Distribution and genetic diversity among Aspergillus flavus isolates across three agro-ecologies essential for maize cultivation in Ghana

Fungal mycotoxins are important contaminants of agricultural commodities that pose serious concerns to producers, consumers and exporters. Aflatoxin is a carcinogenic metabolite produced by the fungi Aspergillus flavus and A. parasiticus. These fungi and other mould species living in the soil contaminate several crops including maize. This study was undertaken with the following objectives: (i) to assess the presence of A. flavus and other moulds in Ghanaian soil; (ii) to determine the distribution and density of A. flavus under three agro-ecologies noted for major maize production; and (iii) to assess the effect of percentage of carbon (C) and nitrogen (N) and soil pH on A. flavus population densities and diversity in the isolates found across the agro-ecologies. The data showed seven moulds that were common across the agro-ecologies. Significant differences (P < 0.05) were observed in the A. flavus density and distribution within and across these agro-ecologies. Fumesua soils recorded the highest levels of A. flavus (1.185 × 10³ cfu g⁻¹) while Akomadan recorded the least (9.76 × 10² cfu g⁻¹). Percentage available C, N and soil pH did not significantly influence A. flavus density. The A. flavus isolates identified in this study varied in genetic sequence within the aflatoxin gene cluster, but these differences were not distinguishable by origin.     

Genetic divergence in two tropical maize composites after four cycles of reciprocal recurrent selection

Two tropical maize composites were subjected to four cycles of reciprocal recurrent selection to develop divergent inbred lines with good combining ability. This study was conducted to examine the extent of genetic diversity, changes in allele composition and genetic structure, of 100 randomly selected S₁ lines each from the original (C₀) and advanced (C₄) selection cycles of TZL COMP3 and TZL COMP4, genotyped using single nucleotide polymorphism (SNP) markers. Results revealed that the proportion of alleles at both low and high frequencies decreased from C₀ to C₄, whereas those at intermediate frequencies increased at C₄ in the two composites. More unique and other alleles were lost at C₄ in TZL COMP3 relative to those in TZL COMP4. The changes in different measures of genetic diversity were either small or negligible with selection in the two composites. The proportion of markers departing from Hardy–Weinberg equilibrium (HWE) decreased with selection, whereas the total number of pairs of markers in linkage disequilibrium increased with selection in the two composites. Examination of changes in population structures using a model‐based approach as well as cluster and multivariate analyses found a high degree of concordance in stratifying the 400 S₁ lines into four non‐overlapping groups corresponding to the two selection cycles each within the reciprocal composites. The observed molecular‐based divergence between cycles within the same composite and the clear differentiation between the complementary composites highlight the importance of reciprocal recurrent selection for preserving genetic diversity for long‐term selection. This increases the potential of the advanced selection cycles to sustain genetic gain in productivity of hybrids adapted to the savannas in West and Central Africa.     

Pharmacokinetics and tissue residues of kitasamycin in healthy and diseased broilers

The pharmacokinetics of kitasamycin after intravenous and oral administration in a dose of 300 mg/kg b.wt. was studied in 18 healthy and 18 Salmonella gallinarum naturally infected chickens. The tissue residue of the studied antibiotic was estimated in 36 normal chickens when it was given orally for 7 successive days. Therapeutic level of kitasamycin was achieved after 15 minutes and persisted for 20-22 hours after its oral administration. Higher serum kitasamycin concentrations were recorded in Salmonella gallinarum infected chickens. The elimination half-life of kitasamycin calculated after single intravenous injection was 9.03 hours in diseased chickens corresponding to 3.74 hours in healthy birds. The body clearance was significantly reduced in diseased chickens (23.86 ml/kg/min) when compared to that in normal ones (62.03 ml/kg/min). Kitasamycin treated broilers should not be slaughtered before 3 days from the last dose as it was detected only in bile and caecum at that time but not in edible tissues.     

Monoclonal antibodies specific to thermostable proteins in animal blood

Bovine plasma proteins are used as high-quality protein supplements in animal feed and as binders or colorants in food for human consumption. Religious observance, as well as recent fears of epidemic bovine spongiform encephalopathy, highlights the need for methods to detect bovine blood in processed food and animal feed for regulatory purposes, as the currently available methods are neither species-specific, blood-specific, nor valid for excessively heat-processed samples. This paper reports the development of monoclonal antibodies (MAbs) raised against bovine thermostable plasma proteins that display a unique species specificity pattern for plasma proteins. Immunoblotting revealed several thermostable antigenic proteins (10, 25, 40, and 60 kDa) in bovine plasma sterilized at 121 degrees C for 15 min. These MAbs can be employed individually or combined in immunoassays for analytical purposes and investigations of the chemical and biological properties of the thermostable plasma proteins identified here.     

Sandwich enzyme-linked immunosorbent assay for the detection of bovine blood in animal feed

The feeding of ruminant proteins to ruminants is prohibited in most countries because the practice is thought to be responsible for the spread of bovine spongiform encephalopathy. However, currently available methods to detect ruminant blood products in rendered feedstuffs are inadequate because they lack species specificity, tissue specificity, and are not based on a thermostable analyte. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for this study that provides reliable and sensitive (0.05-0.5% v/v) detection of bovine blood materials in animal feed. The new sandwich ELISA employs two previously developed monoclonal antibodies (MAbs), Bb6G12 as the capture antibody and biotinylated MAb Bb3D6 as the detecting antibody, and is bovine-specific and blood-specific. The assay is based on the detection of a 60 kDa thermostable protein in bovine blood and provides a useful regulatory tool for monitoring fraudulent labeling or contamination of bovine blood in both heat-processed feedstuffs and unprocessed raw materials. Keywords: Bovine; blood; monoclonal antibody; sandwich ELISA.     

Genome-wide association study of grain iron and zinc concentration in common bean (Phaseolus vulgaris)

Common bean is one of the widely consumed food security crop in Africa, Asia and South America. It is a rich source of protein, minerals and micronutrients. High genotype by environment interaction is one of the main challenge in breeding for high grain micronutrient concentration. The objective of this study was to estimate SNP markers associated with grain Fe and Zn concentration using 289 common bean genotypes and 11,480 SNP markers. The study revealed that 43 quantitative trait loci (QTLs) were associated with grain Fe and Zn concentration. Five quantitative trait nucleotides (QTNs), that is, QTN Fe_1.1, QTN Fe_6.3, QTN Fe_6.5, QTN Fe_10.3 and QTN Fe_11.6 were detected both at Haramaya and Melkassa locations. Two of the markers, that is, QTN Fe_6.3 and QTN Fe_6.5, were located on chromosome 06 while QTN Fe_1.1, QTN Fe_10.3 and QTN Fe_ 11.6 were residing on chromosomes 01, 10 and 11, respectively. Among these, QTN Fe_11.6 had a large and positive consistent effect across locations. The five stable QTNs along with the potential candidate genes could be used for Fe biofortification through marker assisted selection.