Genetic Analyses of Resistance to Fusarium Basal Rot in Onion

Fusarium basal rot (FBR) is a serious disease of onion (Allium cepa). We identified sources of FBR resistance, assessed efficacy of selection for increased resistance, and investigated its genetic control. Onion accessions were evaluated for FBR resistance, and percentage survival ranged from 0% to 78%. Survivors were intercrossed, and progenies from one cycle of selection showed increased survival by 18% to 52%. Selections were crossed to male-sterile lines, and hybrids showed specific combining ability for FBR resistance. Segregating families were produced, and quantitative trait loci (QTLs) were identified on chromosomes 2 and 4 conditioning FBR resistance. A second QTL on chromosome 4 was identified that decreased FBR resistance. Plants from families with different genotypes across the 1.5 logarithm of odds (LOD) regions on chromosomes 2 and 4 were self-pollinated, and resulting families were evaluated for FBR survival. Genomic regions on chromosomes 2 and 4 associated with resistance were validated at p = 0.05 and 0.10, respectively. The region on chromosome 4 associated with increased susceptibility was validated at p = 0.05. These results are in agreement with previous studies reporting high heritability and specific combining ability for FBR resistance and should be useful for selection of FBR-resistant onion.

Studies on the Growth Responses of Common Carp (Cyprinus carpio) Progenies Raised Using Different Starter Diets

200 fry of carp obtained from induced breeding by stripping were fed three times daily ad libitum with Artemia (DT1), commercial starter diet (DT2), and dried egg yolk (DT3)for 8 weeks. Each treatments were in triplicates in indoor hatchery. At the end of the experiment, the progenies mean weight gained (MWG) was highest in the treatments fed Artemia (3.490±0.015a) as compared to others with significant difference (P<0.05).Mean weight gained (MWG) was highest in the treatments fed Artemia. Percentage weight gain (%WG) and specific growth rate (SGR) had no significant differences (P<0.05). Feed conversion ratio (FCR) was highest (P<0.05) in treatment fed with smashed egg yolk. Feed conversion efficiency (FCE) was highest (P<0.05) in the treatment fed with composed feed. Mortality rate was highest in treatment fed with Artemia, and percentage survival was highest (P<0.05) in treatment fed with composed feed. Percentage jumpers per treatment were highest in Artemia diet. In conclusion, it is imperative that Artemia feed (amidst other starter diets researched on) is most preferable for the optimum growth and development of common carp fry.

Impacts on the immune system of Cyprinus carpio exposure with a mixed algal extract against Aeromonas hydrophila

This study evaluates the influence of mixed algal extract (<i>Chlorella vulgaris</i>, <i>Euglena viridis</i> and <i>Spirulina platensis</i>) on common carp <i>Cyprinus Carpio</i>, which infected infect with bacterial pathogen <i>Aeromonas hydrophila</i>. <i>C. carpio</i> was administered intraperitoneally with various doses such as methanol extract (0, 0,1, 1, 10 and 100 mg/kg). The immunological parameters of fish blood and serum samples (Neutrophil activity, Lysozyme activity, Serum myeloperoxidase intensity, Serum bactericidal activity, and Serum antiprotease activity) were investigated at 7, 14, 21, and 28 days of post-immunization. Fish had been tested by virulent<i> A. hydrophila</i> for 30 days after treatment and 14 days after infection were identified with mortalities. The findings showed that neutrophil levels, lysozyme activity, serum bactericidal activity, myeloperoxidase activity, and serum antiprotease activity significantly enhanced (p<0.05) compared to untreated control. Mixed dietary algae at 1 and 10 mg/kg levels demonstrated slightly (p<0.05) higher relative percentage survival (90 percent) than control against <i>A. hydrophila</i> disease infection. Results indicated that mixed algal extract in <i>C. carpio</i> positively impacts non-specific immune parameters and boosts disease tolerance to <i>A. hydrophila</i> infections.

Mitigating multiple stresses in Pangasianodon hypophthalmus with a novel dietary mixture of selenium nanoparticles and Omega-3-fatty acid

Effects of a novel dietary mixture of selenium nanoparticles (Se-NPs) and omega-3-fatty acids i.e., Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on mitigating arsenic pollution, high-temperature stress and bacterial infection were investigated in Pangasianodon hypophthalmus. To aim this, four isocaloric and iso-nitrogenous diets were prepared: control feed (no supplementation), Se-NPs at 0.2 mg kg−1 diet with EPA + DHA at 0.2, 0.4 and 0.6% as supplemented diets. Fish were reared under normal condition or concurrent exposure to arsenic (2.65 mg L−1), and temperature (34 °C) (As + T) stress for 105 days. The experiment was conducted with eight treatments in triplicates. Response to various stresses i.e., primary (cortisol), secondary (oxidative stress, immunity, and stress biomarkers) and tertiary stress response (growth performance, bioaccumulation and mortality due to bacterial infection) were determined. Supplementation of dietary Se-NPs at 0.2 mg kg−1 diet and EPA + DHA at 0.2 and 0.4% reduced the primary stress level. Exposure to arsenic and temperature (As + T) and fed with control diet and EPA + DHA at 0.6% aggravated the cortisol level. Anti-oxidative enzymes (Catalase, superoxide dismutase, glutathione peroxidase and glutathione-s-transferase) and immunity (Nitroblue tetrazolium, total protein, albumin, globulin, A:G ratio, total immunoglobulin and myeloperoxidase) of the fish were augmented by supplementation of Se-NPs and EPA + DHA at 0.2 and 0.4%. Neurotransmitter enzyme, HSP 70, Vitamin C were significantly enhanced (p < 0.01) with supplementation of Se-NPs at 0.2 mg kg−1 and EPA + DHA at 0.2 and 0.4%. Whereas total lipid, cholesterol, phospholipid, triglyceride and very low-density lipoprotein (VLDL) were reduced (p < 0.01) with the supplementation of Se-NPs at 0.2 mg kg−1 diet and EPA + DHA at 0.2 and 0.4%. Tertiary stress response viz. growth performance was also significantly enhanced with supplementation of Se-NPs at 0.2 mg kg−1 and EPA + DHA at 0.2 and 0.4% reared under As + T. Whereas arsenic bioaccumulation in fish tissues was significantly reduced with dietary supplementation of Se-NPs and EPA + DHA. Cumulative mortality and relative percentage survival were reduced with Se-NPs at 0.2 mg kg−1 and EPA + DHA at 0.2 and 0.4%. The investigation revealed that a novel combination of Se-NPs at 0.2 mg kg−1 and EPA + DHA at 0.4% followed by 0.2% has the potential to alleviate temperature stress, bacterial infection and arsenic pollution. Whereas diet containing Se-NPs at 0.2 mg kg−1 diet and EPA + DHA at 0.6% was noticeably enhanced the stress in P. hypophthalmus.

Mapping freezing tolerance QTL in alfalfa: based on indoor phenotyping

Background Winter freezing temperature impacts alfalfa (Medicago sativa L.) persistence and seasonal yield and can lead to the death of the plant. Understanding the genetic mechanisms of alfalfa freezing tolerance (FT) using high-throughput phenotyping and genotyping is crucial to select suitable germplasm and develop winter-hardy cultivars. Several clones of an alfalfa F1 mapping population (3010 x CW 1010) were tested for FT using a cold chamber. The population was genotyped with SNP markers identified using genotyping-by-sequencing (GBS) and the quantitative trait loci (QTL) associated with FT were mapped on the parent-specific linkage maps. The ultimate goal is to develop non-dormant and winter-hardy alfalfa cultivars that can produce extended growth in the areas where winters are often mild. Results Alfalfa FT screening method optimized in this experiment comprises three major steps: clone preparation, acclimation, and freezing test. Twenty clones of each genotype were tested, where 10 samples were treated with freezing temperature, and 10 were used as controls. A moderate positive correlation (r ~ 0.36, P < 0.01) was observed between indoor FT and field-based winter hardiness (WH), suggesting that the indoor FT test is a useful indirect selection method for winter hardiness of alfalfa germplasm. We detected a total of 20 QTL associated with four traits; nine for visual rating-based FT, five for percentage survival (PS), four for treated to control regrowth ratio (RR), and two for treated to control biomass ratio (BR). Some QTL positions overlapped with WH QTL reported previously, suggesting a genetic relationship between FT and WH. Some favorable QTL from the winter-hardy parent (3010) were from the potential genic region for a cold tolerance gene CBF. The BLAST alignment of a CBF sequence of M. truncatula, a close relative of alfalfa, against the alfalfa reference showed that the gene’s ortholog resides around 75 Mb on chromosome 6. Conclusions The indoor freezing tolerance selection method reported is useful for alfalfa breeders to accelerate breeding cycles through indirect selection. The QTL and associated markers add to the genomic resources for the research community and can be used in marker-assisted selection (MAS) for alfalfa cold tolerance improvement.

EMBRYO-LARVAL TOXICITY OF ANTI-FUNGUS CHEMICALS USED AS PROPHYLACTIC AGENTS ON CLARIAS GARIEPINUS (BURCHELL, 1822)

Embryo-larval toxicity test of selected anti-fungus chemicals used as prophylactic agents on the African catfish (Clarias gariepinus) was conducted to assess the comparative efficacy as well as the toxicity of the different chemicals. Measured endpoints included hatching, embryo and larval survival, and larval growth. Iodine (100 mg L-1), hydrogen peroxide (250 mg L-1), malachite green (5 mg L-1) and formalin (250 mg L-1) were used as prophylactic agents. The eggs were spread on an incubating raft and continuously dipped in the treatment chemical for 15 minutes. After 48 hours, the mean hatch rate was highest in formalin-treated embryos (70%). Iodine treatment resulted in a hatch rate of 36.7%, which was slightly less than half that of the formalin treatment. Both hydrogen peroxide and malachite green resulted in 60.0% hatch rates. The results indicate that formalin-treated embryos performed significantly better than all the other chemicals. The highest percentage survival rate of 63.3% was observed in formalin at 72hph and 56.7% after 168hph. The lowest survival rate (10%) was observed in the iodine treatment group and closely followed by the group treated with malachite green (16.7%). Except for malachite green, which inhibited growth, the prophylactic treatments did not affect the growth performance of fry. This study presents evidence that can be used to support the use of formalin and hydrogen peroxide as prophylactic treatments in the early stages of C. gariepinus, although caution should be exercised due to the possibility of toxicity at higher concentrations and for longer periods of exposure.

The Efficacy of Moina micrura Enriched with Probiotic Bacillus pocheonensis in Enhancing Survival and Disease Resistance of Red Hybrid Tilapia (Oreochromis spp.) Larvae

The administration of probiotics via live feeds, such as Artemia and rotifers, has gained significant attention. Moreover, indiscriminate use of antibiotics in conventional aquaculture practices in order to prevent or control disease outbreaks has resulted in the occurrence of residues and antimicrobial resistance. Thus, the application of eco-friendly feed additives, such as probiotics, as a safer alternative has received increasing attention in recent years. However, only minimal information on the administration of probiotics via freshwater cladoceran Moina micrura is available despite being commonly used for larval and post-larval feeding of freshwater crustaceans and fish. Thus, this study aimed to evaluate the application of Bacillus pocheonensis strain S2 administered via M. micrura to red hybrid tilapia (Oreochromis spp.) larvae. Bacillus pocheonensis that has been previously isolated from Spirulina sp. was subjected to preliminary in vitro evaluation of antagonistic properties. The agar well-diffusion assay revealed that this probiont could inhibit the growth of Streptococcus agalactiae and Aeromonas hydrophila. The size of inhibition zones ranged from 8.8 ± 0.2 to 18.2 ± 0.4 mm. Moina micrura was later used as a biological model in preliminary in vivo bacterial challenge assays to evaluate the efficacy of B. pocheonensis in protecting the host from diseases. Moina micrura was pre-enriched with B. pocheonensis at 104 and 106 CFU mL-1 before S. agalactiae and A. hydrophila were introduced into the culture. The study revealed that B. pocheonensis at 104 CFU mL-1 was able to significantly enhance the survival of M. micrura after being challenged with both pathogens (63 ± 3%) in comparison to the control group. The relative percentage survival (RPS) of M. micrura was highest (p < 0.05) when treated with B. pocheonensis at both concentrations 104 and 106 CFU mL-1 (38.33) after being challenged against S. agalactiae. To assess the efficacy of B. pocheonensis in protecting red hybrid tilapia against streptococcosis, the larvae were fed with either unenriched (control) Moina or probiont-enriched Moina daily for 10 days. A significantly (p < 0.05) higher survival rate (77 ± 3%) was observed in larvae fed with probiont-enriched M. micrura compared to other treatments, and the RPS was recorded at 62.90. In addition, the S. agalactiae load was suppressed in larvae fed probiont-enriched M. micrura (6.84 ± 0.39 CFU mL-1) in comparison to the control group (7.78 ± 0.09 CFU mL-1), indicating that the probiont might have contributed to the improvement of tilapia health and survival. This study illustrated that M. micrura was suitable to be used as a vector for probiotics in freshwater fish larvae as an alternative to hazardous antibiotics for disease control.

Efficacy of heat-killed and formalin-killed vaccines against Tilapia tilapinevirus in juvenile Nile tilapia (Oreochromis niloticus)

Tilapia tilapinevirus (also known as tilapia lake virus, TiLV) is considered to be a new threat to the global tilapia industry. The objective of this study was to develop simple cell culture-based heat-killed (HKV) and formalin-killed (FKV) vaccines for the prevention of disease caused by TiLV. The fish were immunized with 100 μL of either HKV or FKV by intraperitoneal injection with each vaccine containing 1.8 × 106 TCID50 inactivated virus. A booster vaccination was carried out at 21-day post vaccination (dpv) using the same protocol. The fish were then challenged with a lethal dose of TiLV at 28 dpv. The expression of five immune genes (IgM, IgD, IgT, CD4 and CD8) in the head kidney and spleen of experimental fish was assessed at 14 and 21 dpv and again after the booster vaccination at 28 dpv. TiLV-specific IgM responses were measured by ELISA at the same time points. The results showed that both vaccines conferred significant protection, with relative percentage survival (RPS) of 71.3% and 79.6% for HKV and FKV, respectively. Significant up-regulation of IgM and IgT was observed in the head kidney of fish vaccinated with HKV at 21 dpv, while IgM, IgD and CD4 expression increased in the head kidney of fish receiving FKV at the same time point. After booster vaccination, IgT and CD8 transcripts were significantly increased in the spleen of fish vaccinated with the HKV, but not with FKV. Both vaccines induced a specific IgM response in both serum and mucus. In summary, this study showed that both HKV and FKV are promising injectable vaccines for the prevention of disease caused by TiLV in Nile tilapia.

Antimicrobial and Anti-Biofilm Peptide Octominin for Controlling Multidrug-Resistant Acinetobacter baumannii

Acinetobacter baumannii is a serious nosocomial pathogen with multiple drug resistance (MDR), the control of which has become challenging due to the currently used antibiotics. Our main objective in this study is to determine the antibacterial and antibiofilm activities of the antimicrobial peptide, Octominin, against MDR A. baumannii and derive its possible modes of actions. Octominin showed significant bactericidal effects at a low minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of 5 and 10 µg/mL, respectively. Time-kill kinetic analysis and bacterial viability tests revealed that Octominin showed a concentration-dependent antibacterial activity. Field-emission scanning electron microscopy (FE-SEM) analysis revealed that Octominin treatment altered the morphology and membrane structure of A. baumannii. Propidium iodide (PI) and reactive oxygen species (ROS) generation assays showed that Octominin increased the membrane permeability and ROS generation in A. baumannii, thereby causing bacterial cell death. Further, a lipopolysaccharides (LPS) binding assay showed an Octominin concentration-dependent LPS neutralization ability. Biofilm formation inhibition and eradication assays further revealed that Octominin inhibited biofilm formation and showed a high biofilm eradication activity against A. baumannii. Furthermore, up to a concentration of 100 µg/mL, Octominin caused no hemolysis and cell viability changes in mammalian cells. An in vivo study in zebrafish showed that the Octominin-treated group had a significantly higher relative percentage survival (54.1%) than the untreated group (16.6%). Additionally, a reduced bacterial load and fewer alterations in histological analysis confirmed the successful control of A. baumannii by Octominin in vivo. Collectively, these data suggest that Octominin exhibits significant antibacterial and antibiofilm activities against the multidrug-resistant A. baumannii, and this AMP can be developed further as a potent AMP for the control of antibiotic resistance.

Effects of Organic and Inorganic Fertilizers on Growth and Nutrient Uptake by Young Cacao (Theobroma cacao L.)

Sustainable cacao cultivation in Ghana has been seriously hampered by high seedling mortality due to the low soil fertility of lands used for the establishment of new cacao farms. Alleviating the low soil fertility to enhance sustainable cacao cultivation requires the use of fertilizers. A study was therefore conducted at the Cocoa Research Institute of Ghana to determine the effect of integrated application of poultry manure (PM) and sulphate of ammonia (SOA) fertilizers on survival, growth, and nutrient uptake by cacao seedlings under marginal field conditions. A randomized complete block design with four replications was used to evaluate the effects of six treatments. Traits assessed were percentage survival, height, trunk cross-sectional area (TCSA), and nutrient uptake. Results from the study showed no significant differences in soil N and K among the treatments. PM-amended plots had significantly higher soil P than nonamended control and SOA alone. Seedling survival rate ranged from 73.3 to 89.3% with a higher proportion of surviving plants in PM amended soils. The application of PM alone and its combination with SOA improved growth. Cacao trees of T4 (75% recommended dosage of SOA + PM) had the largest TCSA increment, which was significantly different from the other soil treatments. Leaf nutrient concentrations in cacao of T4 were higher than the other soil treatments. It was therefore concluded that T4 could be used during cacao establishment in marginal areas for improved plant survival and growth.