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.     

Ecological and Industrial Implications of Dynamic Seaweed-Associated Microbiota Interactions

Seaweeds are broadly distributed and represent an important source of secondary metabolites (e.g., halogenated compounds, polyphenols) eliciting various pharmacological activities and playing a relevant ecological role in the anti-epibiosis. Importantly, host (as known as basibiont such as algae)–microbe (as known as epibiont such as bacteria) interaction (as known as halobiont) is a driving force for coevolution in the marine environment. Nevertheless, halobionts may be fundamental (harmless) or detrimental (harmful) to the functioning of the host. In addition to biotic factors, abiotic factors (e.g., pH, salinity, temperature, nutrients) regulate halobionts. Spatiotemporal and functional exploration of such dynamic interactions appear crucial. Indeed, environmental stress in a constantly changing ocean may disturb complex mutualistic relations, through mechanisms involving host chemical defense strategies (e.g., secretion of secondary metabolites and antifouling chemicals by quorum sensing). It is worth mentioning that many of bioactive compounds, such as terpenoids, previously attributed to macroalgae are in fact produced or metabolized by their associated microorganisms (e.g., bacteria, fungi, viruses, parasites). Eventually, recent metagenomics analyses suggest that microbes may have acquired seaweed associated genes because of increased seaweed in diets. This article retrospectively reviews pertinent studies on the spatiotemporal and functional seaweed-associated microbiota interactions which can lead to the production of bioactive compounds with high antifouling, theranostic, and biotechnological potential.     

Marker‐assisted introgression of rare allele of β‐carotene hydroxylase (crtRB1) gene into elite quality protein maize inbred for combining high lysine,tryptophan and provitamin A in maize

Vitamin A deficiency in humans is a widespread health problem. Quality protein maize (QPM) is a popular food rich in lysine and tryptophan, but poor in provitamin A (proA). Here, we report the improvement of an elite QPM inbred, HKI1128Q for proA using marker‐assisted introgression of crtRB1‐favourable allele. HKI1128 was one of the parental lines of three popular hybrids in India and was converted to QPM in our earlier programme. Severe segregation distortion for crtRB1 was observed in BC₁F₁, BC₂F₁ and BC₂F₂. Background selection by 100 SSRs revealed mean recovery of 91.07% recurrent parent genome varying from 88.78% to 93.88%. Across years, introgressed progenies possessed higher mean β‐carotene (BC: 9.22 µg/g), β‐cryptoxanthin (BCX: 3.05 µg/g) and provitamin A (proA: 10.75 µg/g) compared to HKI1128Q (BC: 2.26 µg/g, BCX: 2.26 µg/g and proA: 3.38 µg/g). High concentration of essential amino acids, viz. lysine (mean: 0.303%) and tryptophan (0.080%) in endosperm, was also retained. Multi‐year evaluation showed that introgressed progenies possessed similar grain yield (1,759–1,879 kg/ha) with HKI1128Q (1,778 kg/ha). Introgressed progenies with higher lysine, tryptophan and proA hold immense potential as donors and parents in developing biofortified hybrids.     

Concentration of selected heavy metals in spices,dry fruits and plant nuts

Different spices, dry fruits and plant nuts commonly consumed in Pakistan were assayed for the heavy metals cadmium, lead, copper, zinc, iron and manganese by the potentiometric stripping analysis and AA spectrophotometry. The results revealed wide variation in heavy metal content among different biological materials. Mixed spices generally exhibited higher value for trace metals specially lead (6.6–9.2 µg/g), cadmium (0.65–1.34 µg/g), iron (142.3–285.0 µg/g) and zinc (64.2–65.8 µg/g). Dry fruits contained relatively lesser amounts of heavy metals than plant nuts. Almonds contained higher levels of lead (1.02 µg/g) and cadmium (0.24 µg/g) than other nuts and dry fruits.     

Nutrient digestibility studies in Heteropneustes fossilis (Bloch), Clarias batrachus (Linnaeus) and C. gariepinus (Burchell)

The apparent digestibility coefficient (ADC %) for protein in catfishes, Clarias batrachus (Linnaeus) (16–18 cm; 45–50 g), C. gariepinus (Burchell) (21–22 cm; 60–65 g) and Heteropneustes fossilis (Bloch) (10–12 cm; 10–15 g) was estimated for nine feedstuffs of plant and animal origin. Fat digestibility was tested for C. batrachus and C. gariepinus, wherein the influence of feeding frequency on protein digestibility was also examined. The digestibility trials were conducted with triplicate groups of 25 fish each in 70‐L polyvinyl indoor flow‐through (1–1.5 L min^?1) circular tanks (water volume=55 L). Fish were fed to satiation daily, at 08:00 and 18:00 hours, and faecal matter was collected through slow siphoning. For studying the effect of feeding frequency, fish were fed the experimental diets at 08:00, 11:00, 14:00 and 17:00 hours. The protein digestibility of the feedstuffs varied significantly within each test species. However, the protein digestibility values for most of the ingredients tested were comparable between the three catfish species. The most efficient utilization of protein in these fishes was noted for soybean meal followed by lab‐prepared fishmeal. The lowest protein digestibility was recorded for rice bran. Similarly, the utilization of fat from the feedstuffs was also significantly different within each of the two species, C. batrachus and C. gariepinus. Fat digestibility of feed ingredients was similar between the two species. With respect to the fat, soybean meal, lab‐prepared fishmeal, full‐fat soybean meal and dried fish were more efficiently utilized, while the lowest fat digestibility was obtained for rice bran. A marked decline in protein utilization was observed in the two species where the feeding frequency was increased from two to four times a day.     

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.     

Assessment of gut microbiota in different developmental stages of Malaysian Mahseer (Tor tambroides)

Malaysian Mahseer (Tor tambroides) has a good prospect for aquaculture because of its high market demand. However, there is a scarce information on gut microbiota associated with Malaysian Mahseer unlike other fish species. Therefore, we constructed and compared gut microbiota in different developmental stages (larval, juvenile, fingerling, yearling, and adult) using culture dependent and PCR‐DGGE fingerprinting technique for better understanding of gut microbiota composition associated with T. tambroides. Culturable gut microbiota composition in all developmental stages were composed of β‐ and γ‐Proteobacteria, and Bacilli. Biodiversity analysis of culturable gut microbiota showed that larval, juvenile, and adult stages have higher diversity than fingerling and yearling stages. Ward's linkage cluster analysis showed that culturable gut microbiota composition in larval and juvenile stages were close to adult stages, whereas fingerling and yearling stage composed same cluster. PCR‐DGGE fingerprinting technique showed that unculturable gut microbiota were constituted by α‐and γ‐Proteobacteria, and Actinobacteria. Ward's linkage cluster analysis showed that unculturable gut microbiota composition in both larval and juvenile stages were distinct from other developmental stages. Our results revealed that gut microbiota composition were varied in different developmental stages of Malaysian Mahseer and continuous shifts of gut microbiota from larval to adult stages.     

Diurnal variations in hypothalamic monoamine levels in the teleostChanna punctatus (bloch) in response to melatonin under two photothermal conditions

Hypothalamic dopamine (DA), noradrenaline (NA) and 5-hydroxytryptamine (5-HT) levels exhibited marked day-night variations under ambient photoperiod and temperature (12L12D; 17±1°C) with peak values at mid-light phase. The 16L8D; 22±1°C treatment reversed the diurnal rhythm of 5-HT, but not that of DA and NA. However, there was an overall increase in the levels of the catecholamines on exposure to the long photoperiod and high temperature. The administration of melatonin in the fish held on 16L8D; 22±1°C regime restored the 5-HT rhythm to that of the 12L12D; 17±1°C control group, but with elevated mid-photophase value. However, there was no effect of the indole treatment on the 5-HT rhythm under the 12L12D; 17±1°C regime. Melatonin causes a significant reduction of NA level in both the groups, while DA level did not change in either group.     

Studies on the ecology of actinomycetes in soil—VIII: Distribution and characteristics of acidophilic actinomycetes

Acidophilic actinomycetes, growing between pH 3.5 and 6.5, were isolated from 17 natural soils and mine wastes. Most isolates were Streptomyces spp. and selected ones were compared with some established neutrophilic species by assessment of overall similarity. Acidophilic and neutrophilic strains clustered separately, differing in a number of cultural and physiological characters, in addition to pH requirements. The possible importance of acidophilic actinomycetes in soil is discussed.     

Boron Induced Oxidative Stress,Antioxidant Defence Response and Changes in Artemisinin Content in Artemisia annua L.

Boron is an essential plant micronutrient and the range between deficient and toxic levels of boron is narrow for most of the plants. Like other elements, boron becomes toxic to growth at high concentrations. High boron concentrations in soil reduce crop productivity in many areas of the world. The effect of increasing levels of boron (0, 0.50, 1.00, 1.50, 2.00 mm ) on oxidative stress, antioxidant defence response and changes in artemisinin content in Artemisia annua were investigated in the present study. Boron toxicity reduced the growth parameters viz. stem height, fresh weight and dry weight. Treatments induced oxidative stress resulting in lower net photosynthetic rate, stomatal conductance, internal CO₂ and total chlorophyll content. The increased activities of antioxidant enzymes like CAT, POX and SOD were also noted in response to increasing levels of boron stress. However, H₂O₂ and artemisinin content were found to be high up to 1.00 mm concentration of boron compared to control, and on applying higher doses, further reduced contents were obtained. Thus, the results suggest that a mild stress of boron can be utilized for enhanced artemisinin production.