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.     

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.     

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.     

Genetic Diversity of European and Chinese Oilseed Brassica rapa Cultivars from Different Breeding Periods

The Brassica oilseed crops went through two major breeding bottlenecks during the introgression of genes for zero erucic acid and low glucosinolate content, respectively, which may lead to reduced genetic biodiversity of the crop. This study investigates the impact of these bottlenecks on the genetic diversity within and across European and Chinese winter B. rapa cultivars. We compared eight cultivars from Europe and China, representing three different seed qualities from three different breeding periods： （1） high erucic acid, high glucosinolates （＋＋）; （2） zero erucic acid, high glucosinolates（0＋）; （3） zero erucic acid, low glucosonolates （00, canola quality）. Diversity was estimated on 32 plants per cultivar, with 16 simple sequence repeat （SSR） markers covering each of the B. rapa linkage groups. The analysis of molecular variance （AMOVA） showed that genetic variations within cultivars, across cultivars and across regions （Europe and China） were significant, with about 60% of the total variation within cultivars. There was a slight, but non-significant loss in genetic diversity within cultivars when comparing the three breeding periods as indicated by effective number of alleles （2.39, 2.23, and 1.99 for breeding periods 1, 2, and 3, respectively）, Shannon information index （0.93, 0.90, 0.75）, and expected heterozygosity （0.51, 0.49, 0.42）. By cluster analysis （UPGMA dendrogram） and principal coordinate analysis, Chinese and European cultivars were clearly divided into two distinct groups. In conclusion, quality improvement did not significantly reduce the genetic diversity of European and Chinese B. rapa cultivars.     

Co-application of compost or inorganic NPK fertilizer with biochar influences soil quality,grain yield and net income of rice

Most agricultural soils in sub-Saharan Africa are degraded, compromising the grain yield of rice and farmers return on investment. A 3-year field study was undertaken to explore the effect of the application of compost or inorganic NPK fertilizer applied alone or in combination with biochar on soil quality, grain yield of rice and net income. The five treatments were laid out using a randomized complete block design with four replications. The treatments were applied to supply approximately 75 kg N ha^–1. The best fertilizer input was compost+biochar which resulted in the greatest improvement in soil physico-chemical properties by reducing bulk density and increasing porosity and moisture retention, organic matter content, percent nitrogen, available phosphorus and cation exchange capacity. Apart from treatment with inorganic fertilizer alone, treated soils showed a decrease in pH. Bacterial and fungal counts and basal respiration decreased in soils in the following order: compost+biochar>compost only>inorganic NPK fertilizer+biochar>inorganic NPK fertilizer>control. The increase in pooled grain yield and net income in response to treatment followed the order: compost+biochar>NPK+biochar>NPK>compost>control. The findings suggest that the use of compost or NPK alone might improve soil quality and increase grain yield and net income, but it is greatly recommended to co-apply these fertilisers with biochar.     

Germination requirements of <Emphasis Type="Italic">Allanblackia parviflora</Emphasis> seeds and early growth of seedlings

Allanblackia parviflora A. Chev, is an indigenous fruit tree species that could be used in agroforestry systems with both environmental and economic benefits. The seed oil is of prime importance as a foreign exchange earner and is being developed as a rural based enterprise in many African countries notably Ghana, Nigeria, Cameroon and Tanzania. Methods for propagation and conservation of the species are therefore of prime importance. The seeds of the species are however difficult to germinate hindering its domestication process. Allanblackia parviflora seeds were collected from seven different populations in Ghana and were subjected to four different seed germination trials at the nursery of CSIR-Forestry Research Institute of Ghana at Fumesua, Kumasi. This was followed by studying the effect of stage of seed germination on establishment after potting. Intact seeds began germination at 7 months after sowing and at 24 months, significant (P < 0.001) variations were observed in the seed germination ability of 74 accessions. Mean seed germination ranged from 0 to 35%. Significant differences in seed germination percentages among populations (P < 0.001) and provenances (P < 0.05) were also observed. Removal of seed coat significantly (P < 0.001) enhanced seed germination. Germination percentages of seeds with seed coat removed ranged from 6 to 43% while seed germination percentages for intact seeds were less or equal to 3%. Incubation of seeds with seed coat removed in polythene bags gave an added advantage. Seeds with seed coat removed and kept in polythene bags started germinating from 2 weeks and within 10 months, 75 and 68% germination were obtained for plane and black polythene bags, respectively. Removal of germinants and potting when shoot length was at least 1 cm gave significantly higher growth (P < 0.001 for shoot height, P < 0.05 for stem diameter, P < 0.001 for number of leaves produced) compared to when only radicle had emerged. It became evident in this study that seeds should be classified as fully germinated only when shoots begin to grow.     

Population genetic structure of Milicia speciescharacterised by using RAPD and nucleotide sequencing L.

Milicia species(M. excelsa andM. regia, Moraceae, theirokos) are among the most useful indigenous rain forest treespecies in Africa. They are not grown in plantations but extractedfrom the natural forest at an unsustainable rate. The majorconstraint on cultivation and afforestation is thegall-forming insect Phytolyma lata, whichdestroys seedlings through gall formation, causing dieback of theshoot. Genetic diversity studies should aid the design ofconservation and resistance-breeding strategies. This study,therefore, aimed to investigate the genetic variability inMilicia from three West African countries andthe power of bulk-family DNA in partitioning of geneticdiversity. Genetic variation within and between 5 natural populationsof Milicia from Ghana, Côte d'Ivoireand Sierra Leone was investigated using random amplified polymorphicDNA (RAPD) and non-coding chloroplast DNAsequencing. RAPD analysis of 41 DNA samples with 8 primers produced atotal of 74 bands with 40 bands (54.1%) beingpolymorphic. Dendrogram analysis produced 2 major clusters separatingdry and moist/wet ecotypes. Sub-clustering furtherseparated accessions by forest type and/or geographic region. Agreater proportion of the total genetic variability was attributed tovariation between populations (62.2%) than withinpopulations (37.8%). Sequencing of chloroplasttRNA^Leu intron (cptrnL) and intergenic spacer(cpIGS) between cptrnL and tRNA^Phe(trnF) did not show any variation. The RAPD analysissuggests that trees from wet/moist sites are mostlyM. regia, while those fromdry sites are mostly M.excelsa. Sequencing results, however, suggestthat the two species are closely related. These results are discussedwith reference to conservation strategies.