in improvement of whole‐body composition and enrichment of n‐3 PUFA associated with the restricted feeding of yellow catfish at slow growth rate.     

Effect of different feeds on the growth,survival and reproductive performance of zebrafish,Danio rerio (Hamilton, 1822)

Present experiment was conducted to determine the effect of different feeds with varying protein levels on the growth, survival and reproductive performance of zebrafish, Danio rerio. The control diet (T1) was wild‐collected zooplankton from local fish ponds, while test diets with 350 g kg^?1 protein (T2), 400 g kg^?1 protein (T3) and 450 g kg^?1 protein (T4) were formulated and fed to fish for a period of 210 days. The significantly (P < 0.05) highest mean weight gain and specific growth rate were observed in T1, which were similar with T3 and T4. The significantly (P < 0.05) highest number of egg production per female and relative fecundity were found in T1, followed by T4 and T3 while T2 produced lowest number of eggs. No significant (P > 0.05) differences were observed in brood survival rate, fertilization and hatching rate among the dietary treatments. The highest (P < 0.05) fry survival rate was recorded in T1, followed by T3 and T4. Thus, it is suggested that control diet i.e. mixed zooplankton exhibited better growth, reproductive performance and fry survival rate. However, diet containing 400 g kg^?1 crude protein also gave comparable results in terms of growth, survival and reproductive performance of zebrafish.