UAV method based on multispectral imaging for field phenotyping

In many countries, particularly in West Africa, there is a strong social demand for increased cereal production. Responding to this demand involves the improvement of cereal varieties. Modern varietal breeding programs in the sub-region need to establish the relationship between plant genotype and phenotype to select high-yielding stress-tolerant plants and to enhance agricultural production. However, in most cases, accurate phenotyping of large mapping populations is a limiting factor. The Regional Study Centre for the Improvement of Drought Adaptation (CERAAS) has developed a robust drone-based data collection and spatial modelling process to better measure cereal crops' traits for the benefit of plant breeding programs. Herein, we report an unmanned aerial vehicle (UAV) driven crop characteristics analysis throughout the crop cycle. We present a fully automatic pipeline based on a multispectral imaging system for the indirect measurement of agronomic and phenological characters of crops in agricultural field trials. The pipeline is made up of different stages including image acquisition, georeferencing, generation of orthoimages, creation of masks to delimit individual plots, and calculation of proxies. The incorporation of the UAV into agricultural field experiments has the potential to fast-track the genetic improvement of adaptation to drought

Effect of three years' seasonal malaria chemoprevention on molecular markers of resistance of Plasmodium falciparum to sulfadoxine-pyrimethamine and amodiaquine in Ouelessebougou, Mali

BackgroundIn 2012, seasonal malaria chemoprevention (SMC) was recommended as policy for malaria control by the World Health Organization (WHO) in areas of highly seasonal malaria transmission across the Sahel sub-region in Africa along with monitoring of drug resistance. We assessed the long-term impact of SMC on Plasmodium falciparum resistance to sulfadoxine-pyrimethamine (SP) and amodiaquine (AQ) over a 3-year period of SMC implementation in the health district of Ouelessebougou, Mali.MethodsIn 8 randomly selected sub-districts of Ouelessebougou, Mali, children aged 0-5 years were randomly selected during cross-sectional surveys at baseline (August 2014) and 1, 2 and 3 years post-SMC, at the beginning and end of the malaria transmission season. Blood smears and blood spots on filter paper were obtained and frequencies of mutation in P. falciparum genes related to resistance to SP and AQ (Pfdhfr, Pfdhps, Pfmdr1, and Pfcrt) were assessed by PCR amplification on individual samples and PCR amplification followed by deep sequencing on pooled (by site and year) samples.ResultsAt each survey, approximately 50-100 individual samples were analysed by PCR amplification and a total of 1,164 samples were analysed by deep sequencing with an average read depth of 18,018-36,918 after pooling by site and year. Most molecular markers of resistance did not increase in frequency over the period of study (2014-2016). After 3 years of SMC, the frequencies of Pfdhps 540E, Pfdhps 437G and Pfcrt K76T remained similar compared to baseline (4.0 vs 1.4%, p = 0.41; 74.5 vs 64.6%, p = 0.22; 71.3 vs 67.4%, p = 0.69). Nearly all samples tested carried Pfdhfr 59R, and this proportion remained similar 3 years after SMC implementation (98.8 vs 100%, p = 1). The frequency of Pfmdr1 N86Y increased significantly over time from 5.6% at baseline to 18.6% after 3 years of SMC (p = 0.016). Results of pooled analysis using deep sequencing were consistent with those by individual analysis with standard PCR, but also indicated for the first time the presence of mutations at the Pfdhps A581G allele at a frequency of 11.7% after 2 years of SMC, as well as the Pfdhps I431V allele at frequencies of 1.6-9.3% following 1 and 2 years of SMC, respectively.ConclusionTwo and 3 years of SMC implementation were associated with increased frequency of the Pfmdr1 N86Y mutation but not Pfdhps 540E, Pfdhps 437G and Pfcrt K76T. The first-time detection of the Pfdhps haplotype bearing the I431V and A581G mutations in Mali, even at low frequency, warrants further long-term surveillance

Using UAV Borne, Multi-Spectral Imaging for the Field Phenotyping of Shoot Biomass, Leaf Area Index and Height of West African Sorghum Varieties under Two Contrasted Water Conditions

International audienceMeeting food demand for the growing population will require an increase to crop production despite climate changes and, more particularly, severe drought episodes. Sorghum is one of the cereals most adapted to drought that feed millions of people around the world. Valorizing its genetic diversity for crop improvement can benefit from extensive phenotyping. The current methods to evaluate plant biomass, leaves area and plants height involve destructive sampling and are not practical in breeding. Phenotyping relying on drone based imagery is a powerful approach in this context. The objective of this study was to develop and validate a high throughput field phenotyping method of sorghum growth traits under contrasted water conditions relying on drone based imagery. Experiments were conducted in Bambey (Senegal) in 2018 and 2019, to test the ability of multi-spectral sensing technologies on-board a UAV platform to calculate various vegetation indices to estimate plants characteristics. In total, ten (10) contrasted varieties of West African sorghum collection were selected and arranged in a randomized complete block design with three (3) replicates and two (2) water treatments (well-watered and drought stress). This study focused on plant biomass, leaf area index (LAI) and the plant height that were measured weekly from emergence to maturity. Drone flights were performed just before each destructive sampling and images were taken by multi-spectral and visible cameras. UAV-derived vegetation indices exhibited their capacity of estimating LAI and biomass in the 2018 calibration data set, in particular: normalized difference vegetative index (NDVI), corrected transformed vegetation index (CTVI), seconded modified soil-adjusted vegetation index (MSAVI2), green normalize difference vegetation index (GNDVI), and simple ratio (SR) (r2 of 0.8 and 0.6 for LAI and biomass, respectively). Developed models were validated with 2019 data, showing a good performance (r2 of 0.92 and 0.91 for LAI and biomass accordingly). Results were also promising regarding plant height estimation (RMSE = 9.88 cm). Regression plots between the image-based estimation and the measured plant height showed a r2 of 0.83. The validation results were similar between water treatments. This study is the first successful application of drone based imagery for phenotyping sorghum growth and development in a West African context characterized by severe drought occurrence. The developed approach could be used as a decision support tool for breeding programs and as a tool to increase the throughput of sorghum genetic diversity characterization for adaptive traits

Using UAV Borne, Multi-Spectral Imaging for the Field Phenotyping of Shoot Biomass, Leaf Area Index and Height of West African Sorghum Varieties under Two Contrasted Water Conditions

Meeting food demand for the growing population will require an increase to crop production despite climate changes and, more particularly, severe drought episodes. Sorghum is one of the cereals most adapted to drought that feed millions of people around the world. Valorizing its genetic diversity for crop improvement can benefit from extensive phenotyping. The current methods to evaluate plant biomass, leaves area and plants height involve destructive sampling and are not practical in breeding. Phenotyping relying on drone based imagery is a powerful approach in this context. The objective of this study was to develop and validate a high throughput field phenotyping method of sorghum growth traits under contrasted water conditions relying on drone based imagery. Experiments were conducted in Bambey (Senegal) in 2018 and 2019, to test the ability of multi-spectral sensing technologies on-board a UAV platform to calculate various vegetation indices to estimate plants characteristics. In total, ten (10) contrasted varieties of West African sorghum collection were selected and arranged in a randomized complete block design with three (3) replicates and two (2) water treatments (well-watered and drought stress). This study focused on plant biomass, leaf area index (LAI) and the plant height that were measured weekly from emergence to maturity. Drone flights were performed just before each destructive sampling and images were taken by multi-spectral and visible cameras. UAV-derived vegetation indices exhibited their capacity of estimating LAI and biomass in the 2018 calibration data set, in particular: normalized difference vegetative index (NDVI), corrected transformed vegetation index (CTVI), seconded modified soil-adjusted vegetation index (MSAVI2), green normalize difference vegetation index (GNDVI), and simple ratio (SR) (r2 of 0.8 and 0.6 for LAI and biomass, respectively). Developed models were validated with 2019 data, showing a good performance (r2 of 0.92 and 0.91 for LAI and biomass accordingly). Results were also promising regarding plant height estimation (RMSE = 9.88 cm). Regression plots between the image-based estimation and the measured plant height showed a r2 of 0.83. The validation results were similar between water treatments. This study is the first successful application of drone based imagery for phenotyping sorghum growth and development in a West African context characterized by severe drought occurrence. The developed approach could be used as a decision support tool for breeding programs and as a tool to increase the throughput of sorghum genetic diversity characterization for adaptive traits.</jats:p

Age-dependent increase in antibodies that inhibit Plasmodium falciparum adhesion to a subset of endothelial receptors

Abstract Background Plasmodium falciparum-infected erythrocytes (IE) sequester in deep vascular beds where their adhesion is mediated by an array of endothelial surface receptors. Because parasite adhesion has been associated with disease, antibodies that block this activity may confer protective immunity. Here, levels of plasma anti-adhesion activity and surface reactivity against freshly collected IEs from malaria-infected children were measured in a Malian birth cohort and related to child age and malaria infection history. Methods Plasma samples from children enrolled at birth in a longitudinal cohort study of mother–infant pairs in Ouelessebougou, Mali were collected at multiple time points during follow-up visits. Anti-adhesion antibodies (i.e., inhibit IE binding to any of several endothelial receptors) and reactivity with surface IE proteins were measured using a binding inhibition assay and by flow cytometry, respectively. Results Levels of antibodies that inhibit the binding of children’s IE to the receptors ICAM-1, integrin α3β1 and laminin increased with age. The breadth of antibodies that inhibit ICAM-1 and laminin adhesion (defined as the proportion of IE isolates whose binding was reduced by ≥ 50%) also significantly increased with age. The number of malaria infections prior to plasma collection was associated with levels of plasma reactivity to IE surface proteins, but not levels of anti-adhesion activity. Conclusions Age is associated with increased levels of antibodies that reduce adhesion of children’s IE to three of the ten endothelial receptors evaluated here. These results suggest that anti-adhesion antibodies to some but not all endothelial receptors are acquired during the first few years of life

Malaria Infection Is Common and Associated With Perinatal Mortality and Preterm Delivery Despite Widespread Use of Chemoprevention in Mali: An Observational Study 2010 to 2014

Abstract Background In malaria-endemic areas, pregnant women and especially first-time mothers are more susceptible to Plasmodium falciparum. Malaria diagnosis is often missed during pregnancy, because many women with placental malaria remain asymptomatic or have submicroscopic parasitemia, masking the association between malaria and pregnancy outcomes. Severe maternal anemia and low birthweight deliveries are well-established sequelae, but few studies have confirmed the relationship between malaria infection and severe outcomes like perinatal mortality in high transmission zones. Methods Pregnant women of any gestational age enrolled at antenatal clinic into a longitudinal cohort study in Ouelessebougou, Mali, an area of high seasonal malaria transmission. Follow-up visits included scheduled and unscheduled visits throughout pregnancy. Blood smear microscopy and polymerase chain reaction (PCR) analysis were employed to detect both microscopic and submicroscopic infections, respectively. Intermittent preventative treatment in pregnancy with sulfadoxine-pyrimethamine (IPTp-SP) was documented and prompt treatment regardless of symptoms given upon malaria diagnosis. Results Of the 1850 women followed through delivery, 72.6% of women received 2 or more IPTp-SP doses, 67.2% of women experienced at least 1 infection between enrollment up to and including delivery. Malaria infection increased the risks of stillbirth (adjusted hazard ratio [aHR] 3.87, 95% confidence interval [CI]: 1.18–12.71) and preterm delivery (aHR 2.41, 95% CI: 1.35–4.29) in primigravidae, and early neonatal death (death within 7 days) in secundigravidae and multigravidae (aHR 6.30, 95% CI: 1.41–28.15). Conclusions Malaria treatment after diagnosis, alongside IPTp-SP, is insufficient to prevent malaria-related stillbirth, early neonatal death and preterm delivery (PTD). Although IPTp-SP was beneficial in Mali during the study period, new tools are needed to improve pregnancy outcomes. Clinical Trials Registration NCT01168271. </jats:sec

Host factors that modify Plasmodium falciparum adhesion to endothelial receptors

Abstract P. falciparum virulence is related to adhesion and sequestration of infected erythrocytes (IE) in deep vascular beds, but the endothelial receptors involved in severe malaria remain unclear. In the largest ever study of clinical isolates, we surveyed adhesion of freshly collected IE from children under 5 years of age in Mali to identify novel vascular receptors, and examined the effects of host age, hemoglobin type, blood group and severe malaria on levels of IE adhesion to a panel of endothelial receptors. Several novel molecules, including integrin α3β1, VE-cadherin, ICAM-2, junctional adhesion molecule-B (JAM-B), laminin, and cellular fibronectin, supported binding of IE from children. Severe malaria was not significantly associated with levels of IE adhesion to any of the 19 receptors. Hemoglobin AC, which reduces severe malaria risk, reduced IE binding to the receptors CD36 and integrin α5β1, while hemoglobin AS did not modify IE adhesion to any receptors. Blood groups A, AB and B significantly reduced IE binding to ICAM-1. Severe malaria risk varies with age, but age significantly impacted the level of IE binding to only a few receptors: IE binding to JAM-B decreased with age, while binding to CD36 and integrin α5β1 significantly increased with age

Effect of three years’ seasonal malaria chemoprevention on molecular markers of resistance of Plasmodium falciparum to sulfadoxine-pyrimethamine and amodiaquine in Ouelessebougou, Mali

Abstract Background In 2012, seasonal malaria chemoprevention (SMC) was recommended as policy for malaria control by the World Health Organization (WHO) in areas of highly seasonal malaria transmission across the Sahel sub-region in Africa along with monitoring of drug resistance. We assessed the long-term impact of SMC on Plasmodium falciparum resistance to sulfadoxine-pyrimethamine (SP) and amodiaquine (AQ) over a 3-year period of SMC implementation in the health district of Ouelessebougou, Mali. Methods In 8 randomly selected sub-districts of Ouelessebougou, Mali, children aged 0–5 years were randomly selected during cross-sectional surveys at baseline (August 2014) and 1, 2 and 3 years post-SMC, at the beginning and end of the malaria transmission season. Blood smears and blood spots on filter paper were obtained and frequencies of mutation in P. falciparum genes related to resistance to SP and AQ (Pfdhfr, Pfdhps, Pfmdr1, and Pfcrt) were assessed by PCR amplification on individual samples and PCR amplification followed by deep sequencing on pooled (by site and year) samples. Results At each survey, approximately 50–100 individual samples were analysed by PCR amplification and a total of 1,164 samples were analysed by deep sequencing with an average read depth of 18,018–36,918 after pooling by site and year. Most molecular markers of resistance did not increase in frequency over the period of study (2014–2016). After 3 years of SMC, the frequencies of Pfdhps 540E, Pfdhps 437G and Pfcrt K76T remained similar compared to baseline (4.0 vs 1.4%, p = 0.41; 74.5 vs 64.6%, p = 0.22; 71.3 vs 67.4%, p = 0.69). Nearly all samples tested carried Pfdhfr 59R, and this proportion remained similar 3 years after SMC implementation (98.8 vs 100%, p = 1). The frequency of Pfmdr1 N86Y increased significantly over time from 5.6% at baseline to 18.6% after 3 years of SMC (p = 0.016). Results of pooled analysis using deep sequencing were consistent with those by individual analysis with standard PCR, but also indicated for the first time the presence of mutations at the Pfdhps A581G allele at a frequency of 11.7% after 2 years of SMC, as well as the Pfdhps I431V allele at frequencies of 1.6–9.3% following 1 and 2 years of SMC, respectively. Conclusion Two and 3 years of SMC implementation were associated with increased frequency of the Pfmdr1 N86Y mutation but not Pfdhps 540E, Pfdhps 437G and Pfcrt K76T. The first-time detection of the Pfdhps haplotype bearing the I431V and A581G mutations in Mali, even at low frequency, warrants further long-term surveillance. </jats:sec

Antibody levels to recombinant VAR2CSA domains vary with Plasmodium falciparum parasitaemia, gestational age, and gravidity, but do not predict pregnancy outcomes

Abstract Background Maternal malaria is a tropical scourge associated with poor pregnancy outcomes. Women become resistant to Plasmodium falciparum pregnancy malaria as they acquire antibodies to the variant surface antigen VAR2CSA, a leading vaccine candidate. Because malaria infection may increase VAR2CSA antibody levels and thereby confound analyses of immune protection, gravidity-dependent changes in antibody levels during and after infection, and the effect of VAR2CSA antibodies on pregnancy outcomes were evaluated. Methods Pregnant women enrolled in a longitudinal cohort study of mother-infant pairs in Ouelessebougou, Mali provided plasma samples at enrollment, gestational week 30–32, and delivery. Antibody levels to VAR2CSA domains were measured using a multiplex bead-based assay. Results Antibody levels to VAR2CSA were higher in multigravidae than primigravidae. Malaria infection was associated with increased antibody levels to VAR2CSA domains. In primigravidae but not in secundigravidae or multigravidae, antibodies levels sharply declined after an infection. A relationship between any VAR2CSA antibody specificity and protection from adverse pregnancy outcomes was not detected. Conclusions During malaria infection, primigravidae acquire short-lived antibodies. The lack of an association between VAR2CSA domain antibody reactivity and improved pregnancy outcomes suggests that the recombinant proteins may not present native epitopes targeted by protective antibodies