Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Coinfection with Malaria in Selected States in Nigeria

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Plasmodium species are the causative agents of coronavirus disease 2019 (COVID-19) and malaria respectively with similar clinical presentations. The objective of this study is to determine the burden of co-infection of SARS-CoV-2 and malaria in the general population. Five (5 mLs) of blood samples were collected for SARS-CoV-2 and malaria parasite test. The malaria test was performed using a commercially available one-step malaria antigen Plasmodium falciparum histidine-rich protein 2 (Pf HRP-II) rapid test kit. The results of the study showed that the participants that were coinfected with SARS-CoV-2 IgG and malaria were 13 (2.5%) in Lagos, 1114 (39.1%) in Delta and 49 (2.3%) in Sokoto States. The prevalence of coinfection of SARS-CoV-2 and malaria in urban areas in Lagos, Delta and Sokoto States were 7 (2.2%), 1373 (48.1%), and 5 (0.2%) respectively. In rural areas, the prevalence of coinfection of SARS-CoV-2 and malaria in Lagos, Delta and Sokoto States were 6 (0.3%), 365 (12.8%), and 44 (2.1%) respectively in this study. This suggests that participants in the urban areas were more prone to co-infections than the rural areas in Lagos and Delta states, while it was otherwise in Sokoto State. In conclusion, the co-infection of SARS-CoV-2 and malaria was very high in Delta State compared to the other States. It is important for clinics to screen for both diseases when patients present with symptoms of malaria. This is because the infections have similar symptoms and the public is quick to assume malaria infection without diagnosing for COVID-19 and vice versa.

Clinical and epidemiological characteristics of norovirus gastroenteritis among hospitalized children in Lebanon

AIM To assess the burden of norovirus(No V) and to determine the diversity of circulating strains among hospitalized children in Lebanon. METHODS Stool samples were collected from children presenting with acute gastroenteritis to six major hospitals in Lebanon. A total of 739 eligible stool samples, testing negative for diarrhea caused by rotavirus as a possible viral pathogen, were collected between January 2011 and June 2013. A standardized questionnaire including demographic, epidemiological and clinical observations was used at the time of hospitalization of children presenting with diarrhea. Viral RNA was extracted from stool samples followed by reverse transcription polymerase chain reaction and nucleotide sequencing of a fragment of the viral protein 1 capsid gene. Multiple sequence alignments were carried out and phylogenetic trees were constructed using the MEGA 6 software.RESULTS Overall, 11.2% of stool samples collected from children aged < 5 years tested positive for No V genogroups Ⅰ(GⅠ) and Ⅱ(GⅡ). GⅡ accounted for 10.6% of the gastroenteritis cases with only five samples being positive for GⅠ(0.7%). The majority of hospitalized children showed symptoms of diarrhea, dehydration, vomiting and fever. Upon sequencing of positive samples and based on their clustering in the phylogenetic tree, 4/5 of GⅠ gastroenteritis cases were designated GⅠ.3 and one case as GⅠ.4. GⅡ.4 was predominantly detected in stool of our study participants(68%). We report a JB-15/KOR/2008 GⅡ.4 Apeldoorn 2008-like variant strain circulating in 2011; this strain was replaced between 2012 and 2013 by a variant sharing homology with the Sydney/NSW0514/2012/AUS GⅡ.4 Sydney 2012 and Sydney 2012/FRA GⅡ.4 strains. We also report the co-circulation of non-GⅡ.4 genotypes among hospitalized children. Our data show that No V gastroenteritis can occur throughout the year with the highest number of cases detected during the hot months.CONCLUSION The majority of No V-associated viral gastroenteritis cases among our participants are attributable to GⅡ.4, which is compatible with results reported worldwide.

Listeriosis Knowledge and Attitude among Pregnant Women Attending a Tertiary Health Institution, South Western Nigeria

Background: Listeriosis affects immunosuppressed individual’s especially pregnant women. Maternal infections are usually mild for the woman, but may have devastating effects on the unborn child, including miscarriage, stillbirth, preterm labour and serious neonatal illness. WHO recommends that pregnant women should be educated to avoid foods with high risk of contamination. This study seeks to evaluate knowledge and practices predisposing to listeria infections during pregnancy. Methodology: This is a cross-sectional study on pregnant women, attending the antenatal clinic of Lagos University Teaching Hospital (LUTH) using semi structured questionnaires. The knowledge score of participants was collated and analyzed using SPSS version 26. Results: The mean age of the participants was 32.07 ± 5.6 years. Forty-three (30.7%) women have had previous miscarriages with the mean no of miscarriage of 1.54. Out of 147 participants, only 20 (13.6%) admitted having heard of listeriosis while 127 (86.4%) had never heard of listeriosis and none of the participants has ever been tested for listeriosis. The Overall knowledge score was poor, and this was irrespective of age and duration of pregnancy. However, women with graduate and/or postgraduate degrees were more likely to have heard of Listeriosis (X2 = 10.88, P = 0.028). Conclusion: Our study shows a low level of knowledge about this food-borne illness, which can lead to severe illnesses in pregnant women and their unborn child. It is necessarily to educate pregnant women on the risk of listeriosis. This would be a key factor in creating and implementing accurate measures of prevention and control.

Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice

Objective: To evaluate the expression of DNA plasmid-harboring modified antibody gene that produces neutralizing human monoclonal antibodies against four serotypes of dengue virus(DENV) without enhancing activity in BALB/c mice. Methods: We constructed pFUSE-based vectors(pFUSE1 G7 C2hVH and pFUSE1 G7 C2hVL) containing genes encoding the variable domains of the heavy or light chain of the anti-dengue virus antibody 1 G7 C2, a human IgG1 that has been characterized for its neutralizing activity to DENV-1-4. Leucine(L) at positions 234 and 235 on the Fc CH2 domain in pFUSE1 G7 C2hVH was mutated to alanine(A)(LALA mutation) by site direct mutagenesis, and the new plasmid was termed pFUSE1 G7 C2hVHLALA. An equal amount of pFUSE1 G7 C2hVL and 1 G7 C2hG1-LALA plasmids were co-transfected into Chinese hamster ovary cells(CHO-K1) and a single dose of 100 μg 1 G7 C2hG1-LALA plasmid was intramuscularly injected, followed by electroporation in BALB/c mice. The secreted 1 G7 C2hG1-LALA antibodies in cell culture supernatant and mouse serum were examined for their biological functions, neutralization and enhancing activity. Results: The co-transfection of heavy-and light-chain 1 G7 C2hG1-LALA plasmids in CHO-K1 cells produced approximately 3 900 ng/mL human IgG and neutralized 90%-100% all four DENV, with no enhancing activity. Furthermore, the modified human IgG was produced more than 1 000 ng/mL in mouse serum on day 7 post plasmid injection and showed cross-neutralization to four DENV serotypes. Subsequently, antibody production and neutralization decreased rapidly. Nevertheless, the secreted neutralizing 1 G7 C2hG1-LALA in mouse serum demonstrated complete absence of enhancing activities to all DENV serotypes. Conclusions: These findings reveal that a new modified 1 G7 C2h G1-LALA expressing plasmid based on gene transfer is a possible therapeutic antibody candidate against DENV infection.

Occurrence of Multi-Drug Resistant <i>Listeria species</i>in Faecal Samples of Poultry Chickens in Rural Farms in Lagos State, Nigeria

Background: Listeriosis is a common zoonotic disease caused by a foodborne pathogen, Listeria monocytogenes. Poultry meat and products have been established as vehicles of transmission of pathogenic Listeria strains to humans. This study evaluates the occurrence of Listeria species in faeces of poultry chicken in Lagos. Methods: One hundred and fourteen pooled fresh faecal samples from cage-reared broiler chickens were collected from 12 farms in three rural areas in Lagos State from May to August 2019. All samples were analysed for Listeria species detection according to ISO11290-1 standard and confirmed using PCR assay. Susceptibility testing was performed using the Kirby-Bauer disc diffusion technique. Results: Twenty-eight (24.6%) Listeria species were detected from 114 faecal samples. The isolated Listeria species were L. monocytogenes 8 (7.0%), L. ivanovii 9 (7.9%), L. grayi 7 (6.1%) and L. innocua 4 (3.5%). There was no significant difference in the frequency of occurrence of Listeria species across the different locations (X2 = 4.98, p = 0.08). The listeria species were susceptible to Augmentin (96.4%), vancomycin (85.7%) and co-trimoxazole (82.1%), but resistant to ceftazidime (100%), tetracycline (75.0%) and ciprofloxacin (71.4%). Conclusion: This study reveals high occurrence of multi-drug resistant Listeria species in faecal samples of poultry chickens in Lagos state which may be an important vector in the contamination of the environment and transmission of antibiotic resistant Listeria species to consumers.

Humoral and cellular immunity against diverse SARS-CoV-2 variants

Since the outbreak of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in late 2019, the virus has rapidly spread worldwide. This has led to an unprecedented global pandemic, marked by millions of COVID-19 cases and a significant number of fatalities. Over a relatively short period, several different vaccine platforms are developed and deployed for use globally to curb the pandemic. However, the genome of SARS-CoV-2 continuously undergoes mutation and/or recombination, resulting in the emergence of several variants of concern (VOC). These VOCs can elevate viral transmission and evade the neutralizing antibodies induced by vaccines, leading to reinfections. Understanding the impact of the SARS-CoV-2 genomic mutation on viral pathogenesis and immune escape is crucial for assessing the threat of new variants to public health. This review focuses on the emergence and pathogenesis of VOC, with particular emphasis on their evasion of neutralizing antibodies. Furthermore, the memory B cell, CD4+, and CD8+ T cell memory induced by different COVID-19 vaccines or infections are discussed, along with how these cells recognize VOC. This review summarizes the current knowledge on adaptive immunology regarding SARS-CoV-2 infection and vaccines. Such knowledge may also be applied to vaccine design for other pathogens.

Virome in immunodeficiency:what we know currently

Over the past few years,the human virome and its complex interactions with microbial communities and the immune system have gained recognition as a crucial factor in human health.Individuals with compromised immune function encounter distinctive challenges due to their heightened vulnerability to a diverse range of infectious diseases.This review aims to comprehensively explore and analyze the growing evidence regarding the role of the virome in immunocompromised disease status.By surveying the latest literature,we present a detailed overview of virome alterations observed in various immunodeficiency conditions.We then delve into the influence and mechanisms of these virome changes on the pathogenesis of specific diseases in immunocompromised individuals.Furthermore,this review explores the clinical relevance of virome studies in the context of immunodeficiency,highlighting the potential diagnostic and therapeutic gains from a better understanding of virome contributions to disease manifestations.

Nonmuscle myosin IIA promotes the internalization of influenza A virus and regulates viral polymerase activity through interacting with nucleoprotein in human pulmonary cells

Influenza A virus(IAV),responsible for seasonal epidemics and recurring pandemics,represents a global threat to public health.Given the risk of a potential IAV pandemic,it is increasingly important to better understand virushost interactions and develop new anti-viral strategies.Here,we reported nonmuscle myosin IIA(MYH9)-mediated regulation of IAV infection.MYH9 depletion caused a profound inhibition of IAV infection by reducing viral attachment and internalization in human lung epithelial cells.Surprisingly,overexpression of MYH9 also led to a significant reduction in viral productive infection.Interestingly,overexpression of MYH9 retained viral attachment,internalization,or uncoating,but suppressed the viral ribonucleoprotein(vRNP)activity in a minigenome system.Further analyses found that excess MYH9 might interrupt the formation of vRNP by interacting with the viral nucleoprotein(NP)and result in the reduction of the completed vRNP in the nucleus,thereby inhibiting subsequent viral RNA transcription and replication.Together,we discovered that MYH9 can interact with IAV NP protein and engage in the regulation of vRNP complexes,thereby involving viral replication.These findings enlighten new mechanistic insights into the complicated interface of host-IAV interactions,ultimately making it an attractive target for the generation of antiviral drugs.

Longitudinal proteomic investigation of cOVID-19 vaccination

Although the development of covID-19 vaccines has been a remarkable success,the heterogeneous individual antibody generation and decline over time are unknown and still hard to predict.In this study,blood samples were collected from 163 participants who next received two doses of an inactivated COvVID-19 vaccine(CoronaVac)at a 28-day interval.Using TMT-based proteomics,we identified 1,715 serum and 7,342 peripheral blood mononuclear cells(PBMCs)proteins.We proposed two sets of potential biomarkers(seven from serum,five from PBMCs)at baseline using machine learning,and predicted the individual seropositivity 57 days after vaccination(AUC=0.87).Based on the four PBMC's potential biomarkers,we predicted the antibody persistence until 180 days after vaccination(AUC=0.79).Our data highlighted characteristic hematological host responses,including altered lymphocyte migration regulation,neutrophil degranulation,and humoral immune response.This study proposed potential blood-derived protein biomarkers before vaccination for predicting heterogeneous antibody generation and decline after coVID-19 vaccination,shedding light on immunization mechanisms and individual booster shot planning.

地黄苷通过稳定RPL27A缓解5-氟尿嘧啶抑制的核糖体生物合成

天然化合物地黄苷(Martynoside,MAR)在5-氟尿嘧啶(5-FU)诱导的“血虚”小鼠中表现出补血活性.然而,MAR的蛋白靶点和分子机制尚不清楚通过md-LED技术平台对MAR进行了体外单轮亲和多肽的筛选,揭示了核糖体蛋白L27a(RPL27A)是MAR的关键直接作用靶蛋白.结构及突变分析明确了MAR与RPL27A第4-5外显子编码区之间的特异性相互作用,并且R87及K116是介导结合的关键氨基酸残基.功能上,MAR减弱了5-FU诱导的骨髓有核细胞细胞毒性,增加了RPL27A蛋白稳定性,并减少了RPL27A在K92和K94位点的泛素化.靶点蛋白K116Y突变降低了其与MAR的结合,并阻断了MAR的蛋白稳定作用.此外,整合非标记定量蛋白质组学、泛素化修饰组学、转录组学和核糖体分析及功能测定,MAR恢复了RPL27A蛋白水平的同时部分挽救了5-FU损害的核糖体生物合成.具体来说,MAR增加了成熟rRNA的丰度,抑制了全局核糖体蛋白降解,并提升了核糖体丰度及翻译功能.这项工作明确了天然化合物MAR的直接作用蛋白靶点,揭示了核糖体生物合成在造血过程中的重要性,并为靶向RPL27A改善造血功能的研究开辟了新的方向.

A CRISPR activation screen identifies genes that enhance SARS-CoV-2 infection

Dear Editor,Identifying the host factors that are utilized for virus infection and mapping their cell-type expression profile can help to understand the viral tissue/organ tropism and pathogenesis.Much effort has been devoted to the identification of SARS-CoV-2 infection-dependent host factors.CRISPR-based activation(Konermann et al.,2015).

Fight or flee,a vital choice for Clostridioides difficile

Clostridioides difficile is a leading cause of healthcare-associated infections,causing billions of economic losses every year.Its symptoms range from mild diarrhea to life-threatening damage to the colon.Transmission and recurrence of c.difficile infection(CDl)are mediated by the metabolically dormant spores,while the virulence of C.difficile is mainly due to the two large clostridial toxins,TcdA and TcdB.Producing toxins or forming spores are two different strategies for C.difficile to cope with harsh environmental conditions.It is of great significance to understand the molecular mechanisms for C.difficile to skew to either of the cellular processes.Here,we summarize the current understanding of the regulation and connections between toxin production and sporulation in C.difficile and further discuss the potential solutions for yet-to-be-answered questions.

新冠病毒N蛋白与G3BP发生相分离从而促进应激颗粒解组装和病毒产生

新型冠状病毒SARS-CoV-2的爆发导致全世界数以百万计人员的感染和死亡,有效的治疗和干预措施对疫情的控制至关重要.然而到目前为止,新冠病毒逃过宿主抗病毒反应的机制尚未明确.病毒感染宿主细胞时,宿主能够迅速启动细胞的压力应答机制,终止细胞内的蛋白翻译,形成应激颗粒.本研究发现新型冠状病毒核衣壳蛋白(N蛋白)—一种在病毒组装过程非常关键的RNA结合蛋白,与宿主的G3BP蛋白(应激颗粒组装过程中关键的RNA结合蛋白)相互作用,并促进应激颗粒的解组装.N蛋白通过与G3BP发生相分离从而渗入应激颗粒中,阻断了G3BP与其他应激颗粒内蛋白的相互作用,最终破坏了应激颗粒的组装.这一N蛋白介导的相分离过程最终会促进病毒逃过宿主的抗病毒反应,从而有利于病毒的产生.

Suppressing fatty acid synthase by type Ⅰ interferon and chemical inhibitors as a broad spectrum anti-viral strategy against SARS-CoV-2

SARS-CoV-2 is an emerging viral pathogen and a major global public health challenge since December of 2019, with limited effective treatments throughout the pandemic.As part of the innate immune response to viral infection, type Ⅰ interferons(IFN-Ⅰ) trigger a signaling cascade that culminates in the activation of hundreds of genes, known as interferon stimulated genes(ISGs), that collectively foster an antiviral state.We report here the identification of a group of type Ⅰ interferon suppressed genes,including fatty acid synthase(FASN), which are involved in lipid metabolism.Overexpression of FASN or the addition of its downstream product, palmitate, increased viral infection while knockout or knockdown of FASN reduced infection.More importantly, pharmacological inhibitors of FASN effectively blocked infections with a broad range of viruses, including SARS-CoV-2 and its variants of concern.Thus, our studies not only suggest that downregulation of metabolic genes may present an antiviral strategy by type Ⅰ interferon, but they also introduce the potential for FASN inhibitors to have a therapeutic application in combating emerging infectious diseases such as COVID-19.

Gut microbiota,inflammation,and molecular signatures of host response to infection

Gut microbial dysbiosis has been linked to many noncommunicable diseases.However,little is known about specific gut microbiota composition and its correlated metabolites associated with molecular signatures underlying host response to infection.Here,we describe the construction of a proteomic risk score based on 20 blood proteomic biomarkers,which have recently been identified as molecular signatures predicting the progression of the COVID-19.We demonstrate that in our cohort of 990 healthy individuals without infection,this proteomic risk score is positively associated with proinflammatory cytokines mainly among older,but not younger,individuals.We further discover that a core set of gut microbiota can accurately predict the above proteomic biomarkers among 301 individuals using a machine learning model and that these gut microbiota features are highly correlated with proinflammatory cytokines in another independent set of 366 individuals.Fecal metabolomics analysis suggests potential amino acid-related pathways linking gut microbiota to host metabolism and inflammation.Overall,our multi-omics analyses suggest that gut microbiota composition and function are closely related to inflammation and molecular signatures of host response to infection among healthy individuals.These results may provide novel insights into the cross-talk between gut microbiota and host immune system.

Genome-wide evolution analysis reveals low CpG contents of fast-evolving genes and identifies antiviral microRNAs

Noncoding RNAs(ncRNAs) play important roles in many biological processes and provide materials for evolutionary adaptations beyond protein-coding genes, such as in the arms race between the host and pathogen. However, currently, a comprehensive high-resolution analysis of primate genomes that includes the latest annotated ncRNAs is not available. Here, we developed a computational pipeline to estimate the selections that act on noncoding regions based on comparisons with a large number of reference sequences in introns adjacent to the interested regions. Our method yields result comparable with those of the established codon-based method and phyloP method for coding genes;thus, it provides a holistic framework for estimating the selection on the entire genome. We further showed that fastevolving protein-coding genes and their corresponding 50 UTRs have a significantly lower frequency of the CpG dinucleotides than those evolving at an average pace, and these fast-evolving genes are enriched in the process of immunity and host defense. We also identified fast-evolving miRNAs with antiviral functions in cells. Our results provide a resource for high-resolution evolution analysis of the primate genomes.

ACE2-targeting monoclonal antibody as potent and broad-spectrum coronavirus blocker

The evolution of coronaviruses,such as SARS-CoV-2,makes broad-spectrum coronavirus preventional or therapeutical strategies highly sought after.Here we report a human angiotensin-converting enzyme 2(ACE2)-targeting monoclonal antibody,3E8,blocked the S1-subunits and pseudo-typed virus constructs from multiple coronaviruses including SARS-CoV-2,SARS-CoV-2 mutant variants(SARS-CoV-2-D614G,B.1.1.7,B.1.351,B.1.617.1,and P.1).

Identification of Aristolactam Derivatives That Act as Inhibitors of Human Immunodeficiency Virus Type 1 Infection and Replication by Targeting Tat-Mediated Viral Transcription

Despite the success of antiretroviral therapy(ART),efforts to develop new classes of antiviral agents have been hampered by the emergence of drug resistance.Dibenzo-indole-bearing aristolactams are compounds that have been isolated from various plants species and which show several clinically relevant effects,including anti-inflammatory,antiplatelet,and antimycobacterial actions.However,the effect of these compounds on human immunodeficiency virus type 1(HIV-1)infection has not yet been studied.In this study,we discovered an aristolactam derivative bearing dibenzo[cd,f]indol-4(5 H)-one that had a potent anti-HIV-1 effect.A structure-activity relationship(SAR)study using nine synthetic derivatives of aristolactam identified the differing effects of residue substitutions on the inhibition of HIV-1 infection and cell viability.Among the compounds tested,1,2,8,9-tetramethoxy-5-(2-(piperidin-1-yl)ethyl)-dibenzo[cd,f]indol-4(5 H)-one(Compound 2)exhibited the most potent activity by inhibiting HIV-1 infection with a half-maximal inhibitory concentration(IC50)of 1.03 lmol/L and a half-maximal cytotoxic concentration(CC50)of 16.91 lmol/L(selectivity index,16.45).The inhibitory effect of the compounds on HIV-1 infection was linked to inhibition of the viral replication cycle.Mode-of-action studies showed that the aristolactam derivatives did not affect reverse transcription or integration;instead,they specifically inhibited Tat-mediated viral transcription.Taken together,these findings show that several aristolactam derivatives impaired HIV-1 infection by inhibiting the activity of Tat-mediated viral transcription,and suggest that these derivatives could be antiviral drug candidates.

Broadly neutralizing antibodies against Omicron-included SARS-CoV-2 variants induced by vaccination

The SARS-CoV-2 Omicron variant shows substantial resistance to neutralization by infection-and vaccination-induced antibodies,highlighting the demands for research on the continuing discovery of broadly neutralizing antibodies(bnAbs).Here,we developed a panel of bnAbs against Omicron and other variants of concern(VOCs)elicited by vaccination of adenovirus-vectored COVID-19 vaccine(Ad5-nCoV).

High multiple mutations of Plasmodium falciparum-resistant genotypes to sulphadoxine-pyrimethamine in Lagos, Nigeria

Background Plasmodium falciparum-resistance to sulphadoxine-pyrimethamine(SP)has been largely reported among pregnant women.However,the profile of resistance markers to SP dihydrofolate reductase(dhfr)and dihydropteroate synthase(dhps)in the general population are varied and not frequently monitored.Currently,SP is used as partner drug for artemisinin combination therapy(SP-artesunate)in some sub-Saharan African countries or as a prophylactic drug in intermittent preventive treatment of malaria during pregnancy and infants and in seasonal malaria chemoprevention(SMC).Profiling of P.falciparum-resistant genotypes to SP is dynamic and critical in providing data that would be useful for malaria control programmes.This study assessed the profile of dhfr and dhps genes genotypes among individuals with malaria in Lagos,Nigeria.Methods Molecular markers of SP resistance were identified by nested PCR and sequenced among malaria positive dried blood spots(DBS)that were collected from individuals attending health facilities from January 2013 to February 2014 and during community surveys from October 2010 to September 2011 across different Local Government Areas of Lagos State,Nigeria.Results A total of 242 and 167 samples were sequenced for dhfr and dhps,respectively.Sequence analysis of dhfr showed that 95.5%(231/242),96.3%(233/242)and 96.7%(234/242)of the samples had N51I,C59R and S108N mutant alleles,respectively.The prevalence of dhps mutation at codons A437G,A613S,S436A,A581G,I431V and K540E were 95.8%(160/167),41.9%(70/167),41.3%(69/167),31.1%(52/167),25.1%(42/167),and 1.2%(2/167)respectively.The prevalence of triple mutations(CIRNI)in dhfr was 93.8%and 44.3%for the single dhps haplotype mutation(SGKAA).Partial SP-resistance due to quadruple dhfr-dhps haplotype mutations(CIRNI-SGKAA)and octuple haplotype mutations(CIRNI-VAGKGS)with rate of 42.6%and 22.0%,respectively has been reported.Conclusions There was increased prevalence in dhfr triple haplotype mutations when compared with previous reports in the same environment but aligned with high prevalence in other locations in Nigeria and other countries in Africa.Also,high prevalence of dhfr and dhps mutant alleles occurred in the study areas in Lagos,Nigeria five to eight years after the introduction of artemisinin combination therapy underscores the need for continuous monitoring.