Lack of quadruple and quintuple mutant alleles associated with sulfadoxine-pyrimethamine resistance in Plasmodium vivax isolates from Brazilian endemic areas

BACKGROUND AND OBJECTIVE Brazil is responsible for a large number of Plasmodium vivax cases in America. Given the emergence of P. vivax parasites resistant to chloroquine and the effectiveness of antifolates in vivax malaria treatment together with a correlation between mutations in P. vivax dhfr and dhps genes and SP treatment failure, the point mutations in these genes were investigated. METHODS Blood samples from 54 patients experiencing vivax malaria symptomatic episodes in the Amazonian Region were investigated. Genomic DNA was extracted using a DNA extraction kit (QIAGENTM). Nested polymerase chain reaction (PCR) amplification was carried out followed by Sanger sequencing to detect single nucleotide polymorphisms (SNPs). FINDINGS All tested isolates showed non-synonymous mutations in pvdhfr gene: 117N (54/54, 100%) and 58R (25/54, 46%). Double mutant allele 58R/117N (FRTNI, 28%) was the most frequent followed by triple mutant alleles (58R/117N/173L, FRTNL, 11%; 58R/61M/117N, FRMNI, 5% 117N/173L, FSTNL, 4%) and quadruple mutant allele (58R/61M/117N/173L, FRMNL, 2%). A single mutation was observed at codon C383G in pvdhps gene (SGKAV, 48%). CONCLUSION No evidence of molecular signatures associated with P. vivax resistance to SP was observed in the Brazilian samples.

Plasmodium infection and its association with biochemical and haematological parameters in free-living Alouatta guariba clamitans (Cabrera, 1940) (Primates: Atelidae) in Southern Brazil

BACKGROUND The influence of Plasmodium spp. infection in the health of Southern brown howler monkey, Alouatta guariba clamitans, the main reservoir of malaria in the Atlantic Forest, is still unknown. OBJECTIVES The aim of this study was to investigate the positivity rate of Plasmodium infection in free-living howler monkeys in an Atlantic Forest fragment in Joinville/SC and to associate the infection with clinical, morphometrical, haematological and biochemical alterations. METHODS Molecular diagnosis of Plasmodium infection in the captured monkeys was performed by Nested-polymerase chain reaction (PCR) (18S rRNA and coxI). Haematological and biochemical parameters were compared among infected and uninfected monkeys; clinical and morphometrical parameters were also compared. FINDINGS The positivity rate of Plasmodium infection was 70% among forty captured animals, the highest reported for neotropical primates. None statistical differences were detected in the clinical parameters, and morphometric measures comparing infected and uninfected groups. The main significant alteration was the higher alanine aminotransferase (ALT) levels in infected compared to uninfected monkeys. MAIN CONCLUSIONS Therefore, Plasmodium infection in howler monkeys may causes haematological/biochemical alterations which might suggest hepatic impairment. Moreover, infection must be monitored for the eco-epidemiological surveillance of malaria in the Atlantic Forest and during primate conservation program that involves the animal movement, such as translocations.

Malaria y hemoglobina S: ¿resistencia o protección? / Malaria and hemoglobin S: resistance or protection?

La malaria (o paludismo) es una enfermedad infecciosa causada por parásitos de la familia Plasmodium y transmitída por los mosquitos Anopheles hembra. En el año 2015, se calcula que hubo 212 millones de nuevos casos de paludismo y 429 000 muertes. La región africana sigue soportando la mayor carga de paludismo, con un estimado del 90 % de los casos y el 92 % de las muertes por esta enfermedad. Muchos años han transcurridos desde los primeros reportes que relacionaban el desorden genético caracterizado por la presencia de hemoglobina S y la infestación por malaria, presumiendo que existía una resistencia al desarrollo de esta infestación y al daño que esta causaba; lo controvertido en aquellos momentos de estos planteamientos justificó las múltiples investigaciones realizadas a posteriori; los resultados obtenidos por los descubrimientos más actuales han propiciado una visión más clara de esta relación y nuevas motivaciones encaminadas a profundizar en el estudio de esta temática. El objetivo de este trabajo es precisamente analizar la relación antes mencionada a la luz de los estudios contemporáneos

Malaria is an infectious disease caused by parasites of the Plasmodium family and transmitted by the female Anopheles mosquitoes. In the year 2015, it ias calculated that there were 212 million new cases and 420 000 deaths. The African region continues bearing the greatest number of cases with an estimate of 90% and the 92% of deaths due to this disease. Many years have passed since the first reports which related the genetic disorder characterized by the presence of hemoglobin S and malaria infection, presuming that there was a resistance to the development of this of infection and it resulting damage. What was controversial about this statement at those times supported the multiple posterior research. The most recent discoveries have allowed a clearer vision of this relationship and new motivations aimed at deepening in the study of this topic. The objective of this work is precisely to analyze the mentioned relationship according to the most recent studies.

Natural Plasmodium infection in neotropical primates in the island of São Luís, state of Maranhão, Brazil / Infecção natural de Plasmodium em primatas neotropicais da Ilha de São Luís, Estado do Maranhão, Brasil

The states that make up the Legal Amazon Region, which include the state of Maranhão, account for 99% of registered cases of human malaria in Brazil. It is also believed that transmission of malaria from nonhuman primates (NHP) to humans occurs in this region, because of current reports of seroepidemiological results from samples from humans and NHP coexisting in the same areas. This study aimed to make morphological, serological and molecular diagnoses of Plasmodium spp. in neotropical primates on the island of São Luís, state of Maranhão, Brazil. The diagnostic techniques used were optical microscopy, the polymerase chain reaction (PCR) and the indirect immunofluorescence assay (IFA). From June 2009 to April 2010, 70 NHP were sampled: 50 at the Wild Animal Screening Center (CETAS), located in the municipality of São Luís and 20 free-living individuals that were caught in a private reserve located in the municipality of São Jose de Ribamar, state of Maranhão. Under an optical microscope, 140 slides (two from each animal) were evaluated and five animals (7.1%) were found to be positive. IFA did not detect anti-Plasmodium spp. From PCR on the 70 animals sampled, amplified Plasmodium spp. products were observed in 13 samples, of which eight (61.5%) were from free-living animals and five (38.5%) were from animals at CETAS.

Os Estados que compõem a Amazônia Legal, entre eles o Estado do Maranhão, respondem a 99% dos casos registrados de malária humana no Brasil. Também se acredita que nessa região ocorra a transmissão de malária de primatas não humanos (PNH) para humanos, devido a relatos atuais de resultados soroepidemiológicos de amostras de humanos e PNH que coexistem nas mesmas áreas. O presente estudo objetivou realizar o diagnóstico morfológico, sorológico e molecular de Plasmodium spp. em primatas neotropicais na Ilha de São Luís, Estado do Maranhão, Brasil. Foram utilizadas como técnicas de diagnóstico: a microscopia de luz, a reação em cadeia pela polimerase (PCR) e a imunofluorescência indireta (RIFI). No período de junho de 2009 a abril de 2010, foram amostrados 70 PNH, sendo 50 provenientes do Centro de Triagem de Animais Silvestres (CETAS), localizado no município de São Luís, e 20 de vida livre, capturados em reserva particular localizada no município de São José de Ribamar, Estado do Maranhão. Foram avaliadas pela microscopia de luz 140 lâminas (duas de cada animal), das quais cinco animais (7,1%) foram positivos. Pela RIFI não se detectou anticorpos anti-Plasmodium spp. Pela PCR, dos 70 animais amostrados, foi possível observar produtos amplificados para Plasmodium spp. em 13 amostras, das quais oito (61,5%) eram de animais de vida livre e cinco (38,5%) de CETAS.

Associação entre bimarcadores imunogenéticos e a gravidade ou resistência na malária por plasmodium vivax / Association between immunogenic markers and the severity and resistance in plasmodium vivax malaria

INTRODUÇÃO: A malária é uma das doenças infecto-parasitárias mais incidentes no mundo com grande morbimortalidade. Dentre as espécies infectivas ao ser humano, o Plasmodium vivax é a espécie predominante no Brasil, quase que exclusivamente na Região Amazônica. O espectro clínico da malária abrange desde uma infecção assintomática até casos moderados,com hiperbilirrubinemia isolada ou graves. A produção de mediadores inflamatórios pelo sistema imune, a via de metabolização do heme e os níveis sistêmicos de hepcidina são importantes mecanismos associados afisiopatologia dos diferentes desfechos clínicos da malária. Além disso, coinfecções podem modular ou intensificar a resposta imune de indivíduos infectados pelo plasmódio. OBJETIVO: Neste ínterim, a identificação de biomarcadores confiáveis tanto de gravidade ou resistência são indispensáveis para o auxílio no seguimento, diagnóstico e terapêutica da malária...

INTRODUCTION: Malaria is one of the most frequent infectious diseases inthe world with high morbidity and mortality. Among the infective species tohumans, Plasmodium vivax is the most predominant species in Brazil, withdisease incidence almost exclusively observed in the Amazon Region. Theclinical spectrum of malaria can range from asymptomatic infection to mildcases, malaria with isolated hyperbilirubinaemia or severe infection. Theimmune system production of inflammatory mediators, the heme metabolismpathway and systemic levels of hepcidin are important mechanismsassociated with pathophysiology of different malaria clinical outcomes. Inaddition, co-infections can modulate or enhance the immune response ofindividuals infected with P. vivax. OBJECTIVE: In this context, the identification of reliable biomarkers for disease severity and resistance are essential for the diagnosis, treatment and follow-up of malaria...

43 kDa and 66 kDa, two blood stage antigens induce immune response in Plasmodium berghei malaria.

The hunt for an effective vaccine against malaria still continues. Several new target antigens as candidates for vaccine design are being explored and tested for their efficacy. In the present study the sera from mice immunized with 24,000 × g fraction of Plasmodium berghei has been used to identify highly immunogenic blood stage antigens. The protective antibodies present in immune sera were covalently immobilized on CNBr activated sepharose 4B and used for affinity chromatography purification of antigens present in blood stages of P. berghei. Two polypeptides of 66 and 43 kDa molecular weights proved to be highly immunogenic. They exhibited a strong humoral immune response in mice as evident by high titres in ELISA and IFA. Protective immunity by these two antigens was apparent by in vivo and in vitro studies. These two proteins could further be analysed and used as antigens in malaria vaccine design.

Apoptosis of non-parasitised red blood cells in Plasmodium yoelii malaria

Recently, while studying erythrocytic apoptosis during Plasmodium yoelii infection, we observed an increase in the levels of non-parasitised red blood cell (nRBC) apoptosis, which could be related to malarial anaemia. Therefore, in the present study, we attempted to investigate whether nRBC apoptosis is associated with the peripheral RBC count, parasite load or immune response. To this end, BALB/c mice were infected with P. yoelii 17XL and nRBC apoptosis, number of peripheral RBCs, parasitaemia and plasmatic levels of cytokines, nitric oxide and anti-RBC antibodies were evaluated at the early and late stages of anaemia. The apoptosis of nRBCs increased at the late stage and was associated with parasitaemia, but not with the intensity of the immune response. The increased percentage of nRBC apoptosis that was observed when anaemia was accentuated was not related to a reduction in peripheral RBCs. We conclude that nRBC apoptosis in P. yoelii malaria appears to be induced in response to a high parasite load. Further studies on malaria models in which acute anaemia develops during low parasitaemia are needed to identify the potential pathogenic role of nRBC apoptosis.

Seroepidemiología del paludismo en un grupo de migrantes en tránsito (Chiapas, 2008)

OBJETIVO: Conocer la prevalencia del paludismo y los factores asociados con la infección de migrantes en la frontera sur de México, durante 2008. MATERIAL Y MÉTODOS: En 706 migrantes, se investigó la infección activa mediante prueba rápida y PCR o pasada, mediante serología y se aplicó un cuestionario para investigar las condiciones asociadas con la infección. RESULTADOS: 85.6% provenía de Centroamérica. Ninguno presentó infección activa; 4.2% fue seropositivo y la mayoría provenía de los países con mayor incidencia de paludismo en la región. La seropositividad se asoció con el número de episodios previos de paludismo (RM=1.44; IC95% 1.04-2.00), años de permanencia en su comunidad de origen (RM=1.03; IC95% 1.00 -1.07) y conocimiento y automedicación con antipalúdicos (RM=3.38; IC95% 1.48-7.67). CONCLUSIONES: La exposición previa de migrantes al paludismo y las dificultades para su detección indican la necesidad de nuevas estrategias para la vigilancia epidemiológica para estas poblaciones.

OBJECTIVE: To know the prevalence of malaria and the factors associated with the infection in migrants in the southern border of Mexico, during 2008. MATERIALS AND METHODS: In 706 migrants, active malaria infection was investigated using a rapid diagnostic test and PCR and past infection using serology. A questionnaire was applied to investigate the conditions associated to infection. RESULTS: 85.6% originated from Central America, none presented an active infection, although 4.2% were seropositive, most of these came from the countries with the highest malaria incidence in the region. Seropositivity was associated with the number of previous malaria episodes (OR=1.44; IC95% 1.04-2.00), years living in their community of origin (OR=1.03; IC95% 1.00-1.07), and knowledge and self-medication with anti-malaria drugs (OR=3.38; IC95% 1.48-7.67). CONCLUSIONS:. The previous exposure of migrants and the difficulties for their detection indicate the need of new strategies for the epidemiological surveillance for these populations.

Hallazgos hematológicos en sangre periférica para sospecha diagnóstica y seguimiento del paludismo / Hematological findings in peripheral blood for diagnosis and monitoring of malaria

Introducción: El Paludismo o la Malaria es una Histohemoparasitosis, causada por Protozoos del género Plasmodium, usando como vector a los mosquitos del género Anopheles. Es la endemia de origen parasitario más importante del mundo, distribuída por diversos países del mundo, incluyendo Argentina. Las alteraciones clínicas se ven reflejadas en diversos cambios bioquímicos durante el curso de la infección. El método diagnóstico más usado y difundido mundialmente sigue siendo el de la Gota Gruesa. Material y Método: En el presente trabajo se estudiaron 40 muestras de sangre anticoagulada con EDTA-K3 de varones adultos, sin comorbilidades asociadas, de diversas nacionalidades, residentes temporarios en Puerto Príncipe, Haití, con diagnóstico de Paludismo. Se analizaron estadísticamente los recuentos celulares y la concentración de hemoglobina al diagnóstico y antes del alta médica, también, se usó un grupo de controles normales. Resultados: Se observó una significativa disminución de los recuentos tanto leucocitarios como plaquetarios en los pacientes al momento del diagnóstico con respecto a un grupo en iguales condiciones, pero sin la infección (controles normales).Discusión: Dichos valores comienzan a aumentar conforme avanza el tratamiento y la negativización de la gota gruesa, pero, en los hemogramas antes del alta, aún siguen siendo significativamente menores con respecto a los de los controles normales. En un subgrupo con la mayor positividad para la gota gruesa, se observa una media aún menor del recuento plaquetario con respecto a los que presentan menor positividad. Conclusiones: Estos patrones que se han identificado en pacientes con infección por Plasmodium, podrían ser útiles, junto con otros parámetros complementarios, para sospechar el diagnóstico de Paludismo aún antes del examen hemoparasitológico de la gota gruesa.

Introduction: Malaria is an Histohemoparasitosis, caused by protozoa of the Plasmodium genus , using Anopheles mosquitoesas vectors. It's the world's largest endemic parasitosis, distributed on different countries, including Argentina.The clinical changes are reflected in various biochemical changes during the course of infection. The most commonly used diagnostic method and worldwide spreaded is still the thick gout.Methods: In this paper we studied 40 blood samples anticoagulated with EDTA-K3 of male adults, without comorbidities, from different nationalities, temporary residents in Port au Prince, Haiti, with Malaria diagnosis. Statistically analyzed cell counts and hemoglobin concentration at moment of diagnosis and prior to discharge also using a group of normal controls.Results: There was a significant decrease in both leukocyte and platelet counts when diagnosed compared with normal controls.Discussion: These values start to increase as treatment progresses and the thick gout becomes negative, but before discharge, blood counts are still significantly lower compared to normal controls. In a subgroup with higher positivity of the thick gout, we observed even lower platelet counts related to those with less positive test.Conclusions: These patterns identified in patients with Plasmodium infection, may be useful, along with other complementary parameters, to suspect diagnosis of malaria even before thick gout examination.

Hb E/beta-thalassaemia: a common & clinically diverse disorder.

Haemoglobin E-beta thalassaemia (Hb E/β-thalassaemia) is the genotype responsible for approximately one-half of all severe beta-thalassaemia worldwide. The disorder is characterized by marked clinical variability, ranging from a mild and asymptomatic anaemia to a life-threatening disorder requiring transfusions from infancy. The phenotypic variability of Hb E/β-thalassaemia and the paucity of long-term clinical data, present challenges in providing definitive recommendations for the optimal management of patients. Genetic factors influencing the severity of this disorder include the type of beta-thalassaemia mutation, the co-inheritance of alpha-thalassaemia, and polymorphisms associated with increased production of foetal haemoglobin. Other factors, including a variable increase in serum erythropoietin in response to anaemia, previous or ongoing infection with malaria, previous splenectomy and other environmental influences, may be involved. The remarkable variation, and the instability, of the clinical phenotype of Hb E beta-thalassaemia suggests that careful tailoring of treatment is required for each patient, and that therapeutic approaches should be re-assessed over-time.

Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale

Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.

Malaria incidence among paramilitary personnel in an endemic area of Tripura.

Background & objectives: Paramilitary operations along the Indo-Bangladesh border are adversely affected by malaria induced morbidity and mortality. Villages surrounding the paramilitary installations often serve as disease reservoirs. Malaria incidence in Tripura State Rifles (TSR) units in Dhalai District of Tripura was studied and the role of the village population in disease transmission was also assessed. Methods: Mass blood surveys were carried out among TSR personnel and villagers during 2007 to 2009. Malaria diagnosis through blood smear examination and rapid detection kits was done, and percentage parasitaemia was determined. Activity of malaria vectors was monitored using CDC light traps. Results: Slide positivity rates (SPR) in the neighbouring villages (51.4%) was significantly higher than that in TSR (27.7%) (P<0.0001). Malaria incidence in villages did not show seasonal variability while it was lowest during post-monsoon season in TSR (P<0.325; OR = 0.74). Per cent Pf parasitaemia was high in TSR (0.29) as compared to villagers (0.20) (P<0.0001). Anopheles minimus and An. dirus were the major malaria vectors observed. Interpretation & conclusions: Paramilitary and public health authorities should adopt targeted measures to reduce the malaria incidence in the villages surrounding the paramilitary installations as the village populations play a major role in disease transmission.

Disturbance in hemoglobin metabolism and in vivo antimalarial activity of azole antimycotics / Alteración en el metabolismo de la hemoglobina y actividad antimalárica in vivo de azoles antimicóticos

Plasmodium parasites degrade host hemoglobin to obtain free amino acids, essential for protein synthesis. During this event, free toxic heme moieties crystallize spontaneously to produce a non-toxic pigment called hemozoin or ß-hematin. In this context, a group of azole antimycotics, clotrimazole (CTZ), ketoconazole (KTZ) and fluconazole (FCZ), were investigated for their abilities to inhibit ß-hematin synthesis (IßHS) and hemoglobin proteolysis (IHbP) in vitro. The ß-hematin synthesis was recorded by spectrophotometry at 405 nm and the hemoglobin proteolysis was determined by SDS-PAGE 12.5 percent, followed by densitometric analysis. Compounds were also assayed in vivo in a malaria murine model. CTZ and KTZ exhibited the maximal effects inhibiting both biochemical events, showing inhibition of β-hematin synthesis (IC50 values of 12.4 ± 0.9 µM and 14.4 ± 1.4 µM respectively) and inhibition of hemoglobin proteolysis (80.1 ± 2.0 percent and 55.3 ± 3.6 percent, respectively). There is a broad correlation to the in vivo results, especially CTZ, which reduced the parasitemia ( percentP) of infected-mice at 4th day post-infection significantly compared to non-treated controls (12.4 ± 3.0 percent compared to 26.6 ± 3.7 percent, p = 0.014) and prolonged the survival days post-infection. The results indicated that the inhibition of the hemoglobin metabolism by the azole antimycotics could be responsible for their antimalarial effect.

Los parásitos del género Plasmodium degradan la hemoglobina hospedera obteniendo aminoácidos libres para su síntesis proteica. Durante este evento, unidades de hemo libre tóxicas cristalizan espontáneamente formando un pigmento no tóxico denominado ß-hematina. En este trabajo, se investigó la capacidad de un grupo de azoles antimicóticos: clotrimazol (CTZ), ketoconazol (KTZ) y fluconazol (FCZ), en inhibir la síntesis de ß-hematina y la proteólisis de la globina. La síntesis de ß-hematina se registro por espectrofotometría a 405 nm y la proteólisis de la hemoglobina se determino por SDS-PAGE 15 por ciento seguido por análisis densitométrico de las bandas de hemoglobina intactas. Los compuestos fueron también ensayados in vivo en un modelo de malaria murina. CTZ y KTZ inhibieron la síntesis de ß-hematina con CI50 entre 10 y 15 µM y bloquearon la proteólisis de la hemoglobina (80.01 ± 2.04 por ciento y 55.33 ± 3.57 por ciento, respectivamente). En relación directa con los resultados encontrados in vitro, el CTZ redujo la parasitemia de ratones infectados en forma significativa, así como prolongó lo días de sobrevivencia post-infección en comparación con animales controles no tratados. Se sugiere así que la inhibición del metabolismo de la hemoglobina por los antimicóticos azólicos pudiera ser el mecanismo responsable de su actividad antimalárica.

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)

Uso de paneles de láminas estandarizadas para la evaluación de competencias en el diagnóstico microscópico de malaria en la Amazonía Peruana / Use of standardized blood smear slide sets for competency assessment in the malaria microscopic diagnosis in the Peruvian Amazon

Objetivos. Evaluar la competencia de los microscopistas en el diagnóstico de la malaria mediante paneles de láminas estandarizados en la Amazonía peruana. Materiales y métodos. Estudio transversal, realizado entre los meses de julio y septiembre de 2007, en 122 establecimientos de salud de primer nivel de atención de la Amazonía peruana. En el marco del Proyecto PAMAFRO, se evaluó las competencias en el diagnóstico de malaria en 68 microscopistas sin experiencia (

Objectives. To assess the competency of microscopists for malaria diagnosis using standardized slide sets in the Peruvian Amazon. Material and methods. Cross-sectional study carried out in 122 first level health facilities of the Peruvian Amazon, between July and September 2007. Within the frame of the project "Control Malaria in the border areas of the Andean Region: A community approach" (PAMAFRO), we evaluated the malaria diagnosis performance in 68 microscopists without expertise (< 1 year of expertise) and 76 microscopists with expertise (> 1 year) using standardized sets of 20 blood smear slides according to the World Health Organization (WHO) recommendations. A correct diagnosis (correct species identification) was defined as "agreement", a microscopist was qualified as an "expert" if they have an agreement ≥90 percent (≥ 18 slides with correct diagnosis), as a "referent" with an agreement between 80 percent and <90 percent, "competent" if they are between 70 and <80 percent and "in training" if they have <70 percent. Results. Microscopists with expertise (68.6 percent) had more agreement than those without expertise (48.2 percent). The competency assessment was acceptable (competent, referent, or experts levels) in 11.8 percent of the microscopists without expertise and in 52.6 percent from those with expertise. The agreement was lower using blood smear slides with P. falciparum with low parasitaemia, with P. malariae and with mixed infections. Conclusions. Is the first assessment, we found only one of three microscopists from the Peruvian Amazon is competent fro malaria diagnosis according to the WHO standards. From this baseline data, we have to continue working in order to improve the competency assessment of the microscopists within the frame of a quality assurance system.

Clinical features, diagnostic techniques and management of dual dengue and malaria infection

To find out clinical features, diagnostic techniques and management outcome of patients having dual dengue and malaria infection. A case series. Medical Unit-III, Ward- 7, Jinnah Postgraduate Medical Centre, Karachi, from September 2007 to January 2008. Patients presented with fever of less than or equal to 10 days duration, severe body aches, rash and bleeding manifestations were included. Patients with obvious features of other diseases like typhoid, hereditary bleeding diathesis and hematological malignancies and only malarial parasite positive with high grade intermittent fever without rash and myalgia were excluded from the study. Diagnosis of dengue and malaria was based on history, clinical features, laboratory parameters and malarial parasite test by thin and thick films. Serological evaluation was done by dengue IgM and IgG by ELISA test kit. Patients were divided into three groups. Group A was dengue IgM positive plus MP positive, group B was dengue IgM positive and MP negative and group C was dengue IgM negative and were clinically suspected dengue and malaria. The clinical manifestations and laboratory parameters of dual dengue and malaria positive patients were compared with malaria and dengue negative patients. One hundred and fourteen patients were seen during the study period. Antibody titer [IgM] tested in all patients was found positive in 78 patients [69.64%]. Among those 78 patients, 26 [23.21%] were concomitantly positive for malarial parasite [Group A]. Plasmodium vivax was positive in 25 patients and falciparum in one patient. Fifty-two patients [46.42%] were dengue IgM positive and MP negative [Group B]. Thirty four [30.35%] patients were MP and dengue IgM negative [Group C] but were strongly suspected for DHF and malaria on clinical and hematological basis. The hemoglobin of 34.61% of patients of group A, 5.76% of group B and 14.7% of group C were low, hematocrit level was also low in group A[92.3%], group B [15.38%] and group C [70.58%] patients. The platelet count was markedly low in 84.61% of patients of group A, 57.69% of group B and 94.11% of group C. Leukopenia was found in 34.61% of patients of group A, 78.84% in group B and 29.411% in group C. The liver function tests were deranged in all groups. The frequency of dual dengue and malaria infection was 23.21%. The serology of the dengue and malaria showed negative results in 30.35%. The diagnosis of dual infections could be made on the basis of history, clinical examination supported by hematological results. It is recommended that all the patients suspected for dual infections should be treated concomitantly for dengue and malaria in malaria endemic areas

Plasma homocysteine levels in cardiovascular disease, malaria and protein-energy malnutrition in Sudan

This study investigated the role of hyperhomocysteinaemia as a risk factor in Sudanese adults suffering from cardiovascular disease or malaria and children with protein-energy malnutrition. Mean total plasma homocysteine levels [micro mol/L] were significantly higher in patients with coronary heart disease [17.64; SD 11.68] recurrent venous thrombosis [5.06; SD 10.55] and recurrent malaria [13.61; SD 4.82] than in healthy adult controls [7.85; SD 3.39]. The mean homocysteine level was also significantly higher in children with protein-energy malnutrition [8.41; SD 1.61] than in healthy control children [5.72; SD 1.99]

Detection & molecular confirmation of a focus of Plasmodium malariae in Arunachal Pradesh, India.

BACKGROUND & OBJECTIVE: During a malaria epidemiological study in Arunachal Pradesh, Plasmodium malariae like human malaria parasites were seen in blood smears from fever cases. The study was undertaken to detect the presence of P. malariae and to confirm its identity through DNA based polymerase chain reaction approach. METHODS: Fever survey was carried out in 22 villages in Indo-Myanmar bordering district of Lohit, Arunachal Pradesh in 2005. Morphologically suspected P. malariae cases were confirmed using nested PCR based on 18S small subunit ribosomal DNA gene sequence. RESULTS: Screening of 1,995 fever cases resulted in 9 probable cases of P. malariae based on morphological identification in Chakma tribe people residing in 2 villages. Nested PCR confirmed the identity of all probable cases of P. malariae by producing diagnostic band of 144 bp. PCR method was able to detect mixed infection of P. malariae with P. vivax and with P. falciparum. INTERPRETATION & CONCLUSION: P. malariae may have been present in Arunachal Pradesh but most probably is being misdiagnosed due to its close resemblance with P. vivax, especially in ring forms. Estimation of actual case load of P. malariae in north-east India is, therefore, important with accurate species identification using molecular methods.

Possible biochemical impact of malaria infection in subjects with HIV co-infection in Anambra state, Nigeria.

BACKGROUND & OBJECTIVES: The present study was designed to determine possible contributory impact of malaria infection on some biochemical markers in subjects with HIV co-infection in order to know if they are adverse or protective. METHODS: Participants were recruited at the Voluntary Counseling and Testing Unit, Nnamdi Azikiwe University Teaching Hospital, Nnewi, Nigeria and grouped into: (i) Malaria and HIV co-infection group (n = 45); and (ii) HIV infected group without concurrent malaria infection (n = 57). Standard laboratory methods were used for the HIV and Plasmodium falciparum antigen screening, malaria parasite density, CD4+ T-cell count, packed cell volume, white blood cell count, serum iron and albumin concentrations. RESULTS: The results showed that serum iron and albumin were significantly reduced and raised respectively in 'Malaria-HIV co-infection group' compared with 'HIV infection group' (p < 0.05 and p < 0.05). A positive association was observed between age and serum iron concentration in malaria and HIV co-infected group (r = 0.580; p < 0.05) while negative associations were observed between PCV and serum iron (r = - 0.388; p < 0.05) and between CD4+ T-cells and serum iron concentration (r = -0.362; p < 0.05) in malaria and HIV co-infected group. The CD4+ T-cell count, WBC count, PCV were not significantly different between the Malaria-HIV co-infection group and HIV infection group. INTERPRETATION & CONCLUSION: In the present study serum iron and albumin concentrations were the most sensitive indicators that showed the contributory impact of malaria infection on biochemical index in HIV co-infected subjects. The findings suggest that at the defined stage of HIV infection in the present study, malaria co-infection may moderate the impact of HIV infection on iron metabolism and hepatic synthesis of albumin.