Nitrate removal efficiency and bacterial community of polycaprolactone-packed bioreactors treating water from a recirculating aquaculture system

A denitrification reactor packed with polycaprolactone (PCL) as a carbon source and biofilm carrier was developed to remove nitrate nitrogen (\( {\mathrm{NO}}_3^{-}-\mathrm{N} \)) from the water of a recirculating aquaculture system for 115 days. The hydraulic retention time was set to 6 h, and the water flow rate was 0.95 L h^?1. The removal rates of \( {\mathrm{NO}}_3^{-}-\mathrm{N} \) ranged from 34.67 to 155.7 g \( {\mathrm{NO}}_3^{-}-\mathrm{N} \) m^?3 day^?1 when the effluent \( {\mathrm{NO}}_3^{-}-\mathrm{N} \) concentrations ranged between 1.62 and 72.25 mg L^?1. No obvious changes in the Fourier transform infrared spectra of the PCL before and after use were observed. Bacterial community structure in the biofilm of the PCL granules was analyzed using Illumina MiSeq sequencing technology. Proteobacteria and Bacteroidetes were predominant in the biofilm, with relative abundances of 63.9 and 27.4%, respectively. Three genera, Acidovorax, Azospira, and Diaphorobacter, were capable of both denitrification and PCL degradation. This study indicates that PCL-packed reactors may be used and optimized for removing nitrate from aquaculture effluents.     

The influence of \upbeta-hydroxy- \upbeta-methylbutyrate (HMB) on cell-mediated immunity in tench Tinca tinca (L.): in vitro and in vivo study

$\upbeta$ -Hydroxy- $\upbeta$ -methyl butyrate (HMB) has been shown to counteract many of the negative effects of intensive fish production methods and results in increased growth and protection against diseases. In the present study, the in vitro and in vivo effect of HMB on the immunocompetence cell activity in tench Tinca tinca (L.) was examined. In the in vitro study pronephric phagocytes and lymphocytes were isolated from the fish and grown in culture medium (RPMI-1640) containing 0, 0.1, 1, 5, 10, 25, 50 or $100\,\upmu$ g HMB cm^?3 of medium. The effects of HMB on the respiratory burst activity (RBA), the potential killing activity (PKA) and lymphocyte proliferation stimulated by concanavalin A (ConA) or lipopolysaccharide (LPS) were examined. The in vitro study showed that HMB increased the RBA and PKA of the phagocytes, compared to the medium over that of cells grown without HMB. Lymphocyte proliferation stimulated by ConA and LPS was also increased approximately when HMB was added to the culture medium at concentrations between 10 and $100\,\upmu$ g HMB cm^?3. In the in vivo study fish were fed daily with pellets containing HMB at doses 0, 10, 25 and 50 mg kg^?1 body weight day^?1. The study showed that HMB statistically increased the RBA and PKA and highest activity at 50 mg kg^?1 body weight day^?1 were observed. Also lymphocyte proliferation stimulated by ConA and LPS were maximally stimulated at dose 50 mg kg^?1 body weight day^?1. In conclusion, the current study shows that HMB could potentially improve immunocompetence cell activity in tench through increased cell proliferation and functionality.     

Chemical composition and physical properties of salted shrimp paste (Kapi) produced in Thailand

Chemical composition and physical properties of 11 salted shrimp pastes (Kapi) obtained from various places of Thailand were determined. Based on proximate composition, protein constituted the major component (29.44–53.27 %, dry wt. basis). All samples contained 22.77–35.47 % NaCl with A _w of 0.695–0.774. Various formal nitrogen contents (11.96–22.87 mg N/g sample) were in agreement with different degrees of hydrolysis (12.68–20.76 %), suggesting the varying cleavage of peptides among the samples. From electrophoretic study, salted shrimp paste contained a large amount of small molecular weight proteins and peptides. Different samples had different colors with \( \Delta E^{*} \) of 47.10–60.43 and \( \Delta C^{*} \) of 9.46–20.76. The samples had total carotenoid content of 0.54–1.97 mg/g sample. Free astaxanthin, astaxanthin diester and canthaxanthin were the major carotenoids in salted shrimp paste. Thus, salted shrimp paste is a good source of protein and serves as the nutritious condiment.     

Seasonal Changes on Gonadosomatic Index and Maturation Stages in Common Pandora Pagellus erythrinus (L.)

A reproductive study was conducted on 305 individuals of common pandora Pagellus erythrinus, from Mazarrón Bay (Murcia, southeast coast of Spain), providing information for a more adequate interpretation of the gonadosomatic index in this species and its relationship with the stage of maturity of the fish. In terms of the other Mediterranean Sparidae, the GSI values obtained rank in the middle of the scale, being 3.9 ± 1.4% $\left( {\bar X \pm SD} \right)$ in spawning females, and 2.0 ± 0.9% in mature males. The GSI of primary males was lower (0.3 ± 0.8), suggesting that there is no sperm competition in this species. Reproductive season in the study area falls within the period between early April and middle July, although the ovotestes may retain some activity until the end of September.     

Stress mitigating and growth enhancing effect of dietary tryptophan in rohu (Labeo rohita,Hamilton, 1822) fingerlings

An experiment was conducted to study the stress mitigation and growth enhancing role of dietary l-tryptophan (TRP) under thermal stress in rohu, Labeo rohita fingerlings for 45 days. Seven hundred and twenty fishes were distributed in three major groups that are ambient temperature (26 °C), 34 and 38 °C in triplicate following a complete randomized design. Acclimation of fishes to 34 and 38 °C over average ambient temperatures were carried out at 1 °C/day. Each group was fed with a diet supplemented with 0, 0.36, 0.72 or 1.42 % l-TRP. Results showed that blood glucose and serum cortisol level were found to be significantly higher (p < 0.05) in the higher temperature groups than the ambient temperature group. Similarly, aminotransferase, lactate dehydrogenase, malate dehydrogenase, CAT, superoxide dismutase activities were found to be significantly higher (p < 0.05) in the control groups (0 % l-TRP) and decreasing activities of these enzymes were observed with the increasing level of dietary l-TRP. In different temperature groups, l-TRP-supplemented groups were found to have higher (p < 0.05) growth, RGR and PER. The results obtained in the present study indicate that dietary l-TRP mitigates thermal stress and enhances growth. From the present study, we can conclude that dietary supplementation of l-TRP at the 0.72 % level in the diet is found to be optimum to reduce thermal stress even up to 38 °C in rohu, L. rohita. The baseline data obtained here could be useful for the farmers to formulate feeds to culture the fish in different agro-climatic zones.     

Dynamics of free amino acids in the hemolymph of Pacific whiteleg shrimp Litopenaeus vannamei exposed to different types of stress

We analyzed the dynamics of amino acids, ammonia-N, and carbohydrates in hemolymph when Pacific whiteleg shrimp Litopenaeus vannamei was subjected to air-exposure and low-salinity stresses. Glycine, l-arginine, and d- and l-alanine levels in hemolymph were increased under both stress conditions. Ammonia-N, a product of amino acid catabolism, also increased in level. These results suggest that the above-mentioned amino acids are used as energy sources. Levels of total carbohydrates, the sum of glucose and other carbohydrates, in the hemolymph showed different dynamics between the two types of stress and were not always high. This suggests that other energy sources, such as amino acids, are important when animals are subjected to stress. It is possible that particular amino acids act as energy sources under various stress conditions.     

Purification and characterization of trypsin from the pyloric ceca of orange-spotted grouper, Epinephelus coioides

Trypsin from the pyloric ceca of orange-spotted grouper, Epinephelus coioides, was purified by fractionation with ammonium sulfate, ionic exchange, and affinity chromatography. The protein was purified 161.85-fold with a yield of 4%. Purified trypsin had an apparent molecular weight of 24 kDa according to an SDS-PAGE analysis. Optimal profiles of temperature and pH of the enzyme were 50°C and 8–10, respectively, using Nα-benzoyl-l-arginine ethyl ester as the substrate. The results of thermal and pH stability assays showed that the enzyme was stable at temperatures of up to 50°C and in the pH range of 6–8. Trypsin activity decreased with an increasing NaCl concentration (0–0.6 M). The activity of purified trypsin was effectively inhibited by a soybean trypsin inhibitor and N-p-tosyl-l-lysine chloromethyl ketone, and was slightly inhibited by iodoacetic acid, ethylenediaminetetraacetic acid, 1-(l-trans-epoxysuccinyl-leucylamino)-4-guanidinobutane, and pepstatin A. Protein identification of the purified protease showed that the sequences of two peptides, LGEHNI and NLDNDIML, were highly homologous to other fish trypsins. The measurement of trypsin activity in different tissues showed that the highest activity was detected in pyloric ceca, followed by anterior intestine, middle intestine, hind intestine and spleen, but very low activities were found in other tissues. An inverse relationship between the trypsin activity in four tissues of pyloric ceca, anterior intestine, middle intestine and hind intestine and fish body weight as a result of increased pepsin in stomach indicated grouper growth status was increased.     

ATP depletion inhibits retraction of chromatophores induced by GABA in cephalopod Todarodes pacificus

Color changes in cephalopods are generated by the expansion or retraction of chromatophores located under the dermis. The behavior of the chromatophores is regulated by neurotransmitters; l-glutamate (l-Glu) is an excitatory transmitter that causes the chromatophores to expand. To date, serotonin [5-hydroxytryptamine (5-HT)] is the only neurotransmitter known to stimulate retraction of chromatophores. We found that the chromatophores in the Japanese squid Todarodes pacificus were regulated by γ-aminobutyric acid (GABA), an inhibitory neurotransmitter, and that GABA caused expanded chromatophores to retract. We also found that adenosine 5′-triphosphate (ATP) concentrations in skin samples remained stable at their initial values for more than 24 h after the death of each squid; therefore, the chromatophores could respond to both l-Glu and GABA during that period. Furthermore, we attempted to reduce the levels of ATP by storing skin sample in sodium azide solution. The chromatophores in sodium azide-treated skin samples were induced to expand by l-Glu, but these expanded chromatophores could not be induced to retract by GABA. Based on these observations, we conclude that ATP is essential for retraction, but not expansion, of chromatophores.     

Culture-independent analysis of the bacterial community during fermentation of pa-som,a traditional fermented fish product in Laos

Pa-som is a traditional salty fermented food made from varieties of freshwater fish with garlic and rice in Laos. A similar product, plaa-som, is made in Thailand. To investigate the fermentation process of pa-som, lactic acid production and pH were monitored during laboratory-scale production of pa-som. Furthermore, the variety of bacteria involved and their succession during fermentation were examined using culture-independent polymerase chain reaction (PCR) denaturing gel gradient electrophoresis targeting the 16S ribosomal RNA (rRNA) gene. d- and l-lactic acid production and a decrease in pH were observed in the samples after 1 day of fermentation. The lactic acid content and pH continued to increase and decrease, respectively, until 4 days of fermentation. Overall, six lactic acid bacteria and eight other indigenous bacteria species were detected during pa-som fermentation. Among the lactic acid bacteria detected, two Lactococcus and one Weissella species were detected in samples after 1–4 days of fermentation, suggesting that these three species play major roles in pa-som fermentation from the initial phase. This is the first report to investigate the fermentation process in pa-som at the microbial level. This research approach should be applicable to a wide variety of pa-som and related fermented fishery products in Laos to help understand their microbial diversity and to identify beneficial bacterial species for improving quality.     

Detection of the first lectin with antimicrobial activity present in serum of the Amazonian fish tambaqui Colossoma macropomum

Tambaqui Colossoma macropomum is the most important Amazonian native species in South American aquaculture. Innate immunity at least partially depends on the recognition of pathogen-associated molecular patterns by receptor-recognizing pathogens (PRRs). Some PRRs have been characterized in fishes, and several studies have focused on the role of lectins in the immune system of various fishes. Lectins are proteins that specifically recognize carbohydrates and which have important biological functions. Tambaqui serum lectin (ComaSeL), which was identified on the basis of its hemagglutinating activity, was pre-purified, biochemically characterized, and used in assays of antibacterial activity against pathogenic bacteria in freshwater fishes. A study of the seasonality of this lectin was performed. Comasel activity was stable at a pH between 4.0 and 9.0 and lost 100?% of its activity at 70?°C. It recognized the carbohydrates d-galactose, 1-O-methyl-d-galactopyranoside, and d-fucose, showing antibacterial activity for Gram-negative bacteria. Its activity showed significant differences between the summer and winter (p?<?0.05, Tukey test), thereby corroborating observations that tambaqui becomes more susceptible to mortality from diseases caused by bacteria and fungi during the winter. With this information, new tools may be developed for gaining a better understanding of the role of these proteins in the immune system of the tambaqui, ultimately resulting in the improved management of this fish by pisciculturists.     

Rhamnose-binding lectins induce respiratory burst activity in macrophage cells from rainbow trout

Rhamnose-binding lectins (RBLs) have been isolated and characterized from various kinds of fish. RBLs recognize the structures of lipopolysaccharides and lipoteichoic acid on microbes, and have an opsonic effect, suggesting that RBLs play an important role as pattern recognition proteins in innate immunity. However, the function of RBLs in the removal of pathogens from the body has not yet been elucidated. In this study, we investigated the effect of three RBLs—named CSL1, 2, and 3—that were isolated from chum salmon Oncorhynchus keta eggs on the respiratory burst activity in a macrophage cell line. The CSLs induced reactive oxygen species in the peritoneal macrophage cell line from rainbow trout Oncorhynchus mykiss (RTM5). However, this activity was not affected by l-rhamnose or dl-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol, an inhibitor for glucosylceramide synthesis, indicating that the interaction between CSLs and Gb3 on cell surfaces was not involved in the induction of respiratory burst activity.     

Metabolic and phylogenetic profiles of microbial communities from a mariculture base on the Chinese Guangdong coast

This study examined the phylogenetic and metabolic features of microbial communities in Baisha Bay mariculture areas of Nan’ao Island, a mariculture base on the Chinese Guangdong coast. Large seaweed (Gracilaria lemaneiformis) cultivation zones (LZ), fish culture zones (FZ), and control zones (CZ) were sampled. According to community-level physiological profiling, d-mannitol, d-mannitol/α-d-lactose, and d-mannitol/α-cyclodextrin were the carbon sources with the highest utilization rates in CZ, LZ, and FZ, respectively (R _s  > 1.9). Denaturing gradient gel electrophoresis analysis showed that the dominant bacteria in CZ and LZ were Alphaproteobacteria (39.9 and 34.5%, respectively), whereas in FZ they were Flavobacteria (32.2%). The Shannon diversity in CZ and LZ was significantly higher than in FZ (P < 0.05). Some opportunistic pathogens, such as Vibrio, Pseudoalteromonas and Loktanella, were abundant in FZ. The microbial communities and most of the dominant bacteria in FZ were more affected by environmental factors including salinity, chemical oxygen demand and total phosphorus. Some bacteria, such as Brevundimonas and Polaribater, have the potential to degrade agar in seaweed, and were isolated from samples rich in algae which are dominant in LZ. The cultivation of G. lemaneiformis created the notable structures and functions of macroalgae-associated bacterial communities that were less affected by the environmental factors examined.     

溶藻弧菌感染后凡纳滨对虾鳃组织免疫相关基因的表达

分别给凡纳滨对虾(Litopenaeus vannamei)注射生理盐水、3×10⁶ CFU/mL(低浓度组)和9×10⁶ CFU/mL(高浓度组)溶藻弧菌(Vibrio alginolyticus)菌液, 采用荧光定量PCR技术, 检测不同处理后凡纳滨对虾鳃组织中Toll受体、IMD和溶菌酶基因表达量随时间的变化。结果表明, 处理42 h内, 注射生理盐水组对虾的Toll受体和IMD mRNA表达量无显著变化, 溶菌酶mRNA表达量在注射36 h后显著升高。急性感染溶藻弧菌后, 凡纳滨对虾鳃组织中Toll受体、IMD和溶菌酶mRNA的表达量峰值分别出现在感染后24、42和36 h; 溶藻弧菌的感染剂量不影响上述基因表达峰值的出现时间, 但显著影响上述基因的表达峰值(P<0.05), 各基因表达量峰值由大到小均依次为高浓度组、低浓度组、生理盐水组。急性感染初期, 对虾鳃组织中Toll受体mRNA表达量呈现显著下调, 而IMD和溶菌酶mRNA表达量在感染初期不存在显著下调现象。与感染前各基因的表达量相比, 高浓度溶藻弧菌感染组Toll受体mRNA表达量在2 h时显著下调, 低浓度溶藻弧菌感染组Toll受体mRNA表达量在3 h显著下调(P<0.05); 高浓度组IMD和溶菌酶mRNA表达量分别在36 h和12 h时开始有显著上调, 而低浓度组IMD和溶菌酶mRNA表达量则分别在42 h和24 h才有显著上调(P<0.05)。表明溶藻弧菌感染对凡纳滨对虾鳃组织中Toll受体、IMD和溶菌酶mRNA表达量有显著影响, 各基因的表达量与感染进程及溶藻弧菌剂量存在一定的相关性。

     

Stochastic modeling of production risk and technical efficiency of Thai koi (Anabas testudineus) farming in Northern Bangladesh

Thai koi is one of the high commercially valued farm fishes which have expanded tremendously in the northern part of Bangladesh. However, despite its impressive growth in the initial years, output of the industry is highly volatile across different years and farms. This research, following the Just and Pope framework, has made use of Kumbhakar’s (2002 Kumbhakar, S.C. (2002) Specification and estimation of production risk, risk preferences, and technical efficiency. American Journal of Agricultural Economics, 84, 8–22.[Crossref], [Web of Science ®] , [Google Scholar]) extended stochastic frontier model (SFM) to estimate production risk and technical efficiency (TE) in Thai koi farming. Results show that feed is the most important input significantly influencing production. Zeolite and pesticide appear to be risk-reducing, while fingerlings, labor, feed and salt turn out to be risk-increasing inputs. The inefficiency model indicates that labor and pond area significantly reduce the technical inefficiency. Feed, fingerling and lime also act as TE-improving factors. Education of farmers improves TE, while experience, training and frequency of water change do not impact the TE. The mean TE scores are 0.73 and 0.96 for the flexible risk estimate and conventional SFM estimates, respectively. TE values appear to become exaggerated when production risk is excluded from the model. SFM recognizes that risk is involved in Thai koi farming that causes lower TE.     

Effects of <Emphasis Type="SmallCaps">l</Emphasis>-carnitine against H<Subscript>2</Subscript>O<Subscript>2</Subscript>-induced oxidative stress in grass carp ovary cells (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>)

This study was designed in vitro to investigate the effects of l-carnitine against H₂O₂-induced oxidative stress in a grass carp (Ctenopharyngodon idellus) ovary cell line (GCO). GCO cells were pre-treated with different concentrations of l-carnitine, followed by incubation with 2.5 mM H₂O₂ for 1 h to induce oxidative damage. The results indicated that adding l-carnitine at concentrations of 0.01–1 mM into the medium for 12 h significantly increased cell viability. Pre-treatment with l-carnitine at concentrations of 0.1–5 mM for 12 h significantly inhibited 2.5 mM H₂O₂-induced cell viability loss. The significant decreases in the level of reactive oxygen species and cell apoptosis were observed in 0.5 mM l-carnitine group compared to the H₂O₂ group. Malondialdehyde values of all of the l-carnitine groups were significantly lower than those of the H₂O₂ group, while total glutathione levels of all of the l-carnitine groups were significantly higher than of the H₂O₂ group. The activity of antioxidant enzymes, such as total superoxide dismutase (0.1 and 0.5 mM l-carnitine), catalase (0.5 mM l-carnitine) and γ-glutamyl cysteine synthetase (0.5 and 1 mM l-carnitine), was significantly increased. In addition, pre-treatment of l-carnitine in GCO cells exposed to 2.5 mM H₂O₂ significantly increased the mRNA expression of copper, zinc superoxide dismutase, catalase (0.5 mM l-carnitine), glutamate cysteine ligase catalytic subunit (0.1–1 mM) and glutathione peroxidase (0.1 mM l-carnitine). In conclusion, l-carnitine promotes GCO cell growth and improves antioxidant function, it plays a protective role against oxidative stress induced by H₂O₂ in GCO cells, and the appropriate supplemental amount of l-carnitine is 0.1–1 mM.     

Production efficiency in New England’s oyster aquaculture industry

In coastal New England, growing oysters is a long-established practice that has exhibited a renaissance over the past few decades (Lapointe, Northeast region ocean council white paper: Overview of the aquaculture sector in New England, 2013). This industry generates millions in annual revenue, significantly boosting the economies of many coastal communities (Augusto &; Holmes, 2015 Augusto, K. &; Holmes, G. (2015). Massachusetts shellfish aquaculture economic impact study. Woods Hole, MA. Retrieved from http://web.whoi.edu/seagrant/wp-content/uploads/sites/24/2015/01/MA-Aquaculture-Economic-Impact-Study-2015.pdf [Google Scholar]). However, oyster aquaculture has received relatively little attention from the field of economics. To our knowledge, this study represents the first evaluation of productive efficiency among oyster producers in the United States and the first comparison of oyster production methods used in this country. Our findings indicate significant potential to increase productivity by reducing inefficiency in oyster production. However, industry-wide production efficiency has been increasing between 2013 and 2015. Further analysis revealed several factors that significantly influence inefficiency in oyster production, including a grower’s education level, location and farm diversification. This study provides insights into inefficiency in oyster production, enabling governance bodies and outreach specialists to make more informed decisions regarding this industry.     

信江发现溯河洄游型刀鲚的实证研究

利用电子探针微区分析技术,对2014年8月1日采自江西省余干县瑞洪镇信江(鄱阳湖水系五大河流之一)江段的刀鲚(Coilia nasus)的矢耳石进行了锶和钙的微化学分析研究。元素定量线分析表明,样品的锶钙比值(Sr/Ca×103)波动显著,不仅具有对应淡水生活的低值(Sr/Ca×1033),而且出现了对应于海水生活的高值(3Sr/Ca×1037),显示出其典型的溯河洄游特征。耳石Sr含量的面分析结果与之相同,均具有对应淡水生活的低值(蓝色)区域和海水生活的高值(绿色)区域。这两种耳石微化学类型首次直观地证实,目前距长江河口约1000 km的信江中存在洄游型刀鲚。     

Effects of dietary inclusion of <Emphasis Type="Italic">Moringa oleifera</Emphasis> leaves on growth and some systemic and mucosal immune parameters of seabream

The effect of the dietary incorporation of drumstick, Moringa oleifera, leaf meal (MOL; 0, 5, 10 and 15%) on the growth, feed utilization, some skin mucus and systemic immune parameters and intestinal immune-related gene expression in gilthead seabream (Sparus aurata) specimens. The experiment lasted 4 weeks. The results revealed that MOL can be incorporated in S. aurata diet up to 10% with no significant negative effect on growth and feed utilization. However, there was a significant decrease with MOL at a level of 15% after 2 weeks of feeding. The systemic immune status of fish fed with the different levels of MOL showed an improvement in head kidney leucocyte phagocytosis, respiratory burst and peroxidase activities. Also, serum humoral components, including protease, ACH₅₀ and lysozyme activities and IgM level, increased with MOL inclusion especially at the 5% level. MOL at 5% improved skin-mucosal immunity such as protease, antiprotease, peroxidase and lysozyme activities. Moreover, the feeding of MOL revealed an upregulation of the intestinal mucosal immunity genes (lyso and c3), tight junction proteins (occludin and zo-1) and anti-inflammatory cytokines (tgf-β) with a downregulation of pro-inflammatory cytokine (tnf-α). Therefore, it is recommended to incorporate MOL in S. aurata diets at a level of 5% for the best immune status or 10% for the high growth performance and acceptable immune surveillance.

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