The use of a real-time PCR primer/probe set to observe infectivity of Yersinia ruckeri in Chinook salmon, Oncorhynchus tshawytscha (Walbaum), and steelhead trout, Oncorhynchus mykiss (Walbaum)

Yersinia ruckeri is the causative agent of enteric redmouth disease (ERM), a common pathogen affecting aquaculture facilities and implicated in large losses of cultured fish. Fisheries scientists continue to gain a greater understanding of the disease and the pathogen by investigating methods of identification and pre- and post-infection treatment. In this study, a real-time PCR probe set for Y. ruckeri was developed to detect daily changes in the bacterial load during pathogen challenges. Two species of fish, Chinook salmon, Oncorhynchus tshawytscha, and steelhead trout, Oncorhynchus mykiss, were exposed to two strains of Y. ruckeri (Hag and SC) during bath challenges. A subset of fish was killed daily for 14 days, and the kidney tissue was biopsied to enumerate copies of pathogen DNA per gram of tissue. While Chinook exposed to either the Hag or SC strains exhibited similar pathogen loads, those exposed to the Hag strain displayed higher mortality (～66%) than fish exposed to the SC strain (～24% mortality). Steelhead exposed to the Hag strain exhibited a greater pathogen load and higher mortality (～42%) than those exposed to the SC strain (<1% mortality). Steelhead challenged with either strain showed lower pathogen loads than Chinook. The study illustrates the efficacy of the probe set to enumerate Y. ruckeri bacterial growth in the kidneys of fish. Also, strains of Y. ruckeri display species-specific growth patterns that result in differential mortality and pathogen load.     

Movements and behaviors of swordfish in the Atlantic and Pacific Oceans examined using pop‐up satellite archival tags

Swordfish are highly specialized top‐level predators that have been challenging to study. In this paper, data from 31 pop‐up satellite archival tags attached to swordfish from (i) the eastern Pacific, (ii) central Pacific, and (iii) western North Atlantic‐Caribbean were analyzed. Common across locations was a pronounced diel vertical pattern with daytime hours spent primarily below the thermocline and nighttime hours spent in warmer waters, close to the surface. One exception to this pattern was periodic daytime basking events which were most common in cooler waters off California. Maximum daytime depths were significantly correlated with light penetration as measured by the diffuse attenuation coefficient at 490 nm. Temperature did not appear to influence daytime depths, and swordfish tolerated both extremely low temperatures (4°C) and rapid and dramatic temperature changes (>20°C). Temperature did appear to influence the nighttime depths in the Pacific where fish typically remained in the surface mixed layer. In contrast, in the warm tropical Atlantic this was not the case, and nighttime depths were much deeper. In all areas, nighttime depth increased around the full moon. Given the parallels between the vertical movement patterns of swordfish and those of the deep sound scattering layer we suggest that swordfish vertical distribution patterns, especially during daytime, are influenced largely by resource availability. At night, when swordfish are typically targeted by fisheries, both ambient light and temperature influence movements. Understanding vertical movement patterns of swordfish can help evaluate gear vulnerability, improve population assessments, and potentially reduce fisheries bycatch.     

Pectin purified from pomelo (Citrus maxima) peel as a natural hatching agent for fish embryos

Pectin is a biodegradable polysaccharide, and it has been recently applied as a gene delivery, drug delivery, wound healing and tissue engineering agent. In this study, pectin was extracted from pomelo (Citrus maxima) peel and characterized. The extraction recovery of pectin form pomelo peel was 14.5%, and it had 72.56% degree of esterification, 1,245.56 equivalent weight, 7.82% methoxyl and 68.27% anhydrouronic acid contents. Use of pomelo pectin as a hatching enhancing agent for fish embryos and its effect on hatching enzyme 1 (ZHE1) was investigated. The pectin‐exposed zebrafish embryos (100 µg/ml) showed significantly (p < .05) higher hatching rate (96.6%) compared with untreated (control) embryos (66.6%) at 60 hpf. The mRNA expression of ZHE1 was also significantly (p < .05) elevated up to 55.6‐fold in pectin‐exposed embryos at 24 hpf. In situ hybridization results revealed remarkably strong expression of ZHE1 in pectin‐exposed embryos compared with the control group. In addition, considerably larger size of the hatching gland was observed in pectin‐exposed larvae than that of the unexposed larvae group. These results clearly indicate that pectin isolated from pomelo peel has an ability to enhance the hatching process of zebrafish embryos via upregulation of ZHE1.     

Diurnal variations in hypothalamic monoamine levels in the teleostChanna punctatus (bloch) in response to melatonin under two photothermal conditions

Hypothalamic dopamine (DA), noradrenaline (NA) and 5-hydroxytryptamine (5-HT) levels exhibited marked day-night variations under ambient photoperiod and temperature (12L12D; 17±1°C) with peak values at mid-light phase. The 16L8D; 22±1°C treatment reversed the diurnal rhythm of 5-HT, but not that of DA and NA. However, there was an overall increase in the levels of the catecholamines on exposure to the long photoperiod and high temperature. The administration of melatonin in the fish held on 16L8D; 22±1°C regime restored the 5-HT rhythm to that of the 12L12D; 17±1°C control group, but with elevated mid-photophase value. However, there was no effect of the indole treatment on the 5-HT rhythm under the 12L12D; 17±1°C regime. Melatonin causes a significant reduction of NA level in both the groups, while DA level did not change in either group.     

Gene Expression in Black Tiger Prawns Following Intramuscular Injection of β-gal Plasmid

Gene expression in black tiger prawns (Penaeus monodon) was studied following intra-muscular injection of CMVGal plasmid into the second abdominal segment. We used an in situ staining technique to detect -gal expression in one- and three-month-old injected prawns. We found that only one of the three-month-old prawns expressed the marker gene (2 days after injection), and the site of expression was confined to the sixth abdominal segment away from the injection site. We repeated the experiment on a new batch of three-month-old prawns, but using fluorometric determination technique. This time we found that -gal expression was detected (6/42) at the site of injection after 2, 7, and 14 days. In two other test samples, transgene expression was detected in the sixth abdominal segment only, further confirming the possibility of injected DNA dispersal. The results of the study also suggest that direct gene transfer is a feasible technique in black tiger prawns.     

Inhibition of hirame rhabdovirus growth by RNA aptamers

RNA aptamers are artificial nucleic acids that specifically bind to a wide variety of targets. They are an effective tool for pharmaceutical research and development of antiviral agents. Here, we describe four Hirame rhabdovirus (HIRRV)‐RNA aptamers (H1, H2, H3 and H4) that we obtained from an in vitro process called the systematic evolution of ligands by exponential enrichment (SELEX). The HIRRV‐RNA aptamers specifically bind to HIRRV. Hirame natural embryo (HINAE) cells treated with virus and the RNA aptamer showed a decrease in appearance of cytopathic effect when compared with control (treated only with virus). Rhodovulum sulfidophilum was transformed with genes for the RNA aptamers, and the aptamers were detected in the culture medium, indicating that they were secreted from the cells. Thus, the recombinant R. sulfidophilum might be a powerful tool for the prevention of HIRRV in aquaculture.     

Dietary n‐3 highly unsaturated fatty acids affect the biological and serum biochemical parameters,tissue fatty acid profile,antioxidation status and expression of lipid‐metabolism‐related genes in grass carp,Ctenopharyngodon idellus

A 95‐day feeding trial was conducted to determine the effects of n‐3 highly unsaturated fatty acids (n‐3 HUFA) on the growth, antioxidation and lipid metabolism in grass carp (Ctenopharyngodon idellus). Two isonitrogenous and isolipidic diets were formulated with either lard oil (LO) or fish oil (FO) as the main lipid source. The results showed that the intraperitoneal fat (IPF) ratio was significantly lower (P < 0.05) in FO group. The concentration of n‐3 HUFA in muscle, hepatopancreas and IPF was significantly higher in FO group (P < 0.05). The serum low‐density lipoprotein (LDL) content was significantly lower (P < 0.05), and glucose (GLU) content was significantly higher (P < 0.05) in FO group. The serum total superoxide dismutase (T‐SOD) activity was significantly higher (P < 0.05) in FO group, consistent with the serum malondialdehyde (MDA) content. The gene expression of IPF fatty acid synthase (FAS), acetyl‐CoA carboxylase (ACC), sterol regulatory element‐binding protein (SREBP‐1) and peroxisome proliferator‐activated receptor γ (PPARγ) was significantly lower (P < 0.05) and that of peroxisome proliferator‐activated receptor α (PPARα) was significantly higher (P < 0.05) in FO group compared with LO group. Similar trends were found in the hepatopancreas, except for PPARγ. It is suggested that n‐3 HUFA could inhibit lipid accumulation in grass carp by affecting the expression of lipid‐metabolism‐related genes.     

Studies on the growth rate of the grass carp Ctenopharyngodon idella (Valenciennes) fed on two aquatic weeds and a terrestrial grass

The growth rate of grass carp fed with hybrid napier was found to be nearly three times the growth attained with Hydrilla and about five times the growth attained with Ceratophyllum. The gross conversion ratio on fresh weight basis with hybrid napier was on average 26.99 : 1, while with Hydrilla and Ceratophyllum it was 93.98 : 1 and 128.43 : 1 respectively. The average net production of grass carp fed on hybrid napier worked out to 1579.3 kg/ha in 182 days.