Is N‐feedback involved in the regulation of nitrogenase activity in Medicago truncatula?

To determine whether senescing leaves provoke an active nitrogen (N) remobilization that results in the reduction of nitrogenase activity, 60% of Medicago truncatula lower leaves were either darkened or individually excised for two weeks. Although a considerable amount of N was remobilized, N₂ fixation activity was found to be increased to maintain the N source/sink balance, indicating an absence of the negative N‐feedback regulation of nitrogenase activity in the senescing M. truncatula.     

Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada)

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.     

Recent update on the prevalence of Vibrio species among cultured grouper in Peninsular Malaysia

Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene.     

Characterization of a Probiotic Lactobacillus fermentum Isolated from Snakehead,Channa striatus,Stomach

This study sought to isolate and characterize lactic acid bacteria (LAB) from stomach of adult snakehead fish, Channa striatus, to be used as probiotics for freshwater fish. A total of 13 strains were isolated from the stomach of 10 fish, and 4 of these belonged to LAB. Strain LAB‐3 showing highest in vitro growth inhibition of Aeromonas hydrophila in a disk diffusion test was identified as Lactobacillus fermentum by conventional and molecular techniques and evaluated in vitro through various tests. The bacterium could grow at pH 3–8; but the optimum growth was observed at pH 6. Moreover, LAB‐3 grew at 0.15 and 0.3% bile salt concentrations, from 15 to 45 C, and at 4% NaCl. L. fermentum showed in vitro inhibitory activity against three fish pathogens, A. hydrophila, Pseudomonas aeruginosa, and Shewanella putrefaciens, tested by disk diffusion and well diffusion methods. Antibiotic sensitivity tests indicated that L. fermentum was resistant to streptomycin, gentamycin, and kanamycin, intermediate to tetracycline, but sensitive to chloramphenicol, amoxicillin, and ampicillin. Challenge test by using A. hydrophila showed that survival of snakehead was significantly (P < 0.05) improved when 2 × 10⁶ LAB‐3/g was supplemented to the diet. Therefore, this study suggests that L. fermentum might be a promising probiotic in snakehead aquaculture .     

Amino Acid Composition of Developing Larval Freshwater Prawn Macrobrachium rosenbergii

Abstract.— Amino acid profiles of freshwater prawn Macrobrachium rosenbergii were determined during early larval stages (I-IX) to provide some baseline biochemical information of the growing larvae. The larvae obtained from several females were stocked into three 250-L tanks at a density of 30 larvae/L. The feeding regime consisted of newly hatched Artemia nauplii and egg custard containing 5% cod liver oil. For each developmental stage, larval samples from each experimental tank were pooled, freeze dried and after acid hydrolysis, the quantification of amino acids was done by a reverse phase high performance liquid chromatography ( N = 2). The highest represented of the total amino acids were for glutamic acid and phenylalanine (with cystine) with ranges from 13.4–16.6 and 9.7–11.5%, respectively, whereas tryptophan (1.4–1.6%). methionine (1.4–2.7%) and histidine (2.9–4.2%) were relatively lower. The levels of the following essential amino acids did not significantly change during larval development: arginine, leucine, phenylalanine (plus cystine), threonine, tryptophan and valine. Despite statistically significant changes that were observed in levels of some amino acids, amino acid composition appeared to be relatively unchanged during the larval growth. The results may suggest that the amino acid requirements of the freshwater prawn is relatively constant during larval life and can be satistied by a suitable protein source that resembles the larval amino acid profile.     

Effects of thyroxine,cod liver oil and potassium iodide on growth and survival of juvenile seahorse,Hippocampus barbouri

Low survival at early stage is the bottleneck in seahorse aquaculture, particularly in the feeding aspect since newborn seahorses must feed immediately upon birth to sustain themselves. Seahorses are visual predator, therefore preferred live feed such as zooplankton. In aquaculture, the most common live feed used is Artemia. In this study, two experiments were conducted to determine the effect of different Artemia enrichment on the growth and survival of newborn Hippocampus barbouri. In the first experiment, six treatments using Artemia enriched with Culture Selco Plus? (SELCO), thyroxine (T4), potassium iodide (KI), cod liver oil (CLO), cod liver oil in combination with thyroxine (CLO + T4) and potassium iodide (CLO+KI) were fed to newborn H. barbouri. Newly hatched Artemia were used as control. At the end of first experiment, treatments using CLO + T4 produced juvenile H. barbouri with the best (p < .05) wet weight (0.142 ± 0.000 g), while juvenile in treatment CLO+KI recorded the highest (p < .05) standard length (3.947 ± 0.014 cm). Subsequently, a second experiment was carried out using the two best enrichment (CLO + T4 and CLO + KI) from the first experiment, but given at different frequency (daily, twice a week, once a week, once in 2 weeks). Daily enrichment using both CLO + T4 and CLO + KI showed no significant (p > .05) difference in growth performance and survival of juvenile of H. barbouri. Interestingly, juvenile fed CLO + T4 enriched Artemia at frequency of twice a week also has no significant difference (p > .05) in survival and growth performance (except for final standard length) when compared with treatment CLO + T4 at daily frequency.     

Current experimental models,assessment and dietary modulations of intestinal permeability in broiler chickens

Maintaining and optimising the intestinal barrier (IB) function in poultry has important implications for the health and performance of the birds. As a key aspect of the IB, intestinal permeability (IP) is mainly controlled by complex junctional proteins called tight junction proteins (TJ) that link enterocytes together. The disruption of TJ is associated with increased gut leakage with possible subsequent implications for bacterial translocation, intestinal inflammation, compromised health and performance of the birds. Despite considerable data being available for other species, research on IP in broiler chickens and in general avian species is still an understudied topic. This paper reviews the available literature with a specific focus on IP in broiler chickens with consideration given to practical factors affecting the IP, current assessment methods, markers and nutritional modulation of IP. Several experimental models to induce gut leakage are discussed including pathogens, rye-based diets, feed deprivation and stress-inducing agents such as exogenous glucocorticoids and heat stress. Although various markers including fluorescein isothiocyanate dextran, expression of TJ and bacterial translocation have been widely utilized to study IP, recent studies have identified a number of excreta biomarkers to evaluate intestinal integrity, in particular non-invasive IP. Although the research on various nutrients and feed additives to potentially modulate IP is still at an early stage, the most promising outcomes are anticipated for probiotics, prebiotics, amino acids and those feed ingredients, nutrients and additives with anti-inflammatory properties. Considerable research gaps are identified for the mechanistic mode of action of various nutrients to influence IP under different experimental models. The modulation of IP through various strategies (i.e. nutritional manipulation of diet) may be regarded as a new frontier for disease prevention and improving the health and performance of poultry particularly in an antibiotic-free production system.