Quality of fresh frozen tilapia from selected supermarkets in Malawi

Fish provides a major source of dietary animal protein to Malawi’s population. Majority of tilapia in supermarkets are of different origins and bought from different suppliers. Fish is highly perishable commodity and its quality degrades even in frozen form due to microbial activity. The quality of frozen tilapia (the most commonly traded and consumed fish in Malawi) sold in some reputable supermarkets in Malawi was determined. Fish were collected from nine (9) reputable supermarkets in three (3) regions of the country (north, central and south) and analysed in the laboratory for sensory quality, microbiological, chemical and proximate analyses. Sensory quality evaluation was performed following guidelines earlier developed for fresh tilapia (Oreochromis spp.) in Malawi. Differences and changes in the fish sensory quality were attributed to the effect of storage duration and conditions within the freezer compartment. Two types of bacteria namely, Salmonella spp. and Escherichia coliwere identified on the frozen tilapia, suggesting poor and unhygienic pre-handling. Despite the presence of bacteria on the fish and differences in sensory quality, the frozen tilapia were within the acceptable range for human consumption. Nutrient composition of frozen tilapia was high despite differences (p<0.05) in moisture, ash and crude fat. Fish from different origin were sold mixed in all supermarkets, poor handling along the fish market chain was identified as the major source of fish contamination. Mechanical damages were reminiscent of the effects of frozen storage. There is a need to establish optimum storage time for frozen tilapia in supermarkets to provide products with good quality in terms of sensory properties, nutrient content and safe microbial loads.&nbsp

Automatic Detection of Open and Vegetated Water Bodies Using Sentinel 1 to Map African Malaria Vector Mosquito Breeding Habitats

Providing timely and accurate maps of surface water is valuable for mapping malaria risk and targeting disease control interventions. Radar satellite remote sensing has the potential to provide this information but current approaches are not suitable for mapping African malarial mosquito aquatic habitats that tend to be highly dynamic, often with emergent vegetation. We present a novel approach for mapping both open and vegetated water bodies using serial Sentinel-1 imagery for Western Zambia. This region is dominated by the seasonally inundated Upper Zambezi floodplain that suffers from a number of public health challenges. The approach uses open source segmentation and machine learning (extra trees classifier), applied to training data that are automatically derived using freely available ancillary data. Refinement is implemented through a consensus approach and Otsu thresholding to eliminate false positives due to dry flat sandy areas. The results indicate a high degree of accuracy (mean overall accuracy 92% st dev 3.6) providing a tractable solution for operationally mapping water bodies in similar large river floodplain unforested environments. For the period studied, 70% of the total water extent mapped was attributed to vegetated water, highlighting the importance of mapping both open and vegetated water bodies for surface water mapping

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

MORPHOMETRIC VARIATIONS AMONG OPSARIDIUM MICROLEPIS (GÜNTHER, 1864) FROM LAKE MALAŴI MIGRATING TO DIFFERENT RIVERS FOR BREEDING

Fish is an important resource in Malawi as a source of food for the majority as it provides affordable source of dietary animal protein as well as income. A number of fish species in the Malawi water bodies have their population dwindling. One of the species under serious threat of extinction is the Opsaridium microlepis - a potamodromous fish species that migrate to the rivers during its spawning period and its management seems a nightmare. A number of studies reveal contrasting results on genetic makeup and morphological aspect of this fish species. With changes in the ecosystems of the rivers connecting Lake Malawi, coupled with absence of strong management measures in the major rivers adjoining the lake, problems have arisen in the conservation of potamodromous fish species. This necessitated the present study to investigate if the morphological features of stocks of O. microlepis are the same or not and if they have changed to adapt to changes in the ecosystems. One hundred and eleven O. microlepis fish samples were collected from Linthipe River (48), Bua River (59) and North Rumphi River (4) monthly from March to August 2020 using trawled and static gillnets. Twenty-four morphometric characteristics were measured to determine if any morphological differences existed among the fish samples from the three rivers. Principal component analysis (PCA) was used to compare morphology of the fish. Results of the study showed no significant morphological differences among stocks from the three rivers, implying that O. microlepis in these rivers belong to same stock morphologically. The study reveals that the species do not differ morphologically even though they migrate to different rivers for breeding. The study further notes that numerous activities taking place along the tributary rivers (as observed during the study) such as modification of fishing gears as well as fishing methods and the deterioration of the spawning grounds due to siltation from soil erosion caused by deforestation and agriculture, are putting the potamodromous fish species such as O. microlepis under serious threat. The study recommends that the populations of O. microlepis from the rivers can be managed equally since they are morphologically similar. Adopting uniform catchment management and sustainable exploitation of O. microlepis (such as regulations on mesh sizes and fishing methods, closing the rivers from fishing activities during spawning period, river bank and catchment management and restoration) with the aim of conserving the stocks from further overexploitation in these rivers is recommended so that the communities and the people at large continue to utilize the resource sustainably and at the same time, sustaining their livelihood

MORPHOMETRIC VARIATIONS AMONG OPSARIDIUM MICROLEPIS (GÜNTHER, 1864) FROM LAKE MALAŴI MIGRATING TO DIFFERENT RIVERS FOR BREEDING

Fish is an important resource in Malawi as a source of food for the majority as it provides affordable source of dietary animal protein as well as income. A number of fish species in the Malawi water bodies have their population dwindling. One of the species under serious threat of extinction is the Opsaridium microlepis - a potamodromous fish species that migrate to the rivers during its spawning period and its management seems a nightmare. A number of studies reveal contrasting results on genetic makeup and morphological aspect of this fish species. With changes in the ecosystems of the rivers connecting Lake Malawi, coupled with absence of strong management measures in the major rivers adjoining the lake, problems have arisen in the conservation of potamodromous fish species. This necessitated the present study to investigate if the morphological features of stocks of O. microlepis are the same or not and if they have changed to adapt to changes in the ecosystems. One hundred and eleven O. microlepis fish samples were collected from Linthipe River (48), Bua River (59) and North Rumphi River (4) monthly from March to August 2020 using trawled and static gillnets. Twenty-four morphometric characteristics were measured to determine if any morphological differences existed among the fish samples from the three rivers. Principal component analysis (PCA) was used to compare morphology of the fish. Results of the study showed no significant morphological differences among stocks from the three rivers, implying that O. microlepis in these rivers belong to same stock morphologically. The study reveals that the species do not differ morphologically even though they migrate to different rivers for breeding. The study further notes that numerous activities taking place along the tributary rivers (as observed during the study) such as modification of fishing gears as well as fishing methods and the deterioration of the spawning grounds due to siltation from soil erosion caused by deforestation and agriculture, are putting the potamodromous fish species such as O. microlepis under serious threat. The study recommends that the populations of O. microlepis from the rivers can be managed equally since they are morphologically similar. Adopting uniform catchment management and sustainable exploitation of O. microlepis (such as regulations on mesh sizes and fishing methods, closing the rivers from fishing activities during spawning period, river bank and catchment management and restoration) with the aim of conserving the stocks from further overexploitation in these rivers is recommended so that the communities and the people at large continue to utilize the resource sustainably and at the same time, sustaining their livelihood

One hundred priority questions for the development of sustainable food systems in Sub-Saharan Africa

Sub-Saharan Africa is facing an expected doubling of human population and tripling of food demand over the next quarter century, posing a range of severe environmental, political, and socio-economic challenges. In some cases, key Sustainable Development Goals (SDGs) are in direct conflict, raising difficult policy and funding decisions, particularly in relation to trade-offs between food production, social inequality, and ecosystem health. In this study, we used a horizon-scanning approach to identify 100 practical or research-focused questions that, if answered, would have the greatest positive impact on addressing these trade-offs and ensuring future productivity and resilience of food-production systems across sub-Saharan Africa. Through direct canvassing of opinions, we obtained 1339 questions from 331 experts based in 55 countries. We then used online voting and participatory workshops to produce a final list of 100 questions divided into 12 thematic sections spanning topics from gender inequality to technological adoption and climate change. Using data on the background of respondents, we show that perspectives and priorities can vary, but they are largely consistent across different professional and geographical contexts. We hope these questions provide a template for establishing new research directions and prioritising funding decisions in sub-Saharan Africa

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

ILC Reference Design Report Volume 4 - Detectors

This report, Volume IV of the International Linear Collider Reference Design Report, describes the detectors which will record and measure the charged and neutral particles produced in the ILC's high energy e+e- collisions. The physics of the ILC, and the environment of the machine-detector interface, pose new challenges for detector design. Several conceptual designs for the detector promise the needed performance, and ongoing detector R&D is addressing the outstanding technological issues. Two such detectors, operating in push-pull mode, perfectly instrument the ILC interaction region, and access the full potential of ILC physics.This report, Volume IV of the International Linear Collider Reference Design Report, describes the detectors which will record and measure the charged and neutral particles produced in the ILC's high energy e+e- collisions. The physics of the ILC, and the environment of the machine-detector interface, pose new challenges for detector design. Several conceptual designs for the detector promise the needed performance, and ongoing detector R&D is addressing the outstanding technological issues. Two such detectors, operating in push-pull mode, perfectly instrument the ILC interaction region, and access the full potential of ILC physics