Structure and predictive metabolic contribution of intestinal microbiota of Longfin yellowtail (Seriola rivoliana) juveniles in aquaculture systems.

Seriola rivoliana intestinal microbiota (IM) was characterised under aquaculture conditions through 16S rRNA amplicon sequencing. Specimens of 30 days after hatching (DAH) were maintained in three tanks and fed under the same environmental conditions for characterisation 15 days prior to sampling. Three fish were randomly taken from each tank; total DNA extraction of the gut microbiota was performed to characterise microbial composition and its metabolic prediction. The V3 hypervariable region of the 16S rRNA was amplified and sequenced with Illumina pair-end technology. The prokaryotic components in the S. rivoliana intestine were dominated mainly by the phyla Proteobacteria, Firmicutes, Bacteroidetes, Cyanobacteria and Actinobacteria. No significant differences in beta diversity were detected in the three samples (tanks). However in alpha diversity, they were detected in juveniles of the same cohort within the same group, as exemplified by enrichment of certain bacterial groups, mainly of the Clostridia class, which were specific in each fish within the same tank. The metabolic prediction analyses suggested that S. rivoliana IM contribute to the metabolism of amino acids, carbohydrates, lipids, and immune system. This study provides the first IM characterisation under rearing conditions of S. rivoliana-a species with broad economic potential-and contributes to novel information for potential use of probiotics in future trials.

Evaluation of Homeopathic Phosphoric Acid, Silica and Pathogenic Vibrio on Digestive Enzyme Activity of Longfin Yellowtail Fish (Seriola rivoliana).

BACKGROUND: This research aimed to observe the effect of homeopathically prepared Vibrio parahaemolyticus (ViP) and V. alginolyticus (ViA) and the commercial homeopathic compound Similia (Phosphoricum acidum and Silicea terra) on the digestive enzyme activities of Seriola rivoliana juveniles under usual culture conditions. MATERIALS AND METHODS: Biochemical analysis was used to study the effect of highly diluted substances (7C potency) prepared from ViP and ViA (Treatment 1: T1) and the homeopathic compound Phosphoricum acidum and Silicea terra (Treatment 2: T2) on changes in the main digestive enzymes on weaning-state fish (WS; 30 days post-hatching [DPH]) and early juveniles (EJ; 62 DPH) versus a reference control group that received no homeopathic medicines. RESULTS: Treatment T2 significantly increased the activity of trypsin and lipase and decreased the activity of amylase, whereas treatment T1 increased the activity of chymotrypsin and reduced the activity of aminopeptidase-N in WS fish. Except for alkaline phosphatase, which was significantly reduced in the intestine, no significant differences in enzymatic activity were found between treated EJ fish and controls. The fish of the WS group had a higher growth rate with the T2 treatment. CONCLUSIONS: T1 treatment stimulated chymotrypsin in EJ fish and T2 promoted intestinal maturation of WS fish. Higher growth rate with the T2 treatment may be associated with the stimulation of trypsin activity. Thus, T2 may be applied, under hatchery conditions, during larval stages with an aim to enhance digestion and assimilation of inert food.

Immune and Antioxidant Enzyme Response of Longfin Yellowtail (Seriola rivoliana) Juveniles to Ultra-diluted Substances Derived from Phosphorus, Silica and Pathogenic Vibrio.

BACKGROUND: This research aimed to observe the effect of homeopathic treatments prepared from Vibrio parahaemolyticus and V. alginolyticus (H1) and commercial homeopathic medication Phosphoricum acidum and Silicea terra (H2) on the immune and antioxidant response in Seriola rivoliana juveniles under usual culture conditions and challenged with V. parahaemolyticus. MATERIALS AND METHODS: Quantitative polymerase chain reaction analysis was used to study changes in the expression of key genes related to immune response, cytokines (interleukin-1ß [IL-1ß]), adapter protein for cytokine release (MyD88) and piscidin and spectrophotometric techniques to analyze the activity of antioxidant superoxide dismutase (SOD) and catalase (CAT) enzymes in Seriola rivoliana juveniles at 30 (weaning stage [WS]) and 60 (early juveniles [EJ]) days post-hatching. RESULTS: The H1 treatment led to over-expression of the IL-1ß and MyD88 genes in fish at WS and EJ with respect to control, contrary to the H2 treatment that led to under-expression of the IL-1ß, MyD88 and piscidin genes at the EJ stage. In fish challenged with V. parahaemolyticus, both H1 and H2 led to over-expression of IL-1ß and MyD88; H2 caused an over-expression of piscidin. The SOD activity was higher in H1 with respect to H2 and the control group. CAT remained relatively stable with both H1 and H2 treatments. CONCLUSIONS: The results suggest that the overall effect of H1 was due to the presence of unknown antigens in low concentrations, while the response to H2-specifically during challenge-may have been due to a stimulating effect of nano-structures, prevailing from mother tincture after sequential dilution/succussion, in a pathway similar to that attributed to nano-vaccines.

Changes in digestive enzyme activities during early ontogeny of Seriola rivoliana.

This study contributes to basic knowledge on the development of larval digestive enzyme activities of Seriola rivoliana, a carnivorous species that has shown high mortality in larval stage. The study was conducted under laboratory-controlled conditions until larvae reached 30 days after hatching (DAH). The specific activity of amylase and pepsin was analysed by spectrophotometric methods, and lipase, trypsin, chymotrypsin and alkaline phosphatase by fluorometry. The results showed that S. rivoliana larvae had different enzyme patterns depending on their developmental stage, and also when exogenous food was administered (rotifers and Artemia). At hatching (2.98 ± 0.14 mm SL), lipase, alkaline phosphatase, trypsin and chymotrypsin activities were detected, but the last two enzymes showed an activity close to zero. Trypsin, chymotrypsin, lipase and alkaline phosphatase sharply increased their activities after starting the first feeding. Amylase activity was not detected until 5 DAH (3.51 ± 0.18 mm SL) and had a peak on day 15 (4.59 ± 0.39 mm SL); no activity was observed from then on. Pepsin activity was detected approximately at 15 DAH, showing a constant increase onwards. Trypsin had a peak on day 15 and then decreased on the following days. Chymotrypsin had the highest peak at 5 DAH, then a sharp decrease on day seven (3.88 ± 0.11 mm SL), and it increased gradually until day 30 (11.71 ± 1.59 mm SL). Lipase showed the highest activity at 15 DAH and then decreased its value. Finally, alkaline phosphatase showed the highest value at 3 DAH (3.62 ± 0.06 mm SL); then, a severe decrease was observed with activity stability on the following days. In conclusion, S. rivoliana larvae had a similar pattern as that observed in other marine-carnivorous fish larvae, so it could be considered as juvenile at 20 DAH in terms of digestive physiology; based on our results and previous study, the weaning process could start after this age, which would allow establishing a co-feeding protocol using inert diet.

Histological study of the gastrointestinal tract in longfin yellowtail (Seriola rivoliana) larvae.

This work contributes basic knowledge on larval development of Seriola rivoliana. A histological study describes the development of the digestive tract and accessory glands in S. rivoliana larvae reared under laboratory conditions at 24 °C from hatching to 30 days post-hatching (DPH). At hatching (2.6 ± 0.12 mm), larvae had an undifferentiated digestive tract with a closed straight tube and a large yolk sac with an oil globule. The liver and pancreas were observed at 1 and 2 days, and the mouth and anus opened at day 2. Enriched rotifers were visible in their digestive tract. At the beginning of the pre-flexion stage, a mixed nutritional period was observed. At day 3, exogenous feeding began; the digestive tract became differentiated into the buccopharynx, esophagus, an undifferentiated stomach, and the intestines. Zymogen granules were visible in the exocrine pancreas. At day 4, supranuclear vacuoles were present in the posterior intestine, indicating the beginning of intracellular digestion. At day 5, goblet cells were present in the esophagus and became functional at day 7 in the esophagus and intestine. The buccopharynx goblet cells developed at day 15. The presence of gastric glands and differentiation of the stomach in the fundic, cardiac, and pyloric regions during the post-flexion stage occurred at day 20. This was the onset of the juvenile period and the beginning of weaning; however, a long co-feeding phase is recommended. Pyloric caeca were observed at day 30 (13.6 ± 1.6 mm). These results provide valuable information on S. rivoliana larvae biology and digestive physiology, which should be useful to improve cultivation techniques and identify ecological features involved in ontogeny.

Ontogeny of the digestive tract of Centropomus parallelus larvae.

Development of the digestive tract and accessory glands of larvae of the fat snook Centropomus parallelus was examined under light microscopy, from hatching to 60 day post-hatching (dph). At hatching, the digestive tract is straight and composed by a cubic cell layer. The exogenous feeding starts at 3 dph, concomitantly with the mouth opening and subdivision of the rudimentary stomach and esophagus. At 4 dph, the intestine has three sectins, and vacuoles are observed in the posterior section, indicating the beginning of protein digestion and absorption. The pharyngeal teeth appear at 9 dph, and goblet cells appear at 13 dph in the esophagus. Gastric glands appear at 30 dph, marking the beginning of weaning. The disappearance of supranuclear vacuoles in the posterior intestine occurs at 35 dph, suggesting efficiency of extracellular digestion. This study shows that C. parallelus larvae is able to start weaning 15 days earlier than reported in earlier studies, increasing the success of larviculture.