Effects of microbial growth inhibition by antibiotics on the production of taste-active components during the processing of heshiko produced by aging salted mackerel with rice bran

To elucidate the role of microbes and endogenous proteases in the formation of the taste-active components during processing of heshiko, a Japanese traditional seafood produced by aging salted mackerel with rice bran, the production of lactic acid, free amino acids, and acid-soluble peptides throughout the aging process was compared in the absence and presence of antibiotics including chloramphenicol, penicillin, and cycloheximide. Although viable counts on fish flesh on a 2.5% NaCl-GYP agar plate increased to 10⁶ cfu/g with the aging process of heshiko in the absence of the antibiotics, adding the antibiotics suppressed the increase in viable counts to less than 10³ cfu/g throughout the aging process. Lactic acid barely increased due to the suppression of microbial growth by the antibiotics. In spite of the inhibition of bacterial growth throughout the processing of heshiko upon the addition of the antibiotics, little difference was observed in the production of free amino acids and acid-soluble peptides. These results suggest that, unlike lactic acid (which is a fermentation product of lactic acid bacteria), free amino acids and acid-soluble peptides are mainly produced through protein hydrolysis by endogenous proteases in mackerel meats during the aging process of heshiko.     

Microbial and chemical properties of <Emphasis Type="Italic">aji-no-susu</Emphasis>, a traditional fermented fish with rice product in the Noto Peninsula,Japan

Aji-no-susu is a Japanese fermented fish product prepared from salted horse mackerel Trachurus japonicus, and cooked rice. We studied the organic acid and free amino acid contents and microflora in 12 aji-no-susu products to clarify their features as a lactic-acid-fermented food. Salinity of the samples was approximately 7.0% (rice portion) and 6.0% (fish portion) (w/w). Water activity was approximately 0.9, and pH was approximately 4.4 and lower. In the rice portions, lactic acid content was very high (57 mg/g sample). The predominant amino acids were alanine (2.3 mg/g rice portion) and lysine (2.1 mg/g). In the case of long-fermented (4 and 12 months) aji-no-susu, a high content of gamma-aminobutyric acid (GABA, 1.5 and 1.4 mg/g) was detected. Total viable counts in rice and fish portions were 7.7 and 7.4 log colony-forming units (cfu)/g, respectively. The number of lactobacilli in the rice and fish portions was 7.3 and 7.1 log cfu/g, respectively. Yeasts were detected in eight samples. Furthermore, acid-tolerant lactic acid bacteria (LAB) (Lactobacillus plantarum), GABA-producing LAB (Lactobacillus sp.), and halophilic or halo-tolerant yeast (Debaryomyces hansenii) were isolated and identified. Results in this study indicate that aji-no-susu is a typical traditional lactic-acid-fermented fish product.     

Bacterial flora associated with the giant freshwater prawn Macrobrachium rosenbergii, in the hatchery system

The objective of this study was to understand the microbial flora associated with the hatchery system of giant fresh water prawn, Macrobrachium rosenbergii during an entire rearing cycle. Bacteriological and physico-chemical analysis was done for different samples of water, larvae, and Artemia. The total bacterial load in well water, seawater and inlet water varied from 10¹ to 10⁵ cfu ml^− 1 with higher counts seen in larval rearing tank (LRT) water. The Vibrio count ranged between 10¹ to 10³ cfu ml^− 1. Larval samples harboured a bacterial load of 10⁶ to 10⁷ cfu/10 larvae. The bacterial load in Artemia hatching water ranged from 4.90 × 10⁴ to 5.63 × 10⁶ cfu ml^− 1 while Artemia had a load ranging from 1.08 × 10⁷ to 2.09 × 10⁹ cfu g^− 1. Vibrio count in the LRT water ranged from 10¹-10³ cfu ml^− 1 while the count in larvae ranged from 10² to 10⁴ cfu/10 larvae. The bacterial genera were predominantly Gram-negative and comprised of Aeromonas spp., Pseudomonas spp., Vibrio spp. and Bacillus spp. and non-spore formers (NSF) were the dominant Gram-positive bacteria. This study documents the bacterial flora associated with Macrobrachium hatchery system during a regular normal run. Knowledge of the qualitative and quantitative aspects of bacterial flora in the hatchery would help to understand disturbances, if any, brought about during disease outbreaks.     

Combined effects of dietary malic acid and Bacillus subtilis on growth,gut microbiota and blood parameters of Nile tilapia (Oreochromis niloticus)

The study investigated effect of dietary supplementation with malic acid, Bacillus subtilis and a mixture of the two (3 × 2 factorial trial) on Nile tilapia (Oreochromis niloticus) health. Treatment groups (T1–T6) were fed diets containing three levels of malic acid (0.0, 5.0 and 10.0 g/kg), each of which was supplemented with 0 and 1.1 × 10⁵ cfu/g B. subtilis, respectively. Each treatment group was assigned randomly to triplicate groups of 35 fish (5.26 ± 0.06 g) in 18 tanks for 84 days. The results indicated that survival was higher in all dietary treatments as compared to the control fed fish. The highest values of FBW, WG and SGR (%), PER, PPV and ER were recorded in groups T4 and T6, which were fed diets supplemented with 5 g malic acid/kg and 1.1 × 10⁵ cfu/g B. subtilis and 10 g malic acid/kg and 1.1 × 10⁵ cfu/g B. subtilis, respectively. The best value of FCR was obtained in groups T4 and T6. The lowest total bacterial count in the gut and faeces was detected in fish from group T6. The values of haematocrit, haemoglobin, red blood cells, white blood cells, total protein, albumin and globulin were significantly higher (p < .05) in fish from groups T4 and T6 (diets supplemented with malic acid and B. subtilis). As compared to the control fed fish, te mixture of these substances are promising as immune enhancher in aquacultured fish.     

Bacterial Change in the Gut and Hepatopancreas of Tiger Shrimp Reared in Bioaugmented System

The study was conducted in nine 150‐L capacity glass aquaria for 192 h to determine the changes in the microflora in the gut and hepatopancreas of tiger shrimp, Penaeus monodon, after bioaugmenting (water treatment and probiotics). Results showed that luminous bacterial counts were significantly lower in bioaugmented than in nonbioaugmented systems (range 0–5.9 × 10³ cfu/g and 0–3.2 × 10³ cfu/g in gut and hepatopancreas, respectively). Biochemical tests of isolates showed that the bioaugmented systems were dominated by Gram‐positive Bacillus and Streptococcus and the Gram‐negative Vibrio dominated the control. In water‐treated and probiotics‐fed bioaugmented system, the change in bacterial dominance to Bacillus became evident 2 h in the gut and 24 h in the hepatopancreas. Bacterial dominance shifted to Vibrio species after 120 h. Shrimp in the control were observed to become lethargic after 72 h as Vibrio species significantly increased in number. Dissolved oxygen and unionized ammonia levels were significantly lower in bioaugmented system compared with the control.     

Effects of Dietary Administration of Probiotic Halomonas sp. B12 on the Intestinal Microflora,Immunological Parameters,and Midgut Histological Structure of Shrimp,Fenneropenaeus chinensis

The experiment was conducted to investigate the effects of oral administration of probiotic Halomonas sp. B12 (previously isolated from the intestine of Fenneropenaeus chinensis) on the intestinal microflora, immunological parameters, and midgut histological structure of F. chinensis. Shrimp (initial weight: 4.00 ± 0.10 g) were fed diets containing Halomonas sp. B12 at 0 (control), 3.68 × 10⁷ (T1), and 7.18 × 10¹⁰ (T2) colony‐forming units per gram for 6 wk, respectively. The results showed that the total bacterial counts significantly increased (P < 0.05) with supplementation of dietary probiotic B12. However, Vibrio spp. counts significantly decreased (P < 0.05) in the intestine of shrimp with increasing dietary probiotic B12. Hemocyte counts in the shrimp fed the diets supplemented with probiotic B12 were significantly higher (P < 0.05) than the control group. Both phenoloxidase (PO) activity in plasma and hemocyte lysate supernatant were higher in the shrimp fed diets supplemented with probiotic B12 compared with the control group. No significant difference was found in PO activity in plasma between the T1 and the control (P > 0.05). PO activity in plasma was higher in T2 than that in T1. Cumulative shrimp mortality after 10‐d white spot syndrome virus (WSSV) challenge test significantly decreased with increasing dietary probiotic B12 (P < 0.05). No significant difference was found between T1 and control in cumulative shrimp mortality after challenge test (P > 0.05). The oral administration of probiotic B12 to F. chinensis was beneficial to protect the integrity of shrimp intestinal mucosal layer. In summary, even though the low dose (T1) had some effects on bacterial counts and immunological parameters, only the high dose (T2) significantly increased the resistance of the shrimp to WSSV.     

Optimization of de‐oiled rice bran inclusion level in the diet of Labeo rohita: Effect on growth performance,nutrient utilization,insulin‐like growth factors I and II gene expression

A 60‐day feeding trial was conducted to optimize the inclusion level of de‐oiled rice bran (DORB) in the diet of Labeo rohita fingerlings. Six iso‐nitrogenous (260g/kg), iso‐lipidic (70g/kg) and iso‐energetic (15 MJ/Kg) diets were prepared with different inclusion level of DORB viz., T33, T38_, T43, T48, T53 and T58. One hundred and eighty fingerlings (9.8 ± 0.5 g) were randomly distributed in seven experimental groups in triplicates following a completely randomized design. The weight gain %, specific growth rate, protein efficiency ratio and feed efficiency ratio were found similar in 33%, 38% and 43% DORB fed groups. The expression of both insulin‐like growth factors I and II (IGF‐I and II) was significantly (p < 0.05) higher in T33, T38 and T43 groups. Based on second order polynomial regression analysis (y = ?0.0059x² + 0.4573x ? 4.8404, R² = 0.95), the expression of IGF‐I was found to be maximum at 39.5% inclusion level of DORB. Significantly (p < 0.05) lower muscle RNA content, RNA‐DNA ratio and higher feed conversion ratio were found in T58 group. Hence, from these results, it can be concluded that the optimum inclusion of DORB in the diet (260g/kg CP and 70g/kg lipid) of L. rohita should be 39.5% to support maximum nutrient utilization and growth performance.     

Behavior of histamine-producing bacteria in shimesaba, raw mackerel salted and marinated in vinegar during processing and storage at various temperatures

To study the behavior of histamine-producing bacteria during the processing and storage of shimesaba, we inoculated histamine-producing bacteria into raw mackerel and then evaluated bacterial counts and histamine content at each shimesaba processing step. Six histamine-producing bacteria—Morganella morganii, Enterobacter aerogenes, Raoultella planticola, Photobacterium damselae (2 strains), and Photobacterium phosphoreum—were inoculated onto the surface of mackerel fillets at various cell densities (no inoculation, 10⁴, and 10⁷ cfu ml^?1). The fillets were then treated according to the common procedure used for processing shimesaba. The concentration of NaCl in brine during the salting procedure was 10 % (w/v), and the main ingredients of the marinade broth were 27 % vinegar, 3.8 % sucrose, 0.3 % sodium glutamate, and 0.1 % sodium succinate. The level of histamine-producing bacteria and of histamine did not increase during the salting and marinating procedures. However, during the storage tests, histamine content increased remarkably in the shimesaba products stored at 10 °C for 3 days, and organoleptical degradation was also observed. However, significant deterioration, based on bacterial count, histamine content, and organoleptic evaluation, was not observed in samples stored at 5 °C. These results indicate that the use of traditional marinade broth during the processing of shimesaba and storage below 5 °C are effective methods for preventing histamine accumulation.     

Use of Bacillus coagulans as a Dietary Probiotic for the Common Carp,Cyprinus carpio

Effect of viable Bacillus coagulans as a dietary probiotic on the growth performance, survivorship, immune response, and meat quality of the common carp, Cyprinus carpio, was investigated. Three treatments (T‐1, T‐2, and T‐3) and a control, each with three replicates, were established. T‐1, T‐2, and T‐3 groups were fed with diets containing different levels of probiotic (1 × 10⁷ cfu/g, 2 × 10⁷ cfu/g, and 4 × 10⁷ cfu/g, respectively). After 45 d, the fish fed with probiotic had significantly higher (P < 0.05) daily weight gain compared to that of the control group. Significant increases (P < 0.05) in final weight, relative weight gain rate, lysozyme, myeloperoxidase, and respiratory burst activities were obtained in the T‐2 and T‐3 compared with those of the control. Fish in the control group showed the lowest (P < 0.05) content of complement component 3. Significantly (P < 0.05) improved contents of crude fat, inosine, and inosinic acid were observed in the T‐2 and T‐3. Our results indicated that the use of B. coagulans as a dietary probiotic for the common carp could improve growth performance, meat quality, and induce a positive modulation on immune response.     

Effects of soy sauce <Emphasis Type="Italic">koji</Emphasis> and lactic acid bacteria on the fermentation of fish sauce from freshwater silver carp <Emphasis Type="Italic">Hypophthalmichthys molitrix</Emphasis>

The effects of soy sauce koji and the lactic acid bacterium, Tetragenococcus halophilus, were studied on the fermentation of fish sauce prepared from Chinese silver carp. The fish sauce prepared without koji and the lactic acid bacterium contained low levels of organic materials, total nitrogen, and organic acids. The use of koji was effective in increasing these qualitative parameters and further improved the amino acid score of the fish sauce. Addition of T. halophilus had an effect on lowering the pH value during the initial period of fermentation when the soy sauce koji was also supplemented. In contrast, T. halophilus-like bacteria were found to be predominant for all tanks fermented under the different starting conditions. Although it was not examined whether the T. halophilus-like bacteria observed after fermentation were the same as the starter-bacterium or not, it was suggested that T. halophilus plays an important role in the successful fermentation of silver carp fish sauce. Sensory evaluation conducted with Japanese and Chinese panelists also suggested the superiority of the use of koji for fermentation of silver carp fish sauce.     

Effect of extracts from heshiko , a fermented mackerel product, on cholesterol metabolism in Wistar rats

Heshiko extract was separated into peptide and amino acid fractions by ion-exchange column chromatography. Heshiko extract and these fractions were administered to rats in a diet enriched with lipid and cholesterol for 30 days. In the heshiko extract group, increase of triglyceride, total cholesterol, and low-density lipoprotein (LDL)-cholesterol levels in plasma and accumulation of total lipids in the liver were suppressed, while amounts of both lipid and cholesterol excreted to feces were increased. Administration of heshiko extract tended to increase fecal bile acids and promoted the activity of cholesterol 7α-hydroxylase, the rate-limiting enzyme in the synthesis of bile acid from cholesterol in the liver. However, activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in the cholesterol synthesis system in the liver, decreased due to regulation by the feedback of lipid transportation from diet to the liver. The same effect was observed in the peptide and free amino acid fraction groups. These data suggest that both excretion of cholesterol and bile acid to feces and promotion of cholesterol 7α-hydroxylase activity are related to the hypocholesterolemic effects of heshiko extract and that both peptides and amino acids contribute to that effect.     

Effects of Pediococcus pentosaceus supplementation on growth performance,intestinal microflora and disease resistance of white shrimp,Litopenaeus vannamei

Litopenaeus vannamei is economically important shrimp species in worldwide aquaculture. This study was conducted to assess the effect of different levels of probiotic Pediococcus pentosaceus (PP) on growth performance, feed utilization, digestive enzyme activity, intestinal microflora count and body composition of L. vannamei. Four diets containing different concentrations [0 (PP0), 10⁶ (PPI), 10⁷ (PPII) and 10⁸ (PPIII) CFU/g] of PP were formulated. After 8 weeks feeding trial, the experimental shrimps were challenged with Vibrio anguillarum and noted the surveillance. At the end of the feeding trial, the obtained results revealed a significant increase (p < .05) in final body weight, final length, weight gain (WG), survival rate, protease and amylase activities, lactobacillus sp. and Bacillus sp. intestinal count, total haemocyte counts (THC) and lysozyme activity in PPIII group when compared with the other groups. Similarly, WG, amylase activity, Bacillus sp. count, THC and lysozyme activity were significantly enhanced in case of PPII compared to the control group (p < .05). Interestingly, FCR and Vibrio sp. counts were significantly decreased in case of PPIII group when compared to the other groups (p < .05). Also, significant level of surveillance was noted in the challenging test with V. anguillarum. These results suggested that the P. pentosaceus improved the growth performance, digestive enzyme activity, immunity and tolerance against V. anguillarum of L. vannamei.     

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.     

Effects of dietary supplementation of probiotics on the growth,activities of digestive and non‐specific immune enzymes in hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀)

Effects of Bacillus cereus BC‐01, Lactobacillus acidophilus LAG01, Clostridium butyricum CBG01 and their combinations as supplementation on the growth performance, digestive enzyme activities and serum non‐specific immunity of hybrid grouper (Epinephelus lanceolatus ♂×E. fuscoguttatus ♀) were assessed. Seven different diets, that is one control diet (basal feed without any probiotics, CT) and six treatment diets containing single B. cereus (Bs), L. acidophilus (Ls) and C. butyricum (Cs) at 1.0 × 10⁹ cfu/kg feed, and also their combinations in equal proportion at 1.0, 2.0 and 3.0 × 10⁹ cfu/kg feed (BLC1, BLC2 and BLC3) were prepared respectively. After 60‐day feeding trial, the final weight, specific growth rate,food consumption, food conversion efficiency and apparent digestibility coefficient of fish in Ls and BLC3 were significantly higher compared with the control (p < .05). The activities of pepsin and trypsin in the intestine of fish for Ls and BLC3 were significantly higher relative to the control (p < .05). Relative to controls, significantly enhanced amylase and lipase activities in proximal intestine except for Cs and BLC1 and lipase activities in distal intestine except for Cs were observed (p < .05). Meanwhile, activities of superoxide dismutase in the serum of fish for all treatments, lysozyme and catalase in Ls and BLC3, and glutathione peroxidase except for Cs were significantly enhanced (p < .05). Based on the above, dietary supplementation of single L. acidophilus at 1.0 × 10⁹ cfu/kg or combination of three strains at 3.0 × 10⁹ cfu/kg for hybrid grouper is recommended.     

Effect of Lactobacillus acidophilussupplementation on growth performances,digestive enzyme activities and gut histomorphology of striped catfish (Pangasianodon hypophthalmusSauvage, 1878) juveniles

The influence of dietary supplementation of Lactobacillus acidophilus, on growth performance, digestive enzyme activities, gut histomorphology and gut microflora were evaluated in juveniles striped catfish (Pangasianodon hypophthalmus). Five experimental diets were formulated by supplementing 0, 10³, 10⁵, 10⁷ and 10⁹ CFU/g L. acidophilus in fishmeal and casein‐based semi‐purified diet. Triplicate groups of striped catfish (21.69 ± 0.18 g) were stocked in 15 fiberglass tanks with stocking density of 25 individuals per tank and fed twice daily at 2.5% of the fish body weight for 12 weeks. Weight gain, specific growth rate, feed conversion ratio and the protein efficiency ratio were significantly higher in fish fed with 10⁵ and 10⁷CFU/g L. acidophilussupplemented diets compared with the other treatment groups. Compared with the control and fish fed low (10³ CFU/g) L. acidophilus supplementation, those fed with 10⁵ and 10⁷ CFU/g had significantly higher (p < 0.05) apparent protein digestibility. Inclusion of L. acidophilus at 10⁷ CFU/g diet significantly increased amylase, protease and lipase activities. Microscopic analysis showed that the villi length in both the anterior and posterior gut and microvilli length in the posterior gut increased significantly in fish fed L. acidophilus supplementation at 10⁵ and 10⁷ CFU/g of diet. The fish fed L. acidophilus supplemented diets significantly increased the total lactic acid bacteria counts in the gut of striped catfish compared with the control‐fed group. Based on gut histomorphology and growth performance, inclusion of L. acidophilus at 10⁵ CFU/g appears to have the most positive effect on striped catfish farming.     

Virulence of Flavobacterium psychrophilum isolates in rainbow trout Oncorhynchus mykiss (Walbaum)

Flavobacterium psychrophilum, the causative agent of bacterial cold‐water disease (BCWD) in freshwater‐reared salmonids, is also a common commensal organism of healthy fish. The virulence potential of F. psychrophilum isolates obtained from BCWD cases in Ontario between 1994 and 2009 was evaluated. In preliminary infection trials of rainbow trout juveniles, significant differences (0% to 63% mortality) in the virulence of the 22 isolates tested were noted following intraperitoneal injection with 10⁸ cfu/fish. A highly virulent strain, FPG 101, was selected for further study. When fish were injected intraperitoneally with a 10⁶, 10⁷ or 10⁸ cfu/fish of F. psychrophilum FPG 101, the 10⁸ cfu/fish dose produced significantly greater mortality (p < 0.05). The bacterial load in spleen samples collected from fish every 3 days after infection was determined using rpoC quantitative polymerase chain reaction amplification and by plate counting. Bacterial culture and rpoC qPCR were highly correlated (R² = 0.92); however, culture was more sensitive than the qPCR assay for the detection of F. psychrophilum in spleen tissue. Ninety‐seven per cent of the asymptomatic and the morbid fish had splenic bacterial loads of <2.8 log₁₀ gene/copies and >3.0 log₁₀ gene copies/reaction, respectively, following infection with 10⁸ cfu/fish.     

Bacterial microflora associated with farmed freshwater prawn Macrobrachium rosenbergii (de Man) and the aquaculture environment

The initial counts of bacteria associated with farmed giant freshwater prawn (Macrobrachium rosenbergii de Man) as well as with the water and sediment from two farms located at Kottayam district in Kerala (India) were determined. A total of 367 randomly selected isolates were characterized and identified. Prawn samples yielded mean microbiological counts of 4.92 log CFU g^?1 of shell with muscle and 7.78 log CFU g^?1 of intestine at 30°C. Bacterial numbers in the intestine of prawn were much higher than those in the pond water. Motile and non‐motile aerobic Gram‐negative bacteria together with Enterobacteriaceae accounted for 60–70% of the mesophiles isolated from water and prawn. However, the microorganisms isolated in significant numbers from shell with muscle and intestine of prawn were not recovered from the growing waters. Among Gram‐positive bacteria, Micrococcus, Bacillus, coryneforms, and Arthrobacter were found. Faecal coliforms and enterococci were found in significant numbers in M. rosenbergii. The rearing practices such as feeding and pond fertilization could have influenced the microflora in prawn. The commensal microflora of freshwater prawn included opportunistic pathogens such as Aeromonas spp. and Streptococcus spp. Storage of prawn at elevated temperatures can permit their survival and growth leading to quality loss. Care should be exercised during handling and processing to prevent contamination of edible meat.     

Bacterial Flora of Artemia Cultured in Earthen Saline Ponds

Aerobic bacteria in pond water, sediments, and Artemia in earthen saline ponds in Saudi Arabia were quantified. Total viable counts (TVC; mean ± SD) in pond water ranged between 7.9 ± 4.8 × 10³ and 1.6 ± 3.7 × 10⁴ colony forming units (cfu)/mL; in sediments between 7.1 ± 4.2 × 10⁶ and 6.5 ± 3.8 × 10⁷ cfu/g; on Artemia surfaces, 3.9 ± 0.9 × 10³ and 1.0 ± 2.9 × 10⁴ cfu/Artemia; and in Artemia homogenate, 4.6 ± 2.4 × 10⁷ and 3.2 ± 3.6 × 10⁸ cfu/g. The bacterial flora was predominantly Gram-negative rods, accounting for 89% of total isolates. Altogether, nine bacterial species of seven genera were identified. Bacteria in pond sediments were the most diverse compared to other populations. In all populations, Vibrio alginolyticus, Vibrio sp., and Staphylococcus sp. were dominant (P < 0.005). The bacterial flora of pond water and sediments were reflected in the bacterial composition on and in Artemia.     

Effects of sodium diformate on growth performance,gut microflora,digestive enzymes and innate immunological parameters of Asian sea bass (Lates calcarifer) juveniles

This study was conducted to evaluate the effects of dietary sodium diformate (NaDF) on growth performance, gut microflora, digestive enzyme activities and immune response parameters of Asian sea bass (Lates calcarifer) juveniles. Fish with initial weight of 12.5 ± 0.4 g were fed with five experimental diets contained 0.0 (control), 5, 10.0, 15.0 and 20 g NaDF kg^?1 in triplicate for 6 weeks. Fish fed diet containing 5 g NaDF kg^?1 had significantly the highest final body weight and feed intake among different treatments. The gut total viable bacterial counts gradually decreased with increasing dietary NaDF level. Specific activity of chymotrypsin improved in fish fed diets administered with NaDF compared to the control group. Fish fed 5 g NaDF kg^?1 diet showed the highest serum lysozyme level among different treatments. The serum classical pathway activity of complement showed higher level in fish fed diets contained 5 and 10 g NaDF kg^?1 than other groups. According to break‐point regression method analysis, the optimum inclusion of dietary NaDF in L. calcarifer juveniles was estimated between 4.6 and 5.1 g/kg, when specific growth rate and feed conversion ratio were plotted against dietary NaDF levels.