Anaplasma phagocytophilum and Other Anaplasma spp. in Various Hosts in the Mnisi Community, Mpumalanga Province, South Africa

DNA samples from 74 patients with non-malarial acute febrile illness (AFI), 282 rodents, 100 cattle, 56 dogs and 160 Rhipicephalus sanguineus ticks were screened for the presence of Anaplasma phagocytophilum DNA using a quantitative PCR (qPCR) assay targeting the msp2 gene. The test detected both A. phagocytophilum and Anaplasma sp. SA/ZAM dog DNA. Microbiome sequencing confirmed the presence of low levels of A. phagocytophilum DNA in the blood of rodents, dogs and cattle, while high levels of A. platys and Anaplasma sp. SA/ZAM dog were detected in dogs. Directed sequencing of the 16S rRNA and gltA genes in selected samples revealed the presence of A. phagocytophilum DNA in humans, dogs and rodents and highlighted its importance as a possible contributing cause of AFI in South Africa. A number of recently described Anaplasma species and A. platys were also detected in the study. Phylogenetic analyses grouped Anaplasma sp. SA/ZAM dog into a distinct clade, with sufficient divergence from other Anaplasma species to warrant classification as a separate species. Until appropriate type-material can be deposited and the species is formally described, we will refer to this novel organism as Anaplasma sp. SA dog.

Genetic Diversity of Merozoite Surface Protein 2 (MSP2) Plasmodium falciparum Clinical Isolate in Wamena General Hospital

The genetic diversity of typical clinical isolated Plasmodium falciparum in the malaria population varies greatly, especially at the location where malaria disease were recorded at high incidence rate. MSP2 is known as glycoprotein expressed on the surface of merozoites, which is an antigenic protein and has a potential to act as vaccine candidate for malaria. The MSP2 gene has two main allelic groups called FC27 and 3D7/IC. Block 3 from MSP2 gene is the most polymorphic to describe the diversity of parasite populations. The P. falciparum parasite population is often characterized by wide genetic diversity in areas of high transmission intensity. Therefore, the study on P. falciparum diversity is useful to describe the level of malaria transmission. The study of genetic diversity focused on clinical isolated species at Wamena General Hospital was aimed to determine the presence of the MSP2 gene, variety of MSP2 gene allele and the dominant frequency of the MSP2 gene allele. This research has been carried out from March 2018 to February 2019 using a cross sectional approach. The research sample was taken and prepared from Wamena Regional Hospital and followed by the analyzing of DNA isolation, PCR, electrophoresis of the research samples was done at the genetic science laboratory in Jakarta, Indonesia. The samples studied were patients who met the inclusion criteria, namely a single P. falciparum infection with an asexual parasite density >1000 parasites/µl or >3+ (1-10 P/Lp), and were agreed to become respondents by signing an informed consent. A total of 26 clinical isolates of P. falciparum were isolated with the MSP2 gene distribution on the FC27 allele with the highest as many as 25 samples (96.2%), 22 samples (84.6%) of the 3D7 / IC allele while the mixture of the two alleles was 22 samples (84.6%). From a total of 26 samples, there were samples with the male gender category counted for 77.3% and female 41%. The results of the identification of clinical isolated P. falciparum at Wamena Hospital with a total of 26 samples were found in productive age, between 15-34 years with a single allele (95.8%), while 23 cases and mix (both alleles 87.5%) about 21 cases, meanwhile in cases of before-productive age, in which ages were 12 and 14 years of age with a single allele 100% (FC27) 2 cases and 50% (3D7/IC) found to be 1 case, The mixture of the two alleles is 50% was only 1 case and there was no sample at non-productive age observed. Key words: Malaria; MSP-2; P. falciparum; Wamena

Genetic diversity of Plasmodium falciparum in Grande Comore Island

Background Despite several control interventions resulting in a considerable decrease in malaria prevalence in the Union of the Comoros, the disease remains a public health problem with high transmission in Grand Comore compared to neighboring islands. In this country, only a few studies investigating the genetic diversity of Plasmodium falciparum have been performed so far. For this reason, this study aims to examine the genetic diversity of P. falciparum by studying samples collected in Grande Comore in 2012 and 2013, using merozoite surface protein 1 ( msp1 ), merozoite surface protein 2 ( msp2 ) and single nucleotide polymorphism (SNP) genetic markers. Methods A total of 151 positive rapid diagnostic test (RDT) samples from Grande Comore were used to extract parasite DNA. Allelic families K1, Mad20 and RO33 of the msp1 gene as well as allelic families IC3D7 and FC37 of the msp2 gene were determined by using nested PCR. Additionally, 50 out of 151 samples were genotyped to study 24 SNPs by using high resolution melting (HRM). Results Two allelic families were predominant, the K1 family of msp1 gene (55%) and the FC27 family of msp2 gene (47.4%). Among 50 samples genotyped for 24 SNPs, 42 (84%) yielded interpretable results. Out of these isolates, 36 (85%) were genetically unique and 6 (15%) grouped into two clusters. The genetic diversity of Plasmodium falciparum calculated from msp gene ( msp1 and msp 2) and SNPs was 0.82 and 0.6 respectively. Conclusion In summary, a large genetic diversity of P. falciparum was observed in Grande Comore. This may favor persistence of malaria, and might be one of the reasons for the high malaria transmission compared to neighboring islands. Further surveillance of P. falciparum isolates, mainly through environmental management / vector control, is warranted until complete elimination is attained.

Distribution of msp1, msp2 and eba175 Allelic Family According to Hemoglobin Genotype and G6PD Type from Children with Uncomplicated Malaria in Banfora Heath District (Burkina Faso)

Aim: The present study aimed to evaluate the Plasmodium falciparum genetic diversity according to the host hemoglobin and G6PD genetic variants during the course of malaria in infected children aged from 2 to 10 years and living in endemic area in Burkina Faso. Study Design: The study was designed as a longitudinal follow up conducted between May 2015 and February 2016 in Banfora health district, Burkina Faso. Methodology: We included 136 subjects (73 males and 63 females; age range from 2-10 years). Blood thick and thin film was done by capillary blood. Venous blood was collected for DNA extraction. Malaria diagnosis was done by microscopy. Human and parasite DNA were extracted based on Qiagen kit procedure. Then, hemoglobin and G6PD were genotyped by RLFP-PCR while the msp1, msp2 and eba175 genes were typed by a nested PCR. All PCR products were analyzed by electrophoresis on a 1.5-2% agarose gel and alleles categorized according to the molecular weight. Results: The prevalence of hemoglobin type was 19.11% for abnormal hemoglobin and 80.9% for normal hemoglobin carriage. The prevalence of G6PD type was 91.18% for normal and 8.82% for G6PD deficiency carriage, respectively. The prevalence of msp1 allelic families was 81.60%, 80.80% and 67.20% for k1, ro33 and mad20 respectively while for msp2 gene, fc27 and 3D7 allelic family the prevalence was 70.53% and 69.64% respectively. The eba175 allelic families’ distribution showed 77.31% and 40.21% for fcr3 and Camp respectively. There was no difference in multiplicity of infection (MOI) according to hemoglobin genotypes and G6PD types. We found that k1 was the predominant allelic family of msp1 in normal hemoglobin genotype (AA) and normal G6PD type. The mixed infection of eba175 was statistically higher in abnormal hemoglobin (p=0.04). There was no statistical difference between fcr3 and camp prevalence excepted in G6PD deficient type. The polymorphism results showed that the prevalence of 450 bp in fc27 was statistically significantly higher in normal hemoglobin variant carriers (AA) than abnormal hemoglobin carriers (p=2.10 -4)). However, the prevalence of 350 bp in fc27 was statistically higher in normal G6PD than deficient G6PD carriers (p=0.034). Conclusion: Our result showed that the distribution of msp1 and eba75 polymorphism could be influenced by hemoglobin and G6PD variants. These results suggest that hemoglobin and G6PD could influence P. falciparum genetic diversity.

Genetic polymorphism of Merozoite Surface Protein 1 (msp1) and 2 (msp2) genes and multiplicity of Plasmodium falciparum infection across various endemic areas in Senegal

Introduction: Despite a significant decline in Senegal, malaria remains a burden in various parts of the country. Assessment of multiplicity of Plasmodium falciparum infection and genetic diversity of parasites population could help in monitoring of malaria control.Objective: To assess genetic diversity and multiplicity of infection in P. falciparum isolates from three areas in Senegal with different malaria transmissions. Methods: 136 blood samples were collected from patients with uncomplicated P. falciparum malaria in Pikine, Kedougou and Thies. Polymorphic loci of msp1 and 2 (Merozoite surface protein-1 and 2) genes were amplified by nested PCR.Results: For msp1gene, K1 allelic family was predominant with frequency of 71%. Concerning msp2 gene, IC3D7 allelic family was the most represented with frequency of 83%. Multiclonal isolates found were 36% and 31% for msp1et msp2 genes respectively. The MOI found in all areas was 2.56 and was statistically different between areas (P=0.024). Low to intermediate genetic diversity were found with heterozygosity range (He=0,394-0,637) and low genetic differentiation (Fst msp1= 0.011; Fst msp2= 0.017) were observed between P. falciparum population within the country.Conclusion: Low to moderate genetic diversity of P.falciparum strains and MOI disparities were found in Senegal.Keywords: Senegal, MOI, Genetic diversity, msp1, msp2.