Susceptibility to ergot in Zimbabwe of sorghums tbat remained uninfected in their native climates in Ethiopia and Rwanda

Forty-four local Ethiopian and Rwandan sorghums (Sorghum bicolor) were observed to remain free of ergot, or had only low incidence, in their natural equatorial latitudes and were potentially of interest, in the design of male-sterile lines for F| hybrid breeding, if they possessed a physiologically based resistance mechanism. These sorghums were therefore also investigated under natural and artificial disease pressures in Zimbabwe where unadapted development and inappropriate long daylengtb prevented flowering in 18 accessions. Of the remaining 16 Ethiopian and 10 Rwandan accessions which flowered, only one from each country remained free of ergot. The susceptibility expressed was ascribed to observed asynchrony of stigma exsertion with anthesis. In the Rwandan accession that persistently remained free of ergot in Zimbabwe, histology of ovules showed pollination before floret gaping, so that a general principle of disease escape due to efficient pollination is proposed for the Ethiopian and Rwandan sorghums in their native climates. The findings emphasize that cleistogamy is a desirable character for avoiding ergot infection in self-fertile sorghums and suggest that the Ethiopian and Rwandan sorghutns may not generally be useful for breeding ergot-resistant male-sterile female lines. However, a few accessions deserve more detailed study as a potential genetic resource, before a firm conclusion that all apparent resistance is disease escape owing to efficient pollination

Development and validation of quantitative PCR assays for HIV-associated cryptococcal meningitis in sub-Saharan Africa: a diagnostic accuracy study

Background: HIV-associated cryptococcal meningitis is the second leading cause of AIDS-related deaths, with a 10-week mortality rate of 25–30%. Fungal load assessed by colony-forming unit (CFU) counts is used as a prognostic marker and to monitor response to treatment in research studies. PCR-based assessment of fungal load could be quicker and less labour-intensive. We sought to design, optimise, and validate quantitative PCR (qPCR) assays for the detection, identification, and quantification of Cryptococcus infections in patients with cryptococcal meningitis in sub-Saharan Africa. Methods: We developed and validated species-specific qPCR assays based on DNA amplification of QSP1 (QSP1A specific to Cryptococcus neoformans, QSP1B/C specific to Cryptococcus deneoformans, and QSP1D specific to Cryptococcus gattii species) and a pan-Cryptococcus assay based on a multicopy 28S rRNA gene. This was a longitudinal study that validated the designed assays on cerebrospinal fluid (CSF) of 209 patients with cryptococcal meningitis at baseline (day 0) and during anti-fungal therapy (day 7 and day 14), from the AMBITION-cm trial in Botswana and Malawi (2018–21). Eligible patients were aged 18 years or older and presenting with a first case of cryptococcal meningitis. Findings: When compared with quantitative cryptococcal culture as the reference, the sensitivity of the 28S rRNA was 98·2% (95% CI 95·1–99·5) and of the QSP1 assay was 90·4% (85·2–94·0) in CSF at day 0. Quantification of the fungal load with QSP1 and 28S rRNA qPCR correlated with quantitative cryptococcal culture (R2=0·73 and R2=0·78, respectively). Both Botswana and Malawi had a predominant C neoformans prevalence of 67% (95% CI 55–75) and 68% (57–73), respectively, and lower C gattii rates of 21% (14–31) and 8% (4–14), respectively. We identified ten patients that, after 14 days of treatment, harboured viable but non-culturable yeasts based on QSP1 RNA detection (without any positive CFU in CSF culture). Interpretation: QSP1 and 28S rRNA assays are useful in identifying Cryptococcus species. qPCR results correlate well with baseline quantitative cryptococcal culture and show a similar decline in fungal load during induction therapy. These assays could be a faster alternative to quantitative cryptococcal culture to determine fungal load clearance. The clinical implications of the possible detection of viable but non-culturable cells in CSF during induction therapy remain unclear. Funding: European and Developing Countries Clinical Trials Partnership; Swedish International Development Cooperation Agency; Wellcome Trust/UK Medical Research Council/UKAID Joint Global Health Trials; and UK National Institute for Health Research

Development and validation of quantitative PCR assays for HIV-associated cryptococcal meningitis in sub-Saharan Africa: a diagnostic accuracy study

Background: HIV-associated cryptococcal meningitis is the second leading cause of AIDS-related deaths, with a 10-week mortality rate of 25–30%. Fungal load assessed by colony-forming unit (CFU) counts is used as a prognostic marker and to monitor response to treatment in research studies. PCR-based assessment of fungal load could be quicker and less labour-intensive. We sought to design, optimise, and validate quantitative PCR (qPCR) assays for the detection, identification, and quantification of Cryptococcus infections in patients with cryptococcal meningitis in sub-Saharan Africa. Methods: We developed and validated species-specific qPCR assays based on DNA amplification of QSP1 (QSP1A specific to Cryptococcus neoformans, QSP1B/C specific to Cryptococcus deneoformans, and QSP1D specific to Cryptococcus gattii species) and a pan-Cryptococcus assay based on a multicopy 28S rRNA gene. This was a longitudinal study that validated the designed assays on cerebrospinal fluid (CSF) of 209 patients with cryptococcal meningitis at baseline (day 0) and during anti-fungal therapy (day 7 and day 14), from the AMBITION-cm trial in Botswana and Malawi (2018–21). Eligible patients were aged 18 years or older and presenting with a first case of cryptococcal meningitis. Findings: When compared with quantitative cryptococcal culture as the reference, the sensitivity of the 28S rRNA was 98·2% (95% CI 95·1–99·5) and of the QSP1 assay was 90·4% (85·2–94·0) in CSF at day 0. Quantification of the fungal load with QSP1 and 28S rRNA qPCR correlated with quantitative cryptococcal culture (R2=0·73 and R2=0·78, respectively). Both Botswana and Malawi had a predominant C neoformans prevalence of 67% (95% CI 55–75) and 68% (57–73), respectively, and lower C gattii rates of 21% (14–31) and 8% (4–14), respectively. We identified ten patients that, after 14 days of treatment, harboured viable but non-culturable yeasts based on QSP1 RNA detection (without any positive CFU in CSF culture). Interpretation: QSP1 and 28S rRNA assays are useful in identifying Cryptococcus species. qPCR results correlate well with baseline quantitative cryptococcal culture and show a similar decline in fungal load during induction therapy. These assays could be a faster alternative to quantitative cryptococcal culture to determine fungal load clearance. The clinical implications of the possible detection of viable but non-culturable cells in CSF during induction therapy remain unclear

Independent and combined effects of improved water, sanitation, and hygiene, and improved complementary feeding, on child stunting and anaemia in rural Zimbabwe: a cluster-randomised trial.

BACKGROUND: Child stunting reduces survival and impairs neurodevelopment. We tested the independent and combined effects of improved water, sanitation, and hygiene (WASH), and improved infant and young child feeding (IYCF) on stunting and anaemia in in Zimbabwe. METHODS: We did a cluster-randomised, community-based, 2 × 2 factorial trial in two rural districts in Zimbabwe. Clusters were defined as the catchment area of between one and four village health workers employed by the Zimbabwe Ministry of Health and Child Care. Women were eligible for inclusion if they permanently lived in clusters and were confirmed pregnant. Clusters were randomly assigned (1:1:1:1) to standard of care (52 clusters), IYCF (20 g of a small-quantity lipid-based nutrient supplement per day from age 6 to 18 months plus complementary feeding counselling; 53 clusters), WASH (construction of a ventilated improved pit latrine, provision of two handwashing stations, liquid soap, chlorine, and play space plus hygiene counselling; 53 clusters), or IYCF plus WASH (53 clusters). A constrained randomisation technique was used to achieve balance across the groups for 14 variables related to geography, demography, water access, and community-level sanitation coverage. Masking of participants and fieldworkers was not possible. The primary outcomes were infant length-for-age Z score and haemoglobin concentrations at 18 months of age among children born to mothers who were HIV negative during pregnancy. These outcomes were analysed in the intention-to-treat population. We estimated the effects of the interventions by comparing the two IYCF groups with the two non-IYCF groups and the two WASH groups with the two non-WASH groups, except for outcomes that had an important statistical interaction between the interventions. This trial is registered with ClinicalTrials.gov, number NCT01824940. FINDINGS: Between Nov 22, 2012, and March 27, 2015, 5280 pregnant women were enrolled from 211 clusters. 3686 children born to HIV-negative mothers were assessed at age 18 months (884 in the standard of care group from 52 clusters, 893 in the IYCF group from 53 clusters, 918 in the WASH group from 53 clusters, and 991 in the IYCF plus WASH group from 51 clusters). In the IYCF intervention groups, the mean length-for-age Z score was 0·16 (95% CI 0·08-0·23) higher and the mean haemoglobin concentration was 2·03 g/L (1·28-2·79) higher than those in the non-IYCF intervention groups. The IYCF intervention reduced the number of stunted children from 620 (35%) of 1792 to 514 (27%) of 1879, and the number of children with anaemia from 245 (13·9%) of 1759 to 193 (10·5%) of 1845. The WASH intervention had no effect on either primary outcome. Neither intervention reduced the prevalence of diarrhoea at 12 or 18 months. No trial-related serious adverse events, and only three trial-related adverse events, were reported. INTERPRETATION: Household-level elementary WASH interventions implemented in rural areas in low-income countries are unlikely to reduce stunting or anaemia and might not reduce diarrhoea. Implementation of these WASH interventions in combination with IYCF interventions is unlikely to reduce stunting or anaemia more than implementation of IYCF alone. FUNDING: Bill & Melinda Gates Foundation, UK Department for International Development, Wellcome Trust, Swiss Development Cooperation, UNICEF, and US National Institutes of Health.The SHINE trial is funded by the Bill & Melinda Gates Foundation (OPP1021542 and OPP113707); UK Department for International Development; Wellcome Trust, UK (093768/Z/10/Z, 108065/Z/15/Z and 203905/Z/16/Z); Swiss Agency for Development and Cooperation; US National Institutes of Health (2R01HD060338-06); and UNICEF (PCA-2017-0002)

Effect of dew point temperature and conidium age on germination, germtube growth and infection of maize and sorghum by Peronosclerospora sorghi

The effect of the environment on the germination, survival and infection of sorghum by conidia of Peronosclerospora sorghi is unknown in Africa. Dew point temperature, and the effect of conidium age was characterized for an isolate of P. sorghi from Zimbabwe. Germination and germ tube growth took place in the range 10–34°C (optimal at 10–34 and 20–33°, respectively). Infection was optimal at 14–30°. Incidence of infection at different temperatures was correlated with germ tube growth (r=0·8, P<0·001). Germination and germ tube growth occurred from 5 h after commencing incubation of infected leaf material, although immature conidia harvested at 3 h caused a low incidence of infection. Plant age also affected the incidence of infection. Container grown sorghum plants older than 20 d, and maize plants older than 15 d were resistant to systemic infection by conidia. The results indicate that germination, germ tube growth and infection take place over a wide temperature range, and that some limited biotypic variation may exist when these data are compared to other reports of the environmental requirements of P. sorghi from the U.S.A. and India

Production of conidia by Peronosclerospora sorghi on sorghum crops in Zimbabwe.

Factors affecting the production of conidia of Peronosclerospora sorghi, causing sorghum downy mildew (SDM), were investigated during 1993 and 1994 in Zimbabwe. In the field conidia were detected on nights when the minimum temperature was in the range 10–19°C. On 73% of nights when conidia were detected rain had fallen within the previous 72 h and on 64% of nights wind speed was < 2.0 m s−1. The time period over which conidia were detected was 2–9 h. Using incubated leaf material, conidia were produced in the temperature range 10–26°C. Local lesions and systemically infected leaf material produced 2.4–5.7 × 103 conidia per cm2. Under controlled conditions conidia were released from conidiophores for 2.5 h after maturation and were shown to be well adapted to wind dispersal, having a settling velocity of 1.5 × 10−4 m s−1. Conditions that are suitable for conidia production occur in Zimbabwe and other semi-arid regions of southern Africa during the cropping seaso

Production of conidia by Peronosclerospora sorghi on sorghum crops in Zimbabwe

Factors affecting the production of conidia of Peronosclerctsporas orghi, causing sorghum downy mildew (SDM), were investigated during 1993 and 1994 in Zimbabwe. In the field conidia were detected on nights when the minimum remperature was in the range l0-19ºC. On 73% of nights when conidia were detected rain had fallen within the previous 72h and on 64% of nights wind speed was < 2.0 m s-1. The time period over which conidia were detected was 2-9h. Using incubated leaf material, conidia were produced in the temperature range 10-26oC. Local lesions and systemically infected leaf material produced 2.4-5.7 x 103 conidia per cm2. Under controlled conditions conidia were released from conidiophores for 2.5h after maturation and were shown to be well adapted to wind dispersal, having a settling velocity of 1.5 x 10-4 m s- 1. Conditions that are suitable for conidia production occur in Zimbabwe and other semi-arid regions of southern Africa during the cropping season