Plasmodium falciparum was the predominant species followed by Plasmodium malariae and then Plasmodium ovale. The RDT and LM had the same sensitivity (90.6%) with a slight difference in their specificity . The RDT also demonstrated higher positive and negative predictive values compared with those of the LM.Malaria is still a threat to public health as it remains the first endemic disease in the world. It is a pervasive parasitic disease in tropical and subtropical regions where asymptomatic malaria infection among humans serves as a significant reservoir for transmission. A rapid and correct diagnosis is considered to be an important strategy in the control of the disease especially in children, who are the most vulnerable group. This study assessed the prevalence of asymptomatic malaria in children at the Nkolbisson health area in Yaoundé, Cameroon. A cross-sectional study design and a convenience sampling plan were used. A total of 127 participants were recruited after informed and signed consent from parents and/or guardians. Blood samples were collected by finger-pricking and venipuncture from children aged 6 months to 10 years and then screened for asymptomatic parasitemia by a rapid diagnostic test (RDT), light microscopy (LM) staining with Giemsa and 18S rRNA polymerase chain reaction (PCR) for speciation. The data were analyzed using SPSS version 20 software. The study identified 85 children who were positive from the PCR, 95 positive from the RDT and 71 from the LM, revealing a malaria prevalence of 66.9%, 74.8% and 55.9%, respectively. The prevalence was not observed to be dependent on the sex and age group of the participants.  Plasmodium species have been identified as causing malaria with Plasmodium falciparum accounting for the greater proportion of the global malaria burden especially in children under the age of 5 years [Plasmodium falciparum infection can result in a disease manifested by the asymptomatic carriage of uncomplicated or severe malaria [Plasmodium falciparum and non-falciparum species in children resident in Nkolbisson, Yaoundé, Cameroon.Malaria is still a threat to public health as it remains the first endemic disease in the world. It is a pervasive parasitic disease in tropical and subtropical regions. It is mostly prevalent in Sub-Saharan Africa, Asia and Latin America ,2. The S 5 years . Malaria malaria ,7. Asymp malaria . About 2 malaria ,7. In ma malaria . Most as malaria . The stu malaria . Evidenc malaria . Underst malaria . To comp2. Yaoundé is located 700 m (2300 feet) above sea level and at 3′52′ N 11°31′ E. The climate is of a Guinean-type with two rainy seasons (March–June and September–November) and two dry seasons (December–February and July–August). The average annual rain fall here is 1643 mm with a relative humidity ranging from 85 to 90% [The study was carried out in the Nkolbisson health area in the Cite-Vert health district in the Centre region of Cameroon where Yaoundé is the capital city. Yaoundé has a population of about 2,500,000 inhabitants and covers a surface are of 183 km5 to 90% ,14,15. MThe study received ethical approval from the National Ethics Committee for Human Health Research (N° 2018/07/1091/CE/CNERSH/SP). Research authorizations were also obtained from the Centre Regional Delegation for Public Health and from the health centers where the study was conducted. Prior to the participant enrollment, written and signed consent from each parent or guardian was obtained based on the informed consent forms designed for the study. The potential risks and benefits as well data privacy and confidentiality were clearly explained to parents or guardians. Only children whose parents or guardians signed and returned the consent form were included in the study.This study was a cross-sectional study and a convenience sampling approach was used. All children resident in the study area aged 6 months to 10 years were eligible. All participants were examined by a physician for malaria. Children who presented signs and symptoms of malaria such as a headache, tiredness or fever were not enrolled. Finger-pricking for blood samples was performed on children whose parents or guardians consented to the study. A questionnaire was used to collect the demographic data and vital signs as well as to assess the knowledge and attitude of the participants toward malaria prevention and control. A total of 127 participants were recruited between November 2019 and February 2020.Plasmodium species-specific PCR were blindly submitted to a second independent microscopist. Venous blood samples were collected from each patient by a syringe. A rapid diagnostic test (RDT) was performed on the spot in the field using a malaria rapid diagnostic kit supplied by SD Bioine (batch number: 05EDD011B) for the National Malaria Control Program in Cameroon to detect two antigens of the malaria parasites (HRP2/pLDH) that were at the different health centers, as described by the WHO . About 6Genomic deoxyribonucleic acid (DNA) was extracted from 200 mL of blood in EDTA tubes using an EZNA DNA extraction blood kit  according to the manufacturer’s instructions. Prior to the extraction, a few materials were sterilized by autoclaving and the equipment was disinfected with 10% bleach and 70% alcohol. The blood samples were briefly thawed at room temperature. In a sterile Eppendorf tube, 100 µL blood was introduced with the help of a micropipette. A total of 150 µL of elution buffer (65 °C) was then added onto it. An OB protease and a BL buffer (lysis buffer) was then added to the mixture and vortexed for 15 s, after which it was incubated at 65 °C for 10 min. During the incubation, the tubes were briefly vortexed twice. A total of 260 µL of 100% ethanol was then added to the mixture, vortexed for 20 s and then centrifuged briefly. The mixture was then transferred to a DNA spin column (DNA HiBind Column) and centrifuged for 1 min at 10,000 rpm. A total of 700 μL of a wash buffer was then introduced into each column. This step was repeated for each tube. A 2 min centrifugation at 10,000 rpm was performed to dry the column matrix. The columns were transferred into a nuclease-free microcentrifuge tube. A total of 150 µL of elution buffer was used to elute the DNA, which was then stored at −20 °C for a further analysis.Plasmodium species was performed in accordance with a protocol described by Putapomtip et al. [Plasmodium genus-specific outer primers . A volume of 25 µL from the first reaction contained 20.25 µL of nuclease-free water, 2.5 µL of 10 X thermopol buffer (New England Biolabs), 0.5 µL of 10 mM dNTPs, 0.25 µL of primer, 0.25 µL of Taq polymerase (New England Biolabs) and 1 μL of amplicons. This second step took place under the following conditions: 95 °C for 15 min, 30 cycles of 58 °C for 2 min, 72 °C for 5 min and 94 °C for 1 min followed by 58 °C for 2 min and 72 °C for 5 min.A PCR was performed on all samples and the 18S rRNA gene was used to detect and identify the different malaria-causing species. The PCR amplification of the p et al. . A neste primers  and 0.25A total of 127 children were enrolled into the study . There wPlasmodium falciparum malaria infection by the RDT, 85 (66.9%) by light microscopy and 71 (55.9%), by the PCR. From the patients screened by the PCR, 66.9% were positive, among which were 51 (75.0%) males and 34 (57.6%) females. A total of 42 (33.1%) children were negative for malaria by the PCR. Of the 95 (74.8%) children diagnosed positive for Plasmodium falciparum malaria by the RDT, 54 (79.4%) were males and 41 (69.5%) females. In addition, 32 (25.2%) of the children were diagnosed negative for Plasmodium falciparum malaria by the RDT, among which were 18 (30.5%) females and 14 (20.6%) males. When the children were diagnosed for Plasmodium malaria using microscopy, 71 (55.9%) of them were positive and 56 (44.1) negative. Among the positive cases 17 (28.8%) were females and 54 (79.4%) were males whereas among the negative cases, 42 (71.2%) were females and 14 (20.6%) were males (p = 0.199) and microscopy (p = 0.282). However, an association was observed between sex and malaria prevalence by the PCR (p = 0.038). No significant association was observed between the age group and malaria prevalence, be it by the RDT (p = 0.508), microscopy (p = 0.682) or by the PCR (p = 0.655).The three diagnostic methods, light microscopy (LM), a rapid diagnostic test (RDT) and a polymerase chain reaction (PCR), detected significant malaria infections in asymptomatic children. A total of 95 (74.8%) from the 127 cases detected positive for a re males . A high re males . No signPlasmodium falciparum, 4 were infected with Plasmodium ovale and 7 were infected with Plasmodium malariae. Of the 85 infected with Plasmodium falciparum, 51 (75.0%) were males and 34 (57.6%) were females. A statistical significance was observed between the gender of the children and infection by Plasmodium falciparum (p = 0.038)  was female and 3 (4.4%) were males. Furthermore, 4 (5.9%) of the 7 children infected by Plasmodium malariae were males and 3 (5.1%) were females. No statistical significance was observed between the gender in children infected with Plasmodium ovale (p = 0.382) as well as Plasmodium malariae (p = 0.844)  . Of the = 0.844) .Plasmodium falciparum malaria by microscopy and who were truly infected with Plasmodium falciparum was 80.2% and that of those who were diagnosed negative and who were truly negative was 74.2%. These were the respective positive and negative predictive values for microscopy. The positive and negative predictive values of the RDT stood at 81.1% and 75.0%, respectively.Overall, when light microscopy (LM) and RDT diagnostic tools were compared with the PCR as the standard, 77 out of the 85 children that were diagnosed positive for malaria by the PCR were also positive by microscopy, giving a sensitivity or true positive rate of 90.6% with a false negative rate of 9.4% . Of the p = 0.532) or between the age groups of the children (p = 0.698).The average parasitemia of each child was 8260.19 ± 20,427.84 parasites/µL . Among tPlasmodium falciparum and non-falciparum species in children resident in Nkolbisson, Yaoundé, Cameroon. LM, RDT and PCR malaria diagnostic techniques were used to detect Plasmodium falciparum and non-falciparum species in children between the ages of 6 months and 10 years. The study compared the diagnosis of malaria by LM, a RDT and a PCR with the assessment of asymptomatic malaria in children. A total of 127 asymptomatic children were tested for malaria. The study identified 85 children as positive by the PCR, 95 positive by the RDT and 71 by LM, revealing a malaria prevalence of 66.9%, 74.8% and 55.9%, respectively.This study assessed the prevalence of asymptomatic malaria in children in Nkolbisson, Yaoundé. Nkolbisson is in a forest malaria transmission geo-ecological zone where malaria transmission is year-round . The conBased on LM (55.9%) and the RDT (74.8%), the study reported a high prevalence of asymptomatic malaria in children. This prevalence was higher compared with that observed in other studies carried out in Cameroon; for instance, in Buea where a prevalence of 40.32% from LM and 34.41% from an RDT was observed and in Yaoundé with an overall prevalence of 6.5% ,24. A siPlasmodium infections was observed [Plasmodium compared with younger children [Based on the PCR, a high prevalence of 75.0% was observed compared with a similar study carried out in Ghana where a prevalence of 36.8% of asymptomatic observed . A similobserved  with a pchildren . Overallchildren . The trechildren .Plasmodium carriage showed no significant association with gender in the case of Plasmodium malariae (p = 0.382) but a significant association was observed with Plasmodium falciparum (p = 0.038) and P. ovale (p = 0.382). The total number of species identified was 96, which was slightly than the 95 positive sample diagnoses by the RDT. This difference is due to the fact that one individual was found to be harboring more than one species. The average parasitemia was not significantly associated with the prevalence of parasitemia in either gender or the age group. However, other studies [The distribution of the  studies ,35 have  studies . The sen studies ,20, 37. Plasmodium falciparum was the predominant species followed by Plasmodium malariae and then Plasmodium ovale. Thus, understanding the malaria burden in children has a great implication on the interruption of malaria transmission. Ultimately, an accurate, prompt and cheap diagnosis is a key step in managing and treating the disease.Our study identified 85 children positive by a PCR, 95 children positive by an RDT and 71 by LM, revealing a malaria prevalence of 66.9%, 74.8% and 55.9%, respectively.


Plasmodium falciparum malaria infection, anemia, and fever between the intervention and control groups. Kaplan-Meier survival analyses were used to compare the time to P. falciparum parasitemia infection between the groups. The community acceptance rate was 96.5% (139 of 144). Significant declines were observed in the prevalence of P. falciparum parasitemia  and anemia  in the intervention group compared with the control group. The cumulative incidence of P. falciparum infections was significantly greater  in the control group compared with the intervention group . This study reveals that expanding SMC to older children is likely feasible, has high community acceptance, and is in reducing uncomplicated malaria and anemia in older children.Seasonal malaria chemoprevention (SMC) was adopted in Mali in 2012 for preventing malaria in children younger than 5 years. Although this strategy has been highly effective in reducing childhood malaria, an uptick in malaria occurrence has occurred in children 5 to 15 years of age. This study aimed to investigate the feasibility of providing SMC to older children. A cohort of 350 children age 5 to 14 years were monitored during the 2019 transmission season in Dangassa, Mali. The intervention group received five monthly rounds of sulfadoxine–pyrimethamine plus amodiaquine, whereas the control group consisted of untreated children. Community acceptance for extending SMC was assessed during the final round. Logistic regression models were applied to compare the risk of  According to the 2018 Mali Demographic Health Survey, the prevalence of malaria parasitemia in Mali was 19% in children younger than 5 years old.–,–Recent studies have shown that successful malaria control in children younger than 5 years of age may have increased the malaria burden in children who are at least 5 years old.This study was carried out in the village of Dangassa, a rural area situated in the Koulikoro region of Mali approximately 82 km southwest of Bamako and 4 km from the Niger River. Malaria transmission occurs year-round in Dangassa, but peak transmission typically occurs between July and December.In 2017, in collaboration with West Africa ICEMR researchers and local surveillance and clinical staff, a dynamic cohort of 1,400 subjects was assembled from the general population to identify epidemiological factors associated with malaria. An open-label, non-randomized study was conducted to estimate the effect of administering five SMC rounds (as opposed to a current standard of four rounds), implemented in children age 5 to 14 years in Dangassa. The intervention group for this study was selected from a cohort within the ICEMR study, and a control group was selected from the general population. SMC provisions were administered through fixed delivery by medical staff at community health centers as part of routine SMC delivery. Photo identification cards were used to authenticate participant enrollment throughout the study.A priori sample size estimates were calculated based on the national malaria prevalence among children in rural areas (23%).Children younger than 5 years old are eligible to receive SMC free of charge throughout Mali as a standard of care. This study provided SMC for children as old as 14 years using SP and AQ over five monthly rounds in the intervention group between July and November 2019. All children residing in Dangassa during the 2019 transmission season were eligible for the study. Exclusion criteria include known allergic reaction to SP or AQ, chronic illness, or clinical malaria confirmed by rapid diagnostic test during the course of treatment. A therapeutic dose of AQ (10 mg/kg body weight/d), combined with one dose of SP (25 mg sulfadoxine and 1.25 mg pyrimethamine/kg body weight on day 1), was delivered by trained community health workers, who administered the initial dose. The remaining two doses of AQ were administered at home by the parents of the enrolled children.In July, prior to the first administration of SP and AQ (baseline), and in December (end of study), data were collected on parasitemia, hemoglobin level, body temperature, and use of insecticide-treated bed nets (ITNs) for both the intervention and control groups. During the SMC campaign, routine monitoring and surveillance was performed for all study variables for both comparison groups. Study participants presenting with fever and those who tested positive using rapid diagnostic tests were treated according to standard NMCP guidelines. Household surveys were carried out between 4 and 7 days after each round of SMC to collect and monitor data on adverse events after SMC administration. Community acceptance was assessed via clinical surveys performed during the final round of SMC. Parents of enrolled children were strongly encouraged to bring children experiencing adverse events to a local health center. Adverse events were monitored and tabulated after each of the five rounds of SMC delivery, and parasitemia in blood smears was determined using microscopy. Thick blood films were stained with 10% Giemsa and examined using ×100 magnification and oil immersion of the objective lens of a light microscope. The number of asexual and sexual parasites were counted per 200 leukocytes and per 1,000 leukocytes, respectively. Hemoglobin levels were measured from whole blood using HemoCue Hb 301 analyzers . Axillary temperature was measured using electronic thermometers.Data responses were captured using electronic case report forms and computer tablets. Daily quality control was performed to detect missing and incorrectly logged data prior to the transfer to a data management center in Bamako, Mali. Range and logic checks were used to ensure all data responses lay within clinically plausible ranges. Data were exported to the STATA statistical package  for data management and statistical analyses.Prevalence of parasitemia was expressed as the proportion of asymptomatic participants with positive parasitemia confirmed by microscopy. Anemia was defined as hemoglobin < 11 g/dL, and fever was defined as a body temperature of more than 37.5°C. Adverse events were considered to be any reported clinical symptom or complaint after treatment administration. SMC coverage rates were defined as the proportion of children receiving the initial dose of SP and AQ during each round of the SMC campaign.Interviews with parents of enrolled children were performed using clinical questionnaires to assess SMC acceptability in children at least 5 years of age. These questionnaires included questions regarding the willingness of the parent or guardian to adhere to the SMC strategy and compliance despite its potential adverse events. SMC was deemed acceptable if the parent or guardian reported that the strategy was “good” and generally agreed with the notion of extending the prevention strategy to older children. SMC was deemed unacceptable if the parent reported that the strategy was “bad” and generally did not agree with the notion for extending the SMC program to older children. The clinical questionnaire is provided in Supplemental Appendix SI.Plasmodium falciparum parasitemia, anemia, and fever at baseline (July) and at the end of the study (December) for both comparison groups. Model predictors for the logistic regression models were age group , use of ITNs (yes or no), P. falciparum parasitemia (positive or negative), gametocyte carriage (positive or negative), anemia (yes or no), and fever (yes or no). Cox regression models and Kaplan-Meier curves were used to compare the time to illness onset for P. falciparum parasitemia between the comparison groups in children with negative blood smears at baseline. The type I error threshold was set at 5%.Data were expressed as proportions and percentages. Logistic regression models were used to estimate the risk of This study was carried out through the ICEMR program, in collaboration with the Malian NMCP. The study protocol was approved by the Ethical Committee of the Faculty of Medicine, Pharmacy and Odontostomatology (FMPOS) of the University of Sciences, Techniques and Technologies of Bamako (No. 2019/04/FMPOS). Written informed consent was obtained from all study participants. All malaria cases were treated free of charge at local community health centers by trained personnel as part of routine care.Treatment compliance rates were 81.7% (143 of 175) in August, 100% (175 of 175) in September, 90.9% (159 of 175) in October, and 100% (175 of 175) in November. Traveling and farming were reported as the main reasons for missed health facility visits Figure . The mosP. falciparum parasitemia , fever , and gametocyte carriage . Anemia prevalence was lower in the intervention group than the control group . At the end of the study (December), there was a 78.0% reduction in the prevalence of P. falciparum parasitemia in the intervention group . A reduction of 86.0% in the likelihood of having anemia was also observed in the intervention group . Presence of fever , gametocyte carriage , and ITN use  were similar between the two groups (Table At baseline (July), no significant differences were observed between the intervention and control groups with respect to P. falciparum parasitemia were observed in the intervention group compared with control group in September ; October ; November ; and December   in October  in the control group versus 28.0% in the intervention group. At the end of the study (December), the cumulative incidence was 75.4%  in the control group compared with 40.7%  in the intervention group , SMC likely averted 34.70% of new P. falciparum parasitemia infections in children who received SP and AQ. The actual effect of SMC on malaria illness and P. falciparum parasitemia is associated with the number of SMC rounds received during the campaign and the doses of SP and AQ needed during each round to obtain an optimal plasma concentration to inhibit parasite growth or avoid a new case of P. falciparum parasitemia.,Among children in the intervention group at baseline, a decrease in the monthly prevalence of parasitemia was observed from September to December. Effective reduction in the prevalence of parasitemia and anemia was reported in children younger than 10 years after 5 months of SMC delivery in malaria-endemic areas, which suggests that providing SMC for 5 months during the transmission season is more effective in reducing malaria occurrence in areas with longer transmission seasons.The limitations of our study include the non-randomization of study participants in the two groups because the intervention group was selected from the ICEMR study cohort and the control group from the general population. The intervention group was monitored at regular time intervals and thus may have been generally healthier than the general population. However, children in the general population (and children in the control group) had similar access to health care and malaria treatment as a result of the national standard of care. Clinical data were not collected on malaria illness cases and malaria mortality, which could provide additional evidence of the effect of SMC intervention in reducing malaria infection outcomes, including severe malaria.It is worth mentioning that extending SMC to older children would increase the cost of the SMC programs. Therefore, it is important to reproduce the results from this study on a larger scale and to evaluate its cost-effectiveness before the findings are used to guide public policy.Supplemental materials


Anopheles gambiae sensu lato (s.l.) primarily feeds indoors at night. Identification of factors which influence mosquito house entry could lead to development of novel malaria vector control interventions. A study was therefore carried out to identify risk factors associated with house entry of An. gambiae s.l. in south-west Burkina Faso, an area of high insecticide resistance.In rural Burkina Faso, the primary malaria vector An. gambiae s.l. were measured, including socio-economic status, caregiver’s education and occupation, number of people sleeping in the same part of the house as the child, use of anti-mosquito measures, house construction and fittings, proximity of anopheline aquatic habitats and presence of animals near the house. Mosquito counts were compared using a generalized linear mixed-effect model with negative binomial and log link function, adjusting for repeated collections.Mosquitoes were sampled monthly during the malaria transmission season using CDC light traps in 252 houses from 10 villages, each house sleeping at least one child aged five to 15 years old. Potential risk factors for house entry of An. gambiae s.l. Of the 6691 An. gambiae s.l. identified to species, 4101 (61.3%) were An. gambiae sensu stricto and 2590 (38.7%) Anopheles coluzzii. Having a metal-roof on the child’s sleeping space  was associated with fewer malaria vectors inside the home.20,929 mosquitoes were caught, of which 16,270 (77.7%) were An. gambiae s.l. was 45% lower in sleeping spaces with a metal roof, compared to those with thatch roofs. Improvements in house construction, including installation of metal roofs, should be considered in endemic areas of Africa to reduce the burden of malaria.This study demonstrated that the rate of The online version contains supplementary material available at 10.1186/s12936-021-03926-5. Anopheles gambiae sensu lato (s.l.), an effective malaria vector that readily feeds on people indoors at night, where about 79% of malaria transmission typically occurs [Despite large reductions in the malaria burden across sub-Saharan Africa from 2000 to 2015 [y occurs . The indy occurs , 5, presy occurs , typology occurs , use of y occurs  An. gambiae s.l., and research conducted in the study area suggests that exposure to ITNs may have no impact on the lifelong survival of malaria vectors [Burkina Faso is an area of intense seasonal malaria transmission, and cases are increasing –13 despi vectors . New tooAn. gambiae s.l. during the malaria transmission season in an area of intense malaria transmission in south-west Burkina Faso. Findings from this study might identify potential opportunities for improving malaria control in Burkina Faso and other countries in sub-Saharan Africa experiencing persistently high malaria transmission.Several studies have demonstrated that malaria mosquito house entry can be reduced through simple changes to house design, such as closing eaves and screening windows and doors . The use2 with an estimated population of 407,073 inhabitants [Plasmodium falciparum accounts for 90% of cases [An. gambiae sensu stricto (s.s.) and Anopheles coluzzii [The study was conducted in Banfora Health District, in the Cascades Region, south-west Burkina Faso Fig. . This isabitants . Malariaabitants . Plasmodof cases . The maicoluzzii . In 2016Compounds typically consist of multiple single room buildings arranged in a circular or semi-circular fashion around a shared open space, with sleeping areas, kitchens and toilets existing as separate structures . PolygynP. falciparum infection in children aged five to 15 years [An. gambiae s.l. in the children’s sleeping space during the peak malaria transmission period from 24 July to 28 December 2017.The study was nested in a cohort study of risk factors for 15 years . This stP. falciparum infection  were included in the cohort study and this current study reports on the entomological surveillance from the children’s sleeping space.Sampling and recruitment of the study cohort is described elsewhere . In brieAn. gambiae s.l. were identified using an enzyme-linked immunosorbent assay [An. gambiae s.l. females were typed to species by PCR [An. gambiae s.l. were caught per house then all were typed to species by PCR, but if the number was greater than 100, a third of the mosquitoes were randomly sampled for PCR analysis.CDC light traps  were used to estimate indoor mosquito densities in the study child’s sleeping space. These traps were placed with the bulb 1.5 m above the floor, approximately 0.5 m from the foot end of a bed with an ITN occupied by the study child. Houses were sampled from 19.00 h to 06.00 h every 4 weeks. Two villages (Nofesso and Ouangolodougou) were inaccessible for two weeks at the start of the study period due to flooding. Mosquitoes were taken to the laboratory in cool boxes and killed by freezing. Mosquitoes were identified morphologically using established keys . The prent assay  and An. s by PCR , 26. If In June, a questionnaire was administered to the caregiver of the study child to collect information on ethnicity, education level and occupation of caregivers, ITN use during the previous night, use of other protective measures , number of people sleeping in the same part of the house as the study child, roof, wall and floor construction of the child’s sleeping space, whether the eaves  were open or closed and presence of mosquito screening. Information was also collected from the head of the child’s household  on asset ownership and household characteristics, following standard procedures used in the Burkina Faso Demographic and Health Survey , Open Source Geospatial Foundation Project) was used to determine distances between the child’s sleeping space and aquatic habitats. Principal component analysis (PCA) was used to calculate the socio-economic status (SES) factor score of the head of the child’s household. SES factor scores were ranked, and households divided into five equal wealth quintiles, from 1, the poorest, to 5, the least poor. The entomological inoculation rate (EIR) or estimated number of infectious bites per study child during the transmission season was calculated using the formula EIR = MaSd where Ma is the human biting rate, estimated from the arithmetic mean number of female An. gambiae s.l. caught per light trap night across the transmission season, where S is the proportion of female An. gambiae s.l. found to be CSP positive by village and d is the number of days in the transmission season. Mean values were compared using a t-test and proportions compared using chi-squared tests. A generalized linear mixed-effect model with a negative binomial distribution, to account for overdispersion, and log link function was used to identify risk factors associated with the mean number of An. gambiae s.l. per catch night per sleeping space each month. Risk factors were selected a priori based on importance for malaria vector house entry. These were SES quintile of the household head, ITN use, use of other protective measures, number of people sleeping in the same part of the house as the child, roof, floor and wall material in the sleeping space, eaves (open or closed), presence of large domesticated animals within 5 m of the sleeping space and presence of habitats positive for anopheline larvae within 300 m of the child’s sleeping space. A random effect for study child ID number was used to account for repeated measures on the same sleeping space and village was included as a fixed effect. Univariate analysis was conducted followed by construction of a simple multivariate model in which every risk factor was included, irrespective of whether the variable was significant in the univariate model. Interactions were tested between a subset of variables that were thought to be biologically relevant to explore. Means and 95% confidence intervals were calculated. Statistical analysis was carried out in Stata 15 . The study is reported following STROBE guidelines [Data were collected on Android personal digital assistants programmed using the KoboCollect system and included drop down boxes and consistency checks to reduce data entry errors. Following cleaning, the dataset was locked and saved in Microsoft Access. The primary outcome was the number of female idelines .An. gambiae s.l. (77.7%). Of the 6691 An. gambiae s.l. identified to species (excluding 924 lost and non-identified samples), 4101 were An. gambiae s.s. (61.3%) and 2590 An. coluzzii (38.7%). Malaria vector abundance rose in July after the start of the rains in May, reaching a peak in August, before declining to low levels in November and December. 3.3% of An. gambiae s.l. were CSP positive and the overall EIR in the study area was 80.4 infective bites/child over the six-month transmission season. The village-level EIR ranged from 40.8 in Timperba to 191.9 in Tondoura.As reported elsewhere , a totalThe ethnic composition of the study population was Gouin , Karaboro , Mossi , Turka , Fulani , Senoufo  and other ethnic groups .Sleeping spaces with metal roofs were more likely to have walls and floors made of finished materials and open eaves than thatch roof sleeping spaces. 81.7% (156/191) of sleeping spaces with a metal roof had a cement or tiled floor compared to 42.3% (22/52) of those with a thatch roof (p < 0.001). Metal roof sleeping spaces were also more likely to have brick or cement walls  compared to thatch roof sleeping spaces . Sleeping spaces with a metal roof were also more likely to have open eaves  than sleeping spaces with a thatch roof . Children living in sleeping spaces with a thatch roof were more likely to share the same part of the house with greater than 12 people , than children living in sleeping spaces with a metal roof . There was no association between metal roof sleeping spaces and distance from the nearest anopheline larvae positive habitat  was associated with fewer malaria vectors indoors, after adjusting for the other risk factors including SES of the household head  and no other significant risk factors were identified.The study findings demonstrate highly intense transmission of malaria in Banfora Health District with a person sleeping without an ITN experiencing a seasonal EIR varying from 40.8 infectious bites per person in Timperba village to 191.9 in Toundoura village . ReporteFinding fewer malaria vectors indoors in metal roof houses compared to thatch roof houses may be a result of the indoor climate of the different typologies of houses. Metal roof houses tend to be hotter and less humid than thatch roof houses which can reduce the survivorship of malaria vectors resting indoors . AlternaAnopheles arabiensis than thatch-roof houses [An. gambiae s.l. in metal-roof houses [Reduced malaria vector density in metal-roof houses compared to thatch-roof houses has been reported in several studies, including a Tanzanian study where metal-roof houses had 33% less f houses , and a Uf houses . Resultsf houses , and in f houses . It may f houses , 32.Ultimately, whether a metal-roof house has more or less mosquitoes than a thatch-roof house will depend on how porous the house is to mosquitoes and the extent of ventilation . FurtherThe study has several limitations. Firstly, ITN use the previous night was assessed by asking the caregiver, which may be prone to social desirability bias . The useP. falciparum infection, with only overnight travel and higher SES factor score being associated with higher rates of P. falciparum infection [P. falciparum infection in children differed, since higher indoor vector density does not automatically imply higher infection risk. The indoor density of malaria vectors may be less important in this study area due to the observation of increasing outdoor biting with some studies suggesting ~ 54% of An. gambiae s.l. host seeking outdoors [The cohort study in which this entomological study was nested did not identify strong risk factors for nfection . It is doutdoors  or more outdoors .What are the implications of the study findings for vector control and future research? The study highlights the potential of improved housing to reduce malaria transmission and supports the results of systematic reviews and multi-country research studies on this topic , 37. HouThis study in south-west Burkina Faso demonstrates a 45% reduction in indoor density of malaria vectors in sleeping spaces with a metal roof compared to those sleeping spaces with thatch roofs. The study adds to the growing evidence base supporting the use of housing improvement against malaria.Additional file 1: Questionnaire administered to head of household on asset ownership and household characeristics.


Plasmodium falciparum, with a positive malaria rapid diagnostic test (MRDT) for P. falciparum malaria. The mother had a negative peripheral film for malaria and a negative MRDT. The neonate was managed with intravenous artesunate with improvement.Congenital malaria is the presence of malaria parasites in a blood smear obtained from a neonate usually within 24 hours to 7 days of life. It has for long been regarded a rare condition. However, recent data indicate that congenital malaria complicates around 35.9% of live births globally, 0–37% in Sub-Saharan Africa and about 4–6.1% in Eastern Uganda. We present a 2-day-old neonate who presented with fever, irritability, and failure to breastfeed. Laboratory tests indicated that the neonate had a positive Giemsa-stained peripheral smear for  Uganda remains a highly malaria-endemic country, one of the 6 most affected countries contributing 5% to the global malaria burden . ChildreCongenital malaria (CM) is defined as a positive blood smear for malaria in a neonate from 24 hours to 7 days of life. This is usually due to maternofoetal transfer of malaria parasites. On the other hand, neonatal malaria is defined by the presence of malaria parasites in peripheral blood within 28 days of life, usually attributable to mosquito bites , 4. The CM presents with nonspecific signs and symptoms of fever, anaemia, jaundice, vomiting, lethargy, convulsions, irritability, tachypnoea, respiratory distress, and hepatosplenomegaly which overlap with sepsis syndrome . Due to Neonates have some level of protection from malaria due to passive immunisation from maternal antibodies, presence of foetal haemoglobin, and low iron levels which do not favour growth of plasmodium . HoweverCurrently, it is estimated that there is a 33.7% global burden of congenital malaria  with Subst hour of life and the neonate was suckling well. He passed meconium and urine within the first 24 hours of life.We report a case of a male neonate delivered at term by spontaneous vaginal delivery from a lower level health centre, with a birthweight of 3.7 kg. Apgar score was not documented, but the neonate cried immediately after birth. Labour lasted about 8 hours and membranes ruptured just before delivery. Breastfeeding was initiated within the 1The neonate was born to a 29-year-old G2P1 + 0 who attended antenatal care 6 times, starting at 3 months. She had been screened for HIV, syphilis, hepatitis B, and urinalysis, and all were negative, with a blood group of A Rhesus D negative. She received tetanus toxoid, Fansidar for malaria prophylaxis, haematinics, and deworming. She reported that the pregnancy was uneventful.About 36 hours after delivery, the neonate developed a high-grade fever, irritability, poor breastfeeding, and yellowing of eyes and skin. The neonate was managed with unknown oral medication before referral to Jinja Regional Referral Hospital for further management.At admission, the neonate was conscious, with an axillary temperature of 38.0°C, and jaundice (Kramer stage 2). There was no respiratory distress, no pallor, no dehydration, and no dysmorphic features noted. The systemic physical examination was unremarkable.P. falciparum was positive; thin blood smear showed P. falciparum malaria species with 1.846∗103 malaria-infected red blood cells.Malaria rapid diagnostic test (MRDT) for ∗103/ul, and platelet count 244∗103/ul.Complete blood count was normal with Hb of 15.2 g/dl, white blood cells 11.79Blood group was A Rhesus D negative.Total serum bilirubin was 200.33 umol/l with direct bilirubin 7.48 umol/l.P. falciparum was negative, and no haemoparasites were seen on the peripheral smear of maternal blood.The rapid diagnostic test for Investigations done were as follows:The neonate was initiated on intravenous artesunate at admission, 12 hr and 24 hr later, then once a day for 5 days. Intravenous empiric antibiotics (cloxacillin and cefotaxime) for presumed neonatal sepsis were also administered. After 48 hours of antibiotics, C-reactive protein (CRP) was done and it was nonreactive, and the antibiotics were stopped. Blood culture and sensitivity was not performed for this patient. A peripheral blood smear done on day 5 of antimalarial treatment revealed no malaria parasites. The neonate was discharged after 5 days of artesunate with great improvement. We were not able to follow up the patient after discharge.P. falciparum being the most predominant in the Sub-Saharan region [P. falciparum as the causative plasmodium species.Malaria is a parasitic disease that is transmitted by an infectious female Anopheles mosquito during a blood meal. This is caused by a parasitic protozoan of the genus plasmodium which has 5 different species, with n region . In thisIn 2019, there were 229 million cases of malaria, with over 94% of an estimated 409,000 deaths globally. More than 94% of all cases and deaths occurred in Sub-Saharan Africa. Children under 5 years are the most vulnerable, and in 2019, they accounted for 67% (274000) of all malaria deaths worldwide .Congenital malaria is defined as the presence of malarial parasites in the peripheral blood smear of the new born from 24 hours to 7 days of life. However, it can occur beyond 28 days of life confounding with neonatal malaria , 6. CM iBabies are believed to have partial protection from malaria in the first few months of life, owing to passively acquired maternal IgG antibodies, the predominance of haemoglobin F (HbF) in their erythrocytes, and the low levels of iron and para-amino benzoic acid (both required for parasite growth) in breast milk . DespiteGlobally, there is a 33.7% prevalence of congenital malaria with Africa having 39.5%. Furthermore, it is estimated that 40 neonates per 1000 live births will experience clinical malaria during the first 7 days of life , 13.The true burden of congenital malaria may be underestimated due to absence of routine screening of newborns with fever, low index of suspicion, absence of specific signs, and symptoms, coupled with the late symptom presentation. Only one-third 34%) of the affected neonates present within 72 hours of life , 13. Thi4% of theIt is recommended therefore that malaria parasite testing be included in the routine screening of febrile babies with suspected septicaemia in malaria-endemic regions . To maxiP. falciparum, respectively. PCR could have been useful in differentiating between congenital or neonatal malaria since it is very sensitive compared to MRDT and peripheral smear.Use of peripheral blood smear with microscopy is the routinely used diagnostic modality for diagnosis of malaria in developing countries , but witP. falciparum should be treated, be it symptomatic or asymptomatic because it is associated with high mortality and morbidity [Malaria due to orbidity . Howeverorbidity , 10. Artorbidity . WHO recorbidity . This hoorbidity , as was The prognosis of congenital malaria is variable, depending on when intervention is initiated. In Burkina Faso, Nagalo and colleagues reported that 11.8% of CM-related deaths occurred within an average of 4.8 days from admission, of which 55% of these deaths occurred within 24 hours of admission . HoweverCongenital malaria should be considered as a differential for sepsis in neonates presenting with unexplained fever and failure to breastfeed. There is need to raise awareness of this condition so as to increase diagnostic suspicion and investigatory habit among patients suspected for neonatal sepsis.


Congenital obstructive nephropathy (CON) leads to renal fibrosis and chronic kidney disease. The aim of the study was to investigate the predictive value of urinary endoglin, periostin, cytokeratin-18, and transforming growth factor-β1 (TGF-β1) for assessing the severity of renal fibrosis in 81 children with CON and 60 controls. Children were divided into three subgroups: severe, moderate scars, and borderline lesions based on 99mTc-ethylenedicysteine scintigraphy results. Periostin, periostin/Cr, and cytokeratin-18 levels were significantly higher in the study group compared to the controls. Children with severe scars had significantly higher urinary periostin/Cr levels than those with borderline lesions. In multivariate analysis, only periostin and cytokeratin-18 were independently related to the presence of severe and moderate scars, and periostin was independently related to borderline lesions. However, periostin did not differentiate advanced scars from borderline lesions. In ROC analysis, periostin and periostin/Cr demonstrated better diagnostic profiles for detection of advanced scars than TGF-β1 and cytokeratin-18  and periostin for detecting borderline lesions than endoglin and periostin/Cr . In conclusion, periostin seems to be a promising, non-invasive marker for assessing renal fibrosis in children with CON. CK-18 and TGF-β1 demonstrated low utility, and endoglin was not useful for diagnosing advanced scars. Congenital anomalies of the kidneys and urinary tract (CAKUT) are the leading cause of chronic kidney disease (CKD) in children, and a large part of them is related to congenital obstructive nephropathy (CON) ,2. The pWidely used imaging studies such as ultrasonography (US) and scintigraphy and currently available laboratory indices of the kidney injury such as serum creatinine (Cr), cystatin C, estimated glomerular filtration rate (GFR), and urinary albumin excretion do not provide information about the early stage of renal fibrosis and are poor predictors of the future course of CON ,7. ThereTGF-β1 is a 25 kDA well-known cytokine that plays a pivotal role in renal fibrosis. It is secreted by epithelial cells, macrophages, and fibroblasts as an inactive latent complex . After aEndoglin is a 180 kDa homodimeric protein consisting of extracellular, transmembrane, and intracellular domain. Endoglin expression was identified in endothelial, smooth muscle and mesangial cells, macrophages, and fibroblasts ,18,19. APeriostin is a 90 kDa matricellular protein with a multi-domain structure ,26,27. ICytokeratin-18 (CK-18) is a 45 kDA cytoskeletal protein consisting of three domains ,35. It iTo the best of our knowledge, there are no studies in the available literature investigating the usefulness of urinary endoglin, periostin, and CK-18 in children with CON. Therefore, the aim of our study was to investigate the predictive value of urinary endoglin, periostin, CK-18, and TGF-β1 for assessing the severity of renal fibrosis in children with congenital obstructive nephropathy.p = 0.05.This single-center cross-sectional study included 81 children with congenital obstructive nephropathy who were diagnosed at the Department of Pediatrics and Nephrology of the Medical University of Warsaw, Poland. The study was performed from July 2016 to April 2019. The inclusion criteria were age from 6 months to 18 years and presence of obstructive nephropathy: ureteropelvic junction obstruction (UPJO), ureterovesical junction obstruction (UVJO), and posterior urethral valves (PUV). The exclusion criteria were occurrence of another CAKUT history of urinary tract infection (UTI) during the last 6 months, surgery in the last 12 months, fibrotic disease of other organs, and acute infections (temporary exclusion). Sixty healthy children were included in the control group. The study design is displayed on the flow diagram in The study was approved by the local Bioethics Committee for Human Research (No. KB/152/2016) before the initiation. The clinical investigation was conducted in accordance ethical standards of the institutional research committee and with the guidelines of the 2013 Declaration of Helsinki. Informed consent was obtained from all participant representatives and participants (≥16 years) prior to study inclusion.In all participants, the following clinical data were evaluated: age (years), sex, body height (cm), body weight (kg), and blood pressure measurement.2), according to the revised 2009 Schwartz formula [In all participants, the following laboratory data were evaluated: serum Cr (mg/dL), serum cystatin C (ng/L), urinary albumin (mg/L), and urinary Cr (mg/dL). Urinalysis and urine culture were performed to rule out urinary tract infection (UTI). Estimated GFR  kits from Reagent Genie, Dublin, Ireland: TGF-β1-Cat. No. HUFI 00248, Human TGF-β1/TGF-beta-1; endoglin-Cat. No. HUFI 00064, Human CD105/Endoglin; periostin-Cat. No. HUDLO 0236 Human Periostin (POSTN); cytokeratin-18-Cat. No. HUFI 02320, Human CK-18/KRT18/Cytokeratin-18. The urinary TGF-β1,endoglin, periostin and CK-18 levels were normalized to the urinary Cr levels, measured from the same urine samples, and expressed as ng/mg Cr.In all patients, abdominal ultrasonography and 99mTechnetium-ethylenedicysteine (99mTc-EC) scintigraphy were evaluated at the study entry. US was performed using Philips Epiq 5G device  in B-mode. Pelvic dilatation was classified based on the anterior–posterior diameter (mm) (APD) as follows: severe APD—>20 mm, moderate APD—10–20 mm, and mild APD—<10 mm ,43.99mTc-EC was conducted to evaluate renal scarring and relative renal function (RRF). Children were injected with an adequate dose of 99mTc-EC for the child′s age and body weight. Two experienced nuclear medicine specialists evaluated radionuclide scans. Renal scars were diagnosed as an abnormal parenchymal layer, irregular renal contour, disorders of tracer placement and uptake . RRF of Voiding cystourethrography (VCUG) was evaluated at the moment of CON diagnosis. Children with VUR were excluded from the study, except those with secondary VUR as a consequence of PUV.t-test or the Mann–Whitney U-test was used to compare two groups of variables. ANOVA test or Kruskal–Wallis test and post hoc test were performed to analyze differences between three subgroups. The relationship between variables was evaluated using the Pearson or the Spearman′s rank correlation. Odds ratio (OR), including 95% confidence interval (CI), were calculated by univariate and multivariate logistic regression analysis to identify variables associated with the presence of renal scars. Variables associated with renal scars in the univariate analysis were included in the multivariate model. Variables that correlated with each other with r > 0.600 were excluded from the regression model to avoid collinearity. Receiver operating curve (ROC) analysis was used to calculate the area under the curves (AUC) for laboratory variables and to find the best cut-off values (including 95% CI), sensitivity, and specificity in the detection of renal scarring for each variable. p values less than 0.05 were considered statistically significant for all tests.Statistical analysis was performed using Statistica 13.3 PL software . The results were presented as the mean ± standard deviation or the median, interquartile ranges based on the Shapiro–Wilk and the Lilliefors normality test results. The Student′s Clinical and laboratory data in the study group and in the control group are shown in Children with CON had significantly higher levels of cystatin C and ACR, significantly lower GFR, and significantly higher urinary periostin, CK-18, and periostin/Cr compared to the controls. Groups did not differ in terms of age, sex, serum Cr and other urinary biomarkers. The levels of serum Cr were slightly elevated in 6 (7.4%) children, cystatin C in 15 (18.5%) children, and ACR in 15 (18.5%) children. Arterial hypertension was diagnosed in four children who were treated with calcium channel blocker (three children) and angiotensin-converting enzyme inhibitor (one child). Fourteen (17%) children were treated surgically because of a significant obstruction in urine outflow.p = 0.012; 0.102  vs. 0.028  p = 0.001, respectively) and significantly lower levels of GFR compared to the control group . The groups did not differ in terms of other laboratory indices of kidney injury and urinary biomarkers (data not shown).Comparison of laboratory variables with the results of renal scintigraphy in the study group is shown in Normal RRF (45–55%) demonstrated 49 (60.5%) of children and decreased RRF < 45% 32 (39.5%) of children. RRF less than 10% was found in 2 (2.5%) of children, RRF 10–19% in 4 (5%) of children, 20–39% in 13 (16%) of children, and 40–44% in 13 (16%) of children. Correlations of urinary biomarkers with RRF in the scintigraphy in the study group are shown in Correlations between urinary biomarkers and widely used laboratory indices of kidney injury in the studied children are shown in Inter-correlations of normalized urinary biomarkers in the studied children are shown in Univariate and multivariate logistic regression analysis of laboratory variables related to the presence of severe and moderate scars are shown in Cystatin C, GFR, periostin, and endoglin/Cr were related to the presence of borderline lesions in univariate analysis, whereas in multivariate analysis, cystatin C and periostin were independently related to borderline lesions. Endoglin/Cr was excluded from multivariate analysis due to collinearity.Diagnostic usefulness of laboratory variables for diagnosing severe and moderate renal scars in the scintigraphy is shown in Our cross-sectional single-center study investigated the predictive value of new biomarkers, such as endoglin, periostin, and CK-18, for assessing the severity of renal scarring in children with CON. We also evaluated TGF-β1, the most recognized fibrotic factor, which served as our reference (“gold standard”). Here, we report that urinary periostin, periostin/Cr, and CK-18 were significantly higher in children with CON compared to the healthy controls. In addition, children with severe scars had significantly higher urinary periostin/Cr levels than those with borderline lesions. Thus, periostin and periostin/Cr demonstrated a better diagnostic profile for diagnosing renal scars than other evaluated biomarkers. However, both periostin and periostin/Cr did not differentiate severe and moderate scars from borderline lesions. Endoglin showed moderate usefulness for diagnosing borderline lesions, and CK-18 and TGF-β1 revealed low utility for diagnosing severe and moderate scars.Multiple clinical studies confirmed the usefulness of urinary TGF-β1 for the diagnosis of urinary tract obstruction and tubulointerstitial fibrosis. Children with obstructive uropathy demonstrated significantly higher TGF-β1 levels than those with non-obstructive hydronephrosis and healthy controls ,45,46. AIn contrast, our study found no differences in urinary TGF-β1 and TGF-β1/Cr levels between children with CON and healthy controls. Palmer et al. reported elevated TGF-β1 levels only in the pelvis of the obstructed kidney but not in the bladder urine . Some auThe importance of endoglin in renal fibrosis was documented in several animal models of renal fibrosis. For example, the model of tubulointerstitial fibrosis induced by unilateral ureteral obstruction (UUO) demonstrated an increase in endoglin mRNA and protein expression in the obstructed kidney ,19,50. MIn our cohort of pediatric patients, we demonstrated no differences in urinary endoglin and endoglin/Cr levels between children with CON and the controls, as well as between subgroups with severe, moderate scars, and borderline lesions. Only endoglin/Cr was related to the presence of borderline lesions. In ROC analysis, endoglin showed moderate utility as a biomarker for diagnosing borderline lesions. However, its specificity was very high. We did not evaluate L-Eng and S-Eng isoforms. Therefore, we do not know if an association of endoglin with borderline lesions is due to the upregulation of pro-fibrotic L-Eng or anti-fibrotic S-Eng, especially in children with borderline lesions.Recent studies have identified periostin as a novel key factor in the progression of kidney disease. Overexpression of periostin following renal injury was described in different models of renal fibrosis, such as UUO, 5/6 nephrectomy, hypertensive renal injury, and renal I-R injury ,51,52. HIn the present study, we demonstrated significantly higher urinary periostin and periostin/Cr levels in children with CON compared to the controls and significantly higher periostin/Cr levels in children with severe scars compared to those with borderline lesions. Both periostin and periostin/Cr demonstrated relatively high utility for diagnosing scars and moderate utility for diagnosing borderline lesions. However, based on diagnostic profile and cut-off in ROC analysis, periostin did not allow for differentiation of severe and moderate scars from borderline lesions.It is difficult to explain why periostin was of no use to differentiate scars from borderline lesions, especially that we showed significantly higher levels of periostin/Cr in children with severe scars compared to those with borderline lesions. Although we confirmed that periostin is strongly related to renal fibrosis, we still know too little about the regulation of periostin expression and its signaling pathways . Recent CK-18 is a cell-protective and stress-responsive protein that seems to be a novel sensitive marker of renal tubular cell stress and tubular injury. Experimental studies demonstrated an increase in CK-18 expression in different models of renal injury. In the model of UUO, upregulation of CK-18 occurred very early after UUO induction and increased with renal injury progression. Additionally, the model of progressive GN with secondary tubulointerstitial injury and fibrosis showed only slight CK-18 expression in the early stage of disease and significantly higher expression in the late stage. Overexpression of CK-18 in renal biopsy was reported in the majority of renal tubules in the cast, diabetic, and lupus nephropathy, as well in the Bowman capsule in crescentic necrotic GN .In this study, we found significantly higher urinary CK-18 levels in children with CON compared to the controls, but no differences were observed in terms of CK-18 and CK-18/Cr between the three subgroups. Urinary CK-18 was associated with the presence of scars, but its diagnostic value was low. In the study by Djudjaj et al., elevated urinary CK-18 levels were found in animals with adenine nephropathy and Alport syndrome and in patients with AKI . Roth etIt is not known whether urinary fibrotic biomarkers are associated with clinical and laboratory indices of kidney injury. In the ESCAPE trial, GFR inversely correlated with urinary TGF-β1 . SimilarWe found positive correlations of the normalized biomarkers with cystatin C and ACR, negative correlations with serum Cr, and no relation with GFR except endoglin/Cr. These negative correlations with serum Cr were probably related to our study′s relatively large number of young children. In infants and young children, low serum Cr levels, due to small muscle mass in this age, and greater Cr tubular reabsorption lead to low urinary Cr excretion . It resuIn our study, we also evaluated the association of widely used laboratory indices of kidney injury with the presence of renal scars and borderline lesions. We found that only ACR was able to differentiate children with scars from those with borderline lesions. However, its diagnostic value was relatively low. The ESCAPE trial also demonstrated inter-correlation of urinary TGF-β1 with other biomarkers that participated in the progression of tissue injury in CKD . In lineSome studies evaluated the correlation of the intensity of renal fibrosis and urinary biomarkers with RRF of obstructed kidney in a nuclear renal scan. Zhang demonstrated a correlation between the intensity of renal fibrosis in renal biopsy performed during pyeloplasty and RRF of affected kidney . In contUrinary tract obstruction leads to a reduction in functioning nephrons and to the compensatory adaptation of the remaining. Therefore, it may affect RRF results . In addiOur study has some limitations. This single-center research may be associated with the bias occurrence. The cross-sectional design of the study prevents from drawing definitive and causative conclusions. The number of analyzed patients in subgroups was relatively low. Heterogenicity of patients′ ages, various causes of obstructive nephropathy, and different patient follow-up may impact the results. In addition, we have not analyzed the fibrosis markers in other groups of patients, e.g., in glomerulopathies.To the best of our knowledge, this study, for the first time, demonstrates that periostin, endoglin, and CK-18 are associated with renal fibrosis in children with congenital obstructive nephropathy. Periostin showed a higher diagnostic profile for diagnosing renal scars than other evaluated biomarkers. However, periostin did not differentiate advanced scars from borderline lesions. CK-18 and TGF-β1 demonstrated low utility, and endoglin was not useful for diagnosing advanced scars. In our opinion, periostin seems to be a promising, non-invasive marker for the assessment of renal fibrosis in children with CON. However, future studies on more patients are needed to confirm our results.


The overlap of malaria and Hepatitis B Virus (HBV) infections present a major threat to public health throughout endemic countries of tropical and sub-Saharan Africa. There is a paucity of data on the prevalence and associated factors of malaria and HBV infections among pregnant women in Ejule, a semiurban area of Nigeria. Therefore, the current study was designed to assess the seroprevalence of malaria and HBV among pregnant women attending antenatal clinics in Ejule Metropolis. Plasmodium falciparum  and HBsAg using histidine-rich protein 2 (HRP2) and hepatitis B surface antigen (HBsAg) rapid diagnostic tests (RDTs), respectively. Relevant sociodemographic and putative risk factor information was obtained with structured questionnaires.  In a hospital-based cross-sectional study, blood samples collected from 200 apparently healthy pregnant women at the Ilemona Clinic were screened for P. falciparum monoinfection and HBV monoinfection and coinfection, respectively. Single and concurrent infections peaked at ages 31–40 years but decreased with older ages. High P. falciparum, 31 (59.62%), and HBV 2 (3.85%) infection were observed among those without formal education. Contrary to ages, occupation, and knowledge of infection, malaria parasitemia differed significantly with lower educational qualification (p ≤ 0.001), being single (p=0.001), and inconsistent use of insecticide-treated bed nets (ITNs) . History of blood donation  and multiple sex partners  were found to be significantly associated with hepatitis B surface antigenemia rate during pregnancy. No evidence of HBV infection was observed in women with a history of HBV vaccination.  The prevalence of the infections was 44 (22%), 5 (2.5%), and 1 (0.5%) for  Malaria is still highly prevalent among pregnant women due to high illiteracy and noncompliance to using ITNs. Therefore, routine screening and educating pregnant mothers are crucial in eliminating malaria in endemic settings. The low rate of hepatitis B and coinfection with malaria shows that further improvement in HBV vaccination could considerably reduce the disease burden among pregnant women. Malaria remains a disease of major public health concern globally. A world health report in 2020 showed malaria was responsible for about 229 million cases and 409 000 deaths worldwide . CountriHepatitis B virus (HBV) is the leading cause of preventable liver disease and death globally , 5. HBV Plasmodium vis-à-vis blood-to-blood contact, sharing needles, and blood transfusions are high. Asymptomatic malaria parasitic infection and full-blown illness with typical febrile paroxysms are the heights of the wide spectrum of clinical diseases caused by malaria parasites. People who live in malaria-endemic areas have a greater chance of developing asymptomatic malaria due to their acquired semi-immunity with submicroscopic levels of parasitemia. Coinfection and pregnancy-induced immunosuppression may likely modulate the clinical progression and severity of malaria [P. falciparum [Malaria and HBV coinfection may pose a serious public health problem among pregnant women in developing countries where both infectious diseases are endemic . Despite malaria . Both malciparum  and rapilciparum . The mutlciparum . In Kogilciparum –15. Howehttps://www.city-facts.com/ejule). Indigenes are predominantly Igala with agriculture as a major occupation. A total of 200 randomly selected apparently healthy consenting antenatal attendees aged 15–51 years were recruited at Ilemona Clinic in Ejule from September to November 2019. The hospital runs weekly services for antenatal women and is the reference hospital in the area. The number of visits to Ilemona Clinic is estimated at 180 patients/week. In this cross-sectional study, all subjects signed an informed consent form before being enrolled in the study and agreed with the publication of the findings related to the study. For participants younger than 18 years of age, their parents or guardian signed the consent form. Ethics approval for the study was obtained from the hospital management board on health issues after the due process was followed .The study was carried out at Ejule, which is situated in the eastern part of Kogi State found in the north-central region of Nigeria. Ejule town has a projected population of 24,831 people in 2020 from the 2006 population census  mL of blood was aseptically collected from each of the 200 consenting subjects by the venepuncture method into a well-labeled anticoagulant tube. The samples were transported to the microbiology department under cold conditions. The blood was spun at 3000 rpm for 8–10 minutes to separate sera from whole blood. Sera were stored at −20°C until screened for HBsAg and malaria parasite antigen. Relevant demographic variables  and putative risk factor information  were obtained as appropriate with a closed-ended structured questionnaire with the assistance of a local language translator.P. falciparum was used to test for malaria parasites in blood samples. The RDT kit for P. falciparum was used since the species is the most predominant in Nigeria [The commercial HBsAg test kit  was employed for the qualitative detection of hepatitis B surface antigen in serum. The rapid diagnostic test (RDT) kit  for  Nigeria . Analysip value of ≤0.05 was considered statistically significant.Statistical Packages for Social Sciences (SPSS) version 16 for windows  was used to analyze data obtained in the study. Chi-square statistics was used to compare the difference between proportions. Logistic regression analysis at a 95% confidence interval was used to determine odds ratios used to measure putative risk factors of HBV and malaria transmission. A p > 0.05). In comparison with the other occupational status, women who practiced farming had a higher prevalence of malaria (25.76%) and HBV (3.03%) infection followed by housewives with HBsAg and malaria parasite positivity of 2.94% and 22.55%, respectively. A dual infection (0.5%) was observed in a woman practicing farming. A statistically significant difference was observed between the occupation of subjects and malaria/HBV coinfection (p=0.03). There were more malaria, 31 (59.62%), and HBV 2 (3.85%) infections among women who lacked formal education. Except HBV and HBV/malaria coinfection , the prevalence of malaria alone differed significantly with the educational level of subjects (p ≤ 0.001). Generally, a decreasing trend in HBsAg and/or malaria parasitemia was observed as the level of education increases. More single women (42.86%) were infected with malaria than married (17.56%) and divorced (0.0%) women. There was a statistically significant difference between marital status and malaria infection (p=0.001). Similarly, more single women (8.57%) were infected with HBV than married (1.21%) and divorced (0%) women, and the difference between marital status and hepatitis B surface antigenemia rates was significant (p=0.03) . Knowledge of HBV infection prevention tends to play a role in the study as about 3% of the seropositive subjects were those who never had prior knowledge of viral infection. Subjects with multiple sex partners had about 12 times more chances of being infected with HBV compared to those with one or two partners . The seropositivity rates to HBV were related to subjects' immunization status as 5 (2.75%) and zero prevalence rates among nonimmunized and immunized subjects, respectively, were observed in the current study   were positive for malaria and 5 (2.5%) for HBsAg, while 1 (0.5%) sample was positive for both malaria parasite and HBsAg . The age(p=0.03) . Previount study . A great10–0.47) .Malaria and hepatitis B virus infections are both common and life-threatening infectious diseases in developing countries including Nigeria, and the endemicity of these infections overlaps frequently due to geographical coincidence. Their high prevalence as monoinfection shows their infectious ability as coinfections. In a coinfected individual, both infections can influence each other from a clinical perspective because of some common developmental stages within the hepatocytes, which had been likened to impaired clearance of the liver stages of malaria parasites due to hepatocytes damage in hepatitis B virus infection , 16.P. falciparum infections observed in the abovementioned literature.In the current study, the prevalence of malaria, hepatitis B, and coinfections was 22.0%, 2.5%, and 0.5%, respectively. The malaria prevalence in our study is comparable with the national prevalence rate of 25% , 22.4% iP. falciparum monoinfection among the less-educated women might be due to their poor lifestyle orchestrated by their inability to take adequate measures in infection control and prevention. These findings corroborate the previous report of Anabire et al. [The high rate of e et al.  in NorthThe rationale for the high malaria infection among women whose occupation was farming could be due to their poor awareness about infection control. Our findings are supported by the assertion of Okolo et al.  and OmatP. falciparum/HBV coinfection rate of 0.5% in the current study is lower than the prevalence rates of 4.3%, 6%, and 0.7% earlier reported in parts of Nigeria [Plasmodium falciparum share their life cycle in the liver suggests they could significantly influence the health and well-being of pregnant women and their fetuses in events of coinfections as has been previously reported [The  Nigeria , Gambia  Nigeria , and Gha Nigeria , respectreported . Most bareported .P. falciparum monoinfection among women aged 31–40 years might be explained by their higher-risk behaviors for infection by both Plasmodium spp. and HBV, including sexual promiscuity, fashionable tattooing, and the lack of consistent mosquito nets used for beds than other age groups [Plasmodium spp. and HBV coinfection suggested in a meta-analysis study of Kotepui and Kotepui [The high prevalence of e groups . These a Kotepui . The dec Kotepui  or decli Kotepui . AnotherP. falciparum and HBV monoinfection among single women in the study area. These could be attributed to the absence of family cover, which can increase the tendency for outdoor adventures  that predisposed people to mosquito bites or unrestricted sexual activities without protection [Plasmodium vis-à-vis blood-to-blood contact, sharing needles, and blood transfusions are not uncommon in the endemic region. The high HBV and malaria antigenemia among women with poor knowledge of HBV or malaria infection prevention in the current study has been documented in the area [P. falciparum monoinfection than consistent users, which has been reported in most studies [We observed significant otection . Women wotection  who posithe area . Consequ studies , 27. ThiP. falciparum infection and occult HBV DNA, was not performed because of financial constraints. The use of less-sensitive methods suggests a higher problem on the ground and more sensitive PCR diagnostic methods should be used. Notwithstanding, the current study provides useful baseline epidemiological information regarding P. falciparum monoinfection and coinfection with HBV that future studies can build on in the study area. Our findings suggest the need for the initiation of routine screening of pregnant women for both hepatitis B and malaria at every antenatal care visit, as this would ensure timely management of cases and prevention of any possible adverse effects that could be associated with P. falciparum/HBV coinfection.One limitation of the study is that PCR, which is a more sensitive means of identification and confirmation of P. falciparum were observed. Women with the previous receipt of HBV vaccines show relative resistance to the viral infection. To considerably and rapidly reduce the burden of malaria and further decrease coinfection with hepatitis B, there is the need for government to strengthen malaria and hepatitis B prevention programs in all socioeconomic facets of the pregnant women population.The high prevalence of malaria in the study area reaffirms its endemicity, which has been reported for most published studies in Nigeria. Low HBV monoinfection and coinfection with


The study was to determine a five-year pattern of malaria infection in Shewarobit, Northcentral Ethiopia.institutional based retrospective study was carried out to determine the prevalence of malaria infection from a five-year examination of malaria cases at Shewarobit Health Center, Ethiopia. The directory of all malaria cases reported between 2013-2017 was carefully examined and recorded. Data were analyzed using SPSS version 20.0 and the results were presented in tables and figures.the results confirmed a total of 33,932 malaria suspects were diagnosed using microscopy over the last 5 years, of which 4705 (13.9%) were positive for malaria infection. Out of 4705 positive individuals, 3074 (65.3%) were males and 1631 (34.7%) were females. Plasmodium vivax, Plasmodium falciparum, and mixed infection (both species) accounted for 44.8%, 44.1%, and 7.1% of the confirmed cases, respectively.the study demonstrated that malaria infection is a public health concern in the study area, and Plasmodium vivax was the predominant species. Thus, the district health bureau and other concerned stakeholders should strengthen evidence-based intervention of malaria control strategies to eliminate malaria infection. Plasmodium [Malaria is a protozoan disease caused by parasites of the genus asmodium . It is oasmodium . Accordiasmodium , 4. It iasmodium . The difasmodium .P. falciparum and P. vivax are the dominant parasites responsible for the majority of malaria cases in Ethiopia [P. falciparum is the dominant parasite species in malaria epidemic situations, and this species causes severe and complicated manifestations and almost all malaria deaths [The burden of malaria continues to cause a substantial number of morbidity and mortality which accounts for most outpatient visits, and it has been one of the main causes of hospitalization and deaths in the country , 8. ApprEthiopia , 10. Thia deaths .Effective malaria control strategies were implemented in different parts of the country , there iP. vivax and P. falciparum exist in the area with widespread throughout the year.The study was conducted in Shewarobit town which is located in North Shewa Zone, Amhara regional state, Northcentral Ethiopia. It is located 225 km to the North of Addis Ababa with geographic coordination of 9°50'16" and 10°01' 32" North latitude and 39° 52' 58", and 39° 54' 9" East longitude with an area of 992.5 hectares in the rift valley with an altitude of 1280 meters above sea level. According to the City Administration Agriculture Office document, the geography of the town is mainly plain (79%), plateau (15.5%), up and down (3%), and swamp area (2.5%). The climate is tropical and has an annual temperature range from 28-37°C and the annual rainfall of the town is 1000 mm. The main rainy season usually occurs from June to September. Demographic data revealed that the town has nine kebeles with a total population of about 50,528 of which 24,638 (48.8%) were men and 25,890 (51.2) were women . Most of the dwellers of Shewarobit town are merchants, employers, daily laborers, and urban farmers. All Kebeles are malarious and infection is the most prevalent seasonal disease in the area in health center and September up to November and from June to August, October to December is the climax malaria infection transmission season in the area. Both A health facility-based retrospective study was conducted to determine the trends of malaria burden during the past 5 years at Shewarobit Health Center. The target populations for the study were all malaria suspected individuals who had visited the health center from January 2013 to December 2017.Inclusion and exclusion criteria: since it was retrospective study, a total of the five years (January 2013 to December 2017) suspected and confirmed individuals with malaria cases reports were included in the study, and all incomplete data and many febrile cases that possibly received treatment either from health extension workers or self-medication were excluded.P. falciparum positive, P. vivax positive, or mixed infection. Patients with positive peripheral blood smears were offered anti-malarial treatment as per national guidelines [A five-year (2013-2017) retrospective data on the trend of malaria prevalence were carefully reviewed from the institutional malaria registration book. The parameters such as date of examination, total clinically treated and confirmed cases in months and years, types of malaria species, and socio-demographic data such as age and sex were collected. Data were collected by experienced medical laboratory technicians. Any data such as socio-demographic, and malaria diagnosis results that were not properly documented were excluded. Throughout the reviewed period, microscopy was used as the gold standard for the detection and species identification of Plasmodium parasites by examination of peripheral smears of stained blood films, as per the WHO protocol. The health center strictly follows the standard operating procedures for capillary and venous blood sample collection, smear preparation, staining, and blood film examination for malaria parasite detection in all phases of quality control. The blood slides were read and then classified qualitatively as either negative, idelines .Malaria suspected individual: any person with a fever or fever with headache, chills, rigor, back pain, sweats, myalgia, nausea, and vomiting arebeing diagnosed clinically as malaria [ malaria .A confirmed malaria case: is any suspected case of malaria confirmed by microscopy for the Plasmodium parasite [parasite .The population at risk: population living in a geographical area in which locally acquired malaria cases occurred in the current and/ or previous years.Malarious area: area which is located at an altitude level below 2,000 meters above sea level [ea level .During malaria data collection, any malaria intervention activities that had been taken in each year to control malaria were collected using a well-prepared checklist. There was increased attention to malaria control and preventive activities by different responsible bodies, increased awareness of the community on the use of insecticide-treated bed nets (ITNs), and other malaria control activities through health education, increased accessibility of ITNs to the community, the increment of a budget for malaria control and prevention activities . But in the study area, there were no special factors that attributed to the increased or decreased occurrence of malaria cases.The completeness of the malaria registration books in the health center was first assessed to ensure the quality of data. Then, a data collection format sheet was prepared and used for data recording. Before data extraction, data collectors were adequately trained about the data extraction. The overall process of data extraction was followed up by the investigators, where a sample of completed data collection forms was randomly selected and checked daily for accuracy, completeness, and consistency. We have also checked the number of confirmed cases with the number of suspected cases throughout the reviewed data.Plasmodium species, gender, age, and distribution by season (months). The Pearson´s Chi-square test was used to describe the associations of variables. Test of significance was estimated assuming a at 0.05 and a p-value less than 0.05 was considered significant. Findings were summarized using tables, line graphs, and bar charts.Data were entered using Epi Data 3.1 and analyzed using Statistical Package for the Social Science (SPSS 20.0). Descriptive statistics were employed to calculate frequencies and percentages of overall malaria cases, trends of malaria transmission in terms of years, Five-year retrospective data were collected from the health centers after ethical clearance was obtained from the College of Natural Sciences Institutional Ethics Review Board, Addis Ababa University, and submitted to Shewarobit Health Center, where all information gathered from the recorded data. After discussing the purpose and method of the study, a permission letter to use the data and conduct the study was obtained from the administrative office of the health center with the reference number S/R/H/C/44/017. Confidentiality of the information gathered from the surveillance database was assured, and all information was used for this study only.Within the last five years (2013-2017), a total of 33,932 blood films were requested for malaria diagnosis in Shewarobit Health Center. From these requested blood films, 4705 (13.9%) outpatients were microscopically confirmed as malaria positive. There were fluctuating trends of malaria within the last five years with the minimum 622 (8.2%) microscopically confirmed malaria positive were reported in 2016 and the maximum 1949 (23.0%) microscopically confirmed malaria positive were reported in 2017 in the study area. The data showed that malaria infection decreased from 18.0% in 2013 to 8.2% in 2016, but the infection increased in 2017 (23.0%) .Even though there were fluctuations in malaria infection and, malaria cases occurred in almost every month of the year in the study area. The highest peak of malaria infection was mainly recorded from September up to November months in all years, followed by the months of June up to August and the minimum numbers of malaria cases were recorded during the months of December up to May in the study area .According to the record review, in the last five years (2013-2017) in the study area, males were more affected than females by malaria infections each year. Out of 4705 positive outpatients, 3074 were males and 1631 were females. The infection rates among males were 65.3% and females were 34.7%. The highest infections of males were occurred in 2014 (72.2%) and females were occurred in 2015 (37.9%) and the least infections of males were occurred in 2015 (62.1%) and females were in 2014 (27.8%) .The age-specific prevalence rate of malaria was 492 (10.5%) for the age group ≤ 4 years old, 1170 (24.9%) for the age group between 5-14 years old, and 3043  and the least infections were occurred in 2016 (6.8%). For the age groups between 5 and 14 years old, the highest infections were occurred in 2013 (30.3%), and the least infections occurred in 2016 that (18.8%), and for the age group =15 the highest infections were occurred in 2016 (74.4%) and the least infections were occurred in 2013 (54.3%) in the study area .P. vivax, followed by 2077 (41.1%) by P. falciparum and 332 (7.1%) were infected by mixed infection. The dominant Plasmodium species in the study area was P. vivax  of the outpatients were infected by P. vivax .P. falciparum, P. vivax, and mixed infections than females. The outpatients were highly infected by P. falciparumin 2015 (63.6%), among these 272 (66.5%) were males and 137 (33.5%) were females. The outpatients were highly infected by P. vivax in 2013 (78.5%), among these 548 (65%) were males and 295 (36%) were females, and the outpatients were highly infected by a mixed infection in 2017 (10.1%) of these 141 (71.9%) were males and 55 (28.1) were females. Concerning the age groups, the age groups ≤ 4 years old were highly infected by P. falciparum in 2014 (12.1%), by P. vivax in 2013 (17.3%), and by mixed infection in 2015 (16.2%). The age groups between 5 to 14 years old were highly infected by P. falciparum in 2014 (27.6%), by P. vivax in 2013 (32.2%), and by mixed infection in 2016 (27.2%). Finally, the age groups ≤ 15 years old were highly infected by P. falciparum, P. vivax, and mixed infection in 2016 with the infection of 269 (76%), 178 (72.3%), and 16 (72.8%) respectively  of which 4705 (13.9%) of the samples were microscopically confirmed as malaria positives. The data confirmed that malaria infection was decreased from 2013 18.2%) to 2016 (8.2%). However, the highest malaria cases were recorded in 2017 (23.0%). This might be due to the proper implementation of malaria control strategies. However, high malaria infection was recorded in 2017 (23.0%) might be due to the creation of mosquito breeding sites (stagnant water) as a result of unfinished construction. The prevalence of malaria infection in Shewarobit Health Center was 13.9%. It is higher than the results of other studies conducted in Woreta Health Center 5.4% , Arsi Ne of which.2% to 20The highest peak of malaria infections was recorded in September, October, and November months of the year followed by June, July, and August, and the minimum numbers of malaria cases were recorded from December to May in the study area. This might be due to the environmental and climatological situations that permit the continual breeding of vectors in permanent breeding sites during these months. The result of this study is in line with the study findings in Aresi Negelle Health Center , Woreta Plasmodium species with the predominant species were P. vivax, which accounted for 2296 (48.8%) of the overall prevalence followed by P. falciparum, accounting for 2077 (44.1%) and mixed infection accounted for 332(7.1%). The results of the study coincide with the study findings reported from Aresi Negelle Health Center [P. vivax (74%), P. falciparum (19.8%) and (6.2%) mixed infection, in Hallaba Health Center [P. vivax (70.41%), P. falciparum (23.08%) and (6.51%) mixed infection, in butajira [P. vivax (86.5%), P. falciparum (12.4%) and (1.1%) mixed infection. The reasons why P. vivax dominates P. falciparum might be due to the relapsing characteristics of P. vivax at the time of relatively malaria-free seasons, climate variability, and developed resistance to the currently used drug chloroquine [P. falciparum 53.7%, P. vivax 42.4%, and 3.9% mixed infection, in Kombolcha which reported the prevalence of P. falciparum 60.2%, P. vivax 35.5% [P. falciparum and P. vivax, in South West Ethiopia which reported the prevalence of P. falciparum 54.6%, P. vivax 41.6% and 3.8% mixed infection (both species) [One of the most important findings relates to the distribution of h Center  P. vivaxh Center , P. vivalciparum .08% and lciparum .8% and (ax 35.5%  which respecies) . The useP. vivax followed by P. falciparum. Notably, there was a slight reduction of malaria infection from 2013 to 2016. However, malaria infection was high in 2017, and males of productive age were mostly affected which results in an economical loss. Thus, these findings call for urgent evidence-based interventions to eliminate the infection.Malaria infection is a serious health problem in Shewarobit with the predominant of Malaria is a serious health problem in most lowland areas of Ethiopia with the predominant species is Plasmodium falciparum followed by Plasmodium vivax;The distribution of malaria infection is seasonal and varies agroecological;The factors associated with malaria infection were varied from place to place and study setting.The present study revealed that Plasmodium vivax is the predominant malaria infection in Shewarobit, Northcentral Ethiopia;The data showed that there was great variation in the distribution of malaria infection within each kebele of the Shewarobit town; The research also highlighted the current situation of the village has an impact on the distribution of malaria infection;The study also emphasized the age of the participants, living status, income level, long-lasting insecticidal nets, Indoor Residual Spray, and mosquito breeding sites are the major determinants of malaria infection.


Plasmodium parasite is dependent on the part of its life cycle that occurs in mosquitoes, to eliminate the possibility of malaria infections, transmission-blocking strategies against the mosquito stage should be the first choice. In fact, after the gametocyte enters the mosquito body, it undergoes a series of transformation processes over a short period, thus providing numerous potential blocking targets. Many research groups have carried out studies based on targeting the blocking of transmission during the mosquito phase and have achieved excellent results. Meanwhile, the direct killing of mosquitoes could also significantly reduce the probability of malaria infections. Microorganisms that display complex interactions with Plasmodium, such as Wolbachia and gut flora, have shown observable transmission-blocking potential. These could be used as a biological control strategy and play an important part in blocking the transmission of malaria.Malaria is still the most widespread parasitic disease and causes the most infections globally. Owing to improvements in sanitary conditions and various intervention measures, including the use of antimalarial drugs, the malaria epidemic in many regions of the world has improved significantly in the past 10 years. However, people living in certain underdeveloped areas are still under threat. Even in some well-controlled areas, the decline in malaria infection rates has stagnated or the rates have rebounded because of the emergence and spread of drug-resistant malaria parasites. Thus, new malaria control methods must be developed. As the spread of the  Malaria is still the most prevalent parasitic disease affecting humans globally, with approximately 228 million cases and 405,000 deaths per year . Althougvia attacking the sporozoites  pre-erythrocytic vaccines targeting the sporozoites and liver stages; (2) blood-stage malaria vaccines targeting the asexual blood stages; and (3) transmission-blocking vaccines (TBVs) targeting the sexual stages and mosquito midgut antigens . Currentrozoites . In addirozoites . Thereforozoites . In addirozoites . TBVs carozoites . The sturozoites .However, it is inevitable that the development of vaccines is an extremely long process that requires time to overcome many difficulties and technical obstacles. In fact, several studies have shown that some biological control strategies could also provide solutions to blocking the transmission of malaria . ThroughWolbachia and the mosquito intestinal flora. We believe that these findings may provide a reference for the development of malaria transmission-blocking strategies in the future.In this review, we mainly focus on transmission-blocking drugs, the results arising from the design of TBVs and the challenges that need to be overcome in the subsequent period, as well as summarizing existing biological blocking strategies, which are mainly based around Although ACT treatment is very effective in killing the asexual blood stage malaria parasites and curing patients, it cannot completely remove mature gametocytes from the blood. Therefore, malaria patients treated with ACT remain infectious to blood-sucking mosquitoes for 1–3 weeks . Primaquvia inhibiting ookinete formation and oocyst maturation  . Atovaquturation . Many otturation . Howeverturation . Moreoveturation .Malaria causes alarming morbidity and mortality in more than 100 countries worldwide. On studying the vaccine development of malaria, it was observed that pre-erythrocytic vaccines can only protect residents in areas of low-endemicity from becoming infected, and blood-stage malaria vaccines have been designed to reduce the severity of the clinical disease . Thus, Tvia mosquitoes. In the erythrocytic stage, < 1% of Plasmodium parasites commit to forming gametocytes in infected red blood cells. After male and female gametocytes are ingested in a mosquito blood meal, they break out of their red blood cells and initiate the sexual stage of parasite development. Therefore, from a life history perspective, blocking malaria gametocytes from transferring to mosquitoes is the first important step in reducing global malaria transmission.Throughout malaria infection progression, the asexual forms are responsible for clinical malaria, while the sexual stages are responsible for continued transmission in vivo now  protein family, and the most studied members of this family are Pfs48/45 and Pfs230 . Knockouvivo now . Producit choice . In addit choice . Similart choice . In addit choice .In addition, this vaccine is unusual in that the antibodies cannot kill the gametocytes in the red blood cells of malaria patients; it is only during the mosquito’s blood meal from an infected host that the antibodies can efficiently act on the gametes after they have emerged from their host red blood cells, thus preventing infection in the mosquito and halting the spread of malaria . In the Furthermore, intact mature gametocytes do not undergo phagocytosis mediated by monocytes, benefiting malaria transmission. However, red blood cells containing gametocytes that are not transmitted to mosquitoes during infection will die and then be eliminated by phagocytic cells. Therefore, the analysis of the molecular mechanisms in this phagocytic process and subsequent immune response may help to identify more targets for transmission-blocking strategies .When gametocytes are ingested by mosquitoes, they transform into gametes as the environment changes . Each fePlasmodium berghei, named Pb22, is located on the plasma membrane of gametes and ookinetes during gamete-to-ookinete development. A recent study showed that the exflagellation of male gametes (~89%) and ookinete numbers (~97%) were significantly reduced after Pb22 was knocked out, and these defects were rescued in parasites when Pb22 was restored. Further analysis showed that the defects of the Pb22 knockout (KO)line were limited to the male gametes, and the female gametes in Pb22-KO line were fertile at the wild-type level, indicating that Pb22 was indeed a key protein in the development of male gametes. Of the Pb22-KO line male gametocytes, 30% failed to assemble axonemes, whereas ~ 48.9% formed flagella but failed to egress from the host erythrocytes , a member of the fibrinogen-related protein family , has been shown to be involved in the infection process of squitoes . Previousquitoes . In thisood meal . This suIn particular, the exploration into blocking strategies during this stage need to consider a particular problem. A recent study showed that when the intensity of mosquito infection was extremely high, starvation conditions reduced the activity of RNA polymerase III in the oocysts, leading to a decrease in the growth and maturation rate of the oocysts. However, when the mosquitoes were given another blood meal, the number of oocysts was completely restored . As the Plasmodium vivax circumsporozoite and P25 proteins was included in a combination vaccine, significant protective effects (43%) and blocking effects (82%) were observed simultaneously  and pathogen inhibition  using CI to kill mosquitoes and (2) relying on the transmission-blocking ability of Wolbachia in the mosquito stage of Plasmodium development. Through CI, Wolbachia-infected females produce viable Wolbachia-infected offspring when they mate with uninfected males or males infected with the same Wolbachia strain. However, Wolbachia-infected males only produce viable offspring when they mate with females infected with the same Wolbachia strain. Meanwhile, the molecular mechanism of CI has been recently elucidated: CI causes embryonic death when a male expressing the prophage WO genes, cifA and cifB, mates with an uninfected female or a female infected with an incompatible Wolbachia strain. In contrast, embryonic development could be rescued in mosquito females harboring a compatible cifA-expressing strain   . Moreovetissues) , suggestWolbachia is controlled by itself but not manipulated by humans; apart from understanding how it infects parasites, we also need to explore whether pathogen escape occurs under infection conditions. To date, although no evidence of pathogen escape was found in wild populations of Wolbachia, pathogens seem to be able to survive under infection by certain laboratory strains, suggesting that we need to remain vigilant and take precautions . This is problematic given the possibility of amplifying prokaryotic 16S rRNA genes from non-living cells , the Scientific Research Project of Army Medical University (2021XJS05), the Graduate Scientific Research Innovation Project of Chongqing China (CYB20191) and the Natural Science Foundation of Chongqing China (cstc2018jcyjAX0182).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.