Biochemical Changes in Freshwater Prawn Macrobrachium rosenbergii During Larval Development

Abstract.— Quantitative changes in the protein, lipid and carbohydrate were studied in the early larval stages of developing freshwater prawn Macrobrachium rosenbergii reared under fed and starved conditions to understand the relative importance of these nutrients in energy metabolism of the growing larvae. 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. Protein was always the major organic constituent followed by lipids and then carbohydrates of both fed and starved larvae. Protein levels of both fed and starved larvae increased during development, suggesting an important role in morphogenesis. The decline of lipid during the larval growth that was more rapid for starved larvae, suggests a probable utilization of lipid as the major metabolic source of energy. Carbohydrates formed less than 5 and 2.4% of the larval dry weight of fed and starved larvae, respectively, suggesting their limited role in larval metabolism.     

Nutritional significance of amino acids,vitamins and minerals as nutraceuticals in poultry production and health – a comprehensive review

Nutraceuticals have gained immense importance in poultry science recently considering the nutritional and beneficial health effects of their constituents. Besides providing nutritional requirements to birds, nutraceuticals have beneficial pharmacological effects, for example, they help in establishing normal physiological health status, prevent diseases and thereby improve production performance. Nutraceuticals include amino acids, vitamins, minerals, enzymes, etc. which are important for preventing oxidative stress, regulating the immune response and maintaining normal physiological, biochemical and homeostatic mechanisms. Nutraceuticals help in supplying nutrients in balanced amounts for supporting the optimal growth performance in modern poultry flocks, and as a dietary supplement can reduce the use of antibiotics. The application of antibiotic growth enhancers in poultry leads to the propagation of antibiotic-resistant microbes and drug residues; therefore, they have been restricted in many countries. Thus, there is a demand for natural feed additives that lead to the same growth enhancement without affecting the health. Nutraceuticals substances have an essential role in the development of the animals’ normal physiological functions and in protecting them against infectious diseases. In this review, the uses of amino acids, vitamins and minerals as well as their mode of action in growth promotion and elevation of immune system are discussed.     

Comparison of pregnancy per AI of heifers inseminated with sex-sorted or conventional semen after oestrus detection or timed artificial insemination

The objective of the study was to compare the fertility after using sex-sorted or conventional semen either with oestrus detection (EST) or timed artificial insemination (TAI) in Holstein heifers. Holstein heifers were randomly assigned to one of the following treatments in a 2 × 2 factorial design. Heifers in the EST group were inseminated with sex-sorted (n = 114) or conventional semen (n = 100) after spontaneous or induced oestrus. Heifers in the TAI, subjected to the 5-day Cosynch+Progesterone protocol (GnRH+P4 insertion-5d-PGF_2α+P4 removal-1d-PGF_2α-2d-GnRH+TAI), were inseminated with sex-sorted (n = 113) or conventional semen (n = 88). Statistical analyses were performed using PROC GLIMMIX procedure of SAS 9.4 (SAS Institute Inc., Cary, NC). Overall P/AI was 60.7% for EST and 54.2% for TAI regardless of types of semen and 68.1% for conventional and 48.9% for sex-sorted semen regardless of insemination strategies. Fertility of heifers inseminated with either sex-sorted (53.5%; 44.2%) or conventional (69.0%; 67.0%) semen did not differ between EST and TAI respectively. Besides, the interaction between the semen type and the insemination strategy was not significant for P/AI. The embryonic loss was significantly greater with sex-sorted semen (17.1%) compared to conventional semen (1.6%). There was no sire effect with sex-sorted semen on P/AI (52.6% vs. 46.2%) and embryonic loss (16.4% vs. 18.0%). As expected, sex-sorted semen resulted in more female calves (89.8% vs. 51.6%) than conventional semen. Thus, sex-sorted semen can be used with 5-day Cosynch+Progesterone protocol to eliminate the inadequate oestrus detection and to increase female calves born in dairy heifers.     

Synovial fluid analysis and bacterial findings in arthritic joints of juvenile male camel (Camelus dromedarius) calves

Eighteen synovial fluid samples from 11 male dromedarian calves, 9-12 month old, were analysed cytologically and bacteriologically. Calves were lame and all joints were grossly swollen. The mean +/- SD of total nucleated cell count was 7970 +/- 5000 cells/microl (range 2800-20,000 cells/microl). Polymorphonuclear (PMN) leucocytes were the predominant cell type. The mean +/- SD of absolute and percentages of each cell type were as follows: PMN leucocytes 5518 +/- 3600 cells/microl and 68 +/- 19%, monocytes/macrophages 1600 +/- 1120 cells/microl and 26 +/- 17%, lymphocytes 830 +/- 140 cells/microl and 8 +/- 7%, and red blood cell 350 +/- 130 cells/microl. The mean +/- SD of total protein concentration was 3.5 +/- 1 g/dl (range 2.5-5 g/dl). The most commonly isolated bacteria were non-haemolytic streptococci spp., followed by Arcanobacterium pyogenes and Staphylococcus aureus. No bacterial growth was obtained in eight samples and non-revealed Mycoplasma spp.     

Comparative pharmacokinetics of marbofloxacin in healthy and Mannheimia haemolytica infected calves

The pharmacokinetics of marbofloxacin were investigated in healthy (n=8) and Mannheimia haemolytica naturally infected (n=8) Simmental ruminant calves following intravenous (i.v.) and intramuscular (i.m.) administration of 2 mg kg(-1) body weight. The concentration of marbofloxacin in plasma was measured using high performance liquid chromatography with ultraviolet detection. Following i.v. administration of the drug, the elimination half-life (t(1/2 beta)) and mean residence time (MRT) were significantly longer in diseased calves (8.2h; 11.13 h) than in healthy ones (4.6 h; 6.1 h), respectively. The value of total body clearance (CL(B)) was larger in healthy calves (3 ml min(-1) kg(-1)) than in diseased ones (1.3 ml min(-1) kg(-1)). After single intramuscular (i.m.) administration of the drug, the elimination half-life, mean residence time (MRT) and maximum plasma concentration (C(max)) were higher in diseased calves (8.0, 12 h, 2.32 microg ml(-1)) than in healthy ones (4.7, 7.4 h, 1.4 microg ml(-1)), respectively. The plasma concentrations and AUC following administration of the drug by both routes were significantly higher in diseased calves than in healthy ones. Protein binding of Marbofloxacin was not significantly different in healthy and diseased calves. The mean value for MIC of marbofloxacin for M. haemolytica was 0.1+/-0.06 microg ml(-1). The C(max)/MIC and AUC(24)/MIC ratios were significantly higher in diseased calves (13.0-64.4 and 125-618 h) than in healthy calves (8-38.33 and 66.34-328 h). The obtained results for surrogate markers of antimicrobial activity (C(max)/MIC, AUC/MIC and T > or = MIC) indicate the excellent pharmacodynamic characteristics of the drug in diseased calves with M. haemolytica, which can be expected to optimize the clinical efficacy and minimize the development of resistance.     

Dihydrooxadiazines: Octopaminergic system as a potential site of insecticidal action

Dihydrooxadiazines are structural analogs of octopamine and were compared with octopamine for their ability to compete with [³H]dihydroergocryptine ([³H]DHE) for binding sites on DHE-sensitive receptors, to stimulate adenylate cyclase activity in nervous system homogenates of Periplaneta americana L., and to modulate the action of the peptide proctolin on the oviducal muscles of Locusta migratoria L. [³H]DHE binding was inhibited by low concentrations (μM range) of octopamine, phentolamine, N-demethylchlordimeform (DCDM) and several dihydrooxadiazines. The tested dihydrooxadiazines acted as aminergic agonists in stimulating cyclic AMP production in cockroach nervous system homogenates and did not show additive effects with octopamine, whereas additivity was observed with 5-hydroxytryptamine. The relative potency of octopaminergic antagonists, including mianserin, cyproheptadine, phentolamine, and gramine, to block octopamine-mediated elevation of cyclic AMP production was similar to the rank-order potency of the same antagonists to inhibit dihydrooxadiazine-mediated elevation of cyclic AMP production. Octopamine, 2-(4-bromophenyl)-5,6-dihydro-4H-1,3,4-oxadiazine (4-Br-PDHO), and 8-Br-cyclic AMP caused increased phosphorylation of proteins that are phosphorylated by exogenously added cyclic AMP-dependent protein kinase. These results indicate that the dihydrooxadiazine-induced rise in cyclic AMP levels in homogenates of the cockroach nervous system results directly in activation of an endogenous cyclic AMP-dependent protein kinase. 4-Br-PDHO behaved similarly to octopamine in modulating the action of proctolin-induced contractions in locust oviducal muscles. These observations suggest that dihydrooxadiazines act as octopamine agonists and have an octopaminergic action in modulating the action of proctolin. Thus, it is proposed that dihydrooxadiazines exert at least part of their insecticidal and miticidal actions through interaction with the octopaminergic system.     

In-vitro Metabolism of a Novel Monocrotophos Derivative by Rat and Japanese Quail

NADPH-dependent inhibition of hepatic microsomal carboxylesterase by a derivative of monocrotophos (coded as RPR-5) was studied in rat and Japanese quail as a measure of monooxygenase-catalysed activation of RPR-5. There was NADPH-dependent inhibition of hepatic microsomal α-naphthyl acetate esterase (carboxylesterase) both in rat and quail, indicating monooxygenase-catalysed formation of an oxon that subsequently phosphorylated α-NaE. The pattern of in-vitro metabolism of ¹⁴C-labelled RPR-5 by 11000g supernatant (11-S), microsomes and 105000g supernatant (105-S) fractions of rat and quail livers suggested the involvement of microsomal monooxygenases and carboxylesterases. A radiolabelled metabolite (M2) was tentatively identified as an acid produced by carboxyl esterase attack. In rat, metabolism by microsomal and cytosolic (105-S) carboxylesterases appeared to predominate with relatively little oxidative metabolism. In quail, putative microsomal carboxylesterase hydrolysis of RPR-5 was much lower than in the rat with almost neglible hydrolysis by cytosolic fractions. Also, production of M₂ by quail microsomes was substantially reduced after addition of NADPH, suggesting inhibition of a carboxyl esterase by the oxon of RPR-5. Differences in this detoxification of RPR-5 between rat and quail may be an important factor in determining selective toxicity and the results underline the importance of relating metabolism to toxicity when selecting animal models for toxicity testing.     

Effects of AG1 and AG2 QTLs on Nonstructural Carbohydrate and Seed Management Options for Rice Seedling Growth and Establishment under Flooding Stress

Rice (Oryza sativa) plants acquired excess photosynthates in the form of nonstructural carbohydrates (NSCs) in their stems and grain. Despite keen interest in rice NSC, the dynamics of NSC accumulation, translocation and re-accumulation have not yet been well investigated. AG1 and AG2 QTLs associated with flooding tolerance through catalyzing starch into soluble sugar in germinating seeds. Here we conducted three experiments, greenhouse and field to lay the groundwork for large-scale diversity studies on grain NSC and some agronomic traits under direct-seeded rice (DSR) system, using elite lines incorporating AG1, AG2 and AG1-AG2 QTLs into the popular varieties PSB Rc82 and Ciherang-Sub1 along with the donors Kho Hlan On (AG1) and Ma-Zhan Red (AG2). In germinating seedlings, soluble sugars increased, while starch concentration decreased gradually especially in the tolerant checks and AG1-AG2 introgression lines under flooded soil. Soluble sugar accumulation in stem started to increase from the vegetative stage and peaked at the panicle initiation stage then gradually decreased towards the maturity stage. But Sub1-AG lines had higher sugar and starch concentrations at different growth stages than other genotypes in wet season 2016 and dry season 2017. Plant survival rate was positively correlated with the stem NSC at the early vegetative stage (21 days after sowing), and stem NSC was positively associated with plant height at different growth stages. Among the tested seeding rate, the most suitable seeding rate, 4 g/m² with shallow burial depth (0.5 cm), resulted in better seedling establishment, relatively higher seedling vigor index and higher leaf area index under flooding in DSR system. Introgression of AG1-AG2 QTLs had no any negative impact on nonstructural carbohydrate, germination rate, and growth and biomass production.     

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.