Effects of Malaria and HIV Infection on Anemia and T-cells Levels in Children in Douala City, Cameroon.

Introduction Human immunodeficiency virus (HIV) infection and malaria are priority health issues for sub-Saharan Africa. Both diseases worsen each other through their effect on the immune and hematological systems. This study aimed to determine the effects of HIV infection and asymptomatic malaria on anemia and T-cells counts in children in the city of Douala in the republic of Cameroon. Method From May to November 2016, 197 HIV infected and 98 HIV-free non-febrile children up to 19 years old (128 male and 167 female) participated in the study. All HIV-infected children were receiving antiretroviral treatment and co-trimoxazole. Malaria diagnosis was performed using Giemsa-stained thick blood film; immunological and hematological parameters were assessed through a flow cytometer and an automated analyzer respectively. Chi-squared or Fischer's exact tests was used to compare the proportions, Mann-Whitney and ANOVA tests were used for the means. Statistical significance was set at p˂0.05. Results The prevalence of malaria was 8.8%, and that of anemia was 40.7%. CD4+-T cells were higher in malaria-infected children, both in HIV positive and negative (p=0.049). No significant association was found between malaria parasitemia and CD8+-T cell levels, both in HIV-positive and negative children (p=0.41). Anemia was higher in HIV-positive children (p=0.019), especially in those with severe immunosuppression (p=0.001) and in younger children (p=0.0083). Children on HIV treatment presented lower malaria prevalence (8.6% versus 10.10%), though the difference was not significant (p=0.7068). Malaria infection was associated with lower hemoglobin levels (10.5±1.7 versus 11.2±1.4; p=0.016). Conclusion Malaria infection may enhance CD4+-T cells. Both malaria and HIV infection lead to a drop in hemoglobin levels. The HIV treatment protocol may reduce malaria prevalence.

Green-synthesized metal nanoparticles for mosquito control: A systematic review about their toxicity on non-target organisms.

Studies capturing the high efficiency of green-synthesized metal nanoparticles (NPs) in targeting mosquito vectors of the world's main infectious diseases suggest the NPs' possible utilization as bio-insecticides. However, it is necessary to confirm that these potential bio-insecticides are not harmful to non-target organisms that are often sympatric and natural enemies of the vectors of these diseases. In this systematic review, we comprehensively analyse the content of 56 publications focused on the potentially deleterious effects of NPs on these non-target organisms. Current research on biosynthesised NPs, characterization, and impact on mosquito vectors and non-target larvivorous organisms is reviewed and critically discussed. Finally, we pinpoint some major challenges that merit future investigation. Plants (87.5%) were mainly used for synthesizing NPs in the studies. NPs were found to be spherical or mainly spherical in shape with a large distribution size. In most of the included studies, NPs showed interesting mosquitocidal activity (LC50 < 50 ppm). Some plant families (e.g., Meliaceae, Poaceae, Lamiaceae) have produced NPs with a particularly high larvicidal and pupicidal activity (LC50 < 10 ppm). Regarding non-target organisms, most of the studies concluded that NPs were safe to them, with boosted predatory activity in NP-treated milieu. In contrast, some studies reported NP-elicited adverse effects (i.e., genotoxic, nuclear, and enzymatic effects) on these non-target organisms. This review outlines the promising mosquitocidal effects of biosynthesized NPs, recognizing that NPs' potential usage is currently limited by the harm NPs are thought pose to non-target organism. It is of utmost importance to investigate green NPs to determine whether laboratory findings have applications in the real world.

Comparison of the fluorescence microscopy Cyscope® with light microscopy for malaria diagnosis in a small and active surveillance in Cameroon.

BACKGROUND: Malaria has a negative impact on the activities of companies in endemic countries especially in Cameroon. In this regard, an increasingly growing number of companies have started to include management of malarious patients in their health policies. In the present study, we will evaluate the diagnostic performances of a fluorescence microscopy (FM), Cyscope® microscope, in the detection of malaria parasites. METHODS: A cross-sectional study was conducted among employees of two companies of the town of Douala on 21 and 22 March 2017. Sociodemographic information of employees was collected using a questionnaire form. Blood samples of ~ 10 µL were collected by venipuncture for the diagnosis of malaria using FM and light microscopy (LM). Performances of FM with respect to sensitivity (Se), specificity (Sp), positive and negative predictive values (PPV and NPV), positive and negative likelihood rates (PLR and NLR), accuracy, reliability, and Kappa index were calculated using LM as gold standard. RESULTS: In total, 442 employees, aged 37.8 ± 9.7 years old on average, were included in the study. Prevalence of malaria using FM and LM was 39.2% and 17%, respectively (p < 0.01). Plasmodium falciparum and P. vivax were the two species involved in malaria infection cases. In terms of developmental stages, 68%, 45.3%, and 1.3% of employees carried gametocytes, trophozoites, and schizonts, respectively. Findings on diagnostic performances of FM were as follows: Se = 84%, Sp = 69.95%, PPV = 63.58%, NPV = 95.5%, accuracy = 89.36%, and reliability = 53.95%. Sensitivity of Cyscope® microscope increased as a function of parasitemia with values ranging from 76.92% at parasitemia between 1 and 500 parasites/µL to 91.11% at parasitemia between 501 and 5000 parasites/µL. The geometric mean parasite density was1850 parasites per µL of blood (range 1600-40,000), and most of employees (60.8%) had moderate parasitemia. The performances of FM were similar between febrile and afebrile patients. CONCLUSIONS: This study showed good performances of Cyscope® microscope and outlines that this diagnostic tool could be used in management of malaria at workplace.

A systematic review on anti-malarial drug discovery and antiplasmodial potential of green synthesis mediated metal nanoparticles: overview, challenges and future perspectives.

BACKGROUND: The recent emergence in Southeast Asia of artemisinin resistance poses major threats to malaria control and elimination globally. Green nanotechnologies can constitute interesting tools for discovering anti-malarial medicines. This systematic review focused on the green synthesis of metal nanoparticles as potential source of new antiplasmodial drugs. METHODS: Seven electronic database were used following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. RESULTS: A total of 17 papers were included in the systematic review. 82.4% of the studies used plant leaves to produce nanoparticles (NPs) while three studies used microorganisms, including bacteria and fungi. Silver was the main metal precursor for the synthesis of NPs. The majority of studies obtained nanoparticles spherical in shape, with sizes ranging between 4 and 65 nm, and reported no or little cytotoxic effect of the NPs. Results based on 50% inhibitory concentration (IC50) varied between studies but, in general, could be divided into three NP categories; (i) those more effective than positive controls, (ii) those more effective than corresponding plant extracts and, (iii) those less effective than the positive controls or plant extracts. CONCLUSIONS: This study highlights the high antiplasmodial potential of green-synthesized metal nanoparticles thereby underscoring the possibility to find and develop new anti-malarial drugs based on green synthesis approaches. However, the review also highlights the need for extensive in vitro and in vivo studies to confirm their safety in humans and the elucidation of the mechanism of action.

Malaria infection and anaemia in HIV-infected children in Mutengene, Southwest Cameroon: a cross sectional study.

BACKGROUND: Malaria is one of the leading causes of morbidity and mortality in children and HIV infection as well as other factors may worsen the situation. This study was aimed at determining the factors influencing malaria parasite prevalence and density as well as anaemia in HIV-infected children in Mutengene, Cameroon from November, 2012 to April, 2013. METHODS: A semi-structured questionnaire was used to record information on socio-demographic factors and use of preventive measures by caregivers of HIV-infected children aged 1-15 years and of both sexes. Venous blood was collected; blood films were prepared and Giemsa-stained for parasite detection and speciation. Haemoglobin concentration was measured and the anaemic status determined. Data was analysed using Epi Info 7 software. RESULTS: A total of 234 children were studied. The overall malaria parasite prevalence was 24.8 % (58) and was significantly higher (31.9 %, P = 0 .004) in females, those who did not implement any preventive measure at all (66.7 %, P = 0.03) and children who used antiretroviral therapy (ART) (28.6 %, P = 0.02) when compared with their respective counterparts. Geometric mean parasite density (GMPD) was significantly higher (3098.4, P = 0.02) in children who presented with fever, had CD4 T cells ≥500 cells/µL (491.3, P = 0.003) and those with moderate anaemia (1658.8, P = 0.03) than their respective counterparts. Although there was no significant difference, GMPD was however higher in males (549.0); those not on ART (635.0) and highest in children <5 years old (633.0) than their respective counterparts. The overall prevalence of anaemia was 49.6 % (116). The value was significantly highest (58.3 %, P = 0.01) in the 11-15 years age group; those with CD4 T cell level 200-499 (72.7 %, P = 0.001) and children with fever (85.7 %, P = 0.01). CONCLUSION: Implementation of proper and integrated malaria preventive measures as well as frequent monitoring of anaemia on prescription of ART could likely improve the health conditions of HIV-infected children thus avoiding malaria-related morbidity and mortality.

Malarial anaemia and anaemia severity in apparently healthy primary school children in urban and rural settings in the Mount Cameroon area: cross sectional survey.

BACKGROUND: This study examines the relative importance of living in an urban versus rural setting and malaria in contributing to the public health problem of malarial anaemia (MA) and anaemia respectively in apparently healthy primary school children. METHODS: A cross-sectional study was conducted among 727 school children aged between four and 15 years living in an urban (302) and rural (425) settings in the Mount Cameroon area. Blood sample collected from each child was used for the preparation of blood films for detection of malaria parasites and assessment of malaria parasite density as well as full blood count determination using an automated haematology analyzer. Based on haemoglobin (Hb) measurements, children with malaria parasitaemia were stratified into MA (Hb<11 g/dL); mild MA (Hb of 8-10.9 g/dL); moderate MA (Hb of 6.1-7.9 g/dL) and severe MA (Hb≤6 g/dL). Evaluation of potential determinants of MA and anaemia was performed by multinomial logistic-regression analysis and odds ratios used to evaluate risk factors. RESULTS: Out of the 727 children examined, 72 (9.9%) had MA. The prevalence of MA and anaemia were significantly higher (χ2 = 36.5, P <0.001; χ2 = 16.19, P <0.001 respectively) in children in the urban (17.9%; 26.8% respectively) than in the rural area (4.2%; 14.8% respectively). Majority of the MA cases were mild (88.9%), with moderate (5.6%) and severe MA (5.6%) occurring in the urban area only. The age group ≤6 years was significantly (P <0.05) associated with both MA and anaemia. In addition, low parasite density was associated with MA while malaria parasite negative and microcytosis were associated with anaemia. CONCLUSIONS: Malarial anaemia and anaemia display heterogeneity and complexity that differ with the type of settlement. The presence of severe MA and the contributions of the age group ≤6 years, low parasite density and microcytosis to the public health problem of MA and anaemia are noteworthy.

Malaria and haematologic parameters of pupils at different altitudes along the slope of Mount Cameroon: a cross-sectional study.

BACKGROUND: Haematologic abnormalities are features in Plasmodium falciparum infection, and anaemia is an inevitable outcome. This study examines the influence of malaria status and altitude on haematologic parameters in school-aged pupils. METHODS: A cross-sectional study was conducted among 728 school pupils aged between four and 15 years at three different altitudinal ranges along the slope of the Mount Cameroon region. The investigative methods included the use of questionnaire, clinical evaluation and laboratory investigations. Blood sample collected from each child was used for the preparation of blood films for detection of malaria parasites and assessment of malaria parasite density as well as full blood count determination using an automated haematology analyzer. RESULTS: The prevalence of malaria in the study population was 33.8% and 64.2% (158/246) of these were asymptomatic (AM). Pupils in lowlands had a significantly (P <0.05) prevalence (95% confidence interval, CI) of malaria (60.6%, CI = 54.6-65.9%) than those in middle belt (29.1%, CI = 23.9-34.8%) and highlands (7.7%, CI = 6.1-9.8%), while those in middle belt had significantly higher geometric mean parasite density (475) than those in lowlands (233) and highlands (388). The prevalence of malaria was significantly higher in children that presented with fever (40.4%, CI = 33.8-47.2%) when compared with afebrile subjects (31%, CI = 27-35.2%). Pupils with AM had a higher prevalence of leucopaenia (43.7%, CI = 35.8-51.8%), microcytosis (27.2%, CI = 20.5-34.9%), hypochromasia (27.8%, CI = 21-35.5%) and thrombocytopaenia (14.9%, CI = 8.9-22.8%) when compared with those with clinical malaria (CM). All mean haematological parameters were comparable in pupils with CM and AM, except for the mean white blood cell (WBC) counts. Pupils with AM had significantly lower (P = 0.02) mean WBC counts (5.1 ± 2.5 × 109/L) than those with CM (5.9 ± 2.3 × 109/L). Age, altitude and malaria parasitaemia was of significant influence on several haematological parameters. CONCLUSION: Altitude influenced the distribution and density of malaria parasites and was of confounding influence on the haematologic profiles. These results highlight the insidious effects of AM on the haematologic components.

Malaria risk factors in women on intermittent preventive treatment at delivery and their effects on pregnancy outcome in Sanaga-Maritime, Cameroon.

Malaria is known to have a negative impact on pregnant women and their foetuses. The efficacy of Sulfadoxine-Pyrimethamine (SP) used for intermittent preventive treatment (IPT) is being threatened by increasing levels of resistance. This study assessed malaria risk factors in women on intermittent preventive treatment with SP (IPTp-SP) at delivery and their effects on pregnancy outcome in Sanaga-Maritime Division, Cameroon. Socio-economic and obstetrical data of mothers and neonate birth weights were documented. Peripheral blood from 201 mothers and newborns as well as placental and cord blood were used to prepare thick and thin blood films. Maternal haemoglobin concentration was measured. The overall malaria parasite prevalence was 22.9% and 6.0% in mothers and newborns respectively. Monthly income lower than 28000 FCFA and young age were significantly associated with higher prevalence of placental malaria infection (p = 0.0048 and p = 0.019 respectively). Maternal infection significantly increased the risk of infection in newborns (OR = 48.4; p<0.0001). Haemoglobin concentration and birth weight were lower in infected mothers, although not significant. HIV infection was recorded in 6.0% of mothers and increased by 5-folds the risk of malaria parasite infection (OR = 5.38, p = 0.007). Attendance at antenatal clinic and level of education significantly influenced the utilisation of IPTp-SP (p<0.0001 and p = 0.018 respectively). Use of SP and mosquito net resulted in improved pregnancy outcome especially in primiparous, though the difference was not significant. Malaria infection in pregnancy is common and increases the risk of neonatal malaria infection. Preventive strategies are poorly implemented and their utilization has overall reasonable effect on malaria infection and pregnancy outcome.

The CyScope® fluorescence microscope, a reliable tool for tuberculosis diagnosis in resource-limited settings.

Poor laboratory equipment and few human resources have made it difficult to implement microscopic diagnosis of pulmonary tuberculosis (TB) on a large scale basis worldwide. Three hundred sputum samples from patients in Cameroon were studied by using the CyScope®, a new light-emitting, diode-based, fluorescence microscope, to compare auramine-rhodamine fluorescence with the conventional Ziehl-Neelsen staining method. Five fluorescence protocols were tested to reduce manipulation time. Smear positivity for acid-fast bacilli with the Ziehl-Neelsen staining method was 27.7% (83 of 300) compared with 33.3% (100 of 300) with the fluorescent method. Staining time with the modified fluorescence protocol could be reduced from 21 minutes to 10 minutes. This study confirmed that the fluorescence staining method is more sensitive than the Ziehl-Neelsen staining method. It is suggested that the training of laboratory technicians on fluorescence microscopy should be scaled up for increased disease control.