Strategies for further development of aquatic feeds

ABSTRACT: To date, aquatic feeds have depended heavily on fishmeal and fish oil as their source of protein and lipid. However, the feed industry is encountering shortfalls in the availability of these ingredients because of a decline in the number of fish captured in the wild and the increased human demand for some of the species currently being used for fishmeal and oil production. Therefore, efforts are now being directed in different parts of the world to finding alternative quality ingredients, which ideally are less expensive and readily available for use in practical diets. The data accrued have shown that a large proportion of both fishmeal and fish oil can be replaced by other protein and lipid sources. However, it is emphasized that an optimal essential amino acid balance be maintained and that the n-3 highly unsaturated fatty acid requirement be satisfied when combining economical protein and lipid ingredients. Newly developed feeds should aim at being nutrient-dense in order to reduce the output of solid, P and N waste. This can be done through improving nutrient availability, optimizing the digestible protein to energy balance of the diet, and replacing dietary fishmeal with alternate ingredients. These diets should also be effective for maintaining good health and improving disease resistance in fish through enhancement of immune responses. A wholesome approach to culturing fish would be to use appropriate feeding standards that are aimed not only at improving economic returns but also at developing a lasting cohabitation of sustainable aquaculture and a cleaner environment. Furthermore, in the 2lst century aquaculture would still retain its place as a prominent source of food protein, signifying that fish feed research remains a forceful area in discovering better feeds for the industry.     

Digesta viscosity, nutrient digestibility and organ weights in African catfish (Clarias gariepinus) fed diets supplemented with different levels of a soluble non-starch polysaccharide

The objective of the present study was to investigate if dietary soluble non‐starch polysaccharides (NSP) increase digesta viscosity, reduce nutrient digestibility and increase organ weights in African catfish. The fish (mean weight 80 g) were fed diets supplemented with the soluble NSP guar gum at three levels, 0 g kg^?1 (GG0), 40 g kg^?1 (GG4) and 80 g kg^?1 (GG8). Guar gum inclusion significantly increased digesta viscosity in the proximal (GG0: 1.7 centipoise or cP; GG4: 84.9 cP; GG8: 98.3 cP) and distal (GG0: 1.9 cP; GG4: 109.8 cP; GG8: 66.4 cP) intestine. Apparent digestibility coefficients (ADC) were significantly lower for the GG8 diet than the GG0 diet (dry matter: 52% versus 69%; protein: 77% versus 90%; ash: 41% versus 54%; energy: 60% versus 77%). The ADC of dry matter and energy were significantly lower for the GG4 diet than the GG0 diet. The relative growth rate of metabolic weight (14.5–15.4 g kg^?0.8 day^?1) and feed conversion ratio (0.8) did not differ between diets. Fish fed the GG8 diet had a significantly higher somatic stomach index than GG0 fish (0.71% versus 0.65% body weight). The intestinal somatic index tended to increase with increasing guar gum supplementation (GG0: 1.08%; GG4: 1.23%; GG8: 1.59%). In conclusion, high digesta viscosities in the guar gum fed fish may explain the observed reduced nutrient digestibilities and increases in digestive organ weights.     

鱼类弹状病毒分子生物学研究动态

鱼类弹状病毒(Fish rhabdovirus)是一类引起鱼及其他水生动物致死性和流行性病害的重要病毒病原。对鱼类弹状病毒基因组的结构和功能研究,是深入探讨这类病毒感染和致病机理、研制病毒疫苗及诊断试剂的理论基础。本文就鱼类弹状病毒的基因组结构,包括N基因、P基因、M基因、G基因、NV基因、L基因及编码的相应蛋白、3′前导区和5′拖尾区、基因连接区等的组成及功能的研究进展作一综述,旨为抗病毒疫苗的研制及建立准确高效的检测方法提供借鉴。     

Characterization of culture supernatant proteins from Brucella abortus and its protection effects against murine brucellosis

In this study, we characterized the secreted proteins of Brucella abortus into the enriched media under the bacterial laboratory growth condition and investigated the pathogenic importance of culture supernatant (CS) proteins to B. abortus infection. CS proteins from stationary phase were concentrated and analyzed using 2D electrophoresis. In MALDI TOF/TOF analysis, more than 27 proteins including CuZn SOD, Dps, Tat, OMPs, Adh, LivF, Tuf, SucC, GroEL and DnaK were identified. Cytotoxic effects of CS proteins were found to increase in a dose-dependent manner in RAW 264.7 cells. Upon B. abortus challenge into phagocytes, however, CS proteins pre-treated cells exhibited lower bacterial uptake and intracellular replication compared to untreated cells. Immunization with CS proteins induced a strong humoral and cell mediated immune responses and exhibited significant higher degree of protection against virulence of B. abortus infection compared to mice immunized with Brucella broth protein (BBP). Taken together, these results indicate that B. abortus secreted a number of soluble immunogenic proteins under laboratory culture condition, which can promote antibody production resulted in enhancing host defense against to subsequently bacterial infection. Moreover, further analysis of CS proteins may help to understand the pathogenic mechanism of B. abortus infection and host–pathogen interaction.     

Induction of immune tolerance to caseins and whey proteins by oral intubation in mouse allergy model

This study was designed to evaluate the effect of oral tolerance of caseins (CSN) and whey proteins (WP) in alleviating the allergic response to cow's milk proteins in Swiss albino mice raised on a milk protein–free diet. Oral tolerance was induced by feeding mice with 20 mg of CSN or WP once in a day for 4 days consecutively before immunization with respective protein by intraperitoneal (i.p.) injections (20 μg 200 per μl of PBS) using 2% of alum Al(OH)₃ as adjuvant. Three weeks later, oral tolerance induction was analysed in humoral and cellular compartments of CSN‐ and WP‐fed versus saline‐fed control mice groups by measuring seric and intestinal antibody responses, mRNA abundance in splenic tissue and cytokine secretion patterns. The specific serum immunoglobulin‐E (IgE) levels were significantly suppressed (p < 0.05), while sIgA was enhanced in these groups when compared with their respective saline‐fed mice. Moreover, the mRNA levels of interferon‐γ (IFN‐γ) and interleukin‐4 (IL‐4) in both CSN‐ and WP‐tolerized mice were found to be significantly decreased, while the abundance of interleukin‐10 (IL‐10) and transforming growth factor‐β (TGF‐β) was increased significantly, as compared to respective control groups. Finally, cytokine profiles indicated a reciprocal decrease in IL‐4 and IFN‐γ versus an increase in IL‐10 secretions in supernatants of cultured splenocytes of tolerized mice. Taken together, these results clearly showed that oral administration of cows' milk caseins and whey proteins can induce significant hyposensitization in mice, with the participation of suppressor cytokines.     

Lipid energy reserves influence life-history decision of Atlantic salmon (Salmo salar) and brown trout (S. trutta) in fresh water

Abstract –  Lipid density appears to influence life-history decisions in salmonid fishes. This study shows that parr and smolts of anadromous Atlantic salmon from a south Norwegian river have on average between 30 and 40% higher energy level than corresponding brown trout in spring and summer, which may explain differences in life-history traits between the two species. The higher energy density of young salmon was chiefly due to a 1.8 times higher lipid density in parr and 2.4 times higher lipid density in smolts. The difference was smaller among immature parr in the autumn, with only 1.4 times higher lipid density in salmon than trout. The reason for the decreased difference was probably that the more energy rich salmon parr had attained maturity at the time. Among mature male parr, the somatic energy density was approximately 10% higher in trout than salmon. However, the gonadal energy content was more than twice as high in salmon than in trout. The higher somatic energy allocation in parr of Atlantic salmon probably influences protein growth of the two species in fresh water, and increases the ability of salmon relative to trout to undertake long distance feeding migrations and make large investments in reproduction.     

Intra- and interspecific phenotypic characteristics of fish-pathogenic Edwardsiella ictaluri and E. tarda

Intra‐ and interspecific characteristics of fish‐pathogenic Edwardsiella ictaluri, and E. tarda were determined by numerical analysis of gel electrophoresed protein profiles, fatty acid methyl esters (FAMEs) and immunoblotting. The 18 E. ictaluri isolates revealed a high degree of homogeneity (70% similarity or higher) in their protein profiles and 95% similarity in their FAME, while the nine E. tarda isolates revealed 30% similarity in their protein profiles and 95% similarity in their FAME. Immunoblots probed for antigenic epitopes with goat antiserum produced against E. ictaluri and E. tarda, respectively, revealed that E. ictaluri were more homogeneous compared with the E. tarda isolates. Overall, there was a considerable degree of relatedness between the two species. Our findings suggest that phenotypically E. ictaluri represents a clonal bacterial population structure compared with the less monomorphic E. tarda.     

Vitellogenin in tilapia (Oreochromis mossambicus): Induction of two forms by estradiol,quantification in plasma and characterization in oocyte extract

Two forms of vitellogenin were isolated by DEAE agarose ion-exchange chromatography from plasma of the tilapia, Oreochromis mossambicus. The monomers have apparent molecular masses of 200 and 130 kDa, as indicated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and a total amount of phosphorus of 1.7 and 0.1%, respectively. Antibodies specific to the two forms, designated tVTG-200 and tVTG-130, were generated in rabbits and used to develop enzyme-linked immunosorbent assays (ELISAs) and in Western blot analyses of plasma and oocyte extract. SDS-PAGE of the oocyte extract showed a major protein band at 106.6, minor bands at 26.6, 24.2, and 23.7 kDa, and very faint bands at 83.4 and 17.5 kDa. Western blots of the oocyte extract revealed that the antiserum to tVTG-200 recognized strongly the protein bands at 24.2 and 23.7 kDa, and less strongly the bands at 25.1 and 22.6 kDa, whereas the antiserum to tVTG-130 recognized mainly the protein band at 106.6 kDa. The presence of both VTGs in untreated male tilapia was detected with the ELISAs using relatively high plasma volumes. Their presence in males was confirmed by VTG-like immunoreactive materials eluting from the ion-exchange column at the same positions as tVTG-200 and tVTG-130. The concentrations of the VTGs in males were several orders of magnitude lower than in vitellogenic females. Treatment of male tilapia with estradiol-17β (E₂) induced both VTGs within 24h. After 7 days, tVTG-130 reached a maximum concentration in plasma, whereas tVTG-200 continued to increase. Our findings demonstrate that the two vitellogenins are biochemically distinct, possibly differentially regulated, and made by both sexes.     

Saliva chromogranin A in growing pigs: A study of circadian patterns during daytime and stability under different storage conditions

Salivary chromogranin A (CgA) is considered to be a biomarker of activation of the sympatho-adrenomedullary system, and has recently been proposed as a useful indicator of the acute stress response in pigs. The aim of the present study was to determinate whether salivary CgA concentrations in healthy growing pigs exhibits any circadian pattern during the daytime, and to evaluate its stability under different storage conditions. A total of 80 pigs (40 in spring and another 40 in autumn) of two different ages and genders were used. To establish the circadian pattern, saliva samples were collected at 07.00, 11.00, 15.00 and 19.00 h on two consecutive days. Pooled samples were used for the stability study and were measured on the day of sampling and periodically for up to 360 days later. Samples were stored at 4 °C, ?20 °C or ?80 °C and the effect of repeated freezing and thawing was also evaluated.No circadian pattern was detected for salivary CgA in either season and there were no significant effects of gender or age. However, mean salivary CgA concentrations were significantly higher (P < 0.0001) in the pigs sampled in autumn, compared to those sampled in the spring. Short term storage at 4 °C is recommended for up to 2 days, whereas frozen samples can be stored for 1 year at ?20 °C or ?80 °C, without substantial reduction in CgA values. In addition, samples can be frozen and thawed up to seven times without significant loss of the biomarker.     

Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

ABSTRACT:   In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells.