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Two trials were conducted to determine the e⁄cacy of ¢sh fed live yeast Debaryomyces hansenii strain CBS 8339 on immune and antioxidant systems in leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila. Juveniles (12 AE 0.5 g) were fed with a control diet or a D. hansenii-supplemented diet (10 6 colony-forming units per gram) for 5 weeks. The live weight of ¢sh was registered on a weekly basis. After 4 weeks, ¢sh from each treatment were immunocompromised with pathogenic A. hydrophila and further fed for1week in order to evaluate the e¡ect on immunological and antioxidant parameters. Generally, the results showed enhanced growth performance in ¢sh fed the diet containing yeast compared with the control. Addition of live yeast had no signi¢cant e¡ect on the immunological parameters after 4 weeks of feeding. However, post infection with A. hydrophila ¢sh fed the yeast-supplemented diet resulted in a sig-ni¢cant increase in the levels of plasmatic immunoglobulin M. Superoxide dismutase and catalase (CAT) activities were signi¢cantly higher in the yeast group. In this ¢sh, CAT and heat shock protein 70 genes were up-regulated before and after infection of A. hydrophila. The present study is the ¢rst one reporting that yeast (D. hansenii) can enhance immunity and resistance against A. hydrophila.

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The leopard grouper is an endemic species of the Mexican Pacific with an important commercial fishery and good aquaculture potential. In order to assess the digestive capacity of this species during the larval period and aid in the formulation of adequate weaning diets, this study aimed to characterize the ontogeny of digestive enzymes during development of the digestive system. Digestive enzymes trypsin, chymotrypsin, acid protease, leucine-alanine peptidase, alkaline phosphatase, aminopeptidase N, lipase, amylase and maltase were quantified in larvae fed live prey and weaned onto a formulated microdiet at 31 days after hatching (DAH) and compared with fasting larvae. Enzyme activity for trypsin, lipase and amylase were detected before the opening of the mouth and the onset of exogenous feeding, indicating a precocious development of the digestive system that has been described in many fish species. The intracellular enzyme activity of leucine-alanine peptidase was high during the first days of development, with a tendency to decrease as larvae developed, reaching undetectable levels at the end of the experimental period. In contrast, activities of enzymes located in the intestinal brush border (i.e., aminopeptidase and alkaline phosphatase) were low at the start of exogenous feeding but progressively increased with larval development, indicating the gradual maturation of the digestive system. Based on our results, we conclude that leopard grouper larvae possess a functional digestive system at hatching and before the onset of exogenous feeding. The significant increase in the activity of trypsin, lipase, amylase and acid protease between 30 and 40 DAH suggests that larvae of this species can be successfully weaned onto microdiets during this period.

The reproductive biology of the Blackfin snook Centropomus medius was studied to expand our current knowledge on its aquaculture potential. From April 2001 to November 2002, 301 females and 193 males were captured. A 1:1.6 (male:female) ratio was found. Gonadosomatic (GSI) and hepatosomatic (HSI) indices suggested that the spawning period (SP) extended from September to November, with peak spawning activity in September (GSI=0.98% and HSI=0.7%). The period of pre‐gametogenesis extended from January to June. During the post‐SP in December, 90% of oocytes were pre‐vitellogenic and 10% were atretic. Histological analysis of gonads (haematoxylin–eosin, Sudan black B, Alcian blue PAS, Cleveland Wolfe) showed five stages of oocyte development, each occurring at a characteristic range of oocyte diameters (μm): (I) pre‐vitellogenic (25–55); (II) cortical alveoli (56–250); (III) vitellogenic (251–340); (IV) maturation (341–645) and (V) atretic (21–100). Five reproductive stages were identified for testis morphology: (I) regressed; (II) early maturation; (III) mid‐maturation; (IV) full maturation and (V) regression. The type of ovarian development in C. medius was identified as a group synchronous.

The aim of this study was to evaluate the single or combined effects of Lactobacillus sakei with inulin suitable for immunological in vivo studies in farmed fish. By in vitro assays, L. sakei strain 5-4 showed antibacterial activities against all assayed fish pathogens (except the Vibrio harveyi strain CAIM-1793). L. sakei was able to survive at high fish bile concentrations. Fermentation of the agave inulin resulted in a large increase in number of lactobacilli. For the in vivo study, fish were fed for 8 weeks four practical diets: control diet (control), L. sakei 5-4 (10(7) CFU/g), inulin (1% or 10 g/kg) and L. sakei + inulin (10(7) CFU/g + 10 g/kg). The weight gain showed clearly the synergistic effect of L. sakei 5-4 and inulin at 6 and 8 weeks of treatments. Leopard grouper fed with L. sakei alone or combined with inulin have significantly increased the assayed physiological and humoral immune parameters. By real-time PCR assays, the mRNA transcripts of immunoglobulin M (IgM) were found to be higher expressed in intestine, head kidney, mucus, gill, spleen and skin. Moreover, mRNA expression levels of IgM in head kidney and anterior intestine were measured by real-time PCR. L. sakei 5-4 and L. sakei + inulin supplemented diet up-regulated the expression of IgM at week 4 and 8 in intestine and head kidney, respectively. These results support the idea that the L. sakei 5-4 alone or combined with agave inulin improved growth performance and stimulates the immune system of leopard grouper.

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