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.     

Co-feeding of live feed and inert diet from first-feeding affects Artemia lipid digestibility and retention in Senegalese sole (Solea senegalensis) larvae

The present study intended to evaluate the effects of early introduction of inert diet in lipid digestibility and metabolism of sole, while larval feed intake, growth and survival were also monitored. Solea senegalensis larvae were reared on a standard live feed regime (ST) and co-feeding regime with inert diet (Art R). Trials using sole larvae fed with Artemia enriched with two different lipid emulsions, containing glycerol tri [1-¹⁴C] oleate (TAG) and L-3-phosphatidylcholine-1,2-di-[1-¹⁴C] oleoyl (PL), were performed at 9 and 17 days after hatching (DAH) to study lipid utilization. Co-feeding did not affect sole survival rates (ST 59.1 ± 15.9%; Art R 69.56 ± 9.3%), but was reflected in significantly smaller final weight at 16 DAH (ST 0.71 ± 0.20; Art R 0.48 ± 0.14 mg). Higher feed intake was observed in sole larvae fed on Artemia enriched with labeled PL at 9 DAH but not at 17 DAH. At 17 DAH, the smaller larvae (Art R treatment) ingested proportionally more Artemia in weight percentage, independently of enrichment. At 9 DAH lipid digestibility was equal among treatments and higher than 90%, while at 17 DAH it was higher in ST treatment (around 73%) compared to the Art R group (around 66%). Lipid retention efficiency at 9 DAH was higher in the Art R treatment, reaching values of 50%, while these values almost duplicated at 17 DAH, ranging up to 80% in both treatments without significant differences. These results show that co-feeding of live feed and inert diet from first-feeding in Senegalese sole has a toll in terms of growth and lipid digestibility but does not seem to compromise lipid metabolic utilization.     

Addition of bacteria bioencapsulated in Artemia metanauplii to a rearing system for halibut larvae

Theaim of this study was to determine whether it is possible to influence thespecies composition of the gut microflora in 70-days old halibut larvae(Hippoglossus hippoglossus L.) by addition of bacteriabioencapsulated in Artemia franciscana in two shortpulses.Two Vibrio strains, PB 1-11 and PB 6-1 were used. Twotreatments received each strain alone, a third treatment received the twobacterial strains in mixture, whereas in a control treatment no bacteria wereadded. Five fish tanks were included in each treatment. The bacteria werebioencapsulated in Artemia franciscana metanauplii duringashort-term incubation in bacterial suspensions of the specific strains, andwereadded in two pulses, on day 1 and day 10 of the experiment. Addition ofbacteriadid not increase the total number of colony-forming units (CFU) in the larvalgut. The total CFU in the water was lower in tanks added bioencapsulatedbacteria than in the control treatment (p < 0.05). Animmunocolony blot assay was used to measure the numbers of the specificbacteriaadded in samples of water, A. franciscana, and fishlarvae.At the end of the experiment on day 13, the strain PB 1-11 reached on average10% of the total CFU when added alone and 25% when added in mixture with PB6-1.Strain PB 6-1 reached at the end of the experiment 26.8% of the total CFU whenadded alone and 24.2% when added in mixture with PB 1-11, whereas thepercentages of PB 1-11 and PB 6-1 in the control fish were 12.7% and 10.8% ofthe total CFU, respectively. However, the differences between the experimentaltreatments compared with the control group were not significant(p > 0.05). The addition of bioencapsulated bacteria intwo pulses within a ten days period was therefore not sufficient to influencethe species composition of the microflora of the halibut larvae.     

Natural carbon stable isotope ratios as indicators of the relative contribution of live and inert diets to growth in larval Senegalese sole (Solea senegalensis)

The relative contributions of live Artemia metanauplii and an inert diet for growth of Senegalese sole larvae and postlarvae were assessed through the analysis of carbon stable isotopes ratios (δ¹³C) in both diets and whole larval tissue. Larvae were reared on four dietary regimes: 100% live prey (rotifers and Artemia), 100% inert formulated diet and two co-feeding regimes of 70:30 and 30:70 ratios of Artemia and inert diet, respectively. Larvae from the live food regime and both co-feeding regimes showed a steep increase in δ¹³C from 10 days after hatching (DAH) as a result of the onset and continuation of Artemia consumption. From 12 DAH fish larvae from all the regimes showed significant isotopic differences as their δ¹³C increased to final asymptotic values of − 15.1, − 15.6 and − 16.3‰ in the live food, 70:30 and 30:70 regimes, respectively. Carbon turnover rates in larvae from both live food and co-feeding regimes were relatively high (0.071 to 0.116 d^− 1) but more than 90% of the observed change in fish tissue isotopic values was accounted for by the retention of carbon in new tissue growth. A two-source, one-isotope mixing model was applied to estimate the nutritional contribution of Artemia and inert diet to postlarvae growth in the co-feeding regimes. At 23 DAH, the relative contribution of live and inert diets to tissue growth in larvae was respectively, 88 and 12% for the 70:30 co-feeding regime and 73 and 27% for the 30:70 co-feeding regime. At 17 DAH, the estimated proportion of tissue carbon derived from the inert diet was higher at 23 and 38% for the 70:30 and 30:70 regimes, respectively. The results suggest that co-feeding regimes in Solea senegalensis larvae may be adjusted to meet ontogenetic changes in the capacity for larvae to utilise inert diets. The contrasting levels of carbon isotope discrimination between diet and tissue in larvae reared on either 100% live feed or 100% inert diet indicate relatively poor utilization of nutrients from the inert diet. The use of isotopic discrimination factors as potential indicators of the digestive physiological performance of a consuming organism in regards to its diet is discussed.     

Iodine‐enriched rotifers and Artemia prevent goitre in Senegalese sole (Solea senegalensis) larvae reared in a recirculation system

To study the effect of dietary supplementation of iodine in Solea senegalensis, larvae were randomly distributed in six tanks. Larvae in three tanks were given rotifers and Artemia enriched with iodine in addition to Rich Advance or Super Selco from 2 days after hatch (DAH) until 31 DAH. Larvae in a second set of three tanks were fed control rotifers and Artemia, enriched only with Rich Advance or Super Selco. Samples were collected at 2, 5, 10, 15 and 31 DAH to determine dry weight, total length, myotome height and thyroid status. Larvae fed the iodine‐enriched diet had significantly higher weight at 31 DAH and higher levels of whole body iodine concentration, compared to control larvae. At 31 DAH, larvae from the control treatment showed typical goitrous thyroid follicles. Thyroid cells of larvae from this treatment appeared columnar or afollicular, with the colloid partly or completely depleted, representative of hyperplasia (goitre). The lower growth rate in fish larvae from the control treatment was possibly a consequence of the hyperplasia, and the iodine enrichment prevented Senegalese sole larvae from developing goitre. This study demonstrates the importance of iodine enrichment of live feed for fish reared in a recirculation system.     

Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Development of hatchery technology for the silver pomfret Pampus argenteus (Euphrasen): effect of microalgal species on larval survival

The silver pomfret Pampus argenteus (Euphrasen) is a new candidate for aquaculture and there is not much information available on its larval rearing. Investigations carried out using microalgae alone in the culture system for the initial feeding of the silver pomfret larvae showed that Chlorella, Isochrysis and Nannochloropsis without rotifers are not conducive to the survival of newly hatched larvae. At 6 days after hatching (DAH), a maximum survival of 3% (1.8 ± 1.69%) was observed with Isochrysis followed by Nannochloropsis (0.35 ± 0.21%) and Chlorella (0.25 ± 0.21%). All control larvae died at 6 DAH without microalgae. Further investigations using the above microalgae with rotifers and a mixture of these same microalgae with rotifers showed that significantly higher (P < 0.05) survival could be achieved in the mixture of microalgae with rotifers in the culture system. At 12 DAH, the larval survival was 9.73 ± 1.39% in mixed species of algae compared with that of Isochrysis (6.93 ± 1.86%), Nannochloropsis (6.83 ± 0.61%), Chlorella (5.93 ± 2.76%) and seawater without microalgae or the control (0.73 ± 0.31%). The first incidence of feeding on rotifers at 4 DAH was significantly higher (P < 0.05) in all treatments with microalgae than that of the control. The incidence of feeding in mixed species of algae at 4 DAH (60.0 ± 0.00%) and in Isochrysis (55.0 ± 35.36%) was significantly higher (P < 0.05) than that of Chlorella (40.0 ± 0.00%) and the control (25.0 ± 7.07%). Prey consumption of individual larvae increased significantly (P < 0.01) at 8 DAH compared with that at 4 DAH. During this period, predation on rotifers by larvae was significantly higher (P < 0.05) in mixed species of algae (12.85 ± 5.73 rotifers larva^?1) than that of the control (6.75 ± 1.20 rotifers larva^?1). The fatty acid composition of rotifers used during this investigation shows that significantly higher (P < 0.05) ω3 HUFA was present in rotifers treated with mixed algae plus commercial enrichment media ‘Super Selco’ and ‘DHA Protein Selco’. Rearing of silver pomfret larvae up to the juvenile stage using mixed species of microalgae in the hatchery has been discussed. During 38 days of the larval rearing period, it was possible to achieve 3.6–4.2% larval survival with a mean of 3.9 ± 0.42%, which was considerably higher than in previous attempts (survival up to 1.5%).     

Improving weaning strategies for Senegalese sole: effects of body weight and digestive capacity

To optimize Senegalese sole‐weaning strategies, three experiments were performed. The first trial tested four weaning strategies with a 10 mg sole. Artemia‐fed sole grew threefold less than fish fed an inert diet. Sudden weaning (abrupt change from Artemia to inert diet) and weaning with co‐feeding produced larger sole than did a late weaning treatment; delayed weaning negatively affected fish growth. In the second experiment, the digestive capacity of early‐weaned 1, 2 and 4 mg sole was investigated. The highest growth was observed in sole weaned at 4 mg. Digestive enzyme profiles suggest that sole have an adaptation period to inert diets, with reduced feed intake. This adaptation period is inversely proportional to post‐larvae weight. The third experiment examined weaning with co‐feeding at different weights (2, 5 and 11 mg). These studies demonstrate that sole of 5–10 mg can be weaned, with high survival rates. On the basis of the digestive enzyme profiles, the early introduction of inert diets in co‐feeding with Artemia seems to affect intestinal processes in smaller postlarvae. This study also suggests that trypsin and alkaline phosphatase may be used as indicators of nutritional status in sole of <5 mg.     

Optimum time for weaning South African <Emphasis Type="Italic">Scylla serrata</Emphasis> (Forskål) larvae from rotifers to <Emphasis Type="Italic">Artemia</Emphasis>

To determine the optimum time at which to wean Scylla serrata larvae from rotifers onto Artemia two experiments were conducted, approximately 1 month apart, using larvae from two different female crabs. In the first experiment, the larvae in three treatment groups, with nine replicates each, were fed rotifers for the first 8 days after hatching. Artemia were introduced on days after hatch (DAH) 0 – during the first zoeal instar (treatment R + A); on DAH 4 – during the second zoeal instar (treatment R4A); on DAH 8 – during the third zoeal instar (treatment R8A). In a control (ROT) larvae were fed with rotifers exclusively for 18 days until the completion of metamorphosis to megalopa. In the second experiment, the same four feeding schedules as in experiment 1 were used with an additional group of larvae (treatment AC) that were fed only on Artemia throughout the rearing period. Similar results were recorded in the two experiments. Larvae in treatments R + A and R4A performed significantly better than those in treatments R8A, ROT and AC. This was particularly evident when examining the proportion of zoeae which successfully completed metamorphosis to megalopa. Poor performance of larvae in treatments AC and ROT implied that rotifers are needed as a first food, but that rotifers alone do not fill the nutritional requirements of S. serrata larvae. Poor performance of larvae in treatment R8A suggested that the diet should be supplemented with Artemia before the end of the zoea 3 stage.     

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 probiotic bacteria on growth parameters and immune defence in Eurasian perch Perca fluviatilis L. larvae under intensive culture conditions

Two experiments were conducted to examine whether a mixture of bacteria composed of Bacillus licheniformis, Bacillus subtilis and Bacillus pumilus has a beneficial impact on survival, growth, lysozyme activity and total immunoglobulins (Ig) in perch larvae. In experiment 1, control fish were compared with fish receiving bacteria (4.93 × 10⁷ CFU L⁻¹ day⁻¹) through tank water or both through tank water and Artemia metanauplii enriched with two bacteria doses (2 × 10¹⁰ or 4 × 10¹⁰ CFU L⁻¹). Treatments were performed in 40 L triplicate tanks of 200 fish [initial body weight (IBW)=2.3 mg] each, placed in separated recirculating water systems. Experiment 2 evaluated the combining effects of bacteria feeding and densities (10 vs. 20 fish L⁻¹, IBW=9.5 mg). In both experiments, the bacteria added to live food or both live food and water induced higher growth rate at the highest dietary dose and this was associated with higher Ig levels on days 14 and 28. In experiment 2, higher survival was associated to lower type‐2 cannibalism (ingestion of whole prey) at the two dietary bacteria doses, and such positive effect was observed for fish fed bacteria at high density. Bacteria administration through water had no effect on growth and survival. In conclusion, the results show that high dietary concentrations of the mixture of bacteria used have potential stimulating impact on growth, survival, lysozyme activity and total Ig. But, further investigations are needed to demonstrate the optimal doses for emphasizing immune defence status.     

Influence of light intensity on feeding, growth, and early survival of leopard coral grouper (Plectropomus leopardus) larvae under mass-scale rearing conditions

This study investigated the effect of different light intensities on feeding, growth and survival of early stage leopard coral grouper Plectropomus leopardus larvae. Four different light intensities (0, 500, 1000 and 3000 lx) were used and larvae were kept under constant light conditions from 0 day after hatching (DAH) to 5 DAH. The larvae were fed a small S-type of Thai strain rotifers at a density of 20 individuals/mL from 2 DAH. The number of rotifers in larval digestive organ and total length of larvae were examined at 3 h intervals between 04:00 and 22:00 h on 3 DAH, and thereafter at 6 h intervals until the end of the experiment (5 DAH). Four experimental trials of the larval rearing were repeated using by 60 kL mass-scale rearing tanks. The results indicate that coral grouper larvae are visual feeders and their food intake increases with increasing light intensity. Food intake of larvae reared at 3000 lx was significantly higher than those reared at 0–1000 lx on 3 DAH despite being the first-feeding day (P < 0.01). On 4 DAH, total length of larvae reared at 3000 lx was significantly larger than those reared at the lower light intensities (0, 500 and 1000 lx), and thereafter light intensity significantly influenced larval feeding and growth until the end of the experiment. Survival on 5 DAH did not show a significant difference between light intensities, but survival rate at 3000 lx and 1000 lx had a tendency to be higher than those reared at the lower light intensities (0 and 500 lx). In contrast, larvae reared at 0 lx exhibited stagnant and/or negative growth. These results indicate that light intensity is significantly the factor affecting larval feeding, growth, and survival in coral grouper larvae under the rearing conditions.     

Effect of probiotic (Bacillus spp.) addition during larvae and postlarvae culture of the white shrimp Litopenaeus vannamei

The effect of the addition of Bacillus probiotic during larvae and postlarvae culture of the white shrimp Litopenaeus vannamei was examined in three separate experiments: (I) Nauplius_4‐5 to Zoea₃, which were exposed to probiotic in the water (P_w), only in the microalgae (P_m), in the water and microalgae (P_wm) and a control with no probiotic (C); (II) Mysis₁ to Mysis₃, which were exposed to probiotic in the water (P_w), only in Artemia (P_a), in the water and Artemia (P_wa) and a control (C); (III) PL₁ to PL₁₀, which were exposed to similar treatments for mysis experiment. The use of probiotic Bacillus spp. resulted in an increase in the survival and growth of zoea and mysis phases, especially when the probiotic was added only in the water. For postlarvae, the use of the probiotic had no influence on the zootechnical parameters, however, there was a reduction in the count of presumptive Vibrio both for water and shrimp.     

Enhancement of growth performance, digestive enzyme activities and disease resistance in black tiger shrimp (Penaeus monodon) postlarvae by potential probiotics

Outbreaks of multidrug-resistant pathogens have been constraint on aquaculture production in Thailand, thereby controlling shrimp pathogens by preventive probiotics being importance to sustain the aquaculture system. In this study, the effect of potential probiotics, Bacillus subtilis and Enterococcus sp. was related to growth, digestive enzyme activities (trypsin and chymotrypsin) and pathogenic resistance by postlarval black tiger shrimp (Penaeus monodon). The experiment was divided into two treatments, in triplicate, with and without supplementation of probiotics as a food additive. Shrimp fed with probiotics for a culture period of 84 days showed a significant increase (P < 0.05) in weight (2.03 ± 0.29 g) and survival (71.91 ± 3.15 %) in comparison with non-treated shrimp (1.53 ± 0.28 g and 65.20 ± 5.68 %, respectively). Trypsin activity of treated shrimp was significantly higher (P < 0.05) than that of control shrimp whereas chymotrypsin activities of the two treatments were not significantly different (P > 0.05). After challenging P. monodon postlarvae with a shrimp pathogen, Vibrio harveyi for 10 days, percentage of mortality of P. monodon postlarvae fed with probiotics was 46.67 ± 1.44 %, significantly lower (P < 0.05) than that of non-treated shrimp (61.67 ± 6.29 %). This study showed that potential probiotics was appropriate for application in P. monodon postlarvae cultivation under laboratory condition due to improvement of shrimp weight and survival, enhancement of trypsin activity and reduced mortality causing by pathogenic V. harveyi. This is the first publication reported the effect of probiotics on trypsin and chymotrypsin activities of P. monodon postlarvae.     

Addition of bacteria bioencapsulated in Artemia metanauplii to a rearing system for halibut larvae

Theaim of this study was to determine whether it is possible to influence thespecies composition of the gut microflora in 70-days old halibut larvae(Hippoglossus hippoglossus L.) by addition of bacteriabioencapsulated in Artemia franciscana in two shortpulses.Two Vibrio strains, PB 1-11 and PB 6-1 were used. Twotreatments received each strain alone, a third treatment received the twobacterial strains in mixture, whereas in a control treatment no bacteria wereadded. Five fish tanks were included in each treatment. The bacteria werebioencapsulated in Artemia franciscana metanauplii duringashort-term incubation in bacterial suspensions of the specific strains, andwereadded in two pulses, on day 1 and day 10 of the experiment. Addition ofbacteriadid not increase the total number of colony-forming units (CFU) in the larvalgut. The total CFU in the water was lower in tanks added bioencapsulatedbacteria than in the control treatment (p < 0.05). Animmunocolony blot assay was used to measure the numbers of the specificbacteriaadded in samples of water, A. franciscana, and fishlarvae.At the end of the experiment on day 13, the strain PB 1-11 reached on average10% of the total CFU when added alone and 25% when added in mixture with PB6-1.Strain PB 6-1 reached at the end of the experiment 26.8% of the total CFU whenadded alone and 24.2% when added in mixture with PB 1-11, whereas thepercentages of PB 1-11 and PB 6-1 in the control fish were 12.7% and 10.8% ofthe total CFU, respectively. However, the differences between the experimentaltreatments compared with the control group were not significant(p > 0.05). The addition of bioencapsulated bacteria intwo pulses within a ten days period was therefore not sufficient to influencethe species composition of the microflora of the halibut larvae.     

饲料中添加潜在益生菌对凡纳滨对虾肠道消化酶活性和菌群组成的影响

在基础饲料中添加不同配伍的坚强芽孢杆菌Bacillus firmus活菌（1.0×108CFU/g）、美人鱼发光杆菌Photobacterium damsela灭活菌（1%）和溶藻弧菌Vibrio alginolyticus灭活菌（1%）作为实验饲料,基础饲料为对照组。实验所用凡纳滨对虾初始质量为3.2±0.26 g,投喂15d后进行WSSV感染实验并继续观察14d。实验期间定期取样测定对虾肠道胃蛋白酶、脂肪酶和淀粉酶的活性变化,并分析可培养肠道细菌总数和优势菌数量。结果表明,实验组对虾肠道中胃蛋白酶、脂肪酶和淀粉酶活性整体水平均显著高于对照组（P<0.05）,肠道细菌数量较对照组有显著增加。其中,坚强芽孢杆菌与美人鱼发光杆菌灭活菌复合菌组具有最高的淀粉酶活性,坚强芽孢杆菌与溶藻弧菌灭活菌复合组具有最高的脂肪酶和胃蛋白酶活性,而坚强芽孢杆菌单一组具有最大的肠道细菌总数和优势菌数。本研究表明,饲料中添加潜在益生菌可以提高凡纳滨对虾肠道消化酶活性并调节肠道菌群变化,利于养分的消化吸收。其中以坚强芽孢杆菌与美人鱼发光杆菌灭活菌复合组的整体效果较佳。     

The proteolytic digestive activity and growth during ontogeny of Parachromis dovii larvae (Pisces: Cichlidae) using two feeding protocols

The proteolytic digestive activity and growth of Parachromis dovii larvae during the ontogeny were evaluated in a recirculation system using two feeding strategies during a 28-day period. Larvae were reared using two feeding protocols (three replicates each): (A) Artemia nauplii (at satiation), fed from exogenous feeding [8 days after hatching (DAH)] until 15 DAH followed by nauplii substitution by formulated feed (20 % day^?1) until 20 DAH and then formulated feed until 28 DAH; (B) formulated feed (100 % BW daily) from exogenous feeding until 28 DAH. Levels of acid (pepsin type) and alkaline digestive proteases as well as growth and survival of larvae were measured along the feeding period. Survival was high and similar between treatments: 98.9 ± 0.0 for Artemia, 97.3 ± 0.0 % for formulated feed. The specific growth rate for length and weight was higher in larvae fed with Artemia nauplii than in larvae reared with formulated feed: 3.4 ± 0.1 versus 1.8 ± 0.1 % day^?1 for body length (P = 0.009) and 12.2 ± 0.1 versus 6.5 ± 0.3 % day^?1 for body weight (P = 0.002). The acid and alkaline proteolytic activity was detected, in both treatments, from the beginning of the experiment, at 8 DAH. The total enzymatic activity (U larva^?1) for acid and alkaline proteases was higher in larvae reared with Artemia after 12 DAH, whereas the specific enzymatic activity was similar for both enzyme types in the two treatments. The results suggest that P. dovii larvae were capable to digest formulated diets from the beginning of exogenous feeding and that they could be reared with formulated feeds. However, the formulated feed used should be nutritionally improved because of the poor growth obtained in this research.     

Colonization of the gut in first feeding turbot by bacterial strains added to the water or bioencapsulated in rotifers

Two bacterial strains, 4:44 and PB52, isolated from turbot(Scophthalmus maximus L.) were used during a first feedingexperiment with turbot larvae. Bacteria were either added directly tothe water on the day of hatching of the larvae (day 0), orbioencapsulated in rotifers (Brachionus plicatilis) distributedon day 2 after hatching. The two bacterial strains were found to bepresent in the water of the rearing tanks throughout the experiment. Theaddition of bacteria influenced the species-composition of themicroflora associated with intensively produced rotifers added to thetanks, and resulted in colonization of the gut of the larvae by theadded bacterial strains. The strain 4:44 showed a peak on day 9 afterhatching, and reached a mean(sem) of 2.5(1.4) × 10⁴bacteria per larva, when added in a mixture with PB52, and 0.4(0.1)× 10⁴ bacteria per larva, when added alone. The strainPB52, on day 12 after hatching, reached 5.2(1.5) × 10⁴bacteria per larva when added in a mixture with strain 4:44, and12.5(0.7) × 10⁴ bacteria per larva, when added alone.The added bacteria colonized the gut turbot larvae, whereas thebacterial land, the survival and growth of the larvae were in most casesnot influenced in a negative way by the addition of bacteria.     

Short‐term effect of the inoculation of probiotics in mature bioflocs: Water quality parameters and abundance of heterotrophic and ammonia‐oxidizing bacteria

The aim of this study was to evaluate the short‐term effect of probiotic inoculation on the abundance of heterotrophic and ammonia‐oxidizing bacteria in mature biofloc, as well as on total suspended solids (TSS), chlorophyll‐a and nitrogenous compounds in water. A completely randomized design consisting of five treatments (three commercial probiotics, one native consortia and one control) was performed. At the beginning of the experiment (day 1), each treatment was inoculated with the respective probiotic: PondPlus^® (PP), Efinol^® PT (EF) and Epicin^® ponds (EP), native consortia UE, whereas the control was not inoculated. Water parameters and bacterial abundance were evaluated at 0, 2, 4, 6 and 8 days. The addition of probiotics, either native or commercial, did not show any significant effect on the TSS, Chl‐a and colony‐forming unit (CFU) of heterotrophic bacteria when they were added to the systems containing mature biofloc. A significant increase in ammonium oxidizing bacteria was registered with the probiotics PP and EP, although the levels of total ammonia nitrogen, NO₃‐N and NO₂‐N were statistically similar among all treatments. Modifications on most of the parameters measured were associated with the factor of time, rather than the inclusion of probiotics. Results suggest that the bacterial conglomerates in mature stage contain well‐established bacterial communities that are difficult to be affected by the addition of probiotics.     

Use of the freshwater rotifer <Emphasis Type="Italic">Brachionus angularis</Emphasis> as the first food for larvae of the Siamese fighting fish <Emphasis Type="Italic">Betta splendens</Emphasis>

Larvae of the Siamese fighting fish Betta splendens were reared on the mass-cultured small freshwater rotifer Brachionus angularis Laos strain (UTAC-Lao), Paramecia sp., and Artemia as live food sources. Larvae fed live food were found to have a significantly high survival rate (97.5–100%) 18 days after hatch (DAH) in comparison to the control unfed larvae, which died by 12 DAH. Rotifer-fed larvae were found to grow faster than paramecia-fed larvae. The fastest growth rate was observed in larvae fed a combination of rotifer and Artemia, with growth in these larvae increasing by 282% by 18 DAH [total length (TL) 11.3 ± 1.2 mm] relative to body measurements taken 3 DAH. The next fastest growth rate was observed in rotifer-fed larvae, with a 158% increase in growth observed by 18 DAH (TL 7.6 ± 0.5 mm). The paramecia-fed larvae were found to grow by only 54.3% (TL 4.6 ± 0.1 mm) during the same period.