Does chromic oxide (Cr2O3) affect faecal lipid and intestinal bacterial flora in Arctic charr. Salvelinus alpinus (L.)?

Abstract. Arctic charr. Salvelinus alpinus (L.), were fed a commercial diet with or without a supplement of 1% chromic oxide (Cr₂O₃) in fresh water at 8°C for 70 days. Supplementation of the diet with 1% chromic oxide did not affect the lipid composition of stomach contents, but did affect that of the faeces.
Highest total lipid content in faeces was observed when fish were fed on the diet supplemented with chromic oxide. The proportion of triacylglycerols and free fatty acids increased in the neutral lipid fraction. The level of the dietary fatty acids 16:0, 16:1 and 18:1 increased, while 20:1 (n-9), 22:1 (n-11) and 18:2 (n-6) decreased significantly (P < 0·05) compared to the unsupplemented diet.
Total viable counts (TVC) of aerobic microorganisms were somewhat higher in stomach contents when the charr were fed on the unsupplemented diet. In faeces from charr fed the unsupplemented diet, TVC were higher, by a factor of 10 compared to fish fed diet supplemented with chromic oxide. Bacterial composition in stomach content of both rearing groups were dominated by Pseudomonas sp. and Agrobacterium sp., but the number of Lactobacillus sp. and Streptococcus sp. enhanced in stomach content when the fish were fed on the chromic oxide enriched diet. The faecal bacterial flora were also different between the rearing groups. The predominant bacterial species present in the faeces of fish fed unsupplemented diet were identified as Pseudomonas sp. and Aeromonas sp. Other bacterial species isolated included Lactobacillus sp., Streptococcus sp., Agrobacterium sp., Flavobacterium sp. and Vibrio sp. In contrast, the bacteria flora in faeces of fish fed the chromic oxide diet was dominated by the Gram-positive bacteria Streptococcus and Lactobacillus.
It is suggested that the use of chromic oxide as a marker in digestibility studies in Arctic charr may not be ideal.     

Identification of bacteria from the normal flora of perch, Perca fluviatilis L., and evaluation of their inhibitory potential towards Aeromonas species

Pathogenic Aeromonas sobria has been identified as a causative agent of ulcerative disease in farmed European perch, Perca fluviatilis L. To study the effect of the normal intestinal bacterial flora of perch against A. sobria, we sampled 193 bacterial isolates from the perch digestive tract. The isolates were identified by sequence analysis of the 16S rRNA gene and their inhibitory potential against A. sobria was evaluated in vitro. Nineteen of the strains isolated showed inhibition and were also tested against other aeromonad and non-aeromonad fish pathogens including Yersinia ruckeri and Vibrio anguillarum. Isolates showing inhibition were primarily Pseudomonas spp.; however, inhibitory Shewanella spp., and Delftia sp. were also identified. A Pseudomonas chlororaphis isolate showed inhibition against all fish pathogens tested.     

A Lactobacillus sp. from diseased female rainbow trout, Salmo gairdneri Richardson, in Newfoundland, Canada

Abstract. Post-spawning mortalities of 2- to 3-year old female rainbow trout, Salmo gairdneri Richardson, at a Newfoundland hatchery were studied. The fish were found to have a mixed bacterial infection involving primarily a Lactobacillus sp. (a Gram-positive, chain-forming coccobacillus) and to a lesser degree Enterobacteriaceae, Aeromonas hydrophila and Pseudomonas fluorescens. The bacteria were isolated from the visceral organs, kidney, and ascitic fluid present in the coelomic cavity. The fish had extensive degeneration and necrosis of the liver, spleen and kidney, and sloughing of intestinal epithelium, but whether these were caused directly by the bacterial infection was not established because the fish appeared to have received substantial stress from the mechanical stripping and from the retention of dead, unshed eggs in the coelomic cavity. Tissue sections suggested that substantial growth of the Lactobacillus sp. was occurring within these unshed eggs. The present study describes this unusual observation and compares the Lactobacillus sp. to two other Lactobacillus isolates from cultured trout.     

Surveillance of health status on eight marine rainbow trout, Oncorhynchus mykiss (Walbaum), farms in Denmark in 2006

The health status of eight marine rainbow trout farms was followed from mid-June to mid-September 2006 by sampling both dead and healthy fish approximately every 2 weeks for bacteriological and virological investigation. No fish pathogenic viruses were detected, but all farms experienced disease and mortality as a result of various bacterial infections. Yersinia ruckeri was found on four and Renibacterium salmoninarum on five of the farms, but only during the first part of the surveillance period. This indicates that the fish carried the infection from fresh water, and cleared the infection in salt water. Aeromonas salmonicida subsp. salmonicida caused mortality on five farms, but persisted throughout the sampling period. Although A. salmonicida was probably carried from fresh water, the fish were not able to clear the infection in the sea. Vibrio anguillarum caused mortality on six of the farms throughout the sampling period, O1 being the dominant serovar, and Photobacterium damselae subsp. damselae was found on seven farms as a cause of disease. During the period of highest water temperatures Vibrio parahaemolyticus and Vibrio vulnificus were detected in dead fish in five and two farms, respectively, although their significance as causative pathogens is questionable. Vibrio vulnificus has not previously been found in rainbow trout in Denmark. Both mortality and number of antimicrobial treatments during the period were considerably higher in unvaccinated compared with vaccinated fish. Resistance to commonly used antimicrobials was low or absent.     

穿心莲对草鱼肠内细菌的影响

给不同组草鱼分别投喂含0％、0.5％、1％和2％穿心莲的人工配合饲料，在不同时期取样检测各组草鱼前、中、后肠需氧和兼性厌氧菌的种类和数量。结果显示，草鱼肠内细菌的种类，特别是优势菌群的种类没有受到穿心莲的影响。各组草鱼肠内各部位的优势菌群均为气单胞菌Aeromonas和肠杆菌Enterobacteriaceae。草鱼肠内细菌数量为105～108CFU·g-1，分析后发现，各组草鱼肠内细菌数量之间没有显著差异(P＞0.05)。但1％及2％的穿心莲能降低气单胞菌的组成。上述结果表明，一定浓度的穿心莲可以通过降低优势菌群气单胞菌的组成而对草鱼肠内微生态系产生影响。     

The bacterial flora of rainbow trout, Salmo gairdneri Richardson, and brown trout, Salmo trutta L., eggs and its relationship to developmental success

Abstract. The bacterial flora of rainbow, Salmo gairdneri Richardson, and brown trout, Salmo trutta L., eggs was studied during incubation. Few bacteria were considered to exist internally, although Pseudomonas species and Aeromonas hydrophila were isolated. During incubation, large numbers of bacteria approaching 500 colony forming units/mm² accumulated around egg surfaces. Both rainbow and brown trout eggs placed in a through-flow system were colonized mainly by Pseudomonas sp. and A. hydrophila , whilst rainbow trout eggs in a separate recycling system were dominated by a Cytophaga species. Glass beads were also incubated (as an inert surface) and found to support significantly lower numbers of bacteria. Egg mortalities were recorded daily and found to differ significantly between both fish species and water conditions. Statistical analysis indicated a potentially significant correlation between hatching success and numbers of surface bacteria.     

Effects of Dietary Short-chain Fructooligosaccharides on Intestinal Microflora, Survival, and Growth Performance of Juvenile White Shrimp, Litopenaeus vannamei

Abstract.— Five groups of juvenile white shrimp, Litopenaeus vannamei , in triplicate were separately fed a compound diet with additional short-chain fructooligosaccharides (ScFOS, Profeed^® 95%) 0, 0.4, 0.8, 1.2, or 1.6 g/kg dried diet for 8 wk to investigate the effects of ScFOS on the intestinal microflora, survival, and growth performance of the shrimp. The juvenile shrimp with around 0.17 g of initial body weight were divided into 30 per tank and reared in a standard water recirculation system at 28 C. The shrimp were fed five times a day. At the end of experiment, the shrimp were weighed and their intestinal samples were analyzed for the amounts of Vibrio parahaemolyticus, Aeromonas hydrophila, Lactobacillus sp. , and Streptococcus faecalis using selective agar. The results showed that the shrimps' weight gain and specific growth rate increased with the increment of dietary ScFOS, while their feed conversion ratio decreased. The result was the best when an additional ScFOS 0.4 g/kg dried diet was used.     

Effect of methionine on intestinal enzymes activities, microflora and humoral immune of juvenile Jian carp (cyprinus carpio var. Jian)

An 8-week feeding experiment was conducted to determine the effect of dietary methionine supplementation on intestinal microflora and humoral immune of juvenile Jian carp (initial weight of 9.9 ± 0.0 g) reared in indoor flow-through and aerated aquaria. Eight amino acid test diets (350 g kg⁻¹ crude protein, CP), using fish meal, soybean-condensed protein and gelatin as intact protein sources supplemented with crystalline amino acids, were formulated to contain graded levels of methionine (0.6–22.0%) at a constant dietary cystine level of 3 g kg⁻¹. Each diet was randomly assigned to three aquaria. Growth performance and feed utilization were significantly influenced by the dietary methionine levels ( P  < 0.05). Maximum weight gain, feed intake occurred at 12 g kg⁻¹ dietary methionine ( P  < 0.05). Methionine supplementation improved hepatopancreas and intestine weight, hepatosomatic and intestine index, intestinal γ-glutamyltransferase and creatine kinase activity, Lactobacillus count, Bacillus count, lysozyme activities, lectin potency, sim-immunoglobulin M content, addiment C3,C4 contents and serum total iron-binding capacity and declined Escherichia coli and Aeromonas counts. Quadratic regression analysis of weight gain against dietary methionine levels indicated that the optimal dietary methionine requirement for maximum growth of juvenile Jian carp is 12 g kg⁻¹ of the dry diet in the presence of 3 g kg⁻¹ cystine.     

健康养殖南美白对虾肠道细菌的抗菌活性

用双层琼脂扩散法检测从健康养殖南美白对虾肠道内分离出的111株菌对8株病原菌的抗菌活性。结果表明:111株菌对白假丝酵母菌,非01群霍乱弧菌,嗜水气单胞菌指示菌无抗菌活性,48.6%的菌株对余下的5株指示菌有抗菌活性,其中气单胞菌属和发光杆菌属对荧光假单胞菌与金黄色葡萄球菌的抑制性最强。各菌属中以气单胞菌属,发光杆菌属的抗菌谱最广,弧菌属、肠杆菌科、黄单胞菌属、芽孢杆菌属、小球菌属次之。土壤杆菌属、棒状杆菌属、黄杆菌属、产碱杆菌属、假单胞菌属和色杆菌属则没有抗菌活性。     

Antibacterial abilities of intestinal bacteria from larval and juvenile Japanese flounder against fish pathogens

The present study was undertaken to examine the antibacterial abilities of intestinal bacteria isolated from juveniles and larvae of Japanese flounder Paralichthys olivaceus . Newly hatched larvae of flounder were held in a 100 L plastic circular tank and fed rotifiers, Artemia nauplii and commercial feeds, depending on the developmental stage of the fish. Genera Aeromonas , Moraxella and Vibrio were predominantly isolated from the intestinal tracts of Japanese flounder at larval and juvenile stages, whereas Aeromonas , Bacillus , coryneforms, Moraxella , Pseudomonas and Vibrio were detected at high densities in live diets and artificial feeds. Antibacterial bacteria accounted for 1.7–24.3% of the intestinal isolates against Lactococcus garvieae , Pasteurella piscicida , Vibrio anguillarum and V. vulnificus . In particular, as much as 53.3% of Vibrio spp. other than Vibrio -swarmer isolated from 197-day-old juveniles inhibited the growth of P. piscicida . These results suggest that intestinal bacteria having antibacterial activity play a role in the prevention of infectious diseases.     

Identification of pathogenic bacteria associated with the occurrence of 'red spot' in sea mullet, Mugil cephalus L., in south-eastern Queensland

Abstract. An investigation was undertaken to determine the bacterial species associated with the fish disease known as 'red spot' which affected wild fish stocks in south-eastern Queensland. Outbreaks of the disease in the Noosa River were examined with particular reference to the sea mullet, Mugil cephalus. The bacterial flora of the skin of healthy fish as well as those exhibiting early and advanced lesions was determined. In the lower Noosa River estuary, Vibrio anguillarum was the sole organism associated with very early lesions. It was later shown that V. alginolyticus was also associated with V. anguillarum in early lesions, but was considered to be a secondary invader. Aeromonas hydrophila was isolated from advanced lesions on fish taken in freshwater reaches of the river. This organism was also regarded as a secondary invader.     

Colonization in the fish intestinal tract and production of inhibitory substances in intestinal mucus and faecal extracts by Carnobacterium sp. strain K1

A non-virulent Carnobacterium sp., designated strain K1, isolated from the gastrointestinal tract of Atlantic salmon, Salmo salar L., which produced inhibitory substances against bacterial fish pathogens, was examined in vitro for characteristics important for the colonization of the fish gastrointestinal tract and in vivo for persistence in the tract after oral dosing. In vitro growth experiments showed that the cells of this strain were metabolically active in both the intestinal mucus and faeces from salmonids. The production of growth inhibitors against the two common fish pathogens Vibrio anguillarum and Aeromonas salmonicida by Carnobacterium sp. strain K1 was demonstrated in vitro in mucus and faecal extracts. Furthermore, it was demonstrated that the Carnobacterium cells remained viable in the gastrointestinal tract for several days and that no detrimental effect to the fish was observed as a result of the presence of the bacterium.     

The disease status of Australian salmonids: bacteria and bacterial diseases

Abstract. Eleven freshwater salmonid hatcheries in southern Australia were surveyed for bacterial pathogens and diseases between 1981 and 1985, Twenty-five populations of fish were examined in the study, representing a total of 2755 fish, from which kidney, liver, spleen, and in some cases peritoneum, blood and faeces were cultured. Bacteria of pathogenic significance isolated included Aeromonas hvdrophila, Streptococcus sp., Lactobacillus piscicola, Yersinia ruckeri, Mycobacterium fortuitum, M. chelonei and a filamentous acid-fast organism of uncertain taxonomic position. Lacto-bacillus piscicola and Streptococcus sp. were associated with clinical and subclinical peritonitis. Mycobacterium chelonei was isolated from visceral granulomas in an externally normal fish. Aeromonas salmonicida, Renibacterium salmoninarum and Edwardsiella tarda were not isolated, indicating that the diseases furunculosis, bacterial kidney disease and edwardsiellosis are exotic to Australian salmonids. Similarly, while Y. ruckeri was isolated, enteric redmouth disease had not been recorded and is considered an exotic disease.     

Smolt characteristics of small first-time migrant, and resident Arctic charr, Salvelinus alpinus (L.), from a river system in northern Norway

Smolt characteristics were investigated in sympatric anadromous and resident Arctic charr. Salvelinus alpinus (L.), of a similar size (11-20 cm), A group of first-time migrant anadromous charr was caught while descending the Hals River and two groups of resident conspecifics were caught in Lake Storvatn. one before, and one after, the sea-run of anadromous fish had terminated. When sampled immediately after capture in fresh water the anadromous group had a higher proportion of fish visually classified as smolts. and these charr had higher gill Na⁺-K⁺ ATPase activity, higher densities of developed chloride cells in the gills, a lower condition factor, and lower plasma osmolality, than resident fish. When exposed to sea water (33%o S), only minor differences in mortality and plasma electrolyte levels were observed between anadromous and resident fish, average values of plasma osmolality and chloride concentrations being 377 mOsm and 169 mM and 387 mOsm and 174 mM, respectively. The results suggest that the first-time migrant Arctic charr had undergone some of the physiological changes that are considered typical for a parr-smolt transformation before they left fresh water. Such changes did not appear to have occurred in sympatric, resident, fish of a similar size. The parr-smolt transformation in the first-time migrants may, however, have been incomplete, because the fish did not appear to have the ability to rapidly re-establish osmo- and ionoregutatory homeostasis after direct transfer to sea water.     

The ability of carnobacteria isolated from fish intestine to inhibit growth of fish pathogenic bacteria: a screening study

In the present study a microtitre plate assay was used to evaluate antagonistic activity of 157 intestinal bacteria belonging to Carnobacterium isolated from Arctic charr (Salvelinus alpinus L.), Atlantic salmon (Salmo salar L.) and wolf fish (Anarhichas lupus L.) against fish pathogenic bacteria. One hundred and forty‐nine strains isolated from Arctic charr fed; (a) different lipid levels and (b) different fatty acids were screened for their ability to inhibit growth of the fish pathogen Aeromonas salmonicida ssp. salmonicida strain AL 2020 (the causative agent of furunculosis). Carnobacterium maltaromaticum and Carnobacterium mobile isolated from fish fed a low‐lipid diet inhibited growth of the pathogen, while none of the Carnobacterium divergens isolated from fish fed the high‐lipid diet had this ability. When Arctic charr was fed different fatty acids, was the frequency of antibacterial ability of C. maltaromaticum highest in strains isolated from fish fed 4%α‐linolenic acid (18:3 n‐3) and lowest in strains isolated from fish fed 4% linoleic acid (18:2 n‐6). Extracellular growth inhibitory compounds harvested in exponential and stationary growth phase from eight carnobacteria strains isolated from three fish species were tested for their ability to inhibit growth of six fish pathogens [A. salmonicida, Vibrio splendidus strain VS11, Vibrio salmonicida strain LFI 315, Vibrio anguillarum strain LFI 317, Moritella (Vibrio) viscosa strain LFI 5000 and C. maltaromaticum (piscicola) CCUG 34645]. The highest antibacterial activity was found when cellular extracts of the producer isolate were harvested in stationary growth phase. Scanning electron microscopy (SEM) investigations of A. salmonicida showed that cell morphology was affected by the inhibitory substance produce by strain 8M851, a Carnobacterium inhibens‐like bacteria.     

Phylogenetic analysis of intestinal bacteria of freshwater salmon Salmo salar and sea trout Salmo trutta trutta and diet

The microbial diversity of culturable intestinal microflora of wild freshwater salmonid fishes salmon Salmo salar and sea trout Salmo trutta trutta juveniles (0+ years old) from the same environmental conditions were investigated by means of molecular identification techniques and analysis of diet. Significant differences in the intestinal microbial diversity were observed in different fish species. The predominant group in the intestinal tract of the salmon comprised representatives of the Enterobacteriaceae family (23%), Plesiomonas (19.2%) and Carnobacterium (15.3%). Predominant microbiota in sea trout intestinal tract were Enterobacteriaceae (52%), Aeromonas (22%) and Pseudomonas (14%). The results show that Enterobacteriaceae were predominant in the intestinal tract of the salmon and the sea trout juveniles raised on diets of different compositions. However, molecular identification of the intestinal microbiota at the species or genus level revealed differences in these fish species. Bacteria in the hindgut of salmon included Pragia and Serratia. However, bacteria in the gut contents of sea trout from the Enterobacteriaceae family were Buttiauxella, Enterobacter, Moellerella, Pantoea, Rahnella and Tiedjeia arctica. A novel phylotype of T. arctica is harbored in the intestinal tract of wild salmon, and may correspond to a previously undescribed species.     

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。     

营养代谢过程中黄条鰤幼鱼消化道菌群的演变趋势

为认识消化道菌群结构在宿主营养代谢过程中的变化规律,并探讨饵料菌群对其的影响,采用16SrDNA高通量测序技术,对黄条鰤(Seriola lalandi)幼鱼一个完整摄食周期内的消化道菌群结构特征进行研究,并解析其与饵料菌群的相关性.结果显示,乳杆菌属(Lactobacillus)、假单胞菌属(Pseudomonas)...     

Inhibition of bacterial fish pathogens by Tetraselmis suecica

Abstract. Supernatants and extracts derived from a commercial heterotrophically grown spray-dried preparation of Tetraselmis suecica were observed to inhibit Aeromonas hydrophila, A. salmonicida, Lactobacillus sp., Serratia liquefaciens, Staphylococcus epidermidis, Vibrio anguillariim, V. salmonicida and Yersinia ruckeri type I, but not Planococcus sp., Pseudomonas fluorescens, Streptococcus sp., V. ordalii or Y. ruckeri type II by in vitro methods. When used as a food supplement, the algal cells inhibited laboratory-induced infections in Atlantic salmon. Moreover, there was a reduction in bacterial numbers in tank water. When used therapcutically, algal cells and their extracts reduced mortalities caused by A, salmonicida, Ser. liquefaciens, V. anguillariim, V. salmonicida and Y. ruckeri type I.     

Evaluation of the potential source of bacterial contamination during cryopreservation process of silver barb (Barbodes gonionotus) sperm

Application of good sanitation practice in cryopreservation process is the key issue to improve the quality of cryopreserved fish sperm. This study implemented standard sanitation protocol with minimal contamination and evaluated the source of bacterial contamination associated with laboratory equipment and related materials across a series of cryopreservation process of silver barb (Barbodes gonionotus) semen. The use of 16s rRNA sequencing and traditional biochemical methods were performed for bacterial identification. Animal origin (anal fin, culture water and semen contaminated with faeces and urine) and non‐animal origin (liquid nitrogen from liquid nitrogen dewar, outer surface of straw, air circulation in cryopreservation laboratory and latex gloves used during cryopreservation procedure) were determined for bacterial contamination. Aeromonas punctata subsp. caviae was the most abundant species in anal fin, latex groves and semen contaminated with faeces and urine. Bacillus safensis and Bacillus sp. were found as frequently recovered species from liquid nitrogen dewar. Aeromonas hydrophila subsp. hydrophila and Pseudomonas fluorescens, fish pathogenic bacteria, were isolated from all animal origin samples. This was the first report indicating that standard sanitation and hygiene methodologies are recommended for sperm cryopreservation of B. gonionotus to prevent bacterial contamination.