Phylogenetically diverse Bradyrhizobium genospecies nodulate Bambara groundnut (Vigna subterranea L. Verdc) and soybean (Glycine max L. Merril) in the northern savanna zones of Ghana.

A total of 102 bacterial strains isolated from nodules of three Bambara groundnut and one soybean cultivars grown in nineteen soil samples collected from northern Ghana were characterized using multilocus gene sequence analysis. Based on a concatenated sequence analysis (glnII-rpoB-recA-gyrB-atpD-dnaK), 54 representative strains were distributed in 12 distinct lineages, many of which were placed mainly in the Bradyrhizobium japonicum and Bradyrhizobium elkanii supergroups. Twenty-four of the 54 representative strains belonged to seven putative novel species, while 30 were conspecific with four recognized Bradyrhizobium species. The nodA phylogeny placed all the representative strains in the cosmopolitan nodA clade III. The strains were further separated in seven nodA subclusters with reference strains mainly of African origin. The nifH phylogeny was somewhat congruent with the nodA phylogeny, but both symbiotic genes were mostly incongruent with the core housekeeping gene phylogeny indicating that the strains acquired their symbiotic genes horizontally from distantly related Bradyrhizobium species. Using redundancy analysis, the distribution of genospecies was found to be influenced by the edaphic factors of the respective sampling sites. In general, these results mainly underscore the high genetic diversity of Bambara groundnut-nodulating bradyrhizobia in Ghanaian soils and suggest a possible vast resource of adapted inoculant strains.

Bacteria related to Bradyrhizobium yuanmingense from Ghana are effective groundnut micro-symbionts.

The identification of locally-adapted rhizobia for effective inoculation of grain legumes in Africa's semiarid regions is strategic for developing and optimizing cheap nitrogen fixation technologies for smallholder farmers. This study was aimed at selecting and characterising effective native rhizobia, from Ghanaian soils for groundnut (Arachis hypogaea L.) inoculation. From surface-disinfected root nodules of cowpea and groundnut plants grown on farmers' fields, 150 bacterial isolates were obtained, 30 of which were eventually found to nodulate groundnut plants. After testing the symbiotic potential of these isolates on groundnut on sterilized substrate, seven of them, designated as KNUST 1001-1007, were evaluated in an open field pot experiment using 15N-labelled soil. Although 15N dilution analyses did not indicate differences among treatments in the proportion of nitrogen (N) derived from the atmosphere (%Ndfa), all seven strains increased total N derived from N2 fixation by inoculated groundnut plants as compared to the non-inoculated control. Inoculation with KNUST 1002 led to total N accumulation as high as that of the groundnut reference strain 32H1. Genetic characterisation of the isolates by sequence analysis of 16S rRNA gene, 16S - 23S rRNA intergenic transcribed spacer (ITS) region and nodC gene revealed that isolates KNUST 1003 and 1007 were related to Rhizobium tropici, a common bean symbiont. The other five isolates, including KNUST 1002 belonged to the Bradyrhizobium genus, being closely related to Bradyrhizobium yuanmingense. Therefore, this study revealed novel native Ghanaian rhizobia with potential for the development of groundnut inoculants.

Maize-grain legume intercropping for enhanced resource use efficiency and crop productivity in the Guinea savanna of northern Ghana.

Smallholder farmers in the Guinea savanna practise cereal-legume intercropping to mitigate risks of crop failure in mono-cropping. The productivity of cereal-legume intercrops could be influenced by the spatial arrangement of the intercrops and the soil fertility status. Knowledge on the effect of soil fertility status on intercrop productivity is generally lacking in the Guinea savanna despite the wide variability in soil fertility status in farmers' fields, and the productivity of within-row spatial arrangement of intercrops relative to the distinct-row systems under on-farm conditions has not been studied in the region. We studied effects of maize-legume spatial intercropping patterns and soil fertility status on resource use efficiency, crop productivity and economic profitability under on-farm conditions in the Guinea savanna. Treatments consisted of maize-legume intercropped within-row, 1 row of maize alternated with one row of legume, 2 rows of maize alternated with 2 rows of legume, a sole maize crop and a sole legume crop. These were assessed in the southern Guinea savanna (SGS) and the northern Guinea savanna (NGS) of northern Ghana for two seasons using three fields differing in soil fertility in each agro-ecological zone. Each treatment received 25 kg P and 30 kg K ha-1 at sowing, while maize received 25 kg (intercrop) or 50 kg (sole) N ha-1 at 3 and 6 weeks after sowing. The experiment was conducted in a randomised complete block design with each block of treatments replicated four times per fertility level at each site. Better soil conditions and rainfall in the SGS resulted in 48, 38 and 9% more maize, soybean and groundnut grain yield, respectively produced than in the NGS, while 11% more cowpea grain yield was produced in the NGS. Sole crops of maize and legumes produced significantly more grain yield per unit area than the respective intercrops of maize and legumes. Land equivalent ratios (LERs) of all intercrop patterns were greater than unity indicating more efficient and productive use of environmental resources by intercrops. Sole legumes intercepted more radiation than sole maize, while the interception by intercrops was in between that of sole legumes and sole maize. The intercrop however converted the intercepted radiation more efficiently into grain yield than the sole crops. Economic returns were greater for intercrops than for either sole crop. The within-row intercrop pattern was the most productive and lucrative system. Larger grain yields in the SGS and in fertile fields led to greater economic returns. However, intercropping systems in poorly fertile fields and in the NGS recorded greater LERs (1.16-1.81) compared with fertile fields (1.07-1.54) and with the SGS. This suggests that intercropping is more beneficial in less fertile fields and in more marginal environments such as the NGS. Cowpea and groundnut performed better than soybean when intercropped with maize, though the larger absolute grain yields of soybean resulted in larger net benefits.

Probabilistic quantitative microbial risk assessment model of norovirus from wastewater irrigated vegetables in Ghana using genome copies and fecal indicator ratio conversion for estimating exposure dose.

The need to replace the commonly applied fecal indicator conversions ratio (an assumption of 1:10-5 virus to fecal indicator organism) in Quantitative Microbial Risk Assessment (QMRA) with models based on quantitative data on the virus of interest has gained prominence due to the different physical and environmental factors that might influence the reliability of using indicator organisms in microbial risk assessment. The challenges facing analytical studies on virus enumeration (genome copies or particles) have contributed to the already existing lack of data in QMRA modelling. This study attempts to fit a QMRA model to genome copies of norovirus data. The model estimates the risk of norovirus infection from the intake of vegetables irrigated with wastewater from different sources. The results were compared to the results of a corresponding model using the fecal indicator conversion ratio to estimate the norovirus count. In all scenarios of using different water sources, the application of the fecal indicator conversion ratio underestimated the norovirus disease burden, measured by the Disability Adjusted Life Years (DALYs), when compared to results using the genome copies norovirus data. In some cases the difference was >2 orders of magnitude. All scenarios using genome copies met the 10-4 DALY per person per year for consumption of vegetables irrigated with wastewater, although these results are considered to be highly conservative risk estimates. The fecal indicator conversion ratio model of stream-water and drain-water sources of wastewater achieved the 10-6 DALY per person per year threshold, which tends to indicate an underestimation of health risk when compared to using genome copies for estimating the dose.

Bradyrhizobium Inoculants Enhance Grain Yields of Soybean and Cowpea in Northern Ghana.

This study evaluated the symbiotic effectiveness and economic evaluation of Rhizobium inoculants with the objective of recommending the most effective inoculant strain for soybean and cowpea production in Northern Ghana. Field experiments were established in three locations using randomized complete block design with five blocks. A total of four treatments (Legumefix, Biofix, 100 kg N ha-1 and uninoculated control for soybean and BR 3267, BR 3262, 100 kg N ha-1 and uninoculated control for cowpea) were applied. At Nyankpala, inoculation of soybean with Legumefix and Biofix led to significant (P < 0.05) increases in nodule number (90-118%), nodule dry weight (>two-folds), and grain yield (12-19%) relative to the control. The Biofix effect on soybean grain yield was 1.5-fold of Legumefix. Similarly, inoculation of cowpea with BR 3262 and BR 3267 significantly (P < 0.05) increased nodule number (41-68%), nodule dry weight (45-65%), and grain yield (11-38%) relative to the control. Strain BR 3267 performed consistently (>two-folds) better than BR 3262 on grain yield. At Nyagli, there was no significant effect of inoculation on cowpea. Wilks lambda values (0.067, 0.039; P = 0.00) indicated that 93.3 and 96.1% of the variations observed in soybean and cowpea, respectively, were due to the applied inoculants. Biofix and BR 3267 were economically profitable with VCR ratio of 8.7 and 4.6, respectively. Based on grain yield and economic returns observed, Biofix and BR 3267 can be recommended in Nyankpala for inoculation of soybean and cowpea, respectively.

A probabilistic assessment of the contribution of wastewater-irrigated lettuce to Escherichia coli O157:H7 infection risk and disease burden in Kumasi, Ghana.

Wastewater use for vegetable production is widespread across the cities of many developing countries. Studies on the microbial health risks associated with the practice have largely depended on faecal indicator organisms with potential underestimation or overestimation of the microbial health risks and disease burdens. This study assessed the Escherichia coli O157:H7 infection risk and diarrhoeal disease burden measured in disability-adjusted life years (DALYs) associated with the consumption of wastewater-irrigated lettuce in Kumasi, Ghana using data on E. coli O157:H7 in ready-to-harvest, wastewater-irrigated lettuce. Two exposure scenarios - best case and worst case - associated with a single consumption of wastewater-irrigated lettuce were assessed. The assessment revealed wastewater-irrigated lettuce is contributing to the transmission of E. coli O157:H7 in Kumasi, Ghana. The mean E. coli O157:H7 infection risk and DALYs in the wet and dry seasons, irrespective of the exposure scenario, were above the World Health Organization tolerable daily infection risk of 2.7 × 10⁻7 per person per day and 10⁻6 DALYs per person per year. It is recommended that legislation with clear monitoring indicators and penalties is implemented to ensure that farmers and food sellers fully implement risk mitigating measures.

Reducing microbial contamination on wastewater-irrigated lettuce by cessation of irrigation before harvesting.

OBJECTIVE: To assess the effectiveness of cessation of irrigation before harvesting in reducing microbial contamination of lettuce irrigated with wastewater in urban vegetable farming in Ghana. METHODS: Assessment was done under actual field conditions with urban vegetable farmers in Ghana. Trials were arranged in completely randomized block design and done both in the dry and wet seasons. Seven hundred and twenty-six lettuce samples and 36 water samples were analysed for thermotolerant coliforms and helminth eggs. RESULTS: On average, 0.65 log units for indicator thermotolerant coliforms and 0.4 helminth eggs per 100 g of lettuce were removed on each non-irrigated day from lettuce in the dry season. This corresponded to a daily loss of 1.4 tonnes/ha of fresh weight of lettuce. As an input for exposure analysis to make risk estimates, the decay coefficient, k, for thermotolerant coliforms was 0.66/day for the wet season and 1.49/day for the dry season. CONCLUSION: In combination with other measures for improving water quality, the measure can significantly reduce faecal contamination of lettuce during the dry season. However, it is not suitable for the wet season due to unfavourable conditions for pathogen die-off and re-contamination by splashes from contaminated soils. The results provide a good basis for risk assessments and for devising more appropriate measures for risk reduction, especially in sub-Saharan Africa.

Effect of low-cost irrigation methods on microbial contamination of lettuce irrigated with untreated wastewater.

OBJECTIVE: To assess the effectiveness of simple irrigation methods such as drip irrigation kits, furrow irrigation and use of watering cans in reducing contamination of lettuce irrigated with polluted water in urban farming in Ghana. METHODS: Trials on drip kits, furrow irrigation and watering cans were conducted with urban vegetable farmers. Trials were arranged in a completely randomised block design with each plot having all three irrigation methods tested. This was conducted in both dry and wet seasons. Three hundred and ninety-six lettuce, 72 soil, 15 poultry manure and 32 water samples were analysed for thermotolerant coliforms and helminth eggs. RESULTS: Lettuce irrigated with drip kits had the lowest levels of contamination, with, on average, 4 log units per 100 g, fewer thermotolerant coliforms than that irrigated with watering cans. However, drip kits often got clogged, required lower crop densities and restricted other routine farm activities. Watering cans were the most popular method. Using watering cans with caps on outlets from a height <0.5 m reduced thermotolerant coliforms by 2.5 log units and helminthes by 2.3 eggs per 100 g of lettuce compared with using watering cans without caps from a height >1 m. CONCLUSION: Simple, cheap and easily adoptable irrigation methods have great potential to reduce crop contamination in low-income areas. When used in combination with other on-farm and post-harvest risk reduction measures, these will help to comprehensively reduce public health risks from using polluted water in vegetable farming.

Biodiversity of Saccharomyces cerevisiae isolated from a survey of pito production sites in various parts of Ghana.

Biodiversity among Saccharomyces cerevisiae predominating the spontaneous fermentation of Dagarti pito in Ghana was assessed. Two hundred and forty-nine isolates obtained from samples of dried yeast taken from commercial pito production sites in eight geographical regions of Ghana were characterized phenotypically by colony and cell morphology as well as carbohydrate assimilation profiling. Yeast populations ranged between 10(6) and 10(8) cfug(-1). Ninety-nine percent of the isolates (247) investigated showed macro-and micro morphological characteristics typical of S. cerevisiae. Of these, 72% (179) had assimilation profiles similar to S. cerevisiae while 28% (68) had assimilation profiles atypical of S. cerevisiae or any other member of the Saccharomyces sensu stricto complex. Amplification of the region spanning the two intergenic transcribed spacers (ITS) and the 5.8S ribosomal gene (ITS1-5.8S rDNA-ITS2), followed by restriction analysis, as well as determination of chromosome length polymorphism by pulsed field gel electrophoresis (PFGE) of 25 representative isolates strongly indicated that all belonged to S. cerevisiae, notwithstanding the phenotypic differences. Sequencing of the mitochondrial cytochrome-c oxidase II gene (COX 2) and the actin-encoding gene (ACT1) of four isolates, confirmed their close relatedness to S. cerevisiae, particularly to the type strain CBS1171 (98.7%), as well as other members of the Saccharomyces sensu stricto complex. Twenty isolates selected from eight geographical regions of Ghana and investigated for their technological properties, showed different patterns of growth and flocculation but otherwise similar technological characteristica. Most of the isolates produced pito having sensory attributes, which compared favourably with commercially produced pito.