In vitro selection and characterization of putative probiotics isolated from the gut of Acipenser baerii (Brandt, 1869)

To select and characterize potential probiotic bacteria from the gut microbiota of Siberian sturgeon (Acipenser baerii), 129 strains isolated from the hindgut were screened for antagonistic activity against five fish pathogens. Ten isolates showed antagonism towards three or more pathogens. Nine of these isolates were Gram‐positive, belonging to Lactococcus (seven) and Bacillus (two), and a single strain belonging to the Gram‐negative Citrobacter. These inhibitory isolates were identified using genetic, phentotypic and biochemical traits, and further characterized by in vitro tests assessing the adhesion and growth in mucus and resistance to gastric and intestinal fluids. The candidate probiotics were determined to be non‐pathogenic through an in vivo study. Based on these assays, Lactococcus lactis ssp. lactis STG45 and STG81 showed the broadest inhibitory potential, a high viability in simulated gastrointestinal juice and the highest adhesion capacity to mucus. They were therefore selected as the most promising candidate probiotics. This is the first study screening probiotics among the gut microflora of Siberian sturgeon.     

Antagonism against fish pathogens by cellular components and verification of probiotic properties in autochthonous bacteria isolated from the gut of an Indian major carp,Catla catla (Hamilton)

Probiotic potential of the autochthonous bacteria in catla, Catla catla has been evaluated through determination of antagonistic activity (in vitro) of the cellular components of gut bacteria against seven fish pathogens. Altogether 208 strains were isolated, inhibitory activity of the isolates was evaluated through cross‐streaking and 16 primarily selected antagonistic strains were confirmed using the double‐layer method. Four bacteria that showed antagonism against ≥4 pathogens were selected as putative probiotics. The intracellular, extracellular, whole‐cell and heat‐killed cell components exhibited bactericidal activity against the pathogens. In addition, the selected strains were capable of producing different extracellular enzymes, competent to grow in intestinal mucus and could tolerate diluted bile juice. Analysis of 16SrRNA partial gene sequence revealed that both the strains CC1FG2 and CC1FG4 were Bacillus methylotrophicus (KF559344 and KF559345), while the isolates CC1HG5 and CC2HG7 were Bacillus subtilis subsp. spizizenii (KF559346) and Enterobacter hormaechei (KF559347) respectively. Bio‐safety evaluation through intra‐peritoneal injection of the isolates did not induce any pathological signs or mortalities in C. catla. The study confirmed probiotic properties of autochthonous gut bacteria in C. catla and demonstrated potential for using them as bio‐control agents. However, in vivo studies are essential to explore their efficacy in the commercial aquaculture.     

Isolation and evaluation of putative probiotic strains from different teleost to prevent Pseudomonas aeruginosa infection in Cyprinus carpio

Probiotics renowned as valuable microbes serve as a potential alternative to control diseases in aquaculture and are considered as an efficient and environment‐friendly approach to reduce the use of antibiotics. The present study aims at the isolation of putative probiotic bacteria from the intestinal tract of different fish species from the Doaba region of Punjab, India. In this study, isolated bacterial strains were characterized based on their morphological, biochemical and molecular characterization by 16S rRNA gene sequencing, followed by in vitro evaluation of different selection parameters described in FAO/WHO guidelines. A total of 169 different bacterial strains were isolated from the gastrointestinal tract of 52 different fish species. After in vitro evaluation, out of 169 bacterial strains only five bacteria (S3, S7, BDK2', BDK7 and BDK9) identified as Enterococcus and Bacillus species showed antagonistic activity against the fish pathogen Pseudomonas aeruginosa (MTCC 4 673). These isolates were screened based on their response towards bile tolerance, pH tolerance, adhesion and drug susceptibility to different antibiotic discs. And, the in vivo results indicated improved growth and survival against the infection (P. aeruginosa) after oral administration of the probiotics. The observations of in vitro and in vivo evaluation indicate that these isolated probiotic strains serve as effective probiotics and can be used as a novel and safe treatment to cure current issues in aquaculture.     

Aqualase®, a yeast‐based in‐feed probiotic,modulates intestinal microbiota,immunity and growth of rainbow trout Oncorhynchus mykiss

Yeast probiotics have great promise, yet they received little attention in fish. This study investigated the influence of Aqualase^®, a yeast‐based commercial probiotic composed of Saccharomyces cerevisiae and Saccharomyces elipsoedas, on health and performance of rainbow trout (Oncorhynchus mykiss). Probiotics were incorporated in the diets at three different inclusion levels (1%, 1.5% and 2%) and administered to the fish for a period of 8 weeks. After the feeding trial, intestinal total viable aerobic bacterial count was significantly higher in fish group that received 2% in‐feed probiotics. In addition, a significant increase in at least 11% in intestinal lactic acid bacteria population was observed in all probiotic‐fed groups. Total protein level and lysozyme activity in skin mucus were significantly elevated following probiotic feeding. Inhibitory potential of skin mucus against fish pathogens was significantly enhanced by at least 50% in probiotic‐fed groups. Humoral and cellular immune parameters were influenced by probiotic feeding and the effects were dependent on inclusion level. Digestive physiology was affected by in‐feed probiotics through improvement of intestinal enzyme activities. All growth performance parameters were significantly improved following probiotic administration specifically at inclusion rate 1.5% and above. Taken together, the results revealed that Aqualase^® is a promising yeast‐based probiotic for rainbow trout with the capability of modulating the intestinal microbiota, immunity and growth.     

Effect of feeding Lactobacillus-based probiotics on the gut microflora, growth and survival of postlarvae of Macrobrachium rosenbergii (de Man)

The present study was conducted to study probiotics treatment in the post‐larval diet of Macrobrachium rosenbergii. Three hundred postlarvae (average weight, 114–118 mg±0.11) were divided in five experimental groups each with four replicates. The experiment was conducted for 60 days. Experimental diets were identical in all the aspects except for variation in the probiotics strain. T1 and T2 groups were fed Lactobacillus acidophilus (140×10¹¹ CFU 100 g^?1) and L. sporogenes (24×10⁷ CFU 100 g^?1) respectively. The T3 group was fed L. sporogenes bioencapsulated in Artemia nauplii. T4 was the control group (without probiotic) whereas T5 was fed Artemia along with control diet. The bacteriological study indicated that the gut microflora of postlarvae are devoid of lactic acid bacteria. The probiotic strains were found to have inhibitory effects against the gram‐negative bacterial flora present in the gut. Growth of the probiotic fed groups was significantly higher (P<0.05) than the control group. Significantly higher growth (P<0.05), per cent weight gain (132.5%), specific growth rate (1.41%), feed efficiency ratio (FER) (0.45), protein efficiency ratio (1.29) and protein gain (161.6%) were recorded in T3 group fed Artemia bioencapsulated L. sporogenes over the control group (P<0.05). Although insignificant (P>0.05), growth‐promoting effects of L. sporogenes were found to be higher than L. acidophillus. Survival of the postlarvae was not affected by probiotics in the diet.     

Improvement of intestinal barrier,immunity, and meat quality in common carp infected by Aeromonas hydrophila using probiotics

The fish pathogen Aeromonas hydrophila causes gastrointestinal tract infections and hemorrhagic septicemia and represents a widespread risk in aquaculture. This study aimed to determine whether compound probiotics could improve the intestinal barrier, immunity, and meat quality of common carp infected by A. hydrophila by feeding them compound probiotics. Carp were assigned to one of four groups: (1) control (no infection, no probiotics); (2) control?+?probiotic; (3) A. hydrophila infected; and (4) the A. hydrophila?+?probiotic group. At the beginning of the experiment, the carp were injected with either saline (0.86%) or A. hydrophila (4.87?×?10⁷ CFU/mL). After 2 weeks of the feeding regime, results suggested that in A. hydrophila infected carp, dietary probiotics regulated the intestinal microflora as evidenced by a reduced abundance of Proteobacteria and increased amounts of Firmicutes and Fusobacteria in the intestine. In addition, dietary probiotics ameliorated both villus swelling and the decrease in villus height induced by A. hydrophila (P?<?0.05). Moreover, probiotics prevented the A. hydrophila-induced decline of Occludin, Claudin-1, and ZO-1 levels in the intestine (P?<?0.05). The addition of probiotics into the feed also increased acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM) activity in the serum (P?<?0.05), in comparison to the A. hydrophila group. Importantly, when compared to A. hydrophila-infected carp, compound probiotics alleviated the decrease in muscle nutrient quality (P?<?0.05). In summary, we show that dietary compound probiotics can prevent the impact of A. hydrophila infection on the intestinal function and disease resistance, while ensuring carp muscle quality.

     

Evaluation and characterization of a novel probiotic Lactobacillus pentosus PL11 isolated from Japanese eel (Anguilla japonica) for its use in aquaculture

In the present study, a potential Lactobacilli probiotics were isolated from Japanese eels (Anguilla japonica) and characterized and evaluated for their possible use in eel farming. Sixteen Lactobacilli were isolated from intestines of Japanese eels, using selective media. The lactobacilli strains (represented as PL1 to PL16) were screened by their ability to produce digestive enzyme. Among these, three strains (PL11, PL13 and PL16) producing four digestive enzymes (amylase, cellulase, protease and phytase) simultaneously were characterized further using API ZYM kit. From these, PL11 (Lactobacillu (L.) pentosus) was identified as potential probiotics candidate producing 15 enzymes among 20 tested. Further examination of biological activities of PL11 revealed tolerance against pH, artificial bile juice and antibacterial activity against several fish pathogenic bacteria. The in vitro competitive exclusion assay also revealed 88.4% reduction in adhesion of fish pathogen (Edwardsiella tarda) by PL11 to host intestinal mucus. In vitro incubation of Japanese eel foregut with Baclight‐labelled PL11 showed colonization of the enterocyte surface by confocal and scanning electron microscopy. In summary, PL11 isolated from eels could serve as a potential probiotics with acid and bile tolerance, production of digestive enzymes, antibacterial activity and inhibition of fish pathogen adhesion to intestinal mucus.     

Autochthonous probiotic Lactobacillus sp. in the diet of bullfrog tadpoles Lithobates catesbeianus improves weight gain,feed conversion and gut microbiota

Dietary supplementation with probiotics in animal production is an alternative to antibiotics. In frog culture, studies involving native strains of probiotic bacteria and their effects on the performance and intestinal histology of farmed animals are scarce. Therefore, this study aimed to evaluate a diet supplemented with Lactobacillus sp. in tadpoles of Lithobates catesbeianus. This randomized test was performed with two dietary treatments: non‐supplemented control diet and diet supplemented with Lactobacillus sp., with nine replications. The growth performance of Lactobacillus sp., including its bacterial enzymatic activity and stability in feed, as well as colonization and histology of the intestinal tract, was evaluated after 42 days of experimentation. Animals fed with a supplemented diet showed higher weight gain and concentration of lactic acid bacteria in the gut and lower feed conversion. No significant difference was observed in survival, total heterotrophic bacterial count or histological change in the gut between the two treatments. The Lactobacillus sp. strain was able to colonize the intestinal tract and feed and remain at a high concentration of 10⁷ and 10⁶ CFU g^?1, respectively. It produced several enzymes, which might have contributed to the greater weight gain and lower feed conversion in the supplemented animals, thus demonstrating its probiotic potential for use as a dietary supplement in bullfrog tadpoles.     

Haematologic and immunologic parameters of bullfrogs,Lithobates catesbeianus,fed probiotics

The effects of two probiotics (P₁–Lactobacillus acidophilus, Bifidobacterium bifidum and Enterococcus faecium and P₂–Bacillus subtilis) supplemented to commercial feed (40% crude protein) on the haematological and immunological parameters of the bullfrog Lithobates catesbeianus were studied. Two doses of each probiotic (5 and 10 g kg^?1 of food) were added to the diets and fed to frogs, totalling five treatments over 112 days. Haematological analyses consisted of total and differential leucocyte counts, erythrocyte and thrombocyte counts, haematocrit, haemoglobin levels and RBC indices (mean corpuscular volume, mean corpuscular haemoglobin – and mean corpuscular haemoglobin concentration) and the immunological parameters included phagocytic capacity and phagocytic index of peritoneal phagocytes. The results showed that the probiotics did not significantly influence any of the haematological parameters measured. However, immunological assays showed that the probiotics had an immunostimulating effect. The greatest effects were seen with probiotic P₁ fed at a dose of 10 g kg^?1 of diet and probiotic P₂ fed at 5 g kg^?1 of diet.     

Effects of dietary administration of two Patagonian probiotics on the zoeae I from the swimming crab Ovalipes trimaculatus

In this study, we analyzed the effects of supplying two alternative Patagonian-autochthonous probiotic strains of Bacillus sp. and Lactococcus sp. or no probiotic at all (control treatment) on the survival, stage duration, molting success to the next larval instar, and speed of vertical displacement of the zoeae I of the southern surf crab Ovalipes trimaculatus, also considering two alternative methods for their administration: (i) direct inoculation into the rearing seawater or (ii) bio-encapsulation in Artemia persimilis nauplii. Both probiotic strains were effectively bio-encapsulated by the nauplii at high CFU x naupli^?1. Probiotic inoculation in the rearing seawater improved the performance of the zoeae over the control treatment and also outperformed bio-encapsulation. Bio-encapsulation in Artemia nauplii resulted in lower survival and molting success to zoeae II compared to the control treatment. No significant differences were observed between the mean effects of either one of the two probiotics. Overall, our results show that the direct inoculation of any of both probiotics into rearing seawater improves the larval culture of the portunid crab.

     

Effects of fish origin probiotics on growth performance,immune response and intestinal health of shrimp (Litopenaeus vannamei) fed diets with fish meal partially replaced by soybean meal

This study was conducted to evaluate the application of four fish origin probiotics to relieve the side effects induced by SBM in shrimp. Shrimps were fed with high fish meal diet (C0, positive control containing 500 g/kg FM and 0 g/kg SBM), low fish meal diet (C, control containing 250 g/kg FM and 340 g/kg SBM) or the control diet (C) supplemented, respectively, with 1.0 × 10⁸ CFU/g Bacillus pumilus SE5 (T1), Psychrobacter sp. SE6 (T2), Enterococcus faecium MM4 (T3) or Bacillus claussi DE5 (T4). After 6 weeks of feeding, the growth performance, hepatopancreatic digestive enzymes, intestinal histological structure and immune parameters were determined. The results showed that compared with high FM group (C0), significantly lower WGR and SGR as well as higher FCR were observed in the low FM group (C) (p < .05). Compared with the control, significantly lower FCR was observed in treatment T1 (p < .05), but not in treatments T2–T4. Protease, amylase and lipase activities in all the treatments (T1‐T4) were significantly higher than the control (p < .05). GOT and GPT activities in the control were significantly lower than the C0 (p < .05), while higher MDA level was recorded in the control (p < .05). Meanwhile, higher GPT activities were observed in treatments T1‐T4 compared with the control (p < .05). Compared with the C0, lower SOD, ACP and AKP activities were observed in the control. Significantly improved SOD and AKP activities were observed in all probiotic feeding groups compared with the control (p < .05). The control diet led to significant reduction of intestinal wall thickness and villus height compared with the C0 (p < .05), while significantly higher intestinal wall thickness and villus height were exhibited in all the probiotic groups compared with the control (p < .05), except intestinal wall thickness in treatment T3. Thus, these results demonstrated that the four fish origin probiotic strains could relieve the side effects induced by high level of SBM in shrimp, while probiotic B. pumilus SE5 showed the best performance.     

Dietary Multispecies Probiotic Supplementation Enhances the Immunohematological Responses and Reduces Mortality by Aeromonas hydrophila in Labeo rohita Fingerlings

A study was conducted to determine the immunomodulatory effect of probiotic mixture on the nonspecific immune response and disease resistance of Labeo rohita. Fish were fed with six different test diets with or without probiotics, namely, T1 (basal feed [BF] without probiotics), T2 (BF + Bacillus subtilis [BS] + Lactococcus lactis [LL]), T3 (BF + LL + Saccharomyces cerevisiae [SC]), T4 (BF + BS + SC), T5 (BF + BS + LL + SC), and T6 (BF + heat‐killed bacteria of BS + LL + SC). During the prechallenge period, significantly higher (P < 0.01) nitroblue tetrazolium assay (NBT), red blood cells (RBC), hemoglobin (Hb), globulin, and albumin content were recorded in the probiotic‐supplemented diet group. Feeding all the three probiotics significantly decreased (P < 0.01) the serum aspartate aminotransferases (AST), alanine aminotransferases (ALT), acid phosphatase (ACP), and alkaline phosphatase (ALP) activity and increased the myeloperoxidase (MPO) and immunoglobulin M (IgM) value. Significant difference in the NBT, Hb, serum albumin, and MPO was noticed in both prechallenged and postchallenged fish. Lowest survivability was recorded in the nonprobiotic fed group, followed by the group fed with heat‐killed probiotics, whereas the highest survivability was noticed in the group fed with all the three probiotics in live form. Thus, a probiotic mixture diet constituting of three probiotics in viable form is found to be optimum to enhance the immunity and postinfection survivability in L. rohita fingerlings.     

Isolation and use of beneficial microbiota from the digestive tract of lions‐paw scallop Nodipecten subnodosus and winged pearl oyster Pteria sterna in oyster aquaculture

We isolated microbiota from the digestive tract of Nodipecten subnodosus and Pteria sterna and determined in vitro their haemolytic activity, antagonism against Vibrio spp., bacterial hydrophobicity, production of extracellular enzymes and molecular identification. Five bacterial strains were selected: RL5 and C3 (Lactobacillus spp.) and PB1‐1, PB1‐5 and PB1‐6 (Bacillus spp.). The RL5 and C3 isolates showed antimicrobial activity against Vibrio spp. and the PB1‐1, PB1‐5 and PB1‐6 isolates showed enzymatic activity for amylase, protease, lipase and cellulose; the C3 and PB1‐5 isolates were highly hydrophobic. The selected strains of bacteria were tested in vivo as probiotics, together with a treatment of ampicillin and a control without bacteria on juvenile Kumamoto oysters Crassostrea sikamea. The strains were provided individually and as mixes of isolates. Survival, growth and biochemical composition of the juveniles were determined as in vivo indicators. Juveniles grew significantly larger and faster when treated with a specific mix of isolates (MIX‐B), compared with the control. The protein, lipid and carbohydrate concentrations were also significantly higher in oysters exposed to probiotic treatments, compared with the control and the antibiotic treatment. The selected microbiota showed probiotic proprieties for cultivating C. sikamea juveniles.     

Biolog-ECO方法探究饲喂益生菌对凡纳滨对虾肠道微生物代谢及有效作用时间的影响

以霍氏肠杆菌(E3)和乳酸菌(R3)2株益生菌对凡纳滨对虾进行为期4周的养殖饲喂实验,饲喂后期利用Biolog-ECO方法对实验组及空白组的凡纳滨对虾肠道微生物菌群多样性的差异进行比较分析,以评价益生菌对凡纳滨对虾肠道微生物菌群代谢功能的影响。结果显示,添加霍氏肠杆菌(E3)或乳酸菌(R3)的实验组,与空白组相比较,平均每孔颜色变化率显著上升,表明益生菌增强了肠道微生物活性;凡纳滨对虾肠道微生物利用各类碳源的整体能力显著增强,表明益生菌可以促进水产动物的代谢功能;肠道微生物多样性指数(包括Shannon、Simpson和McIntosh指数)有明显差异,表明饲喂2株益生菌可以提高凡纳滨对虾肠道菌群的丰富度。其中,停喂霍氏肠杆菌后第1天和第5天取样结果表明,Shannon指数显著降低,Simpson和McIntosh指数显著升高;停喂乳酸菌后的第1天和第5天取样结果表明,Shannon指数无显著差异,Simpson和McIntosh指数显著升高;二者在第10天取样的结果中均无显著差异,表明饲料中添加益生菌可以改变凡纳滨对虾肠道内原有菌群的数量和结构,促进对虾肠道内微生物群落间复杂的相互作用,进而在维持或者促进对虾健康方面发挥着重要的作用,同时也表明此两株益生菌在凡纳滨对虾肠道中停留时间最少为5 d。     

Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae: Effects on growth performance and digestive enzyme activities

In this study, the influence of commercial probiotic, Lactobacillus spp., supplementation was investigated on growth parameters and digestive enzyme activities in gilthead sea bream, Sparus aurata, during larval development. All experiments were triplicated and designed in three different administrations of probiotic from 3 days after hatching (DAH) concurrently with starting of exogenous feeding. In the first group, probiotic was added to live food (rotifer and Artemia). In the second group, probiotic was supplemented directly to both live food and water. In the third group, probiotic was added directly to water. Also, no probiotic treatment was maintained in control group. Total bacterial counts among probiotic probiotic-supplemented groups were significantly different from total bacterial counts in controls in water and digestive tract of larvae (p < 0.05). The mean of total bacterial counts in control was approximately 4 × 10⁴-fold increased from the experimental groups in the sea water (p < 0.05). Besides, mean digestive enzyme activities of all probiotics treatment groups were significantly different (p < 0.05) with that of the control. Except probiotic water supplementation group, in all treatments, the specific activities of pancreatic and intestinal enzymes were significantly higher (p < 0.05) in larvae to which probiotic had been supplemented by live food and live food with water. Also, S. aurata larvae that had probiotic administered by live food with water demonstrated significant (p < 0.05) increases in both survival (13–105% higher) and specific growth rate (2–9% higher) as compared to controls. As a result, supplementation of probiotic to directly tank water could not significantly increase growth parameters and digestive enzyme activities and therefore, administration of probiotics by this method would not be effective in terms of husbandry parameters and nutritional condition.     

Growth,survival and enzyme activity of lions‐paw scallop (Nodipecten subnodosus) spat treated with probiotics at the hatchery

Different types and concentrations of probiotics were evaluated for their effects on growth, survival and enzyme activity of hatchery‐reared Nodipecten subnodosus spat. The treatments included (1) a mix of the microalgae Isochrysis galbana and Pavlova salina as the control group; (2) Diet 1 + a mix of Burkholderia cepacia and Pseudomonas aeruginosa; (3) Diet 1 + the commercial probiotic Epicin; and (4) Diet 1 + the commercial probiotic Bactosafe. Both commercial probiotics were tested at 3, 6 and 10 mg L⁻¹. Scallops receiving Bactosafe and Epicin at 3 and 6 mg L⁻¹ grew ~30% faster and larger than in the control group. Regardless of the treatment, survival at the end was 98.7%. There was no relation between the activity of catalase and superoxide dismutase; however, significantly lower catalase levels occurred at increasing doses of Epicin; significant higher catalase levels occurred in the control group and mix of bacilli. Compared with the control group, lysozyme levels were significantly higher with the Epicin and Bactosafe treatments (3 and 6 mg L⁻¹). More research needs to be conducted with probiotics to determine their real effects on the activity of antioxidant enzymes. We expect that treated scallops will increase their performance at the field.     

鲫肠道乳酸菌的分离及益生特性

为获得鱼源益生乳酸菌,本研究从健康鲫肠道内分离鉴定得到38株乳酸菌,并选择其中8株乳酸菌进行体外益生特性分析。结果显示,8株乳酸菌均能在p H=4.5和10%鲫胆汁环境中存活,对嗜水气单胞菌、豚鼠气单胞菌、无乳链球菌和副溶血弧菌均有较强的抑菌能力,但菌体表面疏水性和自凝集能力具有菌株特异性。选择5株疏水性较高(63%~89%)、自凝集能力强(37%~45%)的乳酸乳球菌进行体外黏附能力测定,结果显示,黏附能力在菌株间具有差异性(4.5%~7.9%),但均能显著降低嗜水气单胞菌NJ-35的体外黏附率,黏附抑制率达30%~35%;最后对筛选出的3株高黏附力的乳酸乳球菌进行安全性评价,发现3株菌对鲫均无致病力。鱼源乳酸菌的筛选,为鲫养殖生产中益生菌的实际应用提供了理论依据。     

Assessing the use of a dietary probiotic/prebiotic as an enhancer of spinefoot rabbitfish Siganus rivulatus survival and growth

The use of prebiotics and probiotics as feed supplements that improve efficiency of intestinal bacteria is becoming de rigueur in animal husbandry in many regions worldwide. We tested the effects of a commercial probiotic (Biogen^®) containing allicin, high unit hydrolytic enzyme, Bacillus subtilis spores and ginseng extracts on survival, growth, carcass composition and feed cost/benefit in rabbitfish Siganus rivulatus. Fifteen net cages (100 × 100 × 40 cm; L × W × H) were stocked with 10 juvenile rabbitfish (10.3 g per fish) each and placed in a large rectangular tank and offered feed at 4% body weight daily. Cages were offered one of five isonitrogenous and isocaloric diets containing 0, 1, 2, 3 and 4 g kg⁻¹ probiotic at three replicates per treatment for 98 days. Fish in all cages were weighed at 2‐week intervals and feed regimen was adjusted accordingly. Rabbitfish offered the control diet exhibited lower growth and feed utilization than all experimental treatments. There was no effect of probiotic inclusion level on survival but growth was better at all inclusion levels than in the control. No significant differences (P > 0.05) in growth were observed among fish groups fed various levels of the probiotic. Carcass composition was not affected by dietary probiotic inclusion. Ultimately, when all variables are considered, Biogen^® inclusion to diets appears to reduce feed cost per unit growth of rabbitfish.     

黏红酵母表面黏附蛋白及肠黏液受体的初步研究

为获得对大菱鲆肠道具有较强黏附能力的益生菌株,了解其黏附机制,采用体外黏液黏附模型及Western-blot技术,对黏红酵母等4株益生菌的大菱鲆肠黏液黏附性能及黏红酵母表面粘附蛋白和肠黏液受体进行了初步研究.结果表明,4株益生菌均可黏附于大菱鲆肠黏液;但各菌株的黏附能力存在较大差异,其中黏红酵母黏附能力最强,鼠李糖乳杆菌次之,假丝酵母和枯草芽孢杆菌相对较低.黏红酵母表面蛋白预先与肠黏液孵育2h后,显著抑制了黏红酵母与肠黏液的结合,其黏附百分率下降了83.6%;但上述处理对其他3株益生菌的黏附没有明显影响.Western-blot显示黏红酵母表面蛋白中分子量为38.5ku和28.6ku的两个蛋白参与对肠黏液的黏附过程,大菱鲆肠黏液中分子量为27.3ku和22.3ku两个蛋白可与黏红酵母表面蛋白特异性结合.糖原染色显示上述4个蛋白均为糖蛋白.     

Evaluating the use of Lactobacillus acidophilus as a biocontrol agent against common pathogenic bacteria and the effects on the haematology parameters and histopathology in African catfish Clarias gariepinus juveniles

This study was carried out to evaluate the use of Lactobacillus acidophilus as a biocontrol agent against some common fish pathogenic bacteria (Staphylococcus xylosus, Aeromonas hydrophila gr.2 and Streptococcus agalactiae) in African catfish, Clarias gariepinus. Eight treatments were designed inclusive of 10 C. gariepinus juveniles (mean weight 190 g) per tank, each in triplicate. Four groups of fish were fed a diet supplemented with L. acidophilus, comprising about 3.01 × 10⁷ colony‐forming units per gram of diet (the probiotics diet), while the other four groups were fed a diet not supplemented with probiotics (the non‐probiotics diet). In the first group, fish were injected with 1 mL physiological saline and fed the non‐probiotic diet (non‐probiotic control); in the second, third and fourth groups, fish were injected with 1 mL each of S. xylosus, A. hydrophila gr.2 and S. agalactiae, respectively, and were all fed the non‐probiotic diet (designated as non‐probiotic treatments; NPsx, NPah and NPsa respectively). In the fifth group, fish were injected with 1 mL physiological saline but fed the probiotic diet (probiotic control), while fish in the sixth, seventh and eighth groups were each injected with 1 mL of S. xylosus, A. hydrophila gr.2 and S. agalactiae, respectively, and were all fed the probiotic diet (and designated as probiotic treatments; Psx, Pah and Psa respectively). Blood samples were collected for haematology analysis, while samples of the liver and kidney were examined for pathohistology after 7 and 21 days of infection. The results showed that the haematology parameters, packed cell volume, haemoglobin, erythrocyte sedimentation rate, red blood cell, white blood cell, total serum protein, Mg²⁺, Ca²⁺, Cl^?, glucose, cholesterol and total immunoglobulin concentrations and the pathohistology of the liver and kidney were better in the challenged fish (infected) maintained on the probiotic diet than those in the groups fed the non‐probiotic diet. It is concluded, based on these results, that L. acidophilus is useful as a probiotic agent in C. gariepinus against these pathogenic bacteria (S. xylosus, A. hydrophila gr.2 and S. agalactiae).