Detection of antimalarial drug resistance polymorphisms in Plasmodium falciparum chloroquine resistance transporter and Plasmodium falciparum multidrug resistance 1 genes of Plasmodium falciparum found in Kano State, Nigeria

Objectives: In 2018, malaria claimed an estimated 380,000 lives in African region, with Nigeria accounting for 24.0% (91,368) of malaria deaths from the region. Mutations in Plasmodium falciparum chloroquine resistance transporter (Pfcrt) and P. falciparum multidrug resistance 1 (Pfmdr-1) genes had reduced the effective use of artemisinin combination therapy through the development of resistance to these antimalarial agents. Our study set out to determine the antimalarial drug resistance polymorphisms in Pfcrt and Pfmdr-1 genes of P. falciparum isolates among patients in Kano State, Nigeria. Material and Methods: Malaria positive samples were collected across the three senatorial districts of Kano State. The samples were amplified using nested polymerase chain reaction to detect the Pfcrt and Pfmdr-1 genes. The amplicons were sequenced and bioinformatic analysis was done using CLC Sequence viewer 8.0 and BioEdit sequence alignment editor to detect the single-nucleotide polymorphisms. Results: In the Pfcrt gene, CVIET haplotype was seen in 26.2% of the samples while only two samples showed the 86Y mutation in the Pfmdr-1 gene. All the 86Y mutations and majority of the CVIET haplotypes were detected in the patients from rural settings where some of them noted that they consumed modern and traditional (herbs) antimalarial agents. One sample was observed to have the CVIET haplotype and N86Y mutation while the other five CVIET haplotypes were seen in five separate samples. A new mutation V62A was found in the Pfmdr-1 gene as observed in one of the sample. Conclusion: It is imperative to ensure the rational use of the right antimalarial agents and employ continuous resistance surveillance/mapping to ensure synergy in malaria containment and elimination strategies.

Co-infection of Parvovirus B19 and Plasmodium falciparum among Sickle Cell Disease Patients in Benin City, Nigeria

Background: Infections by parasites, bacteria, viruses such as human parvovirus B19 amongst others, have been widely reported as contributing to high prevalence of anaemia in many populations. This study was conducted to determine the co-infection of Plasmodium falciparum and human parvovirus B19 among sickle cell disease (SCD) patients in Benin City, Edo State, Nigeria. Methodology: A total of 400 participants consisting 300 SCD patients (134 males, 166 females) and 100 (38 males, 62 females) apparently healthy subjects with haemoglobin AA (which served as control) who were contacted in homes, schools and offices, were enrolled for the study. The age of the participants ranged from 1 to 54 years. Venous blood was collected for detection of P. falciparum using Giemsa stain while parvovirus B19 was detected with enzyme linked immunosorbent assay (ELISA). Full blood count was estimated using Sysmex KX-21N haematology auto-analyzer. Results: An overall prevalence of parvovirus B19 and P. falciparum co-infection observed among SCD patients in this study was 3.0% while single infection was 14.0% for P. falciparum and 26.7% for parvovirus B19. Religion was associated with 0 to 22 fold increased risk of acquiring co-infection of P. falciparum and parvovirus B19. Gender was significantly associated with P. falciparum infection (p=0.0291) while tribal extraction, platelet index and seasonal variation were significantly associated with single parvovirus B19 or co-infection of P. falciparum and parvovirus B19 (p<0.05). Conclusion: The provision of strict regulatory policy concerning the screening of whole blood or pooled plasma before the use of blood products and transfusion of SCD patients is advocated

Efficacité et tolérance de Malacur® (dihydroartémisinine-pipéraquine) dans le traitement du paludisme simple à Plasmodium falciparum chez les sujets âgés de plus de 14 ans au Cameroun

Introduction: Les combinaisons thérapeutiques à base d'artémisinine sont depuis quelques années le traitement antipaludique de première intention dans une grande partie des pays endémiques. Des souches résistantes à différentes combinaisons ont été décelées en Asie du sud-est, ce qui oblige à une surveillance continuelle dans le monde entier. Matériels et méthode : Pour cela, à Yaoundé (Cameroun), a été réalisée une étude prospective,ouverte, non-randomisée pour évaluer l'efficacité clinique et parasitologique de l'association dihydroar- témisinine-pipéraquine (DHA-PQ) dans le traitement du paludisme simple à P. falciparum, chez les sujets âgés de plus de 14 ans. Résultats : Les résultats ont montré que 100% des 47 patients inclus dans cette analyse ont été libres de parasitémie dès le premier jour après la fin du traitement et les résultats se sont maintenus jusqu'à la fin du suivi, le 28ème jour. De la même manière 78,7% des patients ont été apyrétiques le jour après la première prise et 100% après les 3 jours de traitement. Aucun patient n'a montré d'évènement indésirable grave ni n'a abandonné le traitement pour cette raison. Conclusion : Les résultats confirment l'efficacité de l'association DHA-PQ comme traitement de première intention dans le traitement du paludisme non-compliqué à Plasmodium falciparum

Sérologie palustre: quel apport dans un pays d'endémie palustre comme la Côte d'Ivoire?

Introduction: la sérologie palustre semble avoir peu d'intérêt dans les pays d'endémie comme la Côte d'Ivoire. Cependant cet examen a été régulièrement réalisé au laboratoire de Parasitologie de l'Unité de Formation et de Recherche Sciences Médicales d'Abidjan. Le but de notre étude était d'apprécier l'apport de la sérologie palustre dans notre contexte de pays endémique.Méthodes: nous avons réalisé une étude rétrospective portant sur la sérologie palustre qui a utilisé le kit Falciparum spot-IF de Biomérieux à la recherche d'anticorps antiplasmodiaux d'isotype IgG. Elle a concerné les sérologies réalisées de janvier 2007 à février 2011 et dont les résultats étaient disponibles dans le registre.Résultats: au total, 136 patients ont été sélectionnés. L'âge moyen était de 36,3 ans avec des extrêmes de 1 an et 81 ans et un sex-ratio de 0,97. Les indications de sérologie palustre étaient variées, dominées par la splénomégalie (49,3%), les cytopénies (14,7%), la fièvre d'origine indéterminée (13,2%). La quasi-totalité des patients (98,5%) avaient des anticorps antiplasmodiaux avec un titre moyen élevé à 1057,35UI/ml. Il n'existait pas de lien entre l'âge et le titre d'Ac qui était plus élevé pour les cytopénies, les fièvres prolongées et la splénomégalie.Conclusion: la sérologie palustre a peu d'intérêt dans notre pratique courante en zone d'endémie car quelque soit le motif de la prescription, les titres étaient élevés

Association of ABO blood group and plasmodium falciparum malaria among children in the Federal Capital Territory, Nigeria

The ABO and Rhesus blood group systems are very important clinical tools that are commonly used in blood transfusion and their associations with various disease conditions have been widely reported. This study investigated the distribution of these blood group systems and assessed the association of malaria infection with the ABO blood groups among children in Federal Capital Territory; Abuja. Blood specimens from deep finger pricks of 730 children aged between 0-2 years were examined for malaria parasites using Field stains method. ABO and Rhesus blood group antigens tests were also performed using standard tile protocols. Of all the children admitted into the study; 445 were sick while 285 were apparently healthy. The prevalence of malaria parasites was significantly higher (P = 0.00047) among the sick children (69.8%) than the apparently healthy children (30.2%). The most prevalent blood group was O (55.7%) and the Rhesus D antigen was positive for 98.4% of all the children. The prevalence of blood group B among the sick children was significantly lower (P = 0.00373) than the other blood group types. There is no association between malaria infection and ABO blood groups but the prevalence of higher malaria parasite density was significantly greater (P = 0.0404) in children with blood group A (7.7%). In conclusion; blood group O was the most prevalent blood group in the study and children with blood group A appeared to be more susceptible to higher level of malaria parasitemia

Oxidant status of children infected with plasmodium falciparum malaria in Katsina metropolis, Northwestern Nigeria

Background: Malaria is a global menace caused by the transfer of a plasmodium parasite to a host by an infected anopheles mosquito. Upon infection; the overwhelmed host releases free radicals which have the capacity to induce oxidative damage by lipid peroxidation. This study was undertaken to assess the effect of malaria caused by Plasmodium falciparum on some antioxidant markers and lipid peroxidation levels in children attending hospitals in Katsina State; Nigeria. Materials and Methods: Blood samples were collected from untreated subjects upon confirmation of Plasmodium falciparum parasitaemia using the Giemsa stain technique. One hundred and sixty (160) consenting individuals (80 infected patients and 80 uninfected subjects) comprising of both sexes were randomly selected. The levels of antioxidant markers and malondialdehyde (MDA) - a lipid peroxidation marker were determined. Descriptive analysis was employed using SPSS version 16.0 and significance between groups was ascertained using students' T-test. Results: P. falciparum malarial infection significantly (p 0.05) reduced the antioxidant markers [vitamins A; C; et E; and reduced glutathione (GSH)] by 65.4%; 29.7%; 48.1%; 40.4% respectively in males and by 54.2%; 36.6%; 55.7% ; 36.6% in females when compared with values obtained from uninfected; healthy children. Conversely; lipid peroxidation levels were significantly (p 0.05) higher in children with parasitaemia than in nonparasitaemic controls. Males showed greater than 200% increase; while it increased by 138% in females. Conclusion: Our findings indicate a reciprocal relationship; where high levels of lipid peroxidation correspond to low levels of antioxidants; which may be due to over utilization of the antioxidants in order to counteract the effect of free radicals. This may be responsible for oxidative stress and consequently; tissue damage associated with pathology of malaria in Nigerian children

The Prevalence of Multi-Drug Resistance-1 Gene of Plasmodium Falciparum Malaria in Gezira State-Central Sudan

Abstract:Malaria parasite resistance to chloroquine poses a severe and increasing health problem in tropical countries. Implementing molecular markers for monitoring the drug resistance may be essential to overcome the problem. The aim of the present study is to investigate the prevalence of multi-drug resistance of p. falciparum parasite in malaria patients. Blood samples for DNA extraction were collected from the positive malaria patients. The prevalence of mutations in P. falciparum multi-drug resistant gene-1 (pfmdr-1) was detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) methods. Approximately; 74.1 of study populations are adults and 25.9 are children. Regression analysis shows a decrease in malaria incidence with increasing age. The prevalence of malaria is higher in males (58.6) compared to females (41.4). There were no statistical differences between malaria incidence and the socioeconomic level within the study population. The frequency of homozygous N/86 and Y/86 alleles were 51.7 and 37.9; respectively; and the heterozygous N/Y86 allele was 10.3.In conclusion the frequency of Pfmdr-1 N/Y86 allele among P. falciparum multi-drug resistant isolates support the hypothesis that Pfmdr-1 N/Y86 allele could be used as predictive marker to monitor multi-drug susceptibility in epidemiological surveys

Malaria Burden in Pregnancy at Mulago National Referral Hospital in Kampala; Uganda

Pregnancy-associated malaria is a major global health concern. To assess the Plasmodium falciparum burden in pregnancy we conducted a cross-sectional study at Mulago Hospital in Kampala; Uganda. Malaria prevalence by each of three measures-peripheral smear; placental smear; and placental histology was 9(35/391); 11.3(44/389); and 13.9(53/382) respectively. Together; smear and histology data yielded an infection rate of 15.5(59/380) of active infections and 4.5(17/380) of past infections; hence 20had been or were infected when giving birth. A crude parity dependency was observed with main burden being concentrated in gravidae 1 through gravidae 3. Twenty-two percent were afflicted by anaemia and 12.2delivered low birthweight babies. Active placental infection and anaemia showed strong association (OR=2.8) whereas parity and placental infection had an interactive effect on mean birthweight (P=.036). Primigravidae with active infection and multigravidae with past infection delivered on average lighter babies. Use of bednet protected significantly against infection (OR=0.56) whilst increased haemoglobin level protected against low birthweight (OR=0.83) irrespective of infection status. Albeit a high attendance at antenatal clinics (96.8); there was a poor coverage of insecticide-treated nets (32) and intermittent preventive antimalarial treatment (41.5)

Total Immunoglobulin G and IgG1 Subclass Levels Specific for the MSP-1 19 of Plasmodium falciparum are Different in Individuals with either Processing-Inhibitory; Blocking or Neutral Antibodies

Background: Some MSP-1 19 specific antibodies that inhibit merozoite invasion also inhibit the secondary processing of MSP-1. However the binding of these inhibitory antibodies can be blocked by another group of antibodies; called blocking antibodies; which recognize adjacent or overlapping epitopes; but themselves have no effect on either MSP-1 processing or merozoite invasion. These antibodies have been reported to be present in individuals living in a malaria endemic area. Methods: Blood samples were obtained from children shown to have processing inhibitory; blocking; and neutral antibodies in a previous study. Enzyme linked immunosorbent assay (ELISA); was used to determine the total IgG; IgM and IgG subtypes.Results: There was a significant difference in anti-MSP-1 19 IgG; while there was no significant difference in the anti-MSP-1 19 IgM. Only anti MSP-1 19 IgG1; amongst the IgG subtypes was significantly different between the groups. Conclusion: This study shows that antibodies against MSP-1 are different not only in specificity and function but also in the amount of total IgG and IgG subtype produced

In vitro susceptibility of Plasmodium falciparum to Monodesethylamodiaquine; Quinine; Mefloquine and Halofantrine in Abidjan (Cote d'Ivoire)

Background: Malaria is the primary cause of hospitalization in Ctte d'Ivoire. Early treatment is one of the strategies to control this illness. However; the spread of resistance of Plasmodium falciparum to antimalarial drugs can seriously compromise this strategy. Objectives: The aim of this study was to assess the in vitro susceptibility of P. falciparum to monodesethylamodiaquine and aminoalcohols in Abidjan (Ctte d'Ivoire). Methods: We assessed the in vitro susceptibility of isolates collected from patients with uncomplicated malaria by using the WHO optical microtest technique. Results: The proportions of resistance to monodesethylamodiaquine; m?floquine and halofantrine were 12.5; 15.6and 25.9; respectively. For quinine; none of isolates showed evidence of in vitro resistance. However; two isolates (6.1) had IC 50 values above 300 nM. The IC 50 of each drug was positively and significantly correlated to that of the other three drugs; and the correlation was higher between halofantrine and mefloquine. Conclusions: Our results showed that the in vitro chloroquine resistance reported in previous studies has been extended to other antimalarial drugs investigated in this study except for quinine. Therefore; it is necessary to implement a long-term monitoring system of antimalarial drug resistance. 15.6and 25.9; respectively. For quinine; none of isolates showed evidence of in vitro resistance. However; two isolates (6.1) had IC 50 values above 300 nM. The IC 50 of each drug was positively and significantly correlated to that of the other three drugs; and the correlation was higher between halofantrine and mefloquine. Conclusions: Our results showed that the in vitro chloroquine resistance reported in previous studies has been extended to other antimalarial drugs investigated in this study except for quinine. Therefore; it is necessary to implement a long-term monitoring system of antimalarial drug resistance

Plasmodium falciparum-induced kidney and liver dysfunction in malaria patients in Freetown; Sierra Leone

This study was undertaken to investigate the effect of Plasmodium falciparum infection on kidney and liver function parameters in malaria patients in Freetown; Sierra Leone. Blood samples taken from 64 malaria patients and 64 non-malaria volunteers at Abanita and Blue Shield Hospitals; Freetown Sierra Leone between January to April; 2009 were examined. Changes in serum biochemical parameters were analysed using normal range values as baseline. Serum bilirubin; alkaline phosphatase (ALP); alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations were significantly elevated in falciparum malaria patients compared to their non-malaria counterparts which is an indication of defective liver function. Most of patients with falciparum malaria also have significantly high serum concentrations of urea; creatinine; sodium and potassium showing alteration in kidney function. This study suggests that malaria parasites could be responsible for derangement of kidney and liver functions in patients and could therefore contribute to organ damage in affected individuals if not treated

Cytokine Profiles and Antibody Responses to Plasmodium Falciparum Malaria Infection in Individuals Living in Ibadan; Southwest Nigeria

Background: The ability of the host immune system to efficiently clear Plasmodium falciparum parasites during a malaria infection depends on the type of immune response mounted by the host. Study design: In a cross-sectional study; we investigated the cellular-and antibody responses in individuals with P. falciparum infection; in an attempt to identify immunological signs indicative of the development of natural immunity against malaria in Ibadan; Nigeria. Levels of IL-10; IL-12(p70); IFN-a; and IgM; IgG and IgG1-4 subclasses in the serum of 36 symptomatic children with microscopically confirmed malaria parasitaemia and 54 asymptomatic controls were analysed by ELISA. Results: IFN-a and IL-10 were significantly higher in the symptomatic children (p=0.009; p=0.025 respectively) than in the asymptomatic controls but no differences were seen for IL-12(p70). Estimated higher ratios of IFN-a/IL-10 and IFN-a/IL-12 were also observed in the symptomatic children while the asymptomatic controls had higher IL-12/IL-10 ratio. The mean concentration levels of anti-P. falciparum IgG1; IgG2; IgG3 antibodies were statistically significantly higher in the individuals 5 years of age than 5 years while anti-P. falciparum IgG3 antibodies were notably low in 5 years category. Children 5 years had higher IgM antibodies than IgG and the expression of IgG subclasses increased with age. Conclusion: Taken together; malaria infection is on a delicate balance of pro- and anti-inflammatory cytokines. The higher levels of IFN-a seen in the symptomatic children (6months) may be instrumental in immune-protection against malaria by limiting parasite replication. The observed variations in immunoglobulin subclass levels were age- dependent and exposure-related

Transmission Dynamics of Malaria in Four Selected Ecological Zones of Nigeria in the Rainy Season

Background: Two of the problems of malaria parasite vector control in Nigeria are the diversity of Anopheline vectors and large size of the country. Anopheline distribution and transmission dynamics of malaria were therefore compared between four ecotypes in Nigeria during the rainy season. Methods: Polymerase chain reaction (PCR) was used in molecular identification after morphological identification microscopically. Enzyme linked immunorsorbent assay (ELISA) was used for the blood meal analysis and sporozoite detection. Results: Five species were identified out of 16;410 anophelines collected. An. gambiae s.s made up approximately 29.2-36.6of the population in each zone. All five species acted as vectors for P. falciparum . An. gambiae s.s had the highest sporozoite rate. The most infected mosquitoes were found in the rain forest. More blood meals were taken from bovids; except the savannah forest; where 73.3were on humans and Human Blood index (HBI) was 57.3. The Entomological inoculation rate (EIR) was a mean of 13.6 ib/p but was highest in the rainforest zone. Conclusions and limitations: This study demonstrates the complex distribution of anophelines and the considerable variations in the intensity of malaria transmission in Nigeria. We highlight the need to consider diverse epidemiological situations when planning countrywide control programmes

Biphasic Clinical Course among Kenyan Children with Cerebral Malaria

Background: Cerebral malaria is the most severe neurological complication of Falciparum malaria. It is associated with a significant risk of death and neurological sequelae. A biphasic clinical picture is associated with an even greater risk of neurological sequelae. Objective:To examine the incidence and clinical characteristics of a biphasic clinical course in children with cerebral malaria and to study its relationship with outcome. Method : We undertook a retrospective study of children admitted to Kilifi District Hospital with a history of impaired consciousness and Falciparum infection between January 1994 and December 2004. We identified children with a biphasic clinical course and examined their clinical characteristics and outcome against that of those with a single clinical course. Results : Out of 587 children with cerebral malaria; 11 were found to have a biphasic clinical course often heralded by recurrence of seizures. This clinical pattern was associated with a greater incidence of neurological sequelae but no death. Conclusion: We speculate that a biphasic clinical course may occur due to recurrent seizures; co-morbidity and reperfusion of cerebral areas previously clogged by parasitized red blood cells. A prospective examination of this group may shed more light on causality and enlighten further on pathogenesis of cerebral malaria

Biological and Haematological Safety Profile of Oral Amodiaquine and Chloroquine in Healthy Volunteers with or without Plasmodium falciparum Infection in Northeast Tanzania

Amodiaquine (AQ); an effective antimalarial drug for uncomplicated malaria; has been greatly restricted after cases of life-threatening agranulocytosis and hepatic toxicity during prophylactic use. We conducted a hospital based open-label randomised clinical trial in 40 indigenous semi-immune healthy adult male volunteers with and without malaria parasites. The objective was to collect data on biological and haematological safety; tolerability; and parasitological efficacy to serve as baseline in the evaluation of the effectiveness of AQ preventive intermittent treatment against malaria morbidity in infants. Volunteers were stratified according to parasitaemia status and randomly assigned 20 participants each arm to three days treatment with either AQ or chloroquine (CQ). The level of difference of selected haematological and hepatological values pre-and post-trial were marginal and within the normal limits. Clinical adverse effects mostly mild and transient were noticed in 33.3CQ treated-aparasitaemic; 23.8of CQ treated-parasitaemic; 28.6of AQ-treated parasitaemic and 14.3of aparasitaemic receiving AQ. Amodiaquine attained 100parasitological clearance rate versus 70in CQ-treated volunteers. The findings indicate that there was no agranulocytosis or hepatic toxicity suggesting that AQ may pose no public health risk in its wide therapeutic dosage uses. Larger studies are needed to exclude rare adverse effects

Chimio-sensibilité de plasmodium falciparum aux antipaludiques au Niger

Nous avons réalisé un test thérapeutique in vivo à la chloroquine, couplé à une étude in vitro de chimiosensibilité et à une étude des gènes de mutation associés à la résistance aux antipaludiques en Septembre et Octobre 2001 à Niamey au Niger. Au total, 244 enfants ont été inclus. Une réponse clinique adéquate du traitement par chloroquine a été observée chez 78,3% des enfants de 1-15 ans,78,9% des enfants de 1-5 ans, 77,9% des enfants de 6-10 ans et 78% des enfants de 11-15 ans. L'échec thérapeutique a été constaté dans 13,1% des cas dont 9,4% d'échec thérapeutique précoce et 3,7% d'échec thérapeutique tardif. Au vu de ces résultats, le Programme National de Lutte contre le Paludisme a décidé de maintenir la chloroquine comme traitement de première intention du paludisme simple à P. falciparum à Niamey. Concernant les tests in vitro, sur les 244 souches, 26 seulement ont pu être cultivées, par défaut du transporteur. Sur ces 26 souches, 15 étaient sensibles à la chloroquine par les tests in vitro isotopiques. Concernant la sensibilité aux autres antipaludiques, 4 étaient résistantes la pyriméthamine, 5 au cycloguanil, 3 à l'atovaquone. Les tests moléculaires ont été effectués sur les souches qui avaient pu être isolées en cultures. Nous présentons ainsi les résultats de la prévalence des mutations des gènes pfcrt, dhfr et dhps et discutons ces résultats par rapport à ceux des tests classiques