Clinical and haematological parameters in malaria caused by different plasmodium species in children

Background: Malaria is a disease of global importance and affects more than ninety countries in both the tropical and subtropical regions. Clinical and haematological parameters vary with type of malaria, although data relating to different species of malaria in children is limited. This study aims to understand the clinical and haematological profile of malaria and to correlate these with different malarial species among children.Methods: This is a descriptive cross-sectional study involving 130 proven malaria cases done over 18 months from October 2014 to April 2016. A detailed history and clinical examination along with haematological parameters were analysed and correlated with different types of malaria.Results: Among 130 children, 97 children were vivax positive, 4 were falciparum and 27 were mixed malaria. Fever was present in all, whilst other symptoms were chills and rigors (86.15%), vomiting (39.52%), headache (19%), pain abdomen (6.84%), myalgia (4.56%) and convulsions (1.52%). Clinical signs were pallor (29.64%), icterus (0.76%), splenomegaly (65.36%), hepatomegaly (23.56%) and hepatosplenomegaly (21.28%).75% of children with falciparum malaria had splenomegaly and pallor whereas hepatomegaly was observed in 34% of mixed malaria cases. Haematological parameters observed were anaemia (47.6%), severe anaemia (2%), leucocytosis (11.5%), leukopenia (39.2%), thrombocytopenia (87%) and severe thrombocytopenia (30%). Severe thrombocytopenia was seen with vivax malaria (70%). No mortality was noted in the studied population.Conclusions: Fever and splenomegaly are important clinical features, whereas anaemia and thrombocytopenia are the most noted haematological parameters in malaria. The parameters vary with different species of malaria knowledge of clinical and haematological parameters aid us in early diagnosis and prompt initiation of treatment and prevention of associated complications.

Bibliometric analysis of researches on the Plasmodium falciparum repetitive interspersed families of polypeptides protein from 1993 to 2022 / 中国血吸虫病防治杂志

Objective To perform a bibliometric analysis of researches on the Plasmodium falciparum repetitive interspersed families of polypeptides (RIFIN) protein from 1993 to 2022 and identify the hot topics in the RIFIN protein research, so as to provide insights into future researches on RIFIN protein. Methods RIFIN protein-associated publications were retrieved in the Web of Science Core Collection from 1993 to 2022 and all bibliometric analyses were performed using the software CiteSpace 6.2.4.0. The annual number of RIFIN protein-associated publications was analyzed from 1993 to 2022, and country, author and institution collaboration networks were created. Keywords were extracted from RIFIN protein-associated publications for plotting keyword co-occurrence, clustering, burst and timeline maps to identify the hot topics in the RIFIN protein research. Results A total of 745 English RIFIN protein-associated publications were included in the final bibliometric analysis, and there were 18 to 36 publications each year from 1993 to 2022. The top three countries with the highest activity in the RIFIN protein research included the United States, the United Kingdom and France, universities and research institutes were highly active in the RIFIN protein research; however, no authors were identified with a high activity in the RIFIN protein research. There were three keyword clusters in the RIFIN protein-associated publications, including repetitive DNA sequence, molecular epidemiology and antigenic variation. Keyword co-occurrence, burst and timeline analyses showed that previous RIFIN protein-associated publications mainly focused on gene properties and functions, involving keywords of repetitive DNA sequence and evolution, and recent hot topics for the RIFIN protein research shifted to genetic diversity and immune response, involving keywords of genetic diversity, antigenic variation and binding. Conclusions The annual number of RIFIN protein-associated publications was relatively stable from 1993 to 2022. This bibliometric analysis may provide insights into future researches on the RIFIN protein.

Plasmodium falciparum malaria with acute abdominal pain as the first symptom: a case report / 中国血吸虫病防治杂志

Plasmodium falciparum malaria, caused by Plasmodium falciparum infection, is an Anopheles mosquito-transmitted infectious diseases, which predominantly occurs in tropical areas of Africa. P. falciparum malaria is characterized by complex and atypical clinical manifestations, and high likelihood of misdiagnosis and missing diagnosis, and may be life-threatening if treated untimely. This case report presents the diagnosis and treatment of a P. falciparum malaria case with acute abdominal pain as the first symptom.

Aspectos clínico-epidemiológicos en pacientes con malaria grave del Hospital Municipal de Cuimba, Angola / Clinical-epidemiological aspects of severe malaria in patients at the Municipal Hospital of Cuimba, Angola / Aspectos clínico-epidemiológicos da malária grave em pacientes do Hospital Municipal do Cuimba, Angola

Introducción: El Plasmodium falciparum es el causante de más del 90 porciento de los casos de malaria en el mundo. Objetivo: Describir aspectos clínico-epidemiológicos de pacientes con malaria grave, atendidos en el Hospital Municipal de Cuimba, provincia de Zaire, República de Angola. Método: Estudio observacional, descriptivo, de corte transversal y retrospectivo, durante el periodo comprendido entre enero-junio de 2023, en pacientes con diagnóstico de malaria grave. El universo fue conformado por 452 pacientes positivos de malaria, la muestra quedó conformada por 97 pacientes que desarrollaron malaria grave. Se estudiaron variables asociadas como: anemia severa, convulsiones, hiperparasitemia, entre otras. Resultados: La media de edad fue de 14,8 años, el 43,3 porciento menor de cinco años, con predominio del sexo masculino (53,9 porciento). El Plasmodium falciparum estuvo presente en 59 casos (60,8 porciento), con elevadas tasas de parasitemia. Las manifestaciones de disfunción cerebral en asociación con la anemia severa resultaron estar en el cuadro clínico del 31 porciento de los pacientes. El 40,2 porciento de los enfermos no presentó complicaciones en su estadía hospitalaria. El síndrome de dificultad respiratoria aguda (18,6 porciento) fue la complicación más frecuente que sobrellevó al fallecimiento del 12,4 porciento de los pacientes. El artesunato fue usado en 77,3 porciento de los pacientes. Conclusiones: El paludismo representa un problema de salud en el Hospital Municipal de Cuimba, con mayor frecuencia en los menores de cinco años. Prevalece la infección por Plasmodium falciparum en pacientes con anemia severa.(AU)

Introduction: Plasmodium falciparum is responsible for more than 90 percent of malaria worldwide. Objective: Characterization of Clinical-epidemiological aspects of severe malaria in patients treated at the Municipal Hospital of Cuimba, Zaire, Angola. Method: An observational, descriptive, cross-sectional and retrospective study was conducted, during the period January - June 2023, in patients reported with severe malaria. The study involved a total of 452 patients with positive malaria but only 97 of them, who presented a complication of severe malaria, were selected as sample. The variables used were as follow: severe anemia, convulsions, hyperparasitemia, among others. Results: Male sex was predominant, with an average age of 14.8 percent and 43.3 percent of patients under five years of age. Plasmodium falciparum was found in 59 patients (60.8 percent) with a high parasitaemia prevalence. Manifestations of cerebral dysfunction in association with severe anemia were found in the clinical picture of 31 percent of patients. The 40.2 percent of patients had no complications in admission period. Acute Respiratory Distress Syndrome was the most frequent complication (18.6 percent), and it was the leading cause of death in 12.4 percent of patients. Artesunate was used in 77.3 percent of patients. Conclusions: Malaria is a health problem in Municipal Hospital of Cuimba, with a higher incidence in children under five years of age; Plasmodium falciparum infection prevailed in patients with severe anemia.(AU)

Introdução: O Plasmodium falciparum é responsável por mais de 90 porcento da malária em todo o mundo. Objetivo: Caracterização dos aspectos clínico-epidemiológicos da malária grave em pacientes atendidos no Hospital Municipal de Cuimba, Zaire, Angola. Método: Foi realizado um estudo observacional, descritivo, transversal e retrospetivo, durante o período de janeiro a junho de 2023, em doentes notificados com malária grave. O estudo envolveu um total de 452 doentes com malária positiva, mas apenas 97 deles, que apresentavam uma complicação de malária grave, foram seleccionados como amostra. As variáveis utilizadas foram as seguintes: anemia grave, convulsões, hiperparasitemia, entre outras. Resultados: O sexo masculino foi predominante, com uma idade média de 14,8 anos e 43,3 porcento dos doentes com menos de cinco anos de idade. O Plasmodium falciparum foi encontrado em 59 doentes (60,8 porcento) com uma elevada prevalência de parasitemia. Manifestações de disfunção cerebral em associação com anemia grave foram encontradas no quadro clínico de 31 porcento dos doentes. Os 40,2 porcento dos doentes não tiveram complicações no período de admissão. A Síndrome de Angústia Respiratória Aguda foi a complicação mais frequente (18,6 porcento) e foi a principal causa de morte em 12,4 porcento dos doentes. O artesunato foi utilizado em 77,3 porcento dos doentes. Conclusões: A malária é um problema de saúde em Hospital Municipal de Cuimba, com uma maior incidência em crianças com menos de cinco anos de idade; a infeção por Plasmodium falciparum prevaleceu em pacientes com anemia grave.(AU)

Falciparum malaria is associated with risk markers of type 2 diabetes mellitus in individuals with or without COVID-19 exposure

Background: Scientific information on the impact of malaria on the risk of developing type 2 diabetes mellitus (T2DM) after recovery from the coronavirus disease 2019 (COVID-19) is limited in the Ghanaian context. The purpose of this study was to examine the association between selected risk markers of T2DM in falciparum malaria patients post-COVID-19 or not at a tertiary hospital in Ghana. Methodology: This was a descriptive cross-sectional comparative study of 38-recovered COVID-19 adult participants with malaria and 40 unexposed COVID-19 adults with malaria at the Tamale Teaching Hospital, Ghana. Demographic, anthropometric and levels of glucose, insulin, C-reactive protein and lipid profiles were measured in the two groups of participants under fasting conditions. Parasitaemia was assessed microscopically but insulin resistance and beta-cell function were assessed by the homeostatic model. Results: The COVID-19 exposed participants were older (p=0.035) with lower parasitaemia (p=0.025) but higher mean levels of insulin, insulin resistance, and beta-cell function compared with their unexposed counterparts (p<0.05). Parasitaemia correlated positively with a number of the measured indices of diabetogenic risk markers in the COVID-19 exposed group only, and predicted (Adjusted R2=0.751; p=0.031) by beta-cell function, C-reactive protein and triglycerides with the model explaining about 75% of the observed variation. Parasitaemia could only be predicted (Adjusted R2=0.245; p=0.002) by C-reactive protein with the model explaining just about a quarter of the observed variation in the COVID-19 unexposed group. Insulin resistance and sub-optimal beta-cell function were detected in both groups of participants. Conclusion: Falciparum malaria is associated with risk markers for development of T2DM irrespective of COVID-19 exposure. Insulin resistance, inflammation and sub-optimal beta-cell secretory function may drive the risk. The observed diabetogenic risk is higher in the recovered COVID-19 participants.

Changes in the endemic-epidemic pattern of malaria in Colombia, 1978-2021

ABSTRACT Background: Malaria is a major global public health issue with varying epidemiologies across countries. In Colombia, it is a priority endemic-epidemic event included in the national public health policy. However, evidence demonstrating nationwide variations in the disease behavior is limited. This study aimed to analyze changes in the levels and distribution of endemic-epidemic malaria transmission in the eco-epidemiological regions of Colombia from 1978 to 1999 and 2000 to 2021. Methods: We conducted a comprehensive time-series study using official secondary data on malaria-associated morbidity and mortality in Colombia from 1978 to 2021. Temporal-spatial and population variables were analyzed, and the absolute and relative frequency measures of general and regional morbidity and mortality were estimated. Results: We observed an 18% reduction in malaria endemic cases between the two study periods. The frequency and severity of the epidemic transmission of malaria varied less and were comparable across both periods. A shift was observed in the frequency of parasitic infections, with a tendency to match and increase infections by Plasmodium falciparum. The risk of malaria transmission varied significantly among the eco-epidemiological regions during both study periods. This study demonstrated a sustained decrease of 78% in malarial mortality. Conclusions: Although the endemic components of malaria decreased slightly between the two study periods, the epidemic pattern persisted. There were significant variations in the risk of transmission across the different eco-epidemiological regions. These findings underscore the importance of targeted public health interventions in reducing malarial morbidity and mortality rates in Colombia.

Study on clinical presentation of malaria and response to antimalarial treatment in children less than twelve years

Background: In the tropical country of India, malaria in pediatric age group is one of the most important cause of mortality and morbidity. The study aimed to identify the clinicoepidemiologic features and response to antimalarial treatment in the children.Methods: Prospective analytical study conducted for the same over the period of two years in tertiary care hospital (September 2019 to September 2021). All children under twelve years hospitalised with fever and were diagnosed to have malaria by Peripheral smear or RDT were included in the study after ethical clearance and parental consent. Total sample size was 90.Results: Incidence of the malarial disease among total admitted patients in our study period was found to be 3.5%. The highest percentage of patients with malaria belonged to age group of 13m to 5 years (40%) and least being the age group 10-12years (7.7%). In study fever was found to be the major chief complaint followed by chills and rigor. Bleeding diathesis was least common It was found out from our study that overall most common clinical finding in the study patients was anemia or pallor with an overall of 35 patients (38.8%). Almost 73.3% (66) patients had anemia out of which 14.4% (13 patients) overall where found to have severe anemia and 27.7% (25) had moderate anemia and 31.1% (28 patients) had mild anemia. 27.7% (25) cases satisfied one or more WHO criteria for severe malaria.Conclusions: The emergence of severe malaria caused by P. vivax could have significant implications in planning the malaria-control programmes for the community. The widespread use of ACT for severe malaria as per the guidelines of the WHO could result in the emergence of resistant parasite strains. It may be rational to continue the use of chloroquine for malaria due to P. vivax in chloroquine-sensitive areas. In places where both P. vivax and P. falciparum co-exist, measures need to be equally targeted to P. vivax to decrease morbidity and mortality due to severe malaria.

Caracterización molecular del dominio de la hélice del gen k13 de Plasmodium falciparum en muestras de comunidades nativas de Condorcanqui, Amazonas, Perú / Molecular characterization of the Plasmodium falciparum k13 gene helix domain in samples from native communities of Condorcanqui, Amazonas, Perú

Introducción. La resistencia de Plasmodium falciparum a diferentes fármacos antipalúdicos es un obstáculo para eliminar la enfermedad. El genotipo resistente de P. falciparum a la artemisinina puede evaluarse examinando los polimorfismos en el dominio de la hélice del gen Pfk13. La Organización Mundial de la Salud recomienda utilizar estas mutaciones como marcadores moleculares para detectar la resistencia a la artemisinina en países donde la malaria por P. falciparum es endémica. Objetivo. Identificar mutaciones relacionadas con la resistencia a artemisinina presentes en el dominio de la hélice del gen k13 de P. falciparum. Materiales y métodos. Mediante la detección pasiva de casos, se recolectaron 51 muestras positivas por microscopía para Plasmodium, provenientes de seis comunidades del distrito de Río Santiago en Condorcanqui, Amazonas. Se realizó la confirmación molecular de la especie mediante PCR en tiempo real y el dominio de la hélice del gen Pfk13 se amplificó y secuenció por electroforesis capilar. Las secuencias obtenidas se compararon con la cepa de referencia 3D7 de fenotipo silvestre. Resultados. Se confirmó un total de 51 muestras positivas para P. falciparum, provenientes de las comunidades de Ayambis, Chapiza, Palometa, Muchinguis, Alianza Progreso y Caterpiza. Después del alineamiento de las secuencias de ADN, se determinó que las muestras no presentaron mutaciones asociadas con resistencia en el gen K13. Discusión. Los resultados obtenidos son coherentes con estudios similares realizados en otros países de Sudamérica, incluyendo Perú. Estos datos proporcionan una línea base para la vigilancia molecular de resistencia a artemisinina en la región Amazonas y refuerzan la eficacia de la terapia combinada con artemisinina en esta área.

Introduction. Resistance of Plasmodium falciparum to different antimalarial drugs is an obstacle to disease elimination. The artemisinin-resistant genotype of P. falciparum can be assessed by examining polymorphisms in the helix domain of the Pfk13 gene. The World Health Organization recommends these mutations as molecular markers to detect artemisinin-resistant in countries where P. falciparum malaria is endemic. Objective. To identify artemisinin resistance-related mutations present in the helix domain of the P. falciparum k13 gene. Materials and methods. We collected a total of 51 samples through passive case detection, positive for Plasmodium by microscopy, from six communities in the district of Río Santiago in Condorcanqui, Amazonas. Molecular species confirmation was performed by real-time PCR and Pfk13 helix domain was amplified and sequenced by capillary electrophoresis. The obtained sequences were compared with the wild type 3D7 reference strain. Results. A total of 51 positive samples were confirmed for P. falciparum from the communities of Ayambis, Chapiza, Palometa, Muchinguis, Alianza Progreso and Caterpiza. DNA sequences alignment showed the absence of resistance-associated mutations in the k13 gene of the collected samples. Discussion. The obtained results are consistent with similar studies conducted in other South American countries, including Perú, so these data provide a baseline for artemisinin- resistance molecular surveillance in the Amazon region and reinforce the efficacy of artemisinin-based combination therapy in this area.

Sequential analysis of erythrocyte aggregation in P. falciparum malaria with and without ASAQ therapy by optical signal and image analysis

The aggregation of erythrocytes is an important mechanism for blood flow through the cardiovascular system. In malaria, this is complicated by infection caused by P. falciparum and is further complicated by the severity of parasitemia. Hence analysis of this micro-mechanism is essential to know the changes in blood not only in diseased conditions but also after artemisinin combination therapy (ASAQ) to alleviate suffering. For analysis purposes, aggregation of erythrocytes was determined by LED laser aggregometer, represented in terms of various parameters related to the changes in laser transmitted intensity. Formed aggregates are further analyzed by imaging and image-processing methods. For this study blood samples from young adults (18 – 40 years old) infected with P. falciparum (n= 80), without any other serious illness, were performed. These samples were selected based on the severity of parasitemia, and were divided into low (LP), medium1 (MP1), medium 2 (MP2), and high (HP) parasitemia. For three days, the selected individuals were treated with artemisinin-based combination therapy ASAQ (Artesunate 4 mg/kg and amodiaquine 10 mg base/ kg once a day). Healthy subjects (n=20) without any history of the disease were selected as a control group. The results, as obtained by various parameters, show a significant elevation of aggregation of erythrocytes (P< 0.05) in P. falciparum malaria with the increase of parasitemia level. There was a decrease in the aggregation after treatment on day four tending towards normal. Thus the current study shows the potential beneficial role of ASAQ on erythrocytes aggregation, which may contribute to reducing the harmful effects on various organs in P. falciparum-infected blood.

Detection and species determination of malaria parasites by microscopy, antigen detection, polymerase chain reaction, and loop-mediated isothermal amplification assay in a tertiary care hospital

Background: Malaria is a major health issue in tropical and subtropical areas. Out of all subtypes, Plasmodium falciparum (Pf) is the most dangerous form accounting for high mortality and morbidity. It is transmitted by infected female anopheles mosquitoes and infected blood transfusions. Aims and Objectives: The aim of the study is to establish correct diagnosis by direct microscopy, Immunochromatographic test (ICT), and molecular studies. Materials and Methods: This prospective study was conducted in the PG Department of Microbiology, SCB Medical College, Cuttack. Thick blood smears were drawn and then stained with Leishman’s stain to visualize falciparum rings. DNA was extracted from infected blood samples by phenol chloroform method with some modification as described by Sambrook and Russel for molecular analysis. Results: In the present study, 150 cases of malaria were analyzed. The male: female ratio was 1.7:1 and age ranged from 0 to 56 years. The Plasmodium vivax positivity was compared with thin smear to 21 (84%) in ICT, 100% both polymerase chain reaction (PCR) and loop mediated isothermal amplification assay (LAMP) assays followed by the Pf positivity as 76 (92.7%) in ICT, 82 (100%) both PCR and LAMP assays, respectively. The results obtained were statistically significant with P < 0.001. The PCR and LAMP showed 100% response to specificity and positive predictive value. Conclusion: The present study established the role of molecular tests such as PCR and LAMP are highly specific for diagnosis of Plasmodium species whereas they are more or less similar in sensitivity as compared to other diagnostic methods such as ICT and microscopy.

Comparative Anti-plasmodial and Cytotoxic Effect of the Methanolic Extracts of Leaves, Stem Bark and Roots of Annickia affinis

Aims: This study aimed to evaluate and compare the in vitro anti-plasmodial and cytotoxic effect of the methanolic extracts from leaves, stem bark and roots of Annickia affinis. Study Design: This is an experimental study. Place and Duration of Study: The work was conducted at the Pharmacochemistry and Natural Substances Laboratories of the Faculty of Medicine and Pharmaceutical Sciences, University of Douala for the extraction and at the Biomedical Chemistry Research Center of Rhodes University in South Africa for the antimalarial and cytotoxic essay. All the experiments were carried out from the 15th October 2019 to the 31th July 2020. Methodology: The anti-plasmodial test was performed on Plasmodium falciparum sensitive strains 3D7 while cytotoxicity was evaluated on the HeLa cell line. Results: The anti-plasmodial tests revealed that the roots and the stem bark exhibited a moderate anti-plasmodial effect with IC50 of 19.7 � 0.8 and 12.1 � 0.8 礸/ml respectively. The anti-plasmodial effects of the leaves were classified as low (33.7 � 1.9 礸/ml). At up to 50 礸/ml, all the extracts showed a high rate of survival among the HeLa cells. No effect was observed with the leaf extracts (100% of survival).Conclusion: This is the first report on the cytotoxic study and comparative anti-plasmodial effect of Annikia affinis. It highlights the potential of Annickia affinis as an important source of anti-plasmodial drugs with less cytotoxic in vitro. In agreement with the use in traditional medicine, the stem bark was more active than wood, while leaves showed low activity. Keywords: Annickia affinis, anti-plasmodial activity, cytotoxicity, Plasmodium falciparum 3D7, HeLa, medicinal plant

Molecular detection of drug-resistant Plasmodium falciparum mutants in Assam

Background & objectives: The spread of drug-resistant Plasmodium falciparum ( Pf) poses a serious threat to the control and elimination of malaria. The objective of this study was to detect the molecular biomarkers of antimalarial drug resistance in Pf in patients visiting a tertiary care hospital in Assam. Methods: Malaria was first detected in fever cases using microscopy and a rapid diagnostic test (RDT), and then confirmed using PCR. Pf chloroquine resistance transporter (Pfcrt), Pf multidrug resistance-1 (Pfmdr-1), and single-nucleotide polymorphisms linked to delayed parasite clearance after treatment with artemisinin MAL 10-688956 and MAL 13-1718319 and Kelch-13 propeller (PfK-13) genes were evaluated by PCR-restriction fragment length polymorphism (RFLP). Results: Sixty nine cases of malaria were found among 300 cases of fever. Of these, 54 were positive for Pf, 47 of which were confirmed by PCR. Pfcrt-K76T mutation was seen in 96.6 per cent and Pfmdr1-N86Y mutation in 84.2 per cent of cases. Mutation was not detected in MAL10 and MAL13 genes. Sequence analysis of Kelch-13 gene showed the presence of a novel mutation at amino acid position 675. Statistically, no significant association was found between the molecular biomarkers and demographic profile, clinical presentation and outcome of the cases. Interpretation & conclusions: Molecular surveillance is essential to assess the therapeutic efficacy of the drugs against circulating Pf isolates in Assam which are found to be highly resistant to CQ. The role of the new mutation found in the Kelch-13 gene in the development of artemisinin resistance in Assam needs to be thoroughly monitored in future research.

Quality assurance of malaria rapid diagnostic tests: An aid in malaria elimination

Background & objectives: India targets malaria elimination by 2030 in a phased manner, so malaria’s assured diagnosis is crucial. Introduction of rapid diagnostic kits in India in 2010 has revolutionized malaria surveillance. The storage temperature of rapid diagnostic tests (RDTs), kit components and handling in transportations impact the results of RDTs. Therefore, quality assurance (QA) is required before it reaches end-users. The Indian Council of Medical Research-National Institute of Malaria Research (ICMR-NIMR) has a World Health Organization (WHO) recognized lot-testing laboratory facility to assure the quality of RDTs. Methods: The ICMR-NIMR receives RDTs from different manufacturing companies as well as various agencies such as National and State Programmes and Central Medical Services Society. The WHO standard protocol is followed to conduct all the tests, including long-term and post-dispatch testing. Results: A total of 323 lots tested during January 2014-March 2021 were received from different agencies. Amongst them, 299 lots passed the quality of test and 24 failed. In long-term testing, 179 lots were tested and only nine failed. A total of 7741 RDTs were received from end-users for post-dispatch testing of which 7540 qualified the QA test with a score of 97.4 per cent. Interpretation & conclusions: RDTs received for quality testing showed compliance with QA evaluation of malaria RDTs based on the protocol recommended by the WHO. However, continuous monitoring of the quality of RDTs is required under QA programme. Quality-assured RDTs have a major role, especially in areas where low parasitaemia of parasites persists.

A rare case of complicated malaria presenting as warm autoimmune hemolytic anemia

Autoimmune hemolytic anemia (AIHA) has been rarely reported worldwide or from India as the underlying cause of anemia in malaria. We hereby present a case of complicated Plasmodium falciparum malaria with concomitant warm AIHA in a 31-year-old male. Direct Antiglobulin Test (DAT) was positive and elution studies showed pan-agglutination reaction. Clinico-hematological and serological follow-up of the patient was done post artesunate treatment until day 9. We suggest that it is important to establish the immune basis of anemia in malaria patients for guiding the treatment plan for the clinicians and providing packed red blood cell transfusion if required.

Malaria asociada al embarazo: revisión narrativa / Malaria in pregnancy: Narrative review

Las infecciones por malaria incrementan el riesgo de presentar complicaciones en el binomio madre-feto. En Venezuela, la casuística de este grupo vulnerable no se ha actualizado en los últimos años. El objetivo de esta revisión narrativa fue describir exhaustivamente qué es la malaria asociada al embarazo y sus efectos maternos, fetales y neonatales; tratando de contestar la siguiente pregunta de investigación. Todas las formas de la malaria asociada al embarazo, incluyendo malaria gestacional, placentaria y congénita, causan alteraciones materno-fetales y neonatales que, de progresar, podrían llevar a la muerte de este binomio. La fisio-patología e inmuno-patología logran explicar la sintomatología de las gestantes y del feto, así como sus complicaciones; dependiendo de la forma parasitaria afectante. Existen nuevas actualizaciones en el diagnóstico, prevención y tratamiento de esta entidad(AU)

Malaria infections increase the risk of complications in the mother-fetus binomial. In Venezuela, the casuistry of this vulnerable group has not been updated in recent years. The objective of this narrative review was to comprehensively describe what pregnancy-associated malaria is and its maternal, fetal and neonatal effects; trying to answer the following research question. All forms of pregnancy-associated malaria, including gestational, placental and congenital malaria, cause maternal-fetal and neonatal alterations that, if they progress, could lead to the death of this binomial. Physiopathology and immunopathology can explain the symptoms of pregnant women and the fetus, as well as their complications; depending on the parasitic form affecting. There are new updates in the diagnosis, prevention and treatment of this entity(AU)

Molecular characterization of Plasmodium Falciparum Kelch-13 among febrile patients in selected Government Hospitals in Nigeria

@#Malaria is a life-threatening disease caused by protozoan Plasmodium species. Plasmodium falciparum is the deadliest species. Reducing and eliminating malaria burden are linked to most of the Sustainable Development Goals (SDG), central to SDG3 targeting the end of malaria by 2030. This study was aimed at assessing the Management of malaria and prevalence of P. falciparum kelch-13 among febrile patients in selected Government Hospitals in Nigeria. Malaria patients (399) attending outpatient clinics of the Hospitals between August, 2019 and January, 2021, were enlisted in the study, following ethical approval and informed consents. Blood (5mL) was collected from patients for microscopic and molecular investigation of malaria parasite. DNA extraction, PCR amplification, BLAST, and alignment were performed. Plasmodium resistance to Artemether/lumefantrine was determined by PCR amplification of extracted DNA using Kelch-13 gene primer. Data obtained were subjected to One-way Analysis of Variance and Linear Regression. The VapA gene primer amplified 55 (68.75%) out of the 80 DNA extracts tested. Twenty-five strains of P. falciparum belonging to 3 clades phylogenetically were identified and they showed evolutionary relationships with others. Plasmodium falciparum resistant Kelch-13 gene was detected in 70% of the isolates. This study observed a high prevalence of resistant gene to ACT drugs in the study area. Monitoring the effectiveness of ACTs must be done routinely to ensure timely changes in National treatment policies.

Establishment and preliminary evaluation of a fluorescent recombinase-aided amplification/CRISPR-Cas12a system for rapid detection of Plasmodium falciparum / 中国血吸虫病防治杂志

Objective To establish a fluorescent assay for rapid detection of Plasmodium falciparum based on recombinaseaided amplification (RAA) and CRISPR-Cas12a system,and to preliminarily evaluate the diagnostic efficiency of this system.. Methods The 18S ribosomal RNA (rRNA) gene of P. falciparum was selected as the target sequence, and three pairs of RAA primers and CRISPR-derived RNA (crRNA) were designed and synthesized. The optimal combination of RAA primers and crRNA was screened and the reaction conditions of the system were optimized to create a fluorescent RAA/CRISPR-Cas12a system. The plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 was generated, and diluted into concentrations of 1 000, 100, 10, 1 copy/μL for the fluorescent RAA/CRISPR-Cas12a assay, and its sensitivity was evaluated. The genomic DNA from P. vivax, P. malariae, P. ovum, hepatitis B virus, human immunodeficiency virus and Treponema pallidum was employed as templates for the fluorescent RAA/CRISPR-Cas12a assay, and its specificity was evaluated. Fifty malaria clinical samples were subjected to the fluorescent RAA/CRISPR-Cas12a assay and nested PCR assay, and the consistency between two assays was compared. In addition, P. falciparum strain 3D7 was cultured in vitro. Then, the culture was diluted into blood samples with parasite densities of 1 000, 500, 200, 50, 10 parasites/μL with healthy volunteers’ O-positive red blood cells for the RAA/CRISPR-Cas12a assay, and the detection efficiency was tested. Results The Pf-F3/Pf-R3/crRNA2 combination, 2.5 μL as the addition amount of B buffer, 40 min as the RAA reaction time, 37 °C as the reaction temperature of the CRISPR-Cas12a system were employed to establish the fluorescent RAA/CRISPR-Cas12a system. Such a system was effective to detect the plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 at a concentration of 1 copy/μL, and presented fluorescent signals for detection of P. falciparum, but failed to detect P. ovum, P. malariae, P. vivax, T. pallidum, hepatitis B virus or human immunodeficiency virus. The fluorescent RAA/CRISPR-Cas12a system and nested PCR assay showed completely consistent results for detection of 50 malaria clinical samples (kappa = 1.0, P < 0.001). Following 6-day in vitro culture of the P. falciparum strain 3D7, 10 mL cultures were generated and the fluorescent RAA/CRISPR-Cas12a system showed the minimal detection limit of 50 parasites/μL. Conclusion The fluorescent RAA/CRISPR-Cas12a system is rapid, sensitive and specific for detection of P. falciparum, which shows promising value for rapid detection and risk monitoring of P. falciparum.

Analysis laboratory results of three cases of COVID-19 complicated with falciparum malaria / 中国热带医学

@#Abstract: Objective　To analyze the laboratory indexes of patients infected with malaria patients and COVID-19, so as to provide reliable evidence for the diagnosis of mixed infection of both. Methods　The routine clinical laboratory items such as routine blood, biochemistry and lymphocyte subsets were tested in three cases of COVID-19 complicated with falciparum malaria who admitted to Guangzhou Eighth People's Hospital Affiliated to Guangzhou Medical University from July to December 2020 were tested. Laboratory data were stage-wise analyzed in conjunction with changes in the course of disease. Results　Three patients confirmed COVID-19 infection recruited all had malaria infection history. Fever, headache, and other symptoms emerged on the 4rd to 11th day after admission. Malaria parasite was detected by malaria parasite antigen testing and blood smear testing, and all three patients had re-ignition of malaria after being confirmed COVID-19 infection. In the early stage of malaria relapse, lymphocytes decreased, CRP and SAA increased, and gradually returned to normal level after antimalarial treatment. Interestingly, we only found one patient at the initial stage of malaria detection showed PLT decreased, no other unnormal changes in other routine blood results (WBC, ESO) and liver function results (ALT, AST, GGT, TBIL, DBIL, CG) were found from the beginning to end course of the disease. Conclusion　COVID-19 infection may promote the resurgence of malaria, so the relapse of malaria should be monitored especially for the patient with malaria infection history who begin to develop fever and other symptoms a few days after the diagnosis of COVID-19. The inflammatory indicators would be worth able as an auxiliary judgment basis for the effective treatment of the two combined infection.

Research progress on stable gene transfection methods of Plasmodium falciparum / 中国热带医学

@#Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.

Monitoring and analysis of kelch13 C580Y mutation in imported Plasmodium falciparum in Wuhan, China / 中国热带医学

@#Abstract: Objective To investigate the prevalence of C580Y mutation of kelch13 gene in the imported P. falciparum cases in Wuhan City, China, and to provide a reference basis for the prevention and treatment of imported falciparum malaria. Methods From 2009 to 2015, blood samples were collected from returnees who infected with P. falciparum in endemic areas of Africa and Southeast Asia in Wuhan City. The P. falciparum DNA was extracted from the blood samples, and kelch13 gene was amplified by loop-mediated isothermal amplification (LAMP), and the distribution of C580Y mutation was analyzed. Results C580Y mutation was detected in 16 of the 208 cases tested by LAMP. No mutations were detected in 69 cases of imported falciparum malaria from Africa during 2009-2012, while 13 cases of the C580Y mutation were detected in 114 cases from 2013 to 2015, with a mutation rate of 11.4%. The mutation rate in South Africa, West Africa, and Central Africa was 12.5%, 9.6%, and 19.0%, respectively, with no mutations detected in cases from North Africa and East Africa. Among the 25 cases of falciparum malaria from Southeast Asia between 2009 and 2013, three cases were positive for the C580Y mutation, all from Myanmar, with a mutation rate of 14.3% (3/21) in Myanmar and 12.0% (3/25) in Southeast Asia. There was no significant difference in the mutation rate between Africa and Southeast Asia after 2013 (P>0.05). Conclusions Our findings highlight the varying degrees of C580Y mutations of kelch13 gene in imported P. falciparum cases in Wuhan and suggest the need for enhanced monitoring and evaluation of related resistance genes.