A review of intestinal microbes in grass carp Ctenopharyngodon idellus (Valenciennes)

Grass carp, Ctenopharyngodon idellus, harbours complex intestinal bacterial communities, which are important in several physiological processes of their host. Intestinal microbiota of grass carp have been previously described in numerous studies. However, an overview on the bacterial community diversity, including their establishment, their functions in host's nutritional processes and immune‐related responses, and use as probiotics, is absent. This study aimed to summarize the current understanding of the grass carp intestinal microbiota. In this review, we provide general information on the establishment and composition of intestinal microbial communities and factors influencing the diversity of gut microbiota. Also, this review covers the dietary effects of probiotics, prebiotics and/or synbiotics on the grass carp intestinal microbial communities and physiological characteristics. Although our knowledge of the grass carp intestinal microbiota is expanding rapidly, further studies on the factors affecting the diversity of intestinal microbes, interactions between intestinal microbiota and their hosts and application of probiotics/prebiotics/synbiotics in aquaculture industry, are needed.     

Analysing the effect of dietary prebiotics and probiotics on gut bacterial richness and diversity of Asian snakehead fingerlings using T‐RFLP method

This study was designed to evaluate the effects of three prebiotics (β‐glucan, galacto‐oligosaccharide [GOS], mannan‐oligosaccharide [MOS]) and two probiotics (Saccharomyces cerevisiae, Lactobacillus acidophilus) on the microbiome of snakehead during growth of fingerlings. In addition, the experiment evaluated the capacity of Channa striata fingerlings to retain the benefits derived from these supplements after withdrawal. Throughout the study, it was observed that supplementation with dietary prebiotics and probiotics led to significant (p < 0.05) change in gut bacterial profile and improvement in gut morphology. Terminal restriction fragment length polymorphism (T‐RFLP) was used for the comparative analysis of gut communities and all 46 of the T‐RFLP detected phylotypes were present in the Lactobacillus supplemented fish, while significantly fewer were detected in controls and other experimental supplement regimes. Histological studies and electron microscopy revealed that both the prebiotic and probiotic treated fish had significantly longer and wider villi and deeper crypts compared to the controls. The microvilli length, as evaluated with electron microscopy, was also longer in all treated fish compared to controls. Furthermore, this study is the first to report the absence of differences in sustaining the efficacies attained after intake of β‐glucan, GOS, MOS and live yeast upon post‐feeding with an unsupplemented feed, over a prolonged period.     

Modulation of nutrient digestibility and digestive enzyme activities in aquatic animals: The functional feed additives scenario

Considering the costs of feed costs (nearly 60% of production cost), nutrition, feeding and feed utilization are among the most important factors in commercial aquaculture. During the last decade, administration of functional feed additives has been practiced for enhancing nutrient digestibility and digestive enzyme activities of cultured fish and shellfish. Traditionally, antibiotics were used for boosting growth performance and nutrient digestibility in commercial aquaculture. However, emergence of resistance pathogens and possible risk to human health resulted in limitation or prohibition of prophylactic administration of antibiotics. Recently, there was increasing attentions towards dietary administration of functional feed additives that include probiotics, prebiotics and synbiotics for elevation of digestive enzyme activity and nutrient digestibility. The results of those studies revealed contradictory effects of different pro‐, pre‐ or synbiotics on various fish species. It seems that the effects are species specific and related to modulation of the intestinal microbiota. In view of this issue, the present review provides a comprehensive sight on the effects of different pro‐, pre‐ and synbiotics on digestive enzyme activity and nutrient digestibility in different species with special focus on the mode of action. In addition, the present review highlighted the gaps of existing knowledge as well as suggesting the subjects which needs additional studies.     

The Effects of Dietary Yeast Culture or Short‐chain Fructo‐oligosaccharides on the Intestinal Autochthonous Bacterial Communities in Juvenile Hybrid Tilapia,Oreochromis niloticus♀×Oreochromis aureus♂

The effects of dietary yeast culture (YC) or short‐chain fructo‐oligosaccharides (FOS) on intestinal autochthonous bacterial communities in juvenile hybrid tilapia, Oreochromis niloticus♀×Oreochromis aureus♂ were studied by 16S rDNA denaturing gradient gel electrophoresis (DGGE). Ninety Tilapias in tanks (10 fish per tank) were randomly and equally divided into three groups. At the end of an 8‐wk feeding period of CK (the control treatment), YC (3 g/kg), or FOS (1 g/kg), autochthonous gut bacteria were analyzed in intestinal samples of all fish in each tank of a recirculating aquaculture system. The clear differences in the banding patterns indicated the obvious effects of dietary prebiotics on intestinal communities in hybrid tilapia. Higher variation was detected within the dietary YC group. This difference might be due to the effects of certain immune‐stimulating agents in YC on the immunity response of hybrid tilapia. It was concluded that dietary prebiotics, YC, and FOS obviously affected the intestinal bacterial community in hybrid tilapia with different patterns for different kinds.     

Probiotics,immunostimulants, plant products and oral vaccines,and their role as feed supplements in the control of bacterial fish diseases

There is a rapidly increasing literature pointing to the success of probiotics, immunostimulants, plant products and oral vaccines in immunomodulation, namely stimulation of the innate, cellular and/or humoral immune response, and the control of bacterial fish diseases. Probiotics are regarded as live micro‐organisms administered orally and leading to health benefits. However, in contrast with the use in terrestrial animals, a diverse range of micro‐organisms have been evaluated in aquaculture with the mode of action often reflecting immunomodulation. Moreover, the need for living cells has been questioned. Also, key subcellular components, including lipopolysaccharides, have been attributed to the beneficial effect in fish. Here, there is a link with immunostimulants, which may also be administered orally. Furthermore, numerous plant products have been reported to have health benefits, namely protection against disease for which stimulation of some immune parameters has been reported. Oral vaccines confer protection against some diseases, although the mode of action is usually linked to humoral rather than the innate and cellular immune responses. This review explores the relationship between probiotics, immunostimulants, plant products and oral vaccines.     

益生菌与鱼类肠道健康研究进展

集约化、高密度养殖已成为水产养殖业的发展趋势.在这种养殖模式下,如何保障鱼体健康是人们关注的焦点.20世纪80年代,益生菌在水产养殖行业中首次应用,这种环保且有效的水产养殖策略获得了业界的关注.目前,鱼类养殖中常用益生菌有芽孢杆菌属、乳酸杆菌属、乳球菌属和酵母菌属.添加的益生菌通过抑制病原微生物的增殖、改善鱼类肠道屏障...     

“水陆互补”理念下的水产品营养健康功效

水产养殖在全球粮食安全中发挥着至关重要的作用,为人类提供优质蛋白源持续稳定供应提供了重要保障。益生菌是一类可用于水产养殖的有益微生物,不仅能够提高水产动物的生长性能、免疫能力,而且具有抑制致病菌、改善养殖水质等功效。益生菌在水产养殖中的推广应用可促进水产养殖业的健康可持续发展。因此,本文首先介绍了水产养殖中益生菌的定义与来源,详细阐述了益生菌对水产可持续养殖的意义以及益生菌在水产养殖中的实际应用,并对其未来研究方向进行了展望,以期为益生菌在水产养殖可持续发展中的推广和应用提供可靠的理论依据。

     

Effects of dietary supplementation with β‐glucan and synbiotics on growth,haemolymph chemistry,and intestinal microbiota and morphology in the Pacific white shrimp

The goal of this study was to investigate the effects of dietary supplementation with β‐glucan and microencapsulated probiotics (Bacillus subtilis or Pediococcus acidilactici) on growth performance, body composition, haemolymph constituents, and intestinal morphology and microbiota of the Pacific white shrimp Litopenaeus vannamei. Four treatment diets [basal diet (C), β‐glucan‐containing diet (β‐glu), β‐glucan plus B. subtilis‐containing diet (β‐glu+Bs), and β‐glucan plus P. acidilactici‐containing diet (β‐glu+Pa)] were fed to L. vannamei for 90 days. Shrimp fed the β‐glu and β‐glu+Pa diets exhibited similar growth performance and body protein content, which were significantly higher than those of shrimp fed the control diet (P < 0.05). No significant differences in haemolymph triglyceride, cholesterol, protein, haemolymph urea nitrogen or chloride were detected among the experimental diets. However, dietary β‐glucan alone increased the haemolymph glucose level and osmolarity (P < 0.05). Synbiotic supplementation had greater effects on intestinal microbiota and morphology than dietary β‐glucan alone. For example, β‐glu+Bs increased the number of intestinal lactic acid bacteria and decreased the number of Vibrio spp. (P < 0.05), and β‐glu+Pa increased the height of intestinal villi.     

益生菌对幼鱼生长发育的影响及调控机制

近年来,高密度、集约化的水产养殖模式导致养殖鱼类疾病频发,其中幼鱼发病率和死亡率的上升严重影响水产养殖业的健康快速发展。如何保障幼鱼的健康是当前水产养殖业亟需解决的问题。益生菌能够通过多种方式提高幼鱼的抗病力、增强幼鱼对营养素的摄取、促进幼鱼的生长发育。已有大量的研究阐述了益生菌在幼鱼生长发育及免疫调节中的作用,但相关机制还需深入探究。本文综述了近年来益生菌在幼鱼生长发育及抗病方面发挥的有益作用及其可能的机制,讨论了益生菌在幼鱼养殖应用时可能存在的问题以及发展方向,为益生菌在水产动物幼体养殖中的应用提供了理论参考。     

Use of live and dead probiotic cells in tilapia Oreochromis niloticus

ABSTRACT:   To investigate the effect of live and dead probiotic cells on the non-specific immune system of tilapia Oreochromis niloticus , probiotics were introduced by feeding either in the form of live or dead cells, or supplying live cells to the rearing water in a closed recirculating system. The probiotics treatment enhanced non-specific immune parameters such as lysozyme activity, migration of neutrophils and plasma bactericidal activity, resulting in improvement of resistance to Edwardsiella tarda infection. Especially, oral administration of live cells seemed to be more effective compared with other probiotic treatments such as oral administration of dead probiotic cells and supply of live probiotic cells to the rearing water. These results indicate that probiotics treatment is promising as an alternative method to antibiotics for disease prevention in aquaculture, and the viability of probiotic bacteria is a key factor to induce more potential effect of probiotics used for fish production.     

Use of chitin and krill in aquaculture – the effect on gut microbiota and the immune system: a review

Chitin consists of β‐1,4‐linked N‐acetylglucosamine residues and is estimated as the second most abundant biomass in the world after cellulose. However, relatively little chitin is utilized as a material for industrial, agricultural and medical applications and aquacultural purposes. Chitin may be useful as a constitutive material in formulated fish feed, and the interesting effects in fish merit further evaluation. There is evidence that fish and aquatic animals harbour a gut bacterial community that is distinctly different from that reported in the surrounding habitat or in the diet. Thus, the gut environment provides a specific niche, and bacterial activity in the gut is not merely a continuum of that observed in the environment. Today, it is well accepted that the gut microbiota in fish are modulated by dietary manipulations. But to what extent can dietary chitin and krill (chitin‐rich) modulate the intestinal microbiota of fish and how do these dietary components affect the immune system? These questions will be discussed in the present review.     

Application of immunostimulants in aquaculture: current knowledge and future perspectives

Similar to other industries, aquaculture constantly requires new techniques to increase production yields. Modern technologies and different scientific fields, such as biotechnology and microbiology, provide important tools that could lead to a higher quality and a greater quantity of products. New feeding practices in farming typically play an important role in aquaculture, and the addition of various additives to a balanced feed formula to achieve better growth is a common practice of many fish and shrimp feed manufacturers and farmers. As ‘bio‐friendly agents’, immunostimulants, such as biological factors, probiotics and vitamins, can be introduced into the culture environment to control and kill pathogenic bacteria, as well as to promote growth of the cultured organisms. In addition, immunostimulants are non‐pathogenic and non‐toxic and do not produce undesirable side effects when administered to aquatic organisms. In this review, we summarize previous studies performed with both traditional immunostimulants and the most promising new generation of immunostimulants, such as polysaccharides, nutrients, oligosaccharides, herbs, microorganisms, prebiotics and different biological factors. This review primarily focuses on their protective efficacies and on what is known concerning their effects on the immune systems of aquatic organisms when delivered in vivo.     

Probiotics in fish and shellfish culture: immunomodulatory and ecophysiological responses

Aquaculture is emerging as one of the most viable and promising enterprises for keeping pace with the surging need for animal protein, providing nutritional and food security to humans, particularly those residing in regions where livestock is relatively scarce. With every step toward intensification of aquaculture practices, there is an increase in the stress level in the animal as well as the environment. Hence, disease outbreak is being increasingly recognized as one of the most important constraints to aquaculture production in many countries, including India. Conventionally, the disease control in aquaculture has relied on the use of chemical compounds and antibiotics. The development of non-antibiotic and environmentally friendly agents is one of the key factors for health management in aquaculture. Consequently, with the emerging need for environmentally friendly aquaculture, the use of alternatives to antibiotic growth promoters in fish nutrition is now widely accepted. In recent years, probiotics have taken center stage and are being used as an unconventional approach that has numerous beneficial effects in fish and shellfish culture: improved activity of gastrointestinal microbiota and enhanced immune status, disease resistance, survival, feed utilization and growth performance. As natural products, probiotics have much potential to increase the efficiency and sustainability of aquaculture production. Therefore, comprehensive research to fully characterize the intestinal microbiota of prominent fish species, mechanisms of action of probiotics and their effects on the intestinal ecosystem, immunity, fish health and performance is reasonable. This review highlights the classifications and applications of probiotics in aquaculture. The review also summarizes the advancement and research highlights of the probiotic status and mode of action, which are of great significance from an ecofriendly, sustainable, intensive aquaculture point of view.     

Effect of dietary components on the gut microbiota of aquatic animals. A never‐ending story?

It is well known that healthy gut microbiota is essential to promote host health and well‐being. The intestinal microbiota of endothermic animals as well as fish are classified as autochthonous or indigenous, when they are able to colonize the host's epithelial surface or are associated with the microvilli, or as allochthonous or transient (associated with digesta or are present in the lumen). Furthermore, the gut microbiota of aquatic animals is more fluidic than that of terrestrial vertebrates and is highly sensitive to dietary changes. In fish, it is demonstrated that [a] dietary form (live feeds or pelleted diets), [b] dietary lipid (lipid levels, lipid sources and polyunsaturated fatty acids), [c] protein sources (soybean meal, krill meal and other meal products), [d] functional glycomic ingredients (chitin and cellulose), [e] nutraceuticals (probiotics, prebiotics, synbiotics and immunostimulants), [f] antibiotics, [g] dietary iron and [h] chromic oxide affect the gut microbiota. Furthermore, some information is available on bacterial colonization of the gut enterocyte surface as a result of dietary manipulation which indicates that changes in indigenous microbial populations may have repercussion on secondary host–microbe interactions. The effect of dietary components on the gut microbiota is important to investigate, as the gastrointestinal tract has been suggested as one of the major routes of infection in fish. Possible interactions between dietary components and the protective microbiota colonizing the digestive tract are discussed.     

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.     

Immune responses and gut morphology in Senegalese sole (Solea senegalensis) fed dietary probiotic supplementation and following exposure to Photobacterium damselae subsp. piscicida

The aim of this study was to determine the effect of a dietary multi‐species probiotic on growth, gut morphology and immune parameters in Senegalese sole (Solea senegalensis). Fish were fed with two experimental diets, a control diet and the same diet supplemented with the probiotic for 72 days. A sub‐lethal bath challenge with Photobacterium damselae subsp. piscicida was performed after the growth trial. Intestine and blood samples were collected to study gut structure and plasmatic immune parameters. No significant differences were found in growth performance. The analysis of gut morphology showed a significant increase in intestinal villi height of Senegalese sole fed the probiotic. Regarding circulating leucocytes, dietary probiotic supplementation increased thrombocytes levels whereas a decrease in the proportion of lymphocytes was observed. No significant differences were observed in humoural immune parameters. Bath challenge differentially affected leucocyte counts and increased peroxidase activity. This study presents the possibility of using dietary probiotic supplementation to increase Senegalese sole welfare since gut morphology was positively affected. Although the immune response after a bacterial challenge was not modulated by dietary treatment, further studies would be instrumental to unravel eventual dietary benefits on immune mechanisms.     

A Review of Probiotics in Shrimp Aquaculture

Development of shrimp aquaculture has been associated with increases in infectious diseases and environmental degradation. An effective alternative to chemicals and antibiotics used for the prevention of these problems is to administer probiotics into the rearing system. Three bacterial genera, Bacillus, Vibrio, and Pseudomonas, are commonly administered as probiotics in shrimp aquaculture. Candidate probiotics are species specific and need to be tested for their effectiveness for certain species in in vitro and in vivo. Supplementation into feed is more effective in conveying probiotics into animals compared to direct application into rearing systems. Overdosage or prolonged administration of probiotics can induce immunosuppression. A cell-density of 10⁵ colony-forming units (CFU) per ml is widely recommended. A combination of probiotics results in better outcomes for the host than individual probiotics. Probiotics improve water quality while reducing pathogenic bacteria. Probiotics show positive effects through an improvement in the physiological and immune responses of shrimps. Probiotics are increasingly becoming important and more common in any organic shrimp farming.     

1,3‐1‐6 ß‐glucans enhance tissue regeneration in zebrafish (Danio rerio): Potential advantages for aquaculture applications

High aquacultural rearing density and handling of fish may frequently result in skin or gills wounds, thereby facilitating the onset of secondary infections. The capacity of the zebrafish to regenerate tissues, as well as fins and other organs, makes it an ideal animal model for studying the mechanisms of tissue regeneration. Since macrophages are involved in tissue regeneration, a diet including ß‐glucans might positively affect the process through activation of macrophages and other immune pathways. Consequently, the aim of this study was to investigate the effects of the oral administration of 1,3‐1,6 β‐glucans on the regeneration process of the caudal fin after its amputation in zebrafish. One hundred and twenty zebrafish were randomly distributed into four groups with three replicates each: an untreated non‐amputated group (CNA) and an untreated amputated group (CA) fed a control diet; two treated and amputated groups (MI and MII) fed for 14 days the same diet with the addition of two differently extracted 1,3‐1,6 ß‐glucans (MacroGard^® and Experimental MacroGard^®, Biorigin^©). ß‐glucans were added to allowed the administration of 12.5 mg/kg of fish body weight (0.35 g/kg of feed). Results showed that 1,3‐1,6 ß‐glucans decreased fish mortality rate and enhanced both daily and cumulative regenerated fin area, independent of the specific ß‐glucan extraction method used. Based on the mechanisms similarities of the innate immune system and tissue regeneration among different teleost species, these results may likely be extended to species of interest for the aquaculture sector.     

Dietary administration of Bacillus (B. licheniformis and B. subtilis) and isomaltooligosaccharide influences the intestinal microflora,immunological parameters and resistance against Vibrio alginolyticus in shrimp,Penaeus japonicus (Decapoda: Penaeidae)

The experiment was conducted to investigate the effects of oral administration of probiotics (Bacillus licheniformis and Bacillus subtilis) in combination with prebiotic isomaltooligosaccharide (IMO) on the intestinal microflora and immunological parameters of Penaeus japonicus and its resistence against Vibrio alginolyticus. A basal diet was supplemented with 0% probiotics and prebiotic (control), 10⁸ colony forming unit (CFU) g^?1B. licheniformis and 0.2% IMO (T1), 10⁸ CFU g^?1B. subtilis and 0.2% IMO (T2), 10⁸ CFU g^?1B. licheniformis in combination with 10⁸ CFU g^?1B. subtilis and 0.2% IMO (T3). The results showed that the total bacterial counts significantly increased (P<0.05) and Vibrio counts significantly decreased (P<0.05) in the intestine of shrimp with supplementation of dietary synbiotics. Shrimp fed the diet with both Bacillus probiotics and IMO (T3) produced significantly higher immune parameters (phenoloxidase activity, lysozyme activity and nitric oxide synthase activity and superoxide dismutase activity) than the control group (P<0.05). Significant lower cumulative mortality after challenge with V. alginolyticus was also observed in shrimp fed diet with both Bacillus probiotics and IMO (T3) than that in the control group (P<0.05). The results indicated that the oral administration of probiotics (B. licheniformis and B. subtilis) and prebiotic IMO has positive effects on bacterial flora in P. japonicus gut, and can activate non‐specific immunity of shrimp.