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Background: Understanding the impact of malnutrition on innate immune response in Plasmodium falciparum (Pf)-infected subjects is critical for malaria control. Aims and Objectives: This study aims to investigate the nutritional status and innate immune response of Pf -infected subjects in Lagos, Nigeria. Materials and Methods: A total of 1183 patients with a history of fever or axillary temperature ≥37°C were screened microscopically for Pf at Ijede General Hospital, Lagos, Nigeria. Malnutrition was determined according to the U.S National Center for Health Statistics (NCHS) as stunting, wasting, or underweight when the Z -score is <−2 in the participants aged <20 years. Serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), and IL-12 were determined by capture ELISA while hematological parameters were measured using an automated hematology system. Results: A total of 384 volunteers were positive for Pf , of which 114 were <20 years with a median age of 10 years. Overall malaria prevalence was 20.89%. The malnutrition rate was 89.5%; 24 (21.05%) were stunted, 30 (26.32%) were underweight, and 48 (42.11%) were wasted. Pro-inflammatory cytokine responses were not affected by the type of malaria. TNF-α was higher in participants <5 years (P = 0.001) and in malnourished patients (P < 0.05). Conclusion: Together, it could be deduced that nutritional status influences Plasmodium falciparum malaria outcomes and progression pattern. [ABSTRACT FROM AUTHOR]

Objectives: The study evaluated sub-microscopic malaria infections in pregnancy using two malaria Rapid Diagnostic Tests (mRDTs), microscopy and RT-PCR and characterized Plasmodium falciparum dihydrofolate reductase (Pfdhfr) and Plasmodium falciparum dihydropteroate synthase (Pfdhps) drug resistant markers in positive samples. Methods: This was a cross sectional survey of 121 pregnant women. Participants were finger pricked, blood drops were collected for rapid diagnosis with P. falciparum histidine-rich protein 11 rapid diagnostic test kit and the ultra-sensitive Alere Pf malaria RDT, Blood smears for microscopy and dried blood spots on Whatman filter paper for molecular analysis were made. Real time PCR targeting the var acidic terminal sequence (varATS) gene of P. falciparum was carried out on a CFX 96 real time system thermocycler (BioRad) in discriminating malaria infections. For each run, laboratory strain of P. falciparum 3D7 and nuclease free water were used as positive and negative controls respectively. Additionally, High resolution melt analyses was employed for genotyping of the different drug resistance markers. Results: Out of one hundred and twenty-one pregnant women sampled, the SD Bioline™ Malaria Ag P.f HRP2-based malaria rapid diagnostic test (mRDT) detected eight (0.06%) cases, the ultra-sensitive Alere™ malaria Ag P.f rapid diagnostic test mRDT had similar outcome in the same samples as detected by the HRP2-based mRDT. Microscopy and RT-PCR confirmed four out of the eight infections detected by both rapid diagnostic tests as true positive and RT-PCR further detected three false negative samples by the two mRDTs providing a sub-microscopic malaria prevalence of 3.3%. Single nucleotide polymorphism in Pfdhps gene associated with sulphadoxine resistance revealed the presence of S613 mutant genotypes in three of the seven positive isolates and isolates with mixed wild/mutant genotype at codon A613S. Furthermore, four mixed genotypes at the A581G codon were also recorded while the other Pfdhps codons (A436G, A437G and K540E) showed the presence of wild type alleles. In the Pfdhfr gene, there were mutations in 28.6%, 28.6%, and 85.7% at the I51, R59 and N108 codons respectively. Mixed wild and mutant type genotypes were also observed in 28.6% each of the N51I, and C59R codons. For the Pfcrt, two haplotypes CVMNK and CVIET were observed. The SVMNT was altogether absent. Triple mutant CVIET 1(14.3%) and triple mutant + wild genotype CVIET + CVMNK 1(14.3%) were observed. The Pfmdr1 haplotypes were single mutants YYND 1(14.3%); NFND 1(14.3%) and double mutants YFND 4(57.1%); YYDD 1(14.3%). [ABSTRACT FROM AUTHOR]

Background: Routine surveillance for antimalarial drug resistance is critical to sustaining the efficacy of artemisinin-based Combination Therapies (ACTs). Plasmodium falciparum kelch-13 (Pfkelch-13) and non-Pfkelch-13 artemisinin (ART) resistance-associated mutations are uncommon in Africa. We investigated polymorphisms in Plasmodium falciparum actin-binding protein (Pfcoronin) associated with in vivo reduced sensitivity to ART in Nigeria. Methods: Fifty-two P. falciparum malaria subjects who met the inclusion criteria were followed up in a 28-day therapeutic efficacy study of artemetherlumefantrine in Lagos, Nigeria. Parasite detection was done by microscopy and molecular diagnostic approaches involving PCR amplification of genes for Pf18S rRNA, varATS, telomere-associated repetitive elements-2 (TARE-2). Pfcoronin and Pfkelch-13 genes were sequenced bi-directionally while clonality of infections was determined using 12 neutral P. falciparum microsatellite loci and msp2 analyses. Antimalarial drugs (sulfadoxine-pyrimethamine, amodiaquine, chloroquine and some quinolones) resistance variants (DHFR_51, DHFR_59, DHFR_108, DHFR_164, MDR1_86, MDR1_184, DHPS_581 and DHPS_613) were genotyped by high-resolution melting (HRM) analysis. Results: A total of 7 (26.92%) cases were identified either as early treatment failure, late parasitological failure or late clinical failure. Of the four posttreatment infections identified as recrudescence by msp2 genotypes, only one was classified as recrudescence by multilocus microsatellites genotyping. Microsatellite analysis revealed no significant difference in the mean allelic diversity, He, (P = 0.19, Mann-Whitney test). Allele sizes and frequency per locus implicated one isolate. Genetic analysis of this isolate identified two new Pfcoronin SNVs (I68G and L173F) in addition to the P76S earlier reported. Linkage-Disequilibrium as a standardized association index, IA S, between multiple P. falciparum loci revealed significant LD (IA S = 0.2865, P=0.02, Monte-Carlo simulation) around the neutral microsatellite loci. The pfdhfr/ pfdhps/pfmdr1 drug resistance-associated haplotypes combinations, (108T/N/ 51I/164L/59R/581G/86Y/184F), were observed in two samples. Conclusion: Pfcoronin mutations identified in this study, with potential to impact parasite clearance, may guide investigations on emerging ART tolerance in Nigeria, and West African endemic countries. [ABSTRACT FROM AUTHOR]

Introduction: Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2. No drug has been generally approved as safe and effective for the treatment of COVID-19. Several therapeutic agents such as COVID Organics® (CVO) have been explored as treatment options. CVO is an herbal tea composed of 62% of Artemisia annua and 38% of other plants. There is presently no existing scientific report and data on the safety and efficacy of CVO herbal drug. Thus, acute and subacute toxicity studies were undertaken to evaluate the safety and toxicity of CVO on short- and long-term usage in animal models. Materials and Methods: Phytochemical and nutritional compositions of CVO were determined using standard methods. Acute oral toxicity was investigated using female Swiss albino mice (three per group). While subacute oral toxicity was done using female and male Swiss albino rats (five per group). The animals were administered 2000 mg/kg, 5000 mg/kg, therapeutic dose; 5500 mg/kg and supratherapeutic dose; 11,000 mg/kg of CVO herbal product. The control group received water ad libitum. The oral toxicity studies were done in accordance with Organization for Economic Corporation and Development guidelines. The experimental protocol was approved by the Institutional Animal Care and Use Committee, Nigerian Institute of Medical Research (Ethics No. IRB/17/043). Results: CVO is rich in antioxidants: flavonoids (10.3%), tannins (29.1%), and phenolics (434.4 mg). It contains proteins (33.8%), carbohydrates (34.5%), fat (6.8%), and fiber (0.5%). In the acute toxicity study, no mortality was recorded in all the treated and untreated groups. The lethal dose of CVO is >5000 mg/kg body weight. The hematological, biochemical, lipid profile, and histologic parameters were all normal at therapeutic doses when compared to the control group. Conclusion: The acute and subacute oral toxicity studies revealed that CVO is not toxic. The specific organ toxicity evaluations also indicated that CVO has no toxic effects on blood parameters and vital organs structure and function at therapeutic dose. Thus, CVO is safe for short- and long-term usage. We recommend that CVO should be subjected to efficacy studies to investigate whether it is effective for COVID-19 treatment as claimed by the manufacturer. [ABSTRACT FROM AUTHOR]

Introduction: There are genuine concerns that long-term use of anti-retroviral drugs may be associated with reproductive complications in females. This study aimed to ascertain the effect of highly active anti-retroviral drugs on the ovarian reserve and reproductive potential of female Wistar rats and by extension to HIV-positive human females.Methods: A total of 25 female Wistar rats, weighing between 140g and 162g, were randomly allotted into non-intervention and intervention groups, receiving the anti-retroviral drugs, Efavirenz (EFV), Tenofovir Disoproxil Fumarate (TDF), Lamivudine (3TC), and a fixed-dose combination (FDC). The dosage was administered orally at 8 am daily for 4 weeks. Serum concentrations of anti-Mullerian hormone (AMH), follicle-stimulating hormone (FSH), luteinising hormone (LH), and estradiol were measured using standard biochemical techniques (ELISA). Follicular counts were made on fixed ovarian tissue from the sacrificed rats.Results: The mean AMH level for the control group and the EFV, TDF, 3TC, and FDC groups were 11.20, 6.75, 7.30, 8.27, and 6.60 pmol/L, respectively. The EFV and FDC groups had the lowest AMH, compared to the other groups, but there was no statistically significant difference in AMH across the groups. The mean count of antral follicles was significantly lower in the group that received EFV when compared to the other groups. The corpus luteal count was significantly higher in the control group than in the intervention groups.Conclusion: The study demonstrated a disruption in the reproductive hormones of female Wistar rats receiving anti-retroviral regimens containing EFV. Clinical studies are required to determine if these changes are seen in women receiving EFV-based treatment, as this may compromise reproductive function and predispose them to early menopause. [ABSTRACT FROM AUTHOR]

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Background Chemoprevention plays an important role in malaria control strategy. Perennial malaria chemoprevention (PMC) using sulfadoxine/pyrimethamine (SP) is a WHO-approved strategy to combat malaria in young children and may lead to drug pressure. Introducing SP-PMC may therefore be compromised due to the emergence of Plasmodium falciparum resistant to SP, particularly mutation at K540E of the dihydropteroate synthase (dhps) gene. Molecular surveillance of resistance markers can support assessment of antimalarial efficacy and effectiveness. High prevalence of 540E is associated with reduced effectiveness of SP, and areas with more than 50% prevalence are considered unsuitable for intermittent preventative treatment in pregnancy (IPTp) implementation. Assessing 540E prevalence is an important undertaking before implementation of SP-PMC. Methods We conducted a rapid surveillance of dhps- 540E to assess the suitability of SP as PMC in field studies from Ebonyi and Osun states in Nigeria. We used an in-house developed amplicon deep-sequencing method targeting part of the dhps gene. Results Our data reveal that 18.56% of individuals evaluated carried the 540E mutation mixed with the WT K540. Mutant variant 540E alone was not found, and 80% of isolates harboured only WT (K540). Clonal analysis of the sequencing data shows a very low proportion of 540E circulating in both states. Conclusions Our data show that both states are suitable for SP-PMC implementation and, based on this finding, SP-PMC was implemented in Osun in 2022. Continuous monitoring of 540E will be required to ensure the chemoprevention effectiveness of SP in Nigeria. [ABSTRACT FROM AUTHOR]

Elimination of malaria infection with available antimalarial drugs is threatened by resistance developed by Plasmodium species and requires more efficacious and potent drugs. Some rural communities in Nigeria, use Thaumatococcus danielli leaves to cure malaria. However, a scientific investigation has not supported this assertion. In this investigation, swiss albino mice were used to test T. danielli extracts' antiplasmodial efficacy. The plant was subjected to qualitative phytochemical analysis, sohxlet extraction using n-hexane, ethyl acetate, ethanol, and aqueous solvents, including the extracts' acute oral toxicity study. Suppressive test of each extract against Plasmodium berghei (NK-65 strain) was tested at 200, 400 and 800mg/kg. Parasitaemia levels, temperature, weight, packed cell volume, and mean survival time of the mice were monitored during the study. In Windows Statistical Package for Social Sciences (SPSS) 26.0., one-way analysis of variance and Tukey's post hoc test were used to analyze the data. The acute oral toxicity study disclosed that LD50 values of T. danielli extracts were above 2,000mg/kg in mice. The maximum suppressive effect of 67.3% was achieved in a dose-dependent pattern (p < 0.001) by an oral administration of ethyl acetate extract at 800 mg/kg body weight. The extract was an ideal antiplasmodial extract since it minimized the fall in red blood cells, body temperature, and caused increased weight (p > 0.05). The plant has considerable antimalarial efficacy caused by individual or synergistic action of its phytochemicals. Therefore, bioassay of isolated and characterized active compound(s) is necessary for development of new antimalarial(s) from T. danielli leaves. [ABSTRACT FROM AUTHOR]

Recent ethnomedicinal studies on Bridelia ferruginea Benth (family Phyllanthaceae) as an antiplasmodial remedy have established its potency as a strong prophylactic and chemosuppressive agent. Human consumption of medicinal herbs without adequate evaluation of its efficacy and safety can result in grave physiological and pathological consequences. Therefore, this study assessed the antiplasmodial bioactivity, biochemical, hematological, histopathological and toxicity profile of the ethanolic stem bark extract of B. ferruginea in mice. Ethanolic stem bark extract of B. ferruginea (200, 400 and 800 mg/kg) were orally administered to Plasmodium berghei-infected mice in models and were subsequently observed for mortality, behavioral changes and signs of toxicity. Acute evaluation was experimented at 1,000 mg/kg for 28 days. Occult blood obtained from the euthanized mice were subjected to biochemical and hematological assays. A comprehensive assessment of the histology of the liver and kidney was also ascertained. The median lethal dose (LD50) was determined and extrapolated using the regression equation obtained from the plot of the probits of mortalities (y) and the log of doses (log10C). Different concentrations of the phytochemical secondary metabolites were revealed. Antiplasmodial bioactivity was established at the 200, 400 and 800 mg/kg of the herbal extract with a dearth in parasitemia at different days post-treatment. The 800 mg/kg group responded by exhibiting a dose-dependent decrease in parasitemia comparable with the chloroquine bi-phosphate group. Significant alterations in the histology of the liver and kidney of the 1,000 mg/kg group was documented. There was a reduction in the titers of LDH, ALT, AST, and urea in the treated group when compared with the control (p < 0.05). Antioxidant profiles were also highly significant with elevation in SOD, GPx, and CAT, but a reduction in MDA. LD50 was established at 424 mg/kg. B. ferruginea Benth (family Phyllanthaceae) is a potent antiplasmodial, antioxidant, regenerative and ameliorative herbal remedy if administered in controlled dosage. [ABSTRACT FROM AUTHOR]

Abstract: Malaria transmission and prevalence involves a triangular web of interactions between man, vector, and the environment. Any meaningful effort in malaria control, elimination and or eradication should target weakening and or breaking the forces of interactions within the triangle. In sub-Saharan Africa, effective malaria control programme is encumbered by myriad of challenges. The unabated burden of malaria could be ascribed to efficient malaria vectors with strong niche for ecological expression that maintain high levels of transmission at all seasons. Parasite genotypic heterogeneity, multiple expressions of traits of adaptations to parasitism and unpredictable behavioural changes are the smart ways the infectious agent thrives, persists and expresses ecological niche with scaring symphony. Environmental factors and climatic changes, population movement, deteriorated socioeconomic situation, lack of access to effective and timely antimalarial treatment, use of sub-standard and or fake anti-malarial drugs, self medication and non-compliance to drug dosage are the galvanizing factors to poor intervention outcomes. Above all, for ages, in most settings and communities, there is lack of knowledge regarding the causative agent of malaria. Diversionary factors and misconceptions such as eating too much palm oil, standing in the sun, drinking too much of alcohol, jinxing and witchcraft attacks are brands of porous perceptions on causes and risk factors of malaria. And if a thousand and one sophisticated strategies are put in place to eliminate malaria in sub-Saharan Africa where indigenous and factorial perceptions with rhetorical contrast of ideas hold sway, then, a predictable defeated performance is the expected result. For the sustenance of the gains already recorded in the fight against malaria in sub-Saharan Africa, the role of the community as a strong partnership for change cannot be underestimated. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the influence of –250G>A (rs2070895) polymorphism in hepatic lipase gene (LIPC) promoter on plasma lipid parameters of ischemic stroke patients. Methods: A total of 100 stroke patients and 100 control subjects matched for sex (59 men and 41 women) and age were selected. Hepatic lipase activity and lipid profiles were measured while lipoprotein ratios were calculated. Genotyping of the –250G>A promoter polymorphism of the LIPC was performed by the polymerase chain reaction and restriction fragment length polymorphism method combined with 2% gel electrophoresis and then confirmed by direct sequencing. The LIPC promoter gene sequencing data were compared with refseqNG011465.1 LIPC from GenBank. Results: The frequencies of GG, GA and AA genotypes of LIPC rs2070895 polymorphism were 39%, 45% and 16% for the control, 10%, 37% and 53% for the stroke subjects (P<0.000 1), respectively. The frequencies of G and A alleles were 61.5% and 38.5% for the control, and 28.5% and 71.5% for the stroke subjects (P<0.000 1). Our study shows that the mutant allele of the LIPC promoter was associated with dyslipidemia, lower hepatic lipase activity, and this variation contributed to the increased defective plasma high-density lipoprotein-cholesterol (HDL-C), HDL2-C and HDL3-C concentration for both subjects. The control subjects had 6 single nucleotide polymorphism and 6 amino acid substitutions while the stroke subjects had 32 single nucleotide polymorphism and 20 amino acid substitutions. Conclusions: LIPC –250G>A polymorphism can influence plasma lipid profiles and lipoprotein ratios in patients with ischemic stroke. [ABSTRACT FROM AUTHOR]

A cross sectional study was conducted to investigate the prevalence and intensity of gastrointestinal helminths in confined pigs in Ogbomoso, South-west Nigeria. A total of 345 pigs (124 (35.9%) males and 221(64.1%) females) were enrolled. Faecal samples were examined with floatation and sedimentation techniques and McMaster-counting chamber was used for egg count/gram. Our result revealed that 159 (46.0) of the examined pigs were infected with at least one helminth species of Ascarops strongylina (57.4%), Physocepahlus sexalatus (32.2%), Trichuris suis (68.7%), Ascaris suum (31.3%), Hyostrongylus rubidus (23.5%), Strongyloides ransomi (53.0%) and Taenia solium (55.7%). The infection was statistically significant (P<0.001) with sex and highest prevalence was recorded among female pigs. Mean egg count for all the identified gastrointestinal helminths were age dependent except with A. suum and S. ransomi (1742.21 ± 185.15; 95%CI 1374 - 2109.83; P<0.001). Infection in adults and growers were not comparable (36.2%) but comparable with piglets (27.5%) at P<0.001. Pigs raised in cemented floor recorded low prevalence (45.2%) compared to uncemented floor (54.8%), (P<0.001). Pigs housed on plain floor showed striking increase in parasite egg intensities when compared to pigs' house on cemented floors. There is a risk of zoonotic infections. Stakeholders can mitigate morbidity and transmission by regular visits, awareness and drug interventions. [ABSTRACT FROM AUTHOR]

Malariometric surveys generate data on malaria epidemiology and dynamics of transmission necessary for planning and monitoring of control activities. This study determined the prevalence of malaria and the knowledge, attitude, and practice (KAP) towards malaria infection in Ibeshe, a coastal community.The study took place during the dry season in 10 villages of Ibeshe. All the participants were screened for malaria. A semistructured questionnaire was used to capture sociodemographic data and KAP towards malaria. A total of 1489 participants with a mean age of 26.7 ± 20.0 years took part in the study. Malaria prevalence was 14.7% (95% CI 13.0-16.6%) with geometric mean density of 285 parasites/μL. Over 97% of participants were asymptomatic. Only 40 (2.7%) of the participants were febrile, while 227 (18.1%) were anemic. Almost all the participants (95.8%) identified mosquito bite as a cause of malaria, althoughmultiple agents were associated with the cause of malaria. The commonest symptoms associated with malaria were hot body (89.9%) and headache (84.9%). Window nets (77.0%) were preferred to LLIN (29.6%). Malaria is mesoendemic in Ibeshe during the dry season. The participants had good knowledge of symptoms of malaria; however, there were a lot of misconceptions on the cause of malaria. [ABSTRACT FROM AUTHOR]

Objective: Archived malaria rapid diagnostic test strips (mRDTs) serves as an important source of plasmodium Deoxyribose Nucleic Acid (DNA) in epidemiological studies. The presence of Plasmodium falciparum DNA (PfDNA) in mRDTs (yr. 2016-2017) and newly used ones (yr. 2018) were enumerated with a view to establish the parasite's optimum genomic DNA volume. Methods: A retrospective study to determine the yield and purity of used mRDTs was carried out on randomly selected mRDTs (2016 - 2018). Both positive and negative mRDTs samples were analyzed with nested Polymerase chain reaction (nPCR). Dried blood spots (DBS) were obtained from study enrolments and analyzed molecularly. nPCR and Agarose gel electrophoresis were used to determine P. falciparum DNA. Results: Agarose gel electrophoresis results showed that only 26 out of the 50 samples eligible for screening were PCR positive for P. falciparum. The following was observed; yrs.: 2016 - 17(34%) with 2.06 X 103 yield, 1.7235 purity; 2017 - 16(32%) with 1.03 X 103 yield, 1.7619 purity and 2018 - 17(34%) with 1.42 X 103 yield, 1.6194 purity. Molecular analysis (P.f. 18Ss rRNA) was determined to ascertain positive result that appeared negative using mRDTs or microscopy. The DNA yield of the DBS for 2018 was 1.66 X 103 and a purity (Optical Density 260/280) of 1.69. The purity was higher than that of the mRDTs with a DNA yield of 1.42 X 103 and 1.62 purity. Conclusion: PfDNA extraction is an important process for malaria PCR screening and the reliability is dependent on pureness and concentration. [ABSTRACT FROM AUTHOR]

Introduction: Histidine-rich protein 2 (HRP2) antigen kits are widely used for malaria diagnosis in tropical regions due to their heat stability. The Plasmodium Lactate Dehydrogenase (pLDH) used in BIOCREDIT® malaria test kit is claimed to be heat-stable. This study aimed to evaluate the heat stability and field performance of three BIOCREDIT® kits., Methodology: A cross-sectional community survey of malaria infection was carried out in Agbowa and Ikosi communities in Lagos State. Finger-prick blood was used to assess the performance of three BIOCREDIT® kits: Kit 1 (PfpLDH), Kit 2 (PfLDH/HRP2), and Kit 3 (PfLDH/PvLDH). Malaria microscopy and a reference mRDT (SD BIOLINE® PfHRP2) were used as comparators. Heat stability testing was performed using Plasmodium falciparum panels after exposure to 35oC and 45oC for 60 days. A total of 834 participants were recruited., Results: The sensitivity of BIOCREDIT® kits were: Kit 1 (PfLDH) 88.4%; Kit 2 PfHRP2 92% and PfLDH 86.6%; Kit 3 (PfLDH) 88.4%. The specificity of kits was: Kit 1 (Pf LDH) 91.7%; Kit 2 (PfHRP2) 82.3%) and (PfLDH) 92.1%; Kit 3 (PfLDH) 91.6%. The sensitivity and specificity of the reference kit were 92.9% and 85.9%. The three BIOCREDIT® Kits were stable. No potential case of pfhrp2 gene deletion was observed., Conclusions: The sensitivities of the three BIOCREDIT® malaria test kits were similar to the reference kit (SD BIOLINE mRDT). However, the specificities of PfLDH kits were significantly higher than the specificities of PfHRP2 kits. The BIOCREDIT® kits are suitable to be deployed in malaria-endemic tropical regions., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2024 Oluwagbemiga O Aina, Chimere O Agomo, Olusola Ajibaye, Celestina O Okoyenta, Nnenna Ogbulafor, Simon Ijezie, Akwaowo B Orok, Chinedum T Oparaugo, Samuel K Akindele, Damilare J Bamgbose, Susan A Holdbrooke, Olaide K Kareem, Yetunde A Olukosi, Isaac Ola.)

Introduction: Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2. No drug has been generally approved as safe and effective for the treatment of COVID-19. Several therapeutic agents such as COVID Organics ® (CVO) have been explored as treatment options. CVO is an herbal tea composed of 62% of Artemisia annua and 38% of other plants. There is presently no existing scientific report and data on the safety and efficacy of CVO herbal drug. Thus, acute and subacute toxicity studies were undertaken to evaluate the safety and toxicity of CVO on short- and long-term usage in animal models., Materials and Methods: Phytochemical and nutritional compositions of CVO were determined using standard methods. Acute oral toxicity was investigated using female Swiss albino mice (three per group). While subacute oral toxicity was done using female and male Swiss albino rats (five per group). The animals were administered 2000 mg/kg, 5000 mg/kg, therapeutic dose; 5500 mg/kg and supratherapeutic dose; 11,000 mg/kg of CVO herbal product. The control group received water ad libitum. The oral toxicity studies were done in accordance with Organization for Economic Corporation and Development guidelines. The experimental protocol was approved by the Institutional Animal Care and Use Committee, Nigerian Institute of Medical Research (Ethics No. IRB/17/043)., Results: CVO is rich in antioxidants: flavonoids (10.3%), tannins (29.1%), and phenolics (434.4 mg). It contains proteins (33.8%), carbohydrates (34.5%), fat (6.8%), and fiber (0.5%). In the acute toxicity study, no mortality was recorded in all the treated and untreated groups. The lethal dose of CVO is >5000 mg/kg body weight. The hematological, biochemical, lipid profile, and histologic parameters were all normal at therapeutic doses when compared to the control group., Conclusion: The acute and subacute oral toxicity studies revealed that CVO is not toxic. The specific organ toxicity evaluations also indicated that CVO has no toxic effects on blood parameters and vital organs structure and function at therapeutic dose. Thus, CVO is safe for short- and long-term usage. We recommend that CVO should be subjected to efficacy studies to investigate whether it is effective for COVID-19 treatment as claimed by the manufacturer., Competing Interests: None

Background: Recent ethnomedicinal studies on Bridelia ferruginea Benth (family Phyllanthaceae) as an antiplasmodial remedy have established its potency as a strong prophylactic and chemosuppressive agent. Human consumption of medicinal herbs without adequate evaluation of its efficacy and safety can result in grave physiological and pathological consequences. Therefore, this study assessed the antiplasmodial bioactivity, biochemical, hematological, histopathological and toxicity profile of the ethanolic stem bark extract of B. ferruginea in mice., Methods: Ethanolic stem bark extract of B. ferruginea (200, 400 and 800 mg/kg) were orally administered to Plasmodium berghei -infected mice in models and were subsequently observed for mortality, behavioral changes and signs of toxicity. Acute evaluation was experimented at 1,000 mg/kg for 28 days. Occult blood obtained from the euthanized mice were subjected to biochemical and hematological assays. A comprehensive assessment of the histology of the liver and kidney was also ascertained. The median lethal dose (LD 50 ) was determined and extrapolated using the regression equation obtained from the plot of the probits of mortalities (y) and the log of doses (log 10 C)., Results: Different concentrations of the phytochemical secondary metabolites were revealed. Antiplasmodial bioactivity was established at the 200, 400 and 800 mg/kg of the herbal extract with a dearth in parasitemia at different days post-treatment. The 800 mg/kg group responded by exhibiting a dose-dependent decrease in parasitemia comparable with the chloroquine bi-phosphate group. Significant alterations in the histology of the liver and kidney of the 1,000 mg/kg group was documented. There was a reduction in the titers of LDH, ALT, AST, and urea in the treated group when compared with the control (p < 0.05). Antioxidant profiles were also highly significant with elevation in SOD, GPx, and CAT, but a reduction in MDA. LD 50 was established at 424 mg/kg., Conclusion: B. ferruginea Benth (family Phyllanthaceae) is a potent antiplasmodial, antioxidant, regenerative and ameliorative herbal remedy if administered in controlled dosage., (© 2020 Walter de Gruyter GmbH, Berlin/Boston.)

Background: Accurate laboratory diagnosis of suspected malaria is the hallmark to the control of the disease., Aim: The clinical proficiency of commercial Rapid Diagnostic test kits (RDTs) using nested PCR as quality control was evaluated among patients attending two public healthcare providing institutions in Ilorin, Kwara state, North-Central, Nigeria., Method: A cross-sectional evaluation of finger prick blood samples of volunteer patients were accessed for malaria parasites with pLDH, HRP2, Pf, Pf/PAN and nested PCR molecular assays. The data derived were analysed using standard formulae for diagnostic accuracy, and the obtained predictive values were subjected to a comparison with one-way analysis of variance (ANOVA)., Result: Three hundred and sixty-eight (368) patients comprising 203 (55%) females and 165 (45%) males participated in this study. Routine microscopy revealed that 54 (32.7%) males and 80 (39.4%) was infected with Plasmodium falciparum. SD Bioline (pLDH) 47.4%; Carestart Malaria (HRP2) 49.8% recorded low sensitivities. Micropoint (pfPAN) 82.8% and Micropoint (Mal. Pf) 64.4% recorded a high sensitivity. SD Bioline (pLDH) 67.4%; Carestart Malaria (HRP2) 85.9%; Micropoint (PfPAN) 62.2% and Micropoint (Mal. Pf) 86.7% had high specificities. The positive predictive value (PPV) ranged from 67.7% to 85.94%, while the negative predictive values (NPV) of 64.4% for SD Bioline (pLDH); 86.7% for Carestart Malaria (HRP2); 89.3% for Micropoint (pfPAN) and 58.5% for Micropoint (Mal. Pf). Agarose gel analysis of P. falciparum ssrRNA gene (206 bp) for 28 specimens containing 10% concordant and discordant samples showed that all 12 negative specimens for RDTs and routine microscopy were truly negative for nPCR. However, the remaining 16 specimens were positive for nPCR and showed discrepancies with routine microscopy and RDTs. Cohen's interrater diagnostic measure analysis revealed that the weighted kappa for the RDTs was moderate 0.417 (p=0.027), 95%CI (0.756, 0.078) and good for nPCR 0.720 (p < 0.001), 95%CI (0.963, 0.477). The area under the curve (AUC) specify that nPCR has been more effective than the RDTs (nPCRAUC = 0.875; p < 0.001 and RDTsAUC = 0.708; p = 0.063)., Conclusion: A thorough large-scale quality control is advocated on all commercial RDTs being used in most sub-Saharan African countries. This is to avoid double jeopardy consequent upon misdiagnosis on unidentified positive cases serving as pool reservoir for the insect vector and cyclical infection and re-infection of the populace.

Background: Polymorphisms in Plasmodium falciparum merozoite surface protein-2 ( msp -2) and associated parasite genetic diversity which varies between malaria-endemic regions remain a limitation in malaria vaccine development. Pro-inflammatory cytokines are important in immunity against malaria, understanding the influence of genetic diversity on cytokine response is important for effective vaccine design., Methods: P. falciparum isolates obtained from 300 Nigerians with uncomplicated falciparum malaria at Ijede General Hospital, Ijede (IJE), General Hospital Ajeromi, Ajeromi (AJE) and Saint Kizito Mission Hospital, Lekki, were genotyped by nested polymerase chain reaction of msp -2 block 3 while ELISA was used to determine the pro-inflammatory cytokine response to describe the genetic diversity of P. falciparum ., Results: Eighteen alleles were observed for msp -2 loci. Of the 195 isolates, 61 (31.0%) had only FC27-type alleles, 38 (19.7%) had only 3D7-type alleles, and 49.3% had multiple parasite lines with both alleles. Band sizes were 275-625 bp for FC27 and 150-425 bp for 3D7. Four alleles were observed from LEK, 2 (375-425 bp) and 2 (275-325 bp) of FC27-and 3D7-types, respectively; 12 alleles from AJE, 9 (275-625 bp) and 3 (325-425 bp) of FC27-types and 3D7-types, respectively; while IJE had a total of 12 alleles, 9 (275-625 bp) and 3 (325-425 bp) of FC27-types and 3D7-types, respectively. Mean multiplicity of infection (MOI) was 1.54. Heterozygosity ( H E ) ranged from 0.77 to 0.87 and was highest for IJE (0.87). Cytokine response was higher among <5 years and was significantly associated with MOI ( P > 0.05) but with neither parasite density nor infection type., Conclusion: P. falciparum genetic diversity is extensive in Nigeria, protection via pro-inflammatory cytokines have little or no interplay with infection multiplicity., Competing Interests: There are no conflicts of interest.

Background: The accuracy of malaria diagnosis by microscopy has been a challenge in health facilities in Nigeria due to poor competence of microscopists and inability to report on malaria species other than Plasmodium falciparum. Short microscopy courses were conducted to improve the skills of laboratory personnel to perform malaria microscopy in public health facilities in Nigeria., Materials and Methods: Seven-day malaria microscopy courses were conducted annually between 2011 and 2013 for microscopists in public health facilities. The training courses contained theoretical and practical sessions. Impact of the training was evaluated by practical and theoretical pre- and post-training assessments on malaria slide reading, parasite enumeration and basic malariology., Results: The 102 participants who completed the training consisted of medical laboratory scientists (62; 60.8%), medical laboratory technicians (24; 23.5%) and other healthcare workers (16; 15.7%). The knowledge of basic malariology (theory) at pre- and post-tests were 34% (95% CI 31.7-36.3%) and 74.9% (95% CI 71.8-78.0%), respectively (P<0.001). The mean slide reading detection, species and counting agreements in pre-training assessment were 48.9%, 27.9% and 0%, respectively, and in post-training 56.8%, 39.2% and 25%, respectively. The mean species agreements in picture test pre- and post-training were 21.9% and 55.1%, respectively. There were significant differences (P<0.05) in the median pre-test scores in picture tests and basic malariology of the three categories of participants but not in malaria slide reading and parasite counting tests. However, post-training, a significant difference in test scores of the three categories of participants was recorded only for basic malariology (P=0.0003)., Conclusions: The 7-day malaria microscopy courses significantly increased the knowledge and microscopy skills of the trainees and were sufficient to bridge the significant difference in baseline microscopy skills of the different categories of trainees that participated in the training courses., Competing Interests: Competing interests: No competing interests declared., (Copyright © 2015; Olukosi et al.)

Introduction: Toxoplasma gondii infection has been described as the most widespread zoonotic infection of humans and other animals. Information concerning T. gondii infection among schoolchildren is unavailable in Lagos City, Nigeria., Methods: This cross-sectional study investigated the seroprevalence and risk factors associated with T. gondii infection among primary schoolchildren (PSC) from a community located in the center of Lagos, southern Nigeria, from November 2013 to March 2014. A total of 382 PSC were screened for the presence of sera anti-T. gondii antibodies using a latex agglutination test (TOXO Test-MT, Tokyo, Japan). A cutoff titer of ≥ 1:32 was considered positive, while titers ≥ 1:1,024 indicated high responders. Questionnaires were also used to obtain data on possible risk factors from parents/guardians., Results: The overall seroprevalence was 24% (91/382), and 83.5% (76/91) of seropositive PSC were classified as high responders. Among the risk factors tested, including contact with cats and soil, consumption of raw meat and vegetables, and drinking unboiled water, none showed statistical significance after multivariate adjustment. No associations were observed among age, gender, body mass index (BMI), and parents' occupation/educational level., Conclusions: The findings in this study show evidence of active infection, and hence, there is need for urgent preventive measures in this city. Further investigation is required to clarify the transmission routes. Policy makers also need to initiate prevention and control programs to protect pregnant women and immunocompromised patients in particular because they are more severely affected by T. gondii infection.

Background: A recovery in chloroquine efficacy following a period of cessation has raised the possibility of its reintroduction for malaria chemotherapy. We investigated the prevalence of the major markers of chloroquine resistance years after the withdrawal of the drug in Nigeria., Materials and Methods: Finger prick blood samples were collected from participants presenting with symptoms of malaria in two selected health centres each representing Lekki and Ijede communities of Lagos, Nigeria. Thick and thin blood smears were prepared for microscopy and dry blood spots made from malaria-positive participants for parasite DNA extraction. The detection of mutations in the Plasmodium falciparum chloroquine resistance transporter ( pfcrt) and P. falciparum multidrug resistance ( pfmdr1) genes was performed by nested polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP)., Results: Of the 1527 blood samples that were confirmed by PCR to be P. falciparum positive, 412 and 344 were typed for the molecular detection of pfcrt and pfmdr1 gene mutations, respectively. The mutant alleles of pfcrt were present among 290 (70%) parasite carriers while the pfmdr1 mutant allele was found in 117 (34%) of the total population. There were higher distributions of the mutant alleles for the two loci in Ijede than in Lekki. The observed frequencies of pfcrt mutant alleles in the two parasite populations were in agreement with the expected frequencies predicted by Hardy-Weinberg. In comparing data with studies conducted between 2000 and 2002 in Ijede, we observed an increase in the prevalence of mutant type pfcrt against a marginal decline in the pfmdr1 mutant type., Conclusion: The high frequencies of pfcrt mutation are suggestive of a persistent drug pressure and continuing inefficacy of chloroquine as an antimalarial drug., Competing Interests: Competing Interests: No competing interests declared., (Copyright © 2014: Olukosi et al.)