Bacillus subtilis, a probiont isolated from the intestine of Cirrhinus mrigala (Hamilton), was incorporated in fish feed at four different concentrations (5 · 10 8 , 5 · 10 7 , 5 · 10 6 and 5 · 10 5 cells g )1 ) and fed to Poecilia reticulata (Peters), Poecilia sphenops (Valenciennes), Xiphophorus helleri (Heckel) and Xiphophorus maculatus (Gunther) for a period of 90 days. The growth parameters, proximate composition of whole body, digestive enzymes and gut microbiology were monitored at regular monthly intervals. The increments in length and weight and the survival were significantly higher (P < 0.05) and the values of food conversions were significantly lower (P < 0.05) in fishes fed the probiotic feeds. The administration of B. subtilis in diet resulted in an increase (P > 0.05) in body ash and protein content and in the specific activity of protease and amylase in the digestive tract of all the fishes. The counts of total heterotrophs, motile aeromonads, presumptive pseudomonads and total coliforms were significantly lower (P < 0.05) in the gut of the probiotic fed fishes. Bacillus subtilis persisted in the fish intestine and in the feed in high numbers during the feeding period. The high survival and low infectivity recorded in the probiotic feed fed fishes when challenged by immersion assay with the pathogenic Aeromonas hydrophila demonstrate that the probiotic strain confers disease resistance in live-bearing ornamental fishes. A probiotic concentration of 10 6 to 10 8 cells g )1 is adequate for improved growth performance and survival and for healthy gut microenvironment and use of higher concentration of the probiont did not always lead to significantly better results. KEY WORDS
A probiotic bacterial strain, Bacillus subtilis, isolated from the intestine of Cirrhinus mrigala (Hamilton), was incorporated in ¢sh feed at four di¡erent concentrations (5 Â 10 8 cells g À 2 , 5 Â 10 7 cells g À 2 , 5 Â 10 6 cells g À1 and 5 Â 10 5 cells g À1 ) and fed to four species of livebearing ornamental ¢sh, Poecilia reticulata (Peters), Poecilia sphenops (Valenciennes), Xiphophorus helleri (Heckel) and Xiphophorus maculatus (Gunther) for one year duration to observe the effect of dietary probiotic supplementation on their reproductive performance. Sixty virgin females of each species were stocked separately in circular FRP tanks (350 L) and fed diets with varying levels of probiotic cells and control feed. Broodstock performance was evaluated based on gonadosomatic index (GSI), fecundity and fry production of female broodstock. The results showed that supplementation of feed with probiotics increased signi¢cantly (Po0.05) the GSI, fecundity and fry production of spawning females and length and weight of fry in all the four species of ¢sh. The number of dead and deformed fry were also signi¢cantly lower (Po0.05) in ¢sh fed with the probiotic feeds. The use of higher concentration of the probiont in diet did not always lead to sig-ni¢cantly improved reproductive performance of the spawners. Collectively, this study showed that female livebearers bene¢t from inclusion of probiotics in diet during their reproductive stages.
Diabetes increases the possibility of germ cell damage, hypogonadism, and male infertility. Diabetic condition negatively impacts zinc (Zn) and selenium (Se) levels in the body. Zn and Se are among the most important trace elements involved in the regulation of redox reaction, antioxidants enzymes activities, and DNA expression in a germ cell. The present study aimed to elucidate the combined effects of Zn and Se treatment on diabetes-induced germ cell damage in male Sprague Dawley rats. Type 1 diabetes was induced by the single intraperitoneal (i.p.) injection of streptozotocin (55 mg/kg). Zn (3 mg/kg, i.p.) and Se (0.5 mg/kg, i.p.) were administered daily for 8 consecutive weeks. All the animals were provided with normal feed and water throughout the study. The effects on germ cell damage were evaluated by body weight, feed-water intake, organ weight, sperm count, motility, sperm head morphology, biochemical analysis, histology, immunohistochemistry, halo assay, germ cell comet assay, testes terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end-labeling (TUNEL) assay, sperm TUNEL assay, serum protein pattern analysis, and subcellular analysis using transmission electron microscopy. Further, the expressions of nuclear erythroid-derived related factor 2, catalase, glutathione peroxidase 4, and glutathione peroxidase 5 were carried out to ascertain the mechanism of protection. The present results demonstrated that 8 weeks combined treatment of Zn (3 mg/kg, i.p.) and Se (0.5 mg/kg, i.p.) reduced diabetes-induced germ cell damage. This study further highlighted that Zn and Se combination treatment might be a better strategy for the germ cell protection in diabetes and deserve further investigation.
Zinc (Zn) plays an important role in maintaining the process of spermatogenesis and reproductive health. Bisphenol A (BPA), an endocrine disrupting chemical is known to be a reproductive toxicant in different animal models. The present study was designed to study the effect of two of the utmost determinative factors (Zn deficient condition and influence of toxicant BPA) on germ cell growth and overall male reproductive health in the testis, epididymis, and sperm using (a) biochemical, (b) antioxidant, (c) cellular damage, (d) apoptosis, and (e) protein expression measurements. Rats were divided into Control (normal feed and water), BPA (100 mg/kg/d), zinc deficient diet (ZDD; fed with ZDD), and BPA + ZDD for 8 weeks. Body and organ weights, sperm motility and counts, and sperm head morphology were evaluated. The histology of testes, epididymides, and prostate was investigated. Testicular deoxyribonucleic acid (DNA) damage was evaluated by Halo and Comet assay, apoptosis of sperm and testes were quantified by TUNEL assay. Serum protein electrophoretic patterns and testicular protein expressions such as Nrf-2, catalase, PCNA, and Keap1 were analyzed by Western blot analysis. The results showed that BPA significantly increased the testicular, epididymal, and prostrate toxicity in dietary Zn deficient condition due to testicular hypozincemia, hypogonadism, increased cellular and DNA damage, apoptosis, as well as perturbations in protein expression.
Bacillus subtilis isolated from the intestine of Cirrhinus mrigala (Hamilton) was incorporated into the rearing water of Poecilia reticulata (Peters), Poecilia sphenops (Valenciennes), Xiphophorus helleri (Heckel) and Xiphophorus maculatus (Gunther) at four different concentrations (5 9 10 8 cells ml -1 , 5 9 10 7 cells ml -1 , 5 9 10 6 cells ml -1 and 5 9 10 5 cells ml -1 ) and its effect on fish growth performance and survival, water quality parameters and bacterial population of water were assessed. The results showed that the addition of bacterial cells in the rearing water resulted in greater survival and a faster growth rate and, hence, greater length and weight increments of the livebearers. The use of a bioaugmentor in the rearing water of the livebearing fishes resulted in significantly lower (P \ 0.05) concentrations of dissolved organic matter and total ammonium nitrogen. The counts of motile aeromonads and total coliforms recorded in the water of bioaugmented tanks were also lower than that in the control tank. Bioaugmentation between 10 6 and 10 8 cells ml -1 in the rearing water is sufficient in establishing a bioaugmentor and the use of a higher concentration of bacterial cells did not always lead to significantly better results.
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