Cellulase degradation of shrimp chitosan for the preparation of a water-soluble hydrolysate with immunoactivity

ABSTRACT:   Samples of colloidal chitin and chitosans with deacetylation degrees (DD) of 50, 65, 75 and 95% (DD50, DD65, DD75 and DD95, respectively) were prepared from shrimp chitin. The hydrolytic activity of cellulase on these samples increased with the increasing DD of chitosan, with DD95 being the most easily hydrolyzed. Colloidal chitin was hardly hydrolyzed. The optimal reaction temperature and pH for cellulase digestion of chitosan were 55°C and pH 5.2, respectively. During cellulase digestion of chitosan, the 9-h hydrolysate had the highest enhancing effect on the proliferation of a human hybridoma cell, HB4C5. This hydrolysate is composed of low-molecular-weight chitosan (LMWC), with a molecular mass of 20.0 kDa, and chitooligosaccharides, which are composed of sugars with a degree of polymerization of 1–6. In vitro , the 9-h hydrolysate increased both cell proliferation and immunoglobulin (Ig)M secretion of HB4C5 because of the presence of chitooligosaccharides for the former activity and LMWC for the latter activity. In vivo , samples of the chitosan hydrolysate, chitooligosaccharide mixture and LMWC significantly increased the levels of serum IgG and IgM, and enhanced concanavalin A- and lipopolysaccharide-induced proliferation of mouse lymphocytes.     

壳聚糖和益生菌对暗纹东方鲀抗病力和免疫功能的影响

为了解壳聚糖和益生菌刺激和增强免疫应答的可能作用,本试验在基础饲料中添加0.2%、0.5%、1.0%的壳聚糖,0.1%、0.2%、0.4%的益生菌,甘露聚糖与益生菌混合物,壳聚糖与益生菌的混合物,在室内水泥池中喂养暗纹东方鲀(3.15±0.05)g 2个月,根据抗菌活力筛选出免疫增强饲料组(0.2%壳聚糖,0.1%、益生菌,甘露聚糖与益生菌混合物,壳聚糖与益生菌混合物)测定免疫功能。结果表明,甘露聚糖与益生菌混合物仅提高头肾T淋巴细胞转化,壳聚糖与益生菌混合物显著增强头肾T、B淋巴细胞转化和脾脏B淋巴细胞体外培养时分泌的IgM。筛选出的四种免疫增强饲料均能使头肾B淋巴细胞体外培养时分泌的IgM显著增加。无论是脾脏B淋巴细胞还是头肾B淋巴细胞,体外培养时所分泌的IFN-α都不同程度地受抑制。     

Growth, stress tolerance and non-specific immune response of Japanese flounder Paralichthys olivaceus to probiotics in a closed recirculating system

ABSTRACT:   Effects of probiotics on growth, stress tolerance and non-specific immune response in Japanese flounder Paralichthys olivaceus were evaluated in a closed recirculating system. Survival and growth of flounder treated by supplying commercial probiotics either in the diet (the probiotic diet group), or into the rearing water (the water supply group), were higher compared to the untreated group (the control group). Water quality parameters, pH, NH₄-N, NO₂-N and PO₄-P showed lower concentration in the probiotic diet group compared with the control group and the supply group. Plasma lysozyme activity in the probiotic diet group and the water supply group was significantly higher ( P  < 0.05) than that in the control group. In heat shock stress tests, flounder in the probiotics-treated groups showed greater heat tolerance (measured by 50% lethal time, LT50) than the control group. Pathogen challenge tests with Vibrio anguillarum (2 × 10⁷ c.f.u./mL) resulted in significantly higher survival in the probiotics-treated groups than the control group. Results indicated that probiotics supplied in the rearing water and the diet of fish enhanced the stress tolerance and the non-specific immune system of Japanese flounder, providing them a higher resistance against stress conditions and pathogens.     

Effects of a probiotic mixture (Bacillus subtilis YB‐1 and Bacillus cereus YB‐2) on disease resistance and non‐specific immunity of sea cucumber,Apostichopus japonicus (Selenka)

The effect of a commercially available compound probiotics product containing Bacillus subtilis YB‐1 (50%) and Bacillus cereus YB‐2 (50%) fed to sea cucumbers, Apostichopus japonicus (Selenka) on challenge infections and non‐specific immune responses was assessed. Sea cucumbers (were randomly allocated into nine aquariums at a density of 30 sea cucumbers per tank and triplicate groups) were fed diets containing 0 (control), 10⁷ and 10¹⁰ cfu (g diet)^?1 of the probiotics mixture for 32 days. The growth factors and immunological parameters were measured. In addition, the effects on resistance against Vibrio alginolyticus infection were also evaluated. The results indicate that all the immunological parameters (phagocytic activity, superoxide anion production, lysozyme activity, catalase activity and phenoloxidase activity) measured and the growth rate of sea cucumbers fed 10¹⁰ cfu of the probiotics mixture were significantly (P < 0.05) improved than control groups at 16 and 32 days. After challenging, the cumulative mortality for the control was 100%, whereas the cumulative mortality for sea cucumbers fed 10¹⁰ cfu of the probiotics mixture was 47% (P < 0.05). Although the total autochthonous intestinal heterotrophic bacterial counts were not affected by dietary treatment (P > 0.05), Bacillus sp. levels were significantly elevated in sea cucumbers fed the probiotics mixture (P < 0.05). These results confirmed that administration of the probiotics mixture in the diet stimulated non‐specific immune responses and enhanced the growth performance of sea cucumbers, and was effective in controlling infections caused by V. alginolyticus.     

Effects of red clover extract on growth performance and body composition of African catfish <Emphasis Type="Italic">Clarias gariepinus</Emphasis>

ABSTRACT:   The present study was conducted to investigate the effects of the red clover extract on growth, body composition and survival of African catfish Clarias gariepinus . Three concentrations (25, 50 and 75 mg/kg) of red clover at dietary were used for 120 days. The final weights of red clover-treated groups were significantly different from each other and the control group. The best growth was observed in the group receiving the 75 mg/red clover kg diet. Fish fed with all levels of red clover had higher weight gains than the control. The highest values of the specific growth rate and the food conversion rate were 1.14 ± 0.01 and 2.26 ± 0.04, respectively, for the 75 mg/red clover kg diet group. During the red clover administration period, the survival ranged from 97.78% for the 50 p.p.m. group to 95.56% for the 25 and 75 p.p.m. groups. Protein contents of the dosage groups were significantly different from control ( P  < 0.05). The highest value of protein content (19.97%) was observed for the 75 mg/ red clover kg diet. Lipid and ash contents were significantly affected by the red clover levels in the diets. The highest lipid contents were in the 75 mg/red clover kg diet (4.67%), and the highest ash content was 1.39% for the 25 mg/red clover kg diet.     

Isolation and characterization of Bacillus spp. M001 for potential application in turbot (Scophthalmus maximus L.) against Vibrio anguillarum

The aim of this study was to screen Bacillus strains from the guts of Scophthalmus maximus, Paralichthys olivacues, Epinephelus coioides and Clupanodon punctatus, for use as probiotics in aquaculture. Eight Bacillus strains were screened, and strain M001 was selected for probiotic study based on its antagonistic activity against multiple aquatic bacterial pathogens including Vibrio anguillarum, V. campbellii, V. vulnificus, V. parahamolyticus, Streptococcus sp. and Edwardsiella tarda. M001 was identified as B. amyloliquefaciens based on the biochemical tests and 16S rRNA gene analysis. In vitro experiments revealed that M001 was able to grow at a wider range of temperature, pH and salinity and was capable to use turbot mucus as nutrient for growth. Additionally, M001 isolate greatly inhibited the growth of V. anguillarum by producing antibacterial substances and was acid tolerance, non‐antibiotic resistance and non‐harmful. Thereafter, the potential probiotic effect of M001 was tested in turbot by dietary administration of M001 at a dose of 10⁸ CFU g^?1 diet for 42 days. No significant differences of weight gain, specific growth rate and feed ratio were found in the M001‐diet group of fish compared with control fish, but which increased, respectively, by 5.5%, 4.7% and 7.0% after 42 days of feeding. Several digestive enzyme activities were found to increase significantly in the M001‐diet group, including protease and amylase activities in hepatopancreas, protease activity in intestine and lipase activity in stomach (P < 0.05). Sera superoxide dismutase activity and total protein content (P < 0.05) were also increased significantly (P < 0.05) in the M001‐diet group. The challenge experiment showed that the M001‐diet group of fish showed a relative per cent of survival of 62.7% against V. anguillarim infection. The Bacillus M001 identified from this study has good potential to provide vibriosis control as probiotic feed additive for turbot aquaculture.     

Effect of dietary administration of probiotics on growth and intestine functionality of juvenile Senegalese sole (Solea senegalensis, Kaup 1858)

The effects of the dietary administration of two bacterial probiotic strains (Ppd11 and Pdp13) from the Alteromonadaceae family for 60 days, were assessed by measuring growth and feed efficiency, activities of leucine aminopeptidase and alkaline phosphatase and structural changes in the intestine of juvenile Senegalese sole. In addition, the profile of intestinal microbiota was studied by Denaturing Gradient Gel Electrophoresis. Growth and nutrient utilization were significantly higher in fish receiving probiotics than in those fed the control diet. No differences were observed in proximal composition between treatments, though higher lipid muscle content was measured in fish receiving Pdp13. Those fish also exhibited higher activities of AP when compared to Ppd11 and control groups. The profile of intestinal microbiota clearly separated those fish receiving probiotics from those of the control group. Microscopical examination revealed accumulation of lipid droplets in the enterocytes of fish receiving the control diet, but not in those fed on probiotics. Interactions between those structural changes and growth performance are discussed.     

Effect of dietary chitosan on haematology,innate immunity and disease resistance of Asian seabass Lates calcarifer (Bloch)

This study was performed to evaluate the effect of dietary chitosan on haematology, innate immunity and protection against Vibrio anguillarum in Asian seabass, Lates calcarifer. A basal diet supplemented with 0, 5, 10 and 20 g chitosan kg^?1 diet was fed to the four different groups for 60 days. The haematological (total erythrocyte count, total leucocyte count, total serum protein, albumin, globulin and albumin‐globulin ratio) and innate immune parameters (phagocytic ratio, respiratory burst, serum lysozyme and serum bactericidal activities) were monitored at fortnight interval to assess the effect of chitosan feeding in Asian seabass. All the studied haematological and innate immune parameters were increased significantly (P ≤ 0.05) in chitosan‐fed groups in comparison with control. However, the group fed diet containing 10 g chitosan Kg^?1 feed showed highest haematological and innate immune parameters on 45^th day in comparison with other groups. Moreover, the fish fed the diet containing 10 g chitosan Kg^?1 feed had significantly higher post‐challenge survival (75.56 ± 4.44%) on the 30th day following V. anguillarum challenge. Therefore, this study suggests that chitosan at 10 g kg^?1 diet could be used as prophylactic in Asian seabass culture to enhance the protection against any possible infection by V. anguillarum.     

Use of probiotics to enhance growth,stimulate immunity and confer disease resistance to Aeromonas salmonicida in rainbow trout (Oncorhynchus mykiss)

An 8‐week feeding trial was conducted to evaluate the effects of dietary probiotics on growth, non‐specific immune responses and disease resistance in juvenile rainbow trout, Oncorhynchus mykiss. Fish averaging 5.8 ± 0.8 g (mean ± SD) were fed one of the five experimental diets; one control (Cont), and four other diets were prepared by supplementing single probiotics 1 (Bacillus subtilis; SP₁, 0.5%), single probiotics 2 (Bacillus licheniformis; SP₂, 0.5%), multi‐probiotics (B. subtilis + B. licheniformis; MP, 0.5%) and oxytetracycline (OTC) at 5 g OTC kg^?1 diet. After 8 weeks of the feeding trial, weight gain and specific growth rate of fish fed SP₁, SP₂ and OTC diets were significantly higher than those of fish fed Cont diet (P < 0.05). Superoxide dismutase (SOD) and lysozyme activities of fish fed SP₁, SP₂ and MP diets were significantly higher than those of fish fed Cont diet (P < 0.05). There were no significant differences in SOD and lysozyme activities among fish fed SP₁, SP₂, MP and OTC diets. In challenge test with Aeromonas salmonicida for 15 days, fish fed SP₁, SP₂ and MP diets showed significantly higher cumulative survival rate than those of fish fed Cont diet (P < 0.05). However, there were no significant differences in cumulative survival rate among fish fed SP₁, SP₂, MP and OTC diets. Although there was a little advantage in fish fed MP diet in terms of non‐specific immune responses, single or multi‐probiotics are equally effective statistically. These results indicate that single or multi‐probiotics had equal beneficial effects as an antibiotic replacer on growth performance, non‐specific immune responses and disease resistance in juvenile rainbow trout.     

Re-evaluation of total sulphur amino acid requirement and determination of replacement value of cystine for methionine in semi-purified diets of juvenile Nile tilapia, Oreochromis niloticus

Two feeding experiments were conducted to re-evaluate the total sulphur amino acid (TSAA; methionine and cystine) requirement and determine the replacement value of cystine for methionine of juvenile Nile tilapia ( Oreochromis niloticus ). Semi-purified diets used in both experiments contained 3510 kcal gross energy and 280 g of protein per kilogram diet from casein, gelatin and crystalline amino acids. The basal diet of the first experiment contained 3.1 g methionine and 0.4 g cystine per kilogram. l -methionine was added to the seven remaining diets at 1.0 g kg⁻¹ increment to produce methionine levels ranging from 3.1 to 10.1 g kg⁻¹ diet. Each diet was fed to four replicate groups of juvenile Nile tilapia (1.28 g mean weight) in a recirculation system for 8 weeks. Broken-line regression analysis of weight gain data indicated that the TSAA requirement of juvenile Nile tilapia was 8.5 g kg⁻¹ of the diet or 30.4 g kg⁻¹ of dietary protein. In the second experiment, TSAA level was set at 95% of the requirement value determined in the first experiment. Seven diets were made with different ratios of l -methionine and l -cystine (20 : 80, 30 : 70, 40 : 60, 50 : 50, 60 : 40, 70 : 30 and 80 : 20, based on an equimolar sulphur basis). Each diet was also fed to four replicate groups of juvenile Nile tilapia (4.14 g mean weight) in a recirculation system for 8 weeks. Regression analysis of weight gain data using broken-line model indicated that cystine (on a molar sulphur basis) could replace up to 49% of methionine requirement in semi-purified diets for juvenile Nile tilapia.     

Evaluation of dietary chitosan effects on growth performance,immunity, body composition and histopathology of Nile tilapia (Oreochromis niloticus) as well as the resistance to Streptococcus agalactiae infection

The present investigation aimed to evaluate the effect of dietary chitosan supplementation on growth performance, body composition, immune response and histopathology of Nile tilapia, and also the in vitro antibacterial activity of chitosan against Streptococcus agalactiae (S. agalactiae). About 180 fish (average body weight 39.3 ± 0.3 g) were randomly divided into three groups according to chitosan supplementation: control group (basal diet without chitosan), Ch3 group (3 g chitosan/kg diet) and Ch5 group (5 g chitosan/kg diet). Growth performance parameters and body proximate composition were measured before infection but biochemical parameters and lysozyme and antibacterial activities before and after experimental infection. Results of the present investigation showed dietary chitosan (5 g chitosan/kg diet) significantly (p < .05) improved growth performance parameters, body composition (dry matter, crude protein, ether extract, ash, and carbohydrate) and serum biochemistry (total protein, albumin, globulin, with no effect on AST, ALT, urea and creatinine) before infection in Ch5 group than the control. After infection, liver enzymes (serum AST and ALT) were maintained lower in fish fed Ch3 or Ch5 than the control. Serum lysozyme and bactericidal activities significantly increased (p < .05) in chitosan groups before and after the challenge. The mortality rate was markedly reduced in the Ch3 group and prohibited in the Ch5 group after the experimental infection. In conclusion, feeding 3 or 5 g chitosan/kg diet increased the growth rate and improved FCR of Nile tilapia. In addition, it reduced mortality by its antibacterial and immunostimulant effects.     

Cottonseed oil as a complementary lipid source in diets for gilthead seabream Sparus aurata juveniles

The efficacy of using cottonseed oil (CSO) as a fish oil (FO) substitute in gilthead seabream (Sparus aurata) juveniles feed was evaluated. Fish (BW_i 4.0 ± 2.9 g) were fed one of four isoproteic (~48% CP) and isolipidic (~18% L) diets for 9 weeks. Added oil was either FO (control diet, CTRL) or CSO, replacing 50% (CSO50 diet), 60% (CSO60 diet) and 70% (CSO70 diet) of dietary FO. Results indicated that FO replacement by CSO up to 60% level had no detrimental effects on growth or nutritive utilization and composition in fish muscles. Higher CSO intake (CSO70 diet, 56 g kg^?1) led to a 16% reduction in weight gain, 14% in feed utilization (FCR) and 57% in muscle n‐3 long‐chain polyunsaturated fatty acids (lc PUFA) as compared with CTRL and to abundant accumulation of lipid within the hepatocytes. Use of CSO altered fatty acid (FA) profiles of muscle and liver. Data suggested utilization of linoleic acid (LOA) by fish and retain of docosahexaenoic acid (DHA) in muscles. Therefore, limits of CSO inclusion as the main source of supplementary dietary lipid, with no negative effects on fish performance or nutritive composition and utilization in muscles, are: 40–48 g kg^?1 feed for gilthead seabream juveniles.     

Effect of chinese parsley Coriandrum sativum and chitosan on inhibiting the accumulation of cadmium in cultured rainbow trout Oncorhynchus mykiss

ABSTRACT:   Toxic heavy metals such as cadmium (Cd) accumulate in cultured fish from solid-form feed mixtures. In this paper, two natural ingredients, supposed to inhibit accumulation of heavy metals in the fish body, were investigated to develop a functional feed mixture for healthy fish. Three test diets: (i) ♯1, regular diet as negative control; (ii) ♯2, ♯1 + 2% lyophilized Chinese parsley Coriandrum sativum (CP); and (iii) ♯3, ♯1 + 3% chitosan (CT), were each fed to 50 rainbow trout Oncorhynchus mykiss once a day, at the constant rate of 1.0–1.2% of body weight for 12 weeks. No undesirable effect on fish growth was observed by an addition of either CP or CT to regular feed. Then, 10 mg Cd was added to 1 kg each of the three test diets and they were named as: (i) ♯4, ♯1 + Cd; (ii) ♯5, ♯2 + Cd; and (iii) ♯6, ♯3 + Cd, respectively. The fish given positive a control feed (diet ♯4) for 3 weeks accumulated Cd in the liver and kidney at approximately 0.52 mg/kg. However, Cd in ordinary muscle was found to be just above the detection limit. Upon accumulation of Cd, three groups of 50 fish were each fed test diets ♯4, ♯5, and ♯6 for 12 weeks at the same feed rate described above. The concentrations of Cd accumulated in the liver, kidney, and ordinary muscle of the fish were measured every 3 weeks. Contrary to the positive control fish fed diet ♯4, those fish fed test diets ♯5 and ♯6 accumulated 20–30% and 25–40% less Cd in the liver, respectively. Test diets were a little less effective for inhibition of Cd uptake in the kidney than that observed in the liver. These results suggest that addition of CP or CT to fish feed might be used to decrease toxic heavy metal accumulation in fish.     

Preparation and characterization of water-soluble chitosan produced by Maillard reaction

ABSTRACT:   The objective of this research was to improve the solubility of chitosan by Maillard reaction with 1% chitosan and 2% reducing sugar (glucose or glucosamine) dissolved in 0.2 M acetic acid, which was adjusted to pH 6.0, and incubated at either 50°C or 70°C for 1–7 days. The physicochemical and rheological properties of the chitosan–saccharide derivatives were also investigated. Results indicated that the solubility of modified chitosan derivatives was significantly greater than that of native chitosan. The solubility of chitosan–glucosamine was higher than that of chitosan–glucose, and the chitosan–glucosamine derivative remained soluble at pH 10. The degree of deacetylation of the derivatives decreased with increasing reaction time. Rheological investigation revealed that the apparent viscosity of the water-soluble chitosan derivatives in aqueous solution depended upon system conditions such as pH, ionic strength, and solution temperature. The measured apparent viscosity decreased as all system conditions increased. As calculated by the Arrhenius equation, the activation energy ( E _a ) of the derivatives in aqueous solution generally decreased with increasing the extent of Maillard reaction with respect to the reducing sugars used.     

Total sulphur amino acid requirement and maximum cysteine replacement value for methionine for fingerling Catla catla (Hamilton)

Two separate 12 weeks feeding trials were performed to quantify the total sulphur amino acid (TSAA) requirement (experiment I) and cysteine replacement value for methionine (experiment II) of fingerling Catla catla. In experiment I, six casein–gelatin based (33% crude protein; 16.72 kJ g^?1 gross energy) diets with graded levels of TSAA (0.56%, 0.81%, 1.06%, 1.31%, 1.56%, 1.81% dry diet) were fed to triplicate groups of fish (3.55 ± 0.06 cm; 0.65 ± 0.02 g) near to satiation. The TSAA requirement was determined by quadratic regression analysis of absolute weight gain (AWG), protein efficiency ratio (PER), feed efficiency (FE), protein gain (PG) and TSAA gain (TSAAG) against dietary TSAA concentrations at 95% maximum response. Above analysis revealed that inclusion of TSAA at 1.28% dry diet (1.22% methionine + 0.06% cysteine), corresponding to 3.87% of dietary protein is optimum. In experiment II, to determine the replacement value of cysteine for methionine, six diets containing 1.28% TSAA determined in experiment I with different ratios of l ‐methionine and l ‐cysteine (80:20, 70:30, 60:40, 50:50, 40:60, 30:70) on equimolar sulphur basis were fed to fish (3.65 ± 0.08 cm; 0.67 ± 0.04 g). Broken‐line regression analysis of AWG, PER, FE, PG and TSAAG against varying methionine to cysteine ratios yielded the optimum cysteine replacement value at 40.3%. Based on above analysis, it is recommended that inclusion of 1.28% dietary TSAA, corresponding to 3.87% of dietary protein is optimum of which 40.3% could be spared by cysteine. Data generated during this study would be useful in formulating TSAA balanced, cost‐effective feeds for the intensive culture of this fish.     

The effectiveness of commercial probiotics in northern white shrimp <Emphasis Type="Italic">Penaeus vannamei</Emphasis> ponds

ABSTRACT:   The effectiveness on water quality, population density of bacteria, and shrimp production in ponds treated with commercial probiotics was tested in Penaeus vannamei ponds in Hai-yan, China. Six ponds with replicates for treatment and control were used. Results showed that the probiotics could improve the population density of beneficial bacterial flora, reduce concentrations of nitrogen and phosphorus, and increase yields of shrimp. The average counts of Bacillus sp., ammonifying bacteria, and protein mineralizing bacteria were found to be significantly higher in treated ponds compared to control ponds ( P  < 0.05). In control ponds, an increase in presumptive vibrios was observed and the average density was up to 2.09 × 10³ cfu/mL, whereas that was only 4.37 × 10² cfu/mL in treated ponds ( P  < 0.05). The use of probiotics also significantly increased dissolved oxygen ( P  < 0.05) and reduced dissolved reactive-phosphorus, total inorganic nitrogen and chemical oxygen demand ( P  < 0.05). An average of 8215 ± 265 kg shrimp/ha was obtained in treated ponds with a feed conversion ratio (FCR) of 1.13 ± 0.05 and survival rate of 81.00 ± 6.25% compared with 4985 ± 503 kg shrimp/ha, 1.35 ± 0.12 and 48.67 ± 3.51%, respectively, in control ponds. This indicates that the addition of the commercial probiotics had a noticeable influence on water quality of shrimp ponds and shrimp production.     

Effects of diets supplemented with either individual or combined probiotics,Bacillus subtilis E20 and Lactobacillus plantarum 7‐40, on the immune response and disease resistance of the mud crab,Scylla paramamosain (Estampador)

Two trials, including firstly, diets incorporating individual or combined species of probiotics, Bacillus subtilis E20 and Lactobacillus plantarum 7‐40, were fed to the mud crab, Scylla paramamosian, for 28 days to evaluate the growth, immunity and disease resistance, and secondly, the optimal level of B. subtilis E20 in the diet by tracking the growth, immunity and disease resistance after 28 days of feeding were analysed. No significant differences in growth, total haemocyte counts, respiratory bursts, superoxide dismutase, or glutathione peroxidase were found in the two trials. Crabs fed the B. subtilis E20‐containing diet (D20) had significantly increased phenoloxidase (PO) activity, phagocytic activity (PA) and disease resistance compared with crabs fed the control and the mixed probiotics diet (MD). The mortality of crabs fed the D20 was significantly lower than that of crabs fed the L. plantarum 7‐40‐containing diet. L. plantarum 7‐40 had a great inhibitory effect on the growth of B. subtilis E20, which may have led to the decreased probiotic effect of the MD. An analysis of the optimal level of B. subtilis E20 in the diet showed that crabs fed the B. subtilis E20‐containing diet at a level of 10^9–10 cfu kg^?1 had better immune response of PO and PA, and disease resistance in the second trial.     

4株凡纳滨对虾肠道益生菌分离、鉴定及应用研究

为将凡纳滨对虾肠道分离的益生菌用于对虾的养殖,利用乳酸菌培养基分离对虾肠道中的益生菌并对菌株进行初步分类鉴定,利用体外抑菌方法筛选有抗菌活性的菌株,用于考察对虾实验室养殖过程中抗有关致病菌的效果。试验结果显示,自虾肠道分离得到W7、W25、W27、W31菌株,分别属于啤酒酵母、短小芽孢杆菌、枯草芽孢杆菌沙漠亚种和暹罗芽孢杆菌,它们对致病菌副溶血弧菌、金黄色葡萄球菌和沙门氏菌有一定的抗菌活性,其中暹罗芽孢杆菌W31抗菌效果最好。将啤酒酵母W7和暹罗芽孢杆菌W31菌体与饲料混合用于对虾的养殖,能够明显增强凡纳滨对虾抵抗弧菌侵染的能力,提高存活率。     

复合益生菌对眼斑双锯鱼(Amphiprion ocellaris)肠道消化酶、菌群结构以及形态的影响

为探讨饲料中添加复合益生菌对眼斑双锯鱼(Amphiprion ocellaris)肠道消化酶、菌群结构及形态的影响,将135尾初始体重(0.79±0.01)g的眼斑双锯鱼随机分为3组,每组3个重复,分别投喂复合益生菌有效活菌添加量为0%(对照组)、3%(L3组)和6%(L6组)的饲料,进行为期28 d的养殖试验。结果显示,复合益生菌能提升试验鱼肠道蛋白酶的活性,其中L3组肠道蛋白酶显著提升(P<0.05);能显著降低试验鱼肠道淀粉酶活性(P<0.05);能降低脂肪酶活性(P>0.05)。随着饲料中益生菌添加量的增加,试验鱼肠道中变形菌门丰度开始增多,拟杆菌门丰度降低。饲料中益生菌添加量的增加显著降低试验鱼肠道肌层厚度并显著增加试验鱼肠道绒毛高度(P<0.05)。可见,饲料中添加3%复合益生菌可改善眼斑双锯鱼肠道消化酶活性,提高肠道微生物群落多样性,改善肠道组织形态和功能。     

Replacement of Soybean Meal in Channel Catfish, Ictalurus punctatus, Diets with Cottonseed Meal and Distiller's Dried Grains with Solubles

Two pond experiments were conducted to evaluate cottonseed meal (CSM), distiller's dried grains with solubles (DDGS), and supplemental lysine as replacements for soybean meal (SBM) in channel catfish diets. In Experiment 1, fish fed diets in which SBM was totally replaced with CSM gained similar weight as fish fed control diet, but fish fed CSM diet in Experiment 2 had 9.5% lower weight gain than fish fed control diet. In both experiments, feed conversion increased significantly for fish fed CSM diet. There were no consistent trends in body composition of fish fed CSM diet versus control diet. Fish fed the DDGS + SMB diet had higher (Experiment 1) or similar (Experiment 2) weight gain than fish fed control diet. Feed conversion ratio was significantly lower in both experiments for fish fed SBM + DDGS diet than that of fish fed control diet. Body fat tended to be higher in fish fed SBM + DDGS diet compared to fish fed control diet. It appears that about 50% of SBM can be replaced with CSM + lysine in catfish diets without negatively affecting fish performance. Further, DDGS can be used up to at least 30% when the diet is supplemented with lysine.