Cross‐effects of dietary probiotic supplementation and rearing temperature on growth performance,digestive enzyme activities,cumulative mortality and innate immune response in seabass (Dicentrarchus labrax)

Probiotic influence on fish immune response and digestive capacity is extensively discussed in aquaculture. In this experiment, a feeding trial was carried out for 100 days to evaluate the cross‐effects of probiotic supplementation and rearing temperature (17, 20 and 23°C) in juvenile seabass (Dicentrarchus labrax). The experimental diet was supplemented with a commercial probiotic blend (Biomin AquaStar Growout) at 3 g/kg diet (5.23 × 10⁸ CFU/kg diet), and tested against a non‐supplemented diet (control). Growth performance and innate immune responses were analysed at 70 and 100 days of feeding, whereas digestive enzyme activities were determined at 100 days of feeding. At the end of the feeding trial, fish were subjected to a handling stress and cumulative mortality was recorded. Digestive enzyme activities were influenced by temperature, with α‐amylase and lipase activities peaking at the higher temperature (23°C) and trypsin at the lower temperature (17°C). Immune parameters showed a significant temperature versus feeding duration effect, with complement system (ACH50) and peroxidase peaking at 70 and 100 days of feeding, respectively. Poststress cumulative mortality was higher at the lowest temperature (17°C), especially in fish fed the control diet. In conclusion, water temperature was the main variable affecting the studied parameters, whereas the dietary probiotic supplementation had influence on the chymotrypsin activity and survival rate in seabass reared at 17°C.     

放养密度和微生态制剂对施氏鲟养殖水质的影响

将初始体质量(54.86±10.19)g的施氏鲟Acipenser schrenckii饲养在面积16m~2(4m×4m)、水深1.7~1.9m的陆基围隔中,密度分别为2 000尾/667m~2、3 000尾/667m~2、4 000尾/667m~2和5 000尾/667m~2,每个密度组均设3个平行,常规饲养,混合泼洒光合细菌、枯草芽孢杆菌和乳酸菌,第一次泼洒量为光合细菌50m L、枯草芽孢杆菌50g和乳酸菌50g,之后每5d泼洒第一次量的1/2,研究微生态制剂对静水土养殖池塘水质的影响。结果显示:水体中溶解氧量随养殖密度的增加逐渐降低(P0.05),氨氮、亚硝酸盐浓度随养殖密度的增加逐渐升高(P0.05)。在使用微孔增氧的条件下,泼洒微生态制剂对溶解氧量和氨氮浓度的影响不显著(P0.05),但显著降低了水体亚硝酸盐浓度(P0.05),显著增加了浮游动物生物量(P0.05)。     

In vitro and in vivo selection of probiotic purple nonsulphur bacteria with an ability to inhibit shrimp pathogens: acute hepatopancreatic necrosis disease‐causing Vibrio parahaemolyticus and other vibrios

Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticusAHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticusSR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.     

Two biomass preparation methods provide insights into studying microbial communities of intestinal mucosa in grass carp (Ctenopharyngodon idellus)

Animal digestive tract is habitat for a large number of autochthonous microbiota, which play central roles in multiple biological and physiological processes of the host. In this study, two different micro‐biomass preparation methods were employed to evaluate the diversity of intestinal mucosa‐associated microbiota in grass carp (Ctenopharyngodon idellus). Genomic DNAs were isolated either directly from intestinal mucosal samples (group A), or from micro‐biomass after microbial dissociation (group B). Community richness, diversity and evenness indices were all higher in group B, but differences were not statistically significant (P = 0.97, P = 0.33, P = 0.34 respectively). Furthermore, group B samples exhibited an increased ratio of bacterial DNA in comparison with group A samples, but the difference was also not statistically significant (P = 0.74). In addition, there were no statistically significant differences between the two groups (P > 0.05) at the taxonomic level. Our results support previous findings that there exists a great abundance of the intestinal mucosa‐adherent microbiota in the grass carp; among these, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Spirochaetes and Fusobacteria were the most common phyla. Within these microbiota, Paenibacillus, Bacteroides, Bacillus and Cetobacterium genera comprise the majority of the community, implicating their functional importance (e.g. as probiotics) to their host. Our results contribute towards a better understanding of the intestinal microbial profile of grass carp. Both micro‐biomass preparation techniques proved to be feasible for studying mucosa‐adherent microbiota of grass carp; however, the second method (group B) provides a protocol that is somewhat more effective than the first method (group A).     

微生态制剂对葡萄抗病能力及果实品质的影响

以云南、四川主栽的4个葡萄品种为试材,在常规用药方式基础上辅以微生态制剂,测定其对葡萄酸腐病和黑曲霉病防控效果及对果实品质的影响,以期为有效防控葡萄酸腐病和黑曲霉病的发生,解决生产实践中的具体问题提供科学参考依据。结果表明:微生态制剂能提高葡萄果粒大小和百粒质量,同时增加糖酸比。在传统用药基础上使用绿康威,对葡萄酸腐病和黑曲霉病的防效最佳,分别达52.3%和42.4%;在传统用药基础上使用绿地康3号,其防效为31.3%和21.9%,均表现出良好的防效;前期使用化学药剂,后期单独使用微生态制剂对酸腐病的防效不太明显。     

Effects of oral administration of Spirulina platensis and probiotics on serum immunity indexes,colonic immune factors,fecal odor,and fecal flora in mice

To evaluate the effects of Spirulina platensis and probiotics on growth, immunity indexes, fecal flora, and fecal odor in mice, 40 mice were randomly allotted to four groups, and each was administrated with nothing, S. platensis, probiotics, or both for 28 days, respectively. Then, many indexes were measured. The results showed that S. platensis was more effective (P < 0.001) than probiotics in improving mice's feed conversion ration (FCR). In immunity, probiotics administration increased (P < 0.042) serum IgE, IgM, IFN-γ, colonic AHR, TLR4, and NF-κB protein expression and decreased (P < 0.039) serum IL-1α, IL-21, IL-22, and colonic ARNT gene expression. However, the S. platensis showed weaker effect, which increased (P < 0.025) the serum IgE, IgM, TNF-α, and the colonic AHR and NF-κB protein expression, and decreased (P < 0.01) serum IL-21. Probiotics consumption decreased the fecal odor by decreasing (P < 0.02) fecal Escherichia coli, indole-3-acetic acid (IAA), and skatole contents, and the S. platensis decreased (P = 0.04) the IAA. These results indicated that oral administration of probiotics, S. platensis, or both of them in mice probably benefited body's immunity and reduced fecal odor. However, their mechanisms were still unclear and need further study.     

Enhancement of innate immune system,survival and yield in Penaeus monodon reared in ponds using Streptococcus phocae PI80

The objective of this study was to evaluate the effect of dietary and water supplementation of probiotic Streptococcus phocae PI80 on growth, immune response and feed utilization of tiger shrimp Penaeus monodon in earthen ponds. The probiotic bacterium S. phocae PI80 was cultured in large fermenter (50 L) by adding additional carbon source in the form of molasses and glucose along with yeast extract as nitrogen source to enrich S. phocae PI80 biomass. This enriched S. phocae PI80 was administered to shrimp in feed (6.5 × 10¹³ bacterial cells mL^?1) as well as in pond water (5 L/pond). Shrimp growth performance was significantly improved (P < 0.05) in 120 days culture when the average body weight of treated molasses + yeast extract (MY) (28.41 ± 0.874 g), glucose + yeast extract (GY) (27.013 ± 0.698 g) was significantly higher than control (23.63 ± 0.684 g). Food conversion ratio FCR was also found to be reduced significantly in ponds treated with probiotics when compared with control pond (1.89 ± 0.09). Vibrio and luminescent bacteria were found to be lower in the treatment receiving MY group, and we hypothesize that this may lead to greater shrimp survival. Furthermore, fermentation product of S. phocae PI80 added to pond water and feed additives enhanced the shrimp immune system. The results indicated that total haemocytes count (THC), phenoloxidase (PO) activity, NBT reductase assay and phagocytic activity significantly increased in shrimps treated with S. phocae PI80. Our study demonstrates that administration of S. phocae PI80 in the water and feed at 6.5 × 10¹³ colony‐forming units (CFU) mL^?1 bacterial cells induce immune modulation and enhances the immune ability of P. monodon in pond‐reared shrimp and increased the shrimp production.     

生物絮团养殖模式下益生菌添加对异育银鲫生长、消化酶活性及肠道组织结构的影响

前期研究表明,生物絮团技术(biofloc technology,BFT)适于异育银鲫(Carassius auratus gibelio)养殖。为进一步优化BFT养殖模式,本研究设置3个实验组:BFT模式下EM菌添加组(BB组)、枯草芽孢杆菌(Bacillus subtilis)添加组(BI组)和BFT对照组(B组),以均体重(1.60±0.50)g的异育银鲫为研究对象,探讨BFT模式下外源添加益生菌对养殖动物生长、消化酶活性及肠道组织结构的影响。结果表明:(1)益生菌添加组异育银鲫增重率和特定生长率显著高于对照组(P0.05),BB和BI组的增重率分别提高了216.70%和184.04%,特定生长率分别提高了141.18%和125.49%,BB和BI组间差异不显著(P0.05);(2)益生菌添加组(BB组和BI组)的消化酶(淀粉酶、脂肪酶和胃蛋白酶)活性均显著高于对照组(B组)(P0.05)。益生菌添加组间,BB组淀粉酶活性显著高于BI组(P0.05),脂肪酶和胃蛋白酶活性亦高于BI组,但差异不显著(P0.05);(3)益生菌添加组肠道肌层厚度和黏膜下层厚度显著高于对照组(B组)(P0.05),BB组异育银鲫肠道黏膜皱襞高度和皱襞间质宽度与BI和对照组相比,均无显著差异(P0.05)。研究表明,BFT养殖模式下外源添加益生菌可以更好地促进异育银鲫生长。     

Study on Liver-protective Effect of Chinese Herbal Compound Probiotics on Acute Liver Injury Layers

This experiment was conducted to investigate the liver-protective effect of Chinese herbal compound probiotics (CHCP) on acute liver injury layers.One hundred and eight 1 day old hens were divided into 4 groups with 3 replicates in each group and 9 layers per replicate in trial 1.The layers in model groups Ⅰ to Ⅲ were gavaged with 10% (V/V) soybean oil solution of carbon tetrachloride (SCCl₄) according to 1,2 and 4 mL/(kg·BW) at 14,28 and 35 d,respectively.The layers in control group were gavaged with 2 mL/(kg·BW) soybean oil.In trial 2,sixty 1 d layers were divided into 5 groups:Control group (soybean oil),model control group(SCCl₄)and low-dose,middle-dose and high-dose CHCP group (SCCl₄+1‰ CHCP,SCCl₄+2‰ CHCP and SCCl₄+4‰ CHCP respectively).CHCP were used by drinking water since 7 days.SCCl₄ were gavaged according to 2 mL/(kg·BW) at 14 and 28 d.The results showed as follows:The model of layers liver damage could be built by intragastric administration of 2 mL/(kg·BW) 10%(V/V) SCCl₄ at 14,28 d respectively,with the signs of hepatic steatosis,severe vacuolar degeneration,nuclear condensation and necrosis.Compared to the model control group,the serum AST levels in low,medium and high dose CHCP groups were decreased by 4.35% (P > 0.05),7.57% (P > 0.05) and 9.79% (P < 0.05),the serum ALT levels in medium and high dose CHCP groups were decreased by 34.92% (P < 0.01),36.51% (P < 0.01),the serum total bilirubin content in medium and high dose CHCP groups were decreased by 25.49% (P < 0.01),27.45% (P < 0.01).The liver cell congestion was reduced to varying degrees in different dose CHCP groups,and the liver cell had no vacuoles,arranged in neat rows,abundant cytoplasm and uniform in different dose CHCP groups.In conclusion,2‰,4‰ CHCP could reduce hepatocyte necrosis,decrease the serum activities of ALT,AST and total bilirubin levels,and had protective effect on hepatic injury induced by SCCl₄.