Cryopreservation of Persian sturgeon (Acipenser persicus) sperm: effects of cryoprotectants,antioxidant, membrane stabilizer,equilibration time and dilution ratio on sperm motility and fertility

Three experiments were performed to develop protocols for cryopreservation of Persian sturgeon Acipenser persicus, sperm. In the first experiment, sperm from six males was individually split in three subsamples and cryopreserved using Modified Tsvetkova's extender (mT) supplemented with dimethyl sulfoxide (DMSO), methanol (MeOH), glycerol (Gly) and ethylene glycol (EG) at concentration of 5%, 10%, 15% and 20%. In the second set of experiments, the effects of six equilibration times (0, 5, 10, 20, 40 and 60 min) and dilution ratios (volume sperm: volume extender 1:0.5, 1:1, 1:2, 1:3, 1:5 and 1:10) and the additive advantage of bovine serum albumin (BSA; 0, 2.5, 5 and 10 mg mL^?1) and ascorbic acid (0, 2.5, 5 and 10 U mL^?1), on the post‐thaw survival of sperm (triplicate set of six fish) were evaluated. Then, sperm was diluted in 1:1 mT extender with 10 mg mL^?1 BSA with selected cryoprotectants (15% MeOH and 10% DMSO) for 5 min. After a month of storage in liquid nitrogen, post‐thawed sperm motility; fertilization and hatching rate and viability of derived larvae were measured (Exp.3). Evaluation of cryoprotectants efficiency showed that MeOH 15% and DMSO 10% were suitable for cryopreservation of Persian sturgeon sperm. Gly and EG resulted in very low post‐thaw motility rates even at lowest concentration. No significant difference was observed among the four different equilibration times (0, 5, 10, 20 min) (P > 0.05) although higher equilibration times than 20 min resulted low post‐thaw motility (P < 0.05). The motility of frozen–thawed sperm did not significantly change when dilution ratio was increased from 1:0.5 to 1:3 (P > 0.05). However, higher dilution ratios (1:5 and 1:10) reduced the percentage of motile sperm. Supplementation of the cryoprotectant solution with 10 mg mL^?1 BSA significantly improved post‐thaw motility (P < 0.05), but ascorbic acid did not improve post‐thaw motility (P > 0.05). The results of experiment 3 showed that the highest fertilization (30.2 ± 5.75) and hatching rates (28.2 ± 5.25) were observed when samples were frozen with 15% MeOH (P > 0.05). Our study indicates that the use of mT extender consisting of 10 mg mL^?1 BSA in 15% MeOH diluted with sperm at 1:1 ratio for 5 min can be recommended cryopreservation method for Persian sturgeon sperm.     

A fluorescence‐based assay for in vitro screening of Saprolegnia inhibitors

The incidence of fish pathogenic oomycetes, Saprolegnia, has increased significantly in aquaculture since the ban of malachite green. For the efficient characterization of anti‐Saprolegnia therapeutics, simple accurate methods are required. However, the current screening methods are limited by time, and none of them are confirming the viability of treated spores or hyphae. In this study, a modified fluorescence‐based assay for the in vitro screening of Saprolegnia inhibitors has been developed. This method involves the use of FUN‐1 viability dye combined with calcofluor white M2R, and is based on the formation of orange‐red cylindrical intravacuolar structures (CIVS) in metabolically active spores, hyphae and biofilms. Heat‐killed and bronopol‐treated Saprolegnia spores, hyphae and biofilms exhibited diffuse bright green fluorescence which confirms complete loss of viability. For boric acid‐treated spores, no germination was observed. However, tiny CIVS were observed in 50% of treated spores which indicated reduction in their viability. Our results proved that FUN‐1 dye is an efficient tool to distinguish between live and dead Saprolegnia spores, hyphae and biofilms and to monitor the change in Saprolegnia viability during qualitative evaluation of potential anti‐Saprolegnia compounds.     

Nano-calcium carbonate reinforced polypropylene and propylene-ethylene copolymer nanocomposites: Tensile vs. impact behavior

Improvement of both the tensile and impact strength of the same polymeric material has always been a great challenge for the plastic industry. The study focuses on the effect of incorporation of calcium carbonate nanoparticles (0.3 wt% to 15 wt%) into three polypropylene (PP) based matrices viz. PP homopolymer, propylene-ethylene (PP-PE) copolymer and the blend of PP:PP-PE (30:70) to improve their impact behavior without hampering the tensile strength much. A loss in both the tensile and impact properties was observed in PP based nanocomposite. However, PP-PE based nanocomposites showed a significant improvement in impact strength (47 %) at 10 wt% loading with a loss of tensile strength by 22 %. To minimize this loss a blend of PP:PP-PE (30:70) was explored as a matrix. At 10 wt% loading, this matrix showed an improvement of 30 % in impact strength whereas the tensile loss was minimized to 10 %. Further, silane coupling agent which promoted good interfacial adhesion was used for best compositions. The variation of crystalline morphology of the nanocomposites with various formulations was analyzed using differential scanning calorimetry and X-ray diffraction.     

Diversity of root-knot nematodes in Moroccan olive nurseries and orchards: does <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> disperse according to invasion processes?

Background

Root-knot nematodes (RKN) are major pest of olive tree (Olea europaea ssp. europaea), especially in nurseries and high-density orchards. Soil samples were collected from main olive growing areas of Morocco, to characterize Meloidogyne species and to discuss the contribution of biotic and abiotic factors in their spatial distribution.

Results

RKN were found in 159 soil samples out of 305 from nurseries (52.1% occurrence) and in 11 out of 49 soil samples from orchards (23.2% occurrence). Biochemical and molecular characterisation (PAGE esterase and SCAR) revealed the dominance of M. javanica both in nurseries and orchards with minor presence of M. incognita only in nurseries, and M. arenaria in only one nursery. RKN were distributed on aggregated basis. Frequent presence of M. javanica in orchards might have come from nurseries. In contrast, the detection of M. incognita in nurseries alone suggests that this species could not reproduce in orchards because of either the competition with other plant-parasitic nematodes or unfit local habitats. The impact of environmental variables (climate, habitat origin and physicochemical characteristics of the substrates) on the distribution of Meloidogyne species is also discussed.

Conclusion

Olive nurseries in Morocco are not able to guarantee the safety of rooted plants. As a result, olive production systems are exposed to strong RKN invasion risks. Consequently, the use of healthy substrates in nurseries may prevent plant-parasitic nematode induction in orchards.

     

Evaluating the Effects of Dietary Prebiotic Mixture of Mannan Oligosaccharide and Poly‐β‐Hydroxybutyrate on the Growth Performance,Immunity, and Survival of Rainbow Trout,Oncorhynchus mykiss (Walbaum 1792), Fingerlings

This study investigated the effects of dietary poly‐β‐hydroxybutyrate (PHB) and mannan oligosaccharide (MOS) on the growth performance, digestive enzyme activities, and disease resistance of rainbow trout fingerlings. A total of 420 fingerlings, distributed equally into four different groups, each with three replicates, were fed a basal diet (control), 2% PHB (PHB), 0.2% MOS (MOS), or a diet supplemented with a mixture of 2% PHB and 0.2% MOS (PHB+MOS) for 60 d. Results showed that dietary PHB and/or MOS did not cause significant improvement in the growth performance of the fingerlings. The gut pH was markedly reduced; however, the activities of the digestive enzymes (except for pepsin and amylase by dietary PHB) were not significantly improved by the dietary supplements. Results also showed that dietary PHB and/or MOS markedly increased the resistance of the fingerlings toward Yersinia ruckeri infection, while the lowest total antibody was observed in fish fed control and MOS, indicating that MOS exerts its protective effects by modulating other mechanisms. Overall, this study provided a first screening effort to determine the effects of PHB and/or MOS as dietary supplements in rainbow trout fingerlings to develop an optimal prebiotic mix for improving the growth and health status of rainbow trout.     

Effect of transferring lignocellulose-degrading bacteria from termite to rumen fluid of sheep on in vitro gas production,fermentation parameters,microbial populations and enzyme activity

The digestive tract of termite(Microcerotermes diversus) contains a variety of lignocellulose-degrading bacteria with exocellulases enzyme activity, not found in the rumen, which could potentially improve fiber degradation in the rumen. The objectives of the current study were to determine the effect of inoculation of rumen fluid(RF) with three species of bacteria isolated from termite digestive tract, Bacillus licheniformis, Ochrobactrum intermedium, and Microbacterium paludicola, on in vitro gas production(IVGP), fermentation parameters, nutrient disappearance, microbial populations, and hydrolytic enzyme activities with fibrous wheat straw(WS) and date leaf(DL) as incubation substrate. Inoculation of RF with either of three termite bacteria increased(P0.05) ammonia-N concentration compared with the control group(free of termite gut bacteria). Termite bacteria inoculation had no effect(P0.05) on gas production characteristics, dry matter, organic matter and neutral detergent fiber disappearance, pH, and concentration and composition of volatile fatty acids. Population of proteolytic bacteria and protozoa, but not cellulolytic bacteria, were increased(P0.05) when RF was inoculated with termite bacteria with both WS and DL substrates. Inoculation of RF with termite bacteria increased protease activity, while activities of carboxymethyl-cellulase, microcrystalline-cellulase, α-amylase and filter paper degrading activity remained unchanged(P0.05). Overall, the results of this study indicated that transferring lignocellulose-degrading bacteria, isolated from digestive tract of termite, to rumen liquid increased protozoa and proteolytic bacteria population and consequently increased protease activity and ammonia-N concentration in vitro, however, no effect on fermentation and fiber degradation parameters were detected. These results suggest that the termite bacteria might be rapidly lysed by the rumen microbes before beneficial effects on the rumen fermentation process could occur.     

Influence of nanochitosan/zeolite composite on growth performance,digestive enzymes and serum biochemical parameters in rainbow trout (Oncorhynchus mykiss)

Protective effects of nanochitosan/zeolite composite besides zeolite and chitosan/zeolite composite on rainbow trout growth, digestive enzyme activities and biochemical parameters were studied. Nanochitosan/zeolite hybrid composites with three different ratio of nanochitosan:zeolite (35:100, 3.5:100 and 0.35:100) were synthesized and analysed by X‐ray powder diffraction (XRD), thermal gravim (TG) analysis and scanning electron microscope (SEM) methods. Prepared composites as well as zeolite and control diets were fed to rainbow trout (50 g) for a period of 60 days. The addition of treated diets significantly improved growth performance compared to the control diet. Supplemental zeolite could only enhance the amylase activity in fish intestine, whereas other treatment groups could increase the pepsin activity besides intestinal alkaline phosphatase, trypsin and amylase activities. No differences were observed for intestinal acid phosphatase and lipase activities among the experimental diets. Meanwhile, serum total antioxidant activity and lipid peroxidation product, indicated by malondialdehyde (MDA), significantly increased and decreased, respectively, with some doses of administration, indicating the elevated antioxidant status in treatment groups. Serum‐specific marker enzymes, namely aspartate aminotransferase (AST) and alkaline phosphatase (ALP), were not affected in all groups. Meanwhile serum total protein in most treatment groups was significantly higher than the control group. Results showed that synthesized composites especially nanochitosan/zeolite composite at 5 g/kg had potential to enhance growth performance, digestive enzyme activities and some biochemical parameters in rainbow trout.     

Agronomic traits at the seedling stage,yield, and fiber quality in two cotton (Gossypium hirsutum L.) cultivars in response to phosphorus deficiency

ABSTRACT

Cotton is critical for phosphorus demands and very sensitive for its deficiency. However, identifying the effect of low-phosphorus tolerance on cotton growth, yield, and fiber quality by reducing phosphorus consumption. This may help to develop phosphorus-tolerant high-yielding cotton cultivars. In a two-year repeated (2015 and 2016) hydroponic experiment (using 0.01 and 1 mM KH₂PO₄), two cotton cultivars with phosphorus sensitivity (Lu 54; a low-phosphorus sensitive and Yuzaomian 9110; a low-phosphorus tolerant) were screened on the base of agronomic traits and physiological indices through correlation analysis, cluster analysis and principal component analysis from 16 cotton cultivars. Low phosphorus nutrition reduced the plant height, leaf number, leaf area, phosphorus accumulation and biomass in various organs of seedlings. The deficiency negatively affected the morphogenesis of seedlings, as well as yield and fiber quality. Further, these screened cultivars were tested in a pot experiment with 0, 50, 100, 150, 200 kg P₂O₅ ha^?1 during 2016 and 2017. It was found to have a significant (P< 0.05) difference in boll number, lint yield, fiber strength, and micronaire at the harvest. Furthermore, after collectively analyzed the characteristics of Lu 54 and Yuzaomian 9110, there were six key indices that could improve the low phosphorus tolerance of cotton cultivars. These were root phosphorus accumulation, stem phosphorus accumulation percentage, leaf and total biomass of seedlings, seed cotton weight per boll and fiber length.     

Zinc biofortification of maize (Zea mays L.): Status and challenges

Zn deficiency is one of the leading health problems in children and women of developing countries. Different interventions could be used to overcome malnutrition, but biofortification is most impactful, convenient, sustainable and acceptable intervention. Maize is one of the major crops grown and consumed in the regions with prevalent Zn malnutrition; therefore, this is suitable target for Zn biofortification. Zn biofortification of maize could be achieved through agronomic and genetic approaches. Discussion of agronomic approaches with genetic approaches is prerequisite because soils in developing countries are deficit of Zn and availability of Zn in soils is mandatory for estimating the genetic responses of maize genotypes through genetic approaches. Seed priming, foliar and soil applications are agronomic tools for biofortification, but solo and combined applications of these treatments have different effects on Zn enrichment. Genetic approaches include the increase of Zn bioavailability or increase of kernel Zn concentration. Zn bioavailability could be increased by reducing the anti‐nutritional factors or by increasing the bioavailability enhancers. Kernel Zn concentration could be improved through hybridization and selections, whereas genetically engineered attempts for improving Zn uptake from soil, loading in xylem, remobilization in grains and sequestration in endosperm can further improve the kernel Zn concentration. Key challenges associated with dissemination of Zn biofortified maize are also under discussion in this draft. Current review emphasized all of above‐mentioned contents to provide roadmap for the development of Zn biofortified maize genotypes to curb the global Zn malnutrition.