Hepatitis B and asymptomatic malaria coinfection in Sub-Saharan African immigrants: epidemiological and clinical features of HBV infection

Abstract INTRODUCTION: Here, we conducted an epidemiological study of hepatitis B virus (HBV) mono-infected and asymptomatic malaria/HBV coinfected immigrants and further discussed the possibility of malaria disease modifying the clinical presentation of HBV infection. METHODS: A total of 195 African immigrants were examined for HBV infection or coinfection with HBV and asymptomatic malaria. HBV infection was diagnosed using serological tests and confirmed by PCR; furthermore, we performed a pan-Plasmodium-specific-nucleic-acid-sequence-based-amplification (NASBA) assay to detect asymptomatic malaria infection. The stage/grade of the liver disease was determined using echotomography and elastometry. RESULTS: PCR-NASBA results confirmed that 62 of 195 subjects (31.8%) were positive for Plasmodium infection, whereas 41 of 195 subjects (21%) tested positive for HBV chronic hepatitis (HBV-DNA positive). Among the HBV-positive subjects, 26 (63.4%) of them were mono-infected patients (Group A), whereas 15 (36.6%) patients had HBV chronic hepatitis and asymptomatic malaria coinfections (Group B). The HBV-DNA median levels were 1.4×105IU/mL in HBV-mono-infected patients and 2.0×105IU/mL in coinfected patients. Echotomography and hepatic elastometry presented similar findings for both groups of patients. CONCLUSIONS: Coinfected patients seem to present with the same clinical symptoms of the liver disease as HBV mono-infected patients.

Assessment of co-segregated TLR4 genotypes among Nigerian children with asymptomatic and clinical malaria

Objective To assess the occurrence and pattern of Toll-like receptor 4 (TLR4) co-segregated genotypes among children with Plasmodium falciparum malaria in Nigeria. Methods In this case-control study, a total of 79 Plasmodium falciparum infected children aged 2–7 years and 105 age-matched uninfected controls of Yoruba descents in Lagos were studied. The extracted DNA samples were used for TLR4 genotyping at codons 299 (Asp > Gly) and 399 (Thr > Ile) by PCR-restriction fragment length polymorphism. Malaria infection was diagnosed by blood smear microscopy and infected children were stratified into asymptomatic, uncomplicated and severe malaria sub-groups. Malnutrition was determined by measuring the mid upper arm circumference and anemia was defined as hemoglobin  0.05) in frequency between infected and non-infected children. However, low and high occurrences of the TLR4 Asp299Asp/Thr399Thr and Asp299Gly/Thr399Thr genotypes were observed in the severe malaria subgroup. Conclusions This study reveals a protective role for TLR4 Asp299Gly/Thr399Ile and Asp299Asp/Thr399Thr genotypes against severe malaria in Nigerian children.

Assessment of co-segregated TLR4 genotypes among Nigerian children with asymptomatic and clinical malaria

Objective: To assess the occurrence and pattern of Toll-like receptor 4 (TLR4) co-segregated genotypes among children with Plasmodium falciparum malaria in Nigeria. Methods: In this case-control study, a total of 79 Plasmodium falciparum infected children aged 2–7 years and 105 age-matched uninfected controls of Yoruba descents in Lagos were studied. The extracted DNA samples were used for TLR4 genotyping at codons 299 (Asp> Gly) and 399 (Thr >Ile) by PCR-restriction fragment length poly-morphism. Malaria infection was diagnosed by blood smear microscopy and infected children were stratified into asymptomatic, uncomplicated and severe malaria sub-groups. Malnutrition was determined by measuring the mid upper arm circumference and anemia was defined as hemoglobin0.05) in frequency between infected and non-infected children. However, low and high occur-rences of the TLR4 Asp299Asp/Thr399Thr and Asp299Gly/Thr399Thr genotypes were observed in the severe malaria subgroup. Conclusions: This study reveals a protective role for TLR4 Asp299Gly/Thr399Ile and Asp299Asp/Thr399Thr genotypes against severe malaria in Nigerian children.

Prevalence of asymptomatic Plasmodium vivax infections in the north-eastern focus of malaria of Venezuela / Prevalencia de infecciones asintomáticas por Plasmodium vivax en el foco malárico oriental de Venezuela

Malaria remains as a public health problem in Venezuela. In 2015 there were 136,402 cases reported by the Ministry of Popular Power for Health, being the parasite prevalence 73.95% for Plasmodium vivax, 17.6% for Plasmodium falciparum, 0.0095% for Plasmodium malariae and 8.42% mixed infections (P. vivax + P. falciparum). During the period 1999-2002 the number of cases in Venezuela ranged between 21,685 and 29,337, being the Sucre State with highest levels of malaria prevalence, with Plasmodium vivax as the unique specie in this region. In 2002 the Municipality of Cajigal had the highest Annual Parasite Incidence (API) of country, being 260 cases per 1000 inhabitants. In view of the difficulty in controlling malaria in this area, the prevalence of asymptomatic carriers was investigated as one of the epidemiological factors contributing to the persistence of malaria transmission. One hundred fifty people were included in the study, with no history of recent malaria infection, or any symptom and also, not having used antimalarial drugs during the 30 days prior to study entry. To do this, a malaria Rapid Diagnostic Test (mRDTs) was used for the determination of antigenemic (OptiMAL®) and PCR (polymerase chain reaction) in conjunction with the reference "Gold Standard", the conventional thick and thin blood smears (TTBS). It was found a prevalence of infection of 1.33% by mRDTs and TTBS and 8% by PCR which allowed the detection of 10 asymptomatic cases in addition, with a sensitivity and specificity of 100% and 93.4% respectively. The presence of asymptomatic carriers in this area reveals the difficulties that face the Malaria Control Program in the eventual elimination of this specific malaria foci. It is necessary reinforces the maintenance of the epidemiological surveillance using more sensitive diagnostic techniques, as well as to adapt the control measures based on the current findings.

La malaria sigue siendo un problema de salud pública en Venezuela. Para el año 2015 el Ministerio del Poder Popular para la Salud reportó 136.402 casos, siendo la fórmula parasitaria 73,95% para Plasmodium vivax, 17,6% para Plasmodium falciparum, 0,0095 para Plasmodium malariae y 8,42% para infecciones mixtas (P. vivax + P. falciparum). Durante el período 1999-2002, el número de casos en Venezuela estuvo entre 21.685 y 29.337, siendo el Estado Sucre el que mostró los niveles más altos de prevalencia de malaria, con P. vivax como única especie en la región. En el año 2002 el Municipio Cajigal registró el Índice Parasitario Anual (IPA) más alto del país, siendo 260 casos por 1000 habitantes. En vista de las dificultades para controlar la malaria en esta área, se investigó la prevalencia de portadores asintomáticos como uno de los factores contribuyentes en la persistencia de la transmisión malárica. Ciento cincuenta personas fueron incluidas en el estudio sin historia de infección reciente por malaria o ningún síntoma, así como no haber consumido drogas antimaláricas durante los 30 días anteriores de ingresar al estudio. Para ello, se usó la prueba rápida de diagnóstico para malaria (PRDxm) para la determinación de antígenemia (OptiMAL®) y la técnica de biología molecular basada en la Reacción en Cadena de la Polimerasa (PCR), conjuntamente con la "prueba oro," como método convencional, la Gota Gruesa y Extendido de Sangre (GGES). Se encontró una prevalencia de infección de 1,33% por GGES y por prueba rápida de diagnóstico OptiMAL® y 8% mediante PCR. La técnica de PCR permitió la detección adicional de 10 casos asintomáticos con una sensibilidad y especificidad del 100% y 93,4% respectivamente. La presencia de portadores asintomáticos en esta área revela las dificultades que enfrenta el Programa de Control de la Malaria en la eliminación eventual de esta parasitosis en este foco. Es necesario reforzar el mantenimiento de la vigilancia epidemiológica usando técnicas de diagnóstico más sensibles, así como adoptar medidas de control basadas en estos hallazgos.

Anopheles (Kerteszia) cruzii (DIPTERA: CULICIDAE) IN PERIDOMICILIARY AREA DURING ASYMPTOMATIC MALARIA TRANSMISSION IN THE ATLANTIC FOREST: MOLECULAR IDENTIFICATION OF BLOOD-MEAL SOURCES INDICATES HUMANS AS PRIMARY INTERMEDIATE HOSTS / Anopheles (Kerteszia) cruzii (Diptera: Culicidae) em área peridomiciliar durante transmissão de malária assintomática na Mata Atlântica: identificação molecular das fontes de repasto sanguíneo indica humanos como hospedeiros intermediários primários

Anopheles (Kerteszia) cruzii has been implicated as the primary vector of human and simian malarias out of the Brazilian Amazon and specifically in the Atlantic Forest regions. The presence of asymptomatic human cases, parasite-positive wild monkeys and the similarity between the parasites infecting them support the discussion whether these infections can be considered as a zoonosis. Although many aspects of the biology of An. cruzii have already been addressed, studies conducted during outbreaks of malaria transmission, aiming at the analysis of blood feeding and infectivity, are missing in the Atlantic Forest. This study was conducted in the location of Palestina, Juquitiba, where annually the majority of autochthonous human cases are notified in the Atlantic Forest of the state of São Paulo. Peridomiciliary sites were selected for collection of mosquitoes in a perimeter of up to 100 m around the residences of human malaria cases. The mosquitoes were analyzed with the purpose of molecular identification of blood-meal sources and to examine the prevalence of Plasmodium. A total of 13,441 females of An. (Ker.) cruzii were collected. The minimum infection rate was calculated at 0.03% and 0.01%, respectively, for P. vivax and P. malariae and only human blood was detected in the blood-fed mosquitoes analyzed. This data reinforce the hypothesis that asymptomatic human carriers are the main source of anopheline infection in the peridomiciliary area, making the probability of zoonotic transmission less likely to happen.

Anopheles (Kerteszia) cruzii é o vetor primário das malárias humana e simiana fora da Amazônia Brasileira e especificamente nas regiões de Mata Atlântica. A presença de casos humanos assintomáticos, macacos silvestres positivos para Plasmodium e a similaridade entre os parasitas que os infectam suportam a discussão se essas infecções podem ser consideradas como zoonoses. Embora muitos aspectos da biologia de An. cruzii já tenham sido abordados, estudos conduzidos durante surtos de transmissão de malária, visando a análise de repasto sanguíneo e infectividade, são ausentes na Mata Atlântica. Este estudo foi conduzido na localidade de Palestina, Juquitiba, Mata Atlântica do Estado de São Paulo, onde anualmente a maioria dos casos humanos autóctones é notificada. Locais em peridomicílio foram selecionados para coleta de mosquitos em um perímetro de até 100 m em torno das residências de casos humanos de malária e da floresta circundante. Os mosquitos foram analisados com o objetivo de identificação molecular das fontes de repasto sanguíneo e para examinar a prevalência de Plasmodium. Um total de 13.441 fêmeas de An. (Ker.) cruzii foi coletado. A taxa de infecção mínima foi calculada a 0,03% e 0,01%, respectivamente, para P. vivax e P. malariae e somente sangue humano foi detectado nos mosquitos analisados que se alimentaram com sangue. Nossos dados reforçam a hipótese de que os portadores humanos assintomáticos são a principal fonte de infecção para os anofelinos na área do peridomicílio, tornando a transmissão zoonótica improvável.

Socio-demographic and Environmental Factors Influencing asymptomatic Malaria and Anaemia Incidence among School Children in Fako Division, South West Cameroon.

Aim: This work was aimed to assess the influence of socio-demographic and environmental factors on the incidence of asymptomatic malaria and anaemia among pupils in Fako Division, southwest Cameroon. Experimental Design: The study was a cross-sectional survey. Place and Duration of Study: The study was carried out in Fako Division, southwest Cameroon -Bolifamba, Dibanda and Mutengene from February to March, 2013. Methodology: A total of 316 pupils aged 4–15 years were studied. Data on sociodemographic and environmental factors was obtained from a semi-structured questionnaire. Blood samples were collected. Malaria parasite incidence and densitywere determined from Giemsa-stained thin and thick blood smears respectively. Haemoglobin (Hb) levels were determined using a haemoglobinometer. Results: The overall incidence of asymptomatic malaria was 43.4% (CI=38-48.9). Malaria incidence was significantly highest (χ2=7,P=0.03) in pupils of 6-10 years age group (49.0%, CI=42.1-59.9) when compared with their counterparts. Although not significant, malaria parasite incidence was higher in males, pupils with fever, highest in pupils of Bolifamba and poor social status than their respective counterparts. Geometric mean parasite density (GMPD) was significantly highest (Kruskal Wallis test, *χ2=6.4, P=0.04) in Dibandathan other sites. Anaemia incidence was higher among inhabitants of Dibanda (56.7%) than other sites. Anaemia incidence was statistically higher (χ2=5.6, P=0.02) in malaria positive pupils, highest in Dibanda (χ2=27.244, P<0.001) and the middle class when compared with their respective counterparts. Mean HB was significantly higher in malaria negative (t=1-8, P=0.02), highest in the poor class (χ2=13.4, P=0.001) and Mutengene (F=21.2, P=0.0001) when compared with their respective counterparts. Conclusion: Sensitization on effective malaria control strategies needs to be emphasized so that a reduction in malaria burden can be achieved.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is associated with asymptomatic malaria in a rural community in Burkina Faso

Objective: To investigate 4 combinations of mutations responsible for glucose-6-phosphate dehydrogenase (G6PD) deficiency in a rural community of Burkina Faso, a malaria endemic country. Methods: Two hundred individuals in a rural community were genotyped for the mutations A376G, G202A, A542T, G680T and T968C using TaqMan single nucleotide polymorphism assays and polymerase chain reaction followed by restriction fragment length polymorphism. Results: The prevalence of the G6PD deficiency was 9.5% in the study population. It was significantly higher in men compared to women (14.3% vs 6.0%, P=0.049). The 202A/376G G6PD A- was the only deficient variant detected. Plasmodium falciparum asymptomatic parasitaemia was significantly higher among the G6PD-non-deficient persons compared to the G6PD-deficient (P<0.001). The asymptomatic parasitaemia was also significantly higher among G6PD non-deficient compared to G6PD-heterozygous females (P<0.001). Conclusions: This study showed that the G6PD A- variant associated with protection against asymptomatic malaria in Burkina Faso is probably the most common deficient variant.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is associated with asymptomatic malaria in a rural community in Burkina Faso

Objective: To investigate 4 combinations of mutations responsible for glucose-6-phosphate dehydrogenase (G6PD) deficiency in a rural community of Burkina Faso, a malaria endemic country. Methods: Two hundred individuals in a rural community were genotyped for the mutations A376G, G202A, A542T, G680T and T968C using TaqMan single nucleotide polymorphism assays and polymerase chain reaction followed by restriction fragment length polymorphism. Results: The prevalence of the G6PD deficiency was 9.5% in the study population. It was significantly higher in men compared to women (14.3%vs 6.0%, P=0.049). The 202A/376G G6PD A-was the only deficient variant detected. Plasmodium falciparum asymptomatic parasitaemia was significantly higher among the G6PD-non-deficient persons compared to the G6PD-deficient (P Conclusions:This study showed that the G6PD A-variant associated with protection against asymptomatic malaria in Burkina Faso is probably the most common deficient variant.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is associated with asymptomatic malaria in a rural community in Burkina Faso

<p><b>OBJECTIVE</b>To investigate 4 combinations of mutations responsible for glucose-6-phosphate dehydrogenase (G6PD) deficiency in a rural community of Burkina Faso, a malaria endemic country.</p><p><b>METHODS</b>Two hundred individuals in a rural community were genotyped for the mutations A376G, G202A, A542T, G680T and T968C using TaqMan single nucleotide polymorphism assays and polymerase chain reaction followed by restriction fragment length polymorphism.</p><p><b>RESULTS</b>The prevalence of the G6PD deficiency was 9.5% in the study population. It was significantly higher in men compared to women (14.3% vs 6.0%, P=0.049). The 202A/376G G6PD A- was the only deficient variant detected. Plasmodium falciparum asymptomatic parasitaemia was significantly higher among the G6PD-non-deficient persons compared to the G6PD-deficient (P<0.001). The asymptomatic parasitaemia was also significantly higher among G6PD non-deficient compared to G6PD-heterozygous females (P<0.001).</p><p><b>CONCLUSIONS</b>This study showed that the G6PD A- variant associated with protection against asymptomatic malaria in Burkina Faso is probably the most common deficient variant.</p>

Ausencia de malaria asintomática en escolares de Quibdó, Chocó / Absence of asymptomatic malaria in schoolchildren of Quibdó, Chocó

La malaria asintomática es considerada común en zonas de alta transmisión en África y prácticamente ausente en zonas de baja transmisión en Latinoamérica. Sin embargo, algunos reportes sugieren que el hallazgo de casos asintomáticos en zonas de alta y moderada transmisión en Latinoamérica es relativamente frecuente. La malaria asintomática tiene consecuencias graves no sólo para el individuo portador de parásitos, sino también para la comunidad donde vive. Los individuos crónicamente infectados se constituyen en un reservorio de la enfermedad que es difícil de identificar por medio de la vigilancia rutinaria de los programas de control. En el marco de un estudio de la epidemiología de la malaria en Quibdó, Chocó, se estimó la prevalencia de malaria asintomática en escolares. El objetivo fue establecer si los individuos con infección asintomática podrían constituir un reservorio importante de infección, capaz de mantener la transmisión local en esta ciudad. Se seleccionaron 255 estudiantes de cinco escuelas de Quibdó, de los cuales, 223 se incluyeron en el análisis. Algunos niños refirieron cefalea (34%), tos (32%) y diarrea (9%). Todos los resultados de gota gruesa fueron negativos. Además, el análisis por inmunofluorescencia indirecta de anticuerpos contra Plasmodium en una submuestra de 25 niños fue negativo. La prevalencia estimada de malaria asintomática en los escolares de Quibdó es, entonces, de 0% (IC95%: 0-1,4). La presencia de malaria asintomática en adultos no se puede descartar aunque es probable que, si existe, sea en una proporción muy baja.

Asymptomatic malaria is characteristic of high intensity transmission areas in Africa but unusual in low transmission areas in Latin America. Nevertheless, asymptomatic malaria has been reported to be frequent in areas in Latin America with high and moderate intensity of transmission. Asymptomatic malaria can affect both individuals who carry parasites and are cryptic carrier reservoirs for the community. Individuals chronically infected with malaria parasites are usually unidentifiable by most malaria control programmes. In order to identify whether asymptomatic individuals harboring malaria parasites are an important reservoir of infection in Quibdó, Chocó, the prevalence of asymptomatic malaria was assessed in schoolchildren. This study was part of a major study of the epidemiology of malaria in Quibdó. A total of 255 children from 5 schools were examined, of which 223 were included in the analysis. Children reported headache (34%), cough (32%), and diarrhoea (9%). None of the children presented a positive thick smear. In addition, IFA tests in a subsample of 25 children were negative. By these criteria, the prevalence of asymptomatic malaria in Quibdó schoolchildren is 0% (95%C.I.: 0.0-1.4). Although asymptomatic malaria in adults possibly occurs, a very low prevalence is predicted.