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Malaria Journal Aug 2023Anopheles funestus, which is considered as secondary vector of malaria in Ethiopia, is known to have several morphologically indistinguishable (sibling) species....
BACKGROUND
Anopheles funestus, which is considered as secondary vector of malaria in Ethiopia, is known to have several morphologically indistinguishable (sibling) species. Accurate identification of sibling species is crucial to understand their biology, behaviour and vector competence. In this study, molecular identification was conducted on the Ethiopian An. funestus populations. Moreover, insecticide resistance mechanism markers were detected, including ace N485I, kdr L1014F, L1014S, and CYP6P9a TaqMan qPCR was used to detect the infective stage of the parasite from field collected adult female An. funestus populations.
METHODS
Adult female mosquito collection was conducted from Lare, Gambella Regional State of Ethiopia between June 2018 to July 2020 using CDC light traps and HLC. Sub-samples of the morphologically identified An. funestus mosquitoes were molecularly identified using species-specific PCR, and the possible presence of insecticide resistance alleles was investigated using TaqMan qPCR (N485I-Ace-1), PCR-Sanger sequencing (L1014F-kdr), and PCR-RFLP (CYP6P9a resistance allele). Following head/thorax dissection, the TaqMan qPCR assay was used to investigate the presence of the infective stage Plasmodium parasite species.
RESULTS
A total of 1086 adult female An. funestus mosquitoes were collected during the study period. All sub-samples (N = 20) that were morphologically identified as An. funestus sensu lato (s.l.) were identified as An. funestus sensu stricto (s.s.) using species- specific PCR assay. The PCR-RFLP assay that detects the CYP6P9a resistance allele that confers pyrethroid resistance in An. funestus was applied in N = 30 randomly selected An. funestus s.l.
SPECIMENS
None of the specimens showed a digestion pattern consistent with the presence of the CYP6P9a resistance allele in contrast to what was observed in the positive control. Consequently, all samples were characterized as wild type. The qPCR TaqMan assay that detects the N485I acetylcholinesterase-1 mutation conferring resistance to organophosphates/carbamates in An. funestus was used in (N = 144) samples. All samples were characterized as wild type. The kdr L1014F and L1014S mutations in the VGSC gene that confer resistance to pyrethroids and DDT were analysed with direct Sanger sequencing after PCR and clean-up of the PCR products were also characterized as wild type. None of the samples (N = 169) were found positive for Plasmodium (P. falciparum/ovale/malariae/vivax) detection.
CONCLUSION
All An. funestus s.l. samples from Lare were molecularly identified as An. funestus s.s. No CYP6P9, N485I acetylcholinesterase 1, kdr L1014F or L1014S mutations were detected in the An. funestus samples. None of the An. funestus samples were positive for Plasmodium. Although the current study did not detect any insecticide resistant mechanism, it provides a reference for future vector monitoring programmes. Regular monitoring of resistance mechanisms covering wider geographical areas of Ethiopia where this vector is distributed is important for improving the efficacy of vector control programs.
Topics: Animals; Female; Anopheles; Acetylcholinesterase; Alleles; Ethiopia; Mosquito Vectors; Insecticides; Pyrethrins; Insecticide Resistance; Malaria
PubMed: 37573300
DOI: 10.1186/s12936-023-04667-3 -
Parasites & Vectors Aug 2023Infections with Plasmodium ovale are widely distributed but rarely investigated, and the resulting burden of disease has been underestimated. Plasmodium ovale curtisi...
BACKGROUND
Infections with Plasmodium ovale are widely distributed but rarely investigated, and the resulting burden of disease has been underestimated. Plasmodium ovale curtisi Duffy binding protein domain region II (PocDBP-RII) is an essential ligand for reticulocyte recognition and host cell invasion by P. ovale curtisi. However, the genomic variation, antigenicity and immunogenicity of PocDBP-RII remain major knowledge gaps.
METHODS
A total of 93 P. ovale curtisi samples were collected from migrant workers who returned to China from 17 countries in Africa between 2012 and 2016. The genetic polymorphism, natural selection and copy number variation (CNV) were investigated by sequencing and real-time PCR. The antigenicity and immunogenicity of the recombinant PocDBP-RII (rPocDBP-RII) protein were further examined, and the humoral and cellular responses of immunized mice were assessed using protein microarrays and flow cytometry.
RESULTS
Efficiently expressed and purified rPocDBP-RII (39 kDa) was successfully used as an antigen for immunization in mice. The haplotype diversity (Hd) of PocDBP-RII gene was 0.105, and the nucleotide diversity index (π) was 0.00011. No increased copy number was found among the collected isolates of P. ovale curtisi. Furthermore, rPocDBP-RII induced persistent antigen-specific antibody production with a serum IgG antibody titer of 1: 16,000. IFN-γ-producing T cells, rather than IL-10-producing cells, were activated in response to the stimulation of rPocDBP-RII. Compared to PBS-immunized mice (negative control), there was a higher percentage of CD4CD44CD62L T cells (effector memory T cells) and CD8CD44CD62L T cells (central memory T cells) in rPocDBP-RII‑immunized mice.
CONCLUSIONS
PocDBP-RII sequences were highly conserved in clinical isolates of P. ovale curtisi. rPocDBP-RII protein could mediate protective blood-stage immunity through IFN-γ-producing CD4 and CD8 T cells and memory T cells, in addition to inducing specific antibodies. Our results suggested that rPocDBP-RII protein has potential as a vaccine candidate to provide assessment and guidance for malaria control and elimination activities.
Topics: Animals; Mice; Plasmodium ovale; Interferon-gamma; CD8-Positive T-Lymphocytes; DNA Copy Number Variations; Protein Domains; Malaria
PubMed: 37553591
DOI: 10.1186/s13071-023-05897-9 -
The American Journal of Tropical... Sep 2023Plasmodium vivax is the second-most common malaria pathogen globally, but is considered very rare in the predominantly Duffy-negative sub-Saharan African population. In...
Plasmodium vivax is the second-most common malaria pathogen globally, but is considered very rare in the predominantly Duffy-negative sub-Saharan African population. In 259 malaria patients from highland southern Rwanda, we assessed Plasmodium species and Duffy blood group status by polymerase chain reaction (PCR). Plasmodium falciparum, P. vivax, Plasmodium malariae, and Plasmodium ovale were seen in 90.7%, 8.1%, 11.6%, and 5.0%, respectively. Plasmodium vivax occurred more frequently as a monoinfection than in combination with P. falciparum. All P. vivax-infected individuals showed heterozygous Duffy positivity, whereas this was the case for only 3.1% of patients with P. falciparum monoinfection and malaria-negative control subjects (P < 0.01). Based on PCR diagnosis, P. vivax is not rare in southern Rwanda. All episodes of P. vivax were observed in heterozygous Duffy-positive patients, whereas elsewhere in Africa, P. vivax is also reported in Duffy-negative individuals. Refined mapping of Plasmodium species is required to establish control and elimination strategies including all malaria species.
Topics: Humans; Malaria, Vivax; Rwanda; Malaria; Plasmodium vivax; Malaria, Falciparum; Plasmodium falciparum; Plasmodium malariae; Duffy Blood-Group System
PubMed: 37549894
DOI: 10.4269/ajtmh.23-0143 -
Genome Biology and Evolution Aug 2023Ubiquitin is an extraordinarily highly conserved 76 amino acid protein encoded by three different types of gene, where the primary translation products are fusions...
Ubiquitin is an extraordinarily highly conserved 76 amino acid protein encoded by three different types of gene, where the primary translation products are fusions either of ubiquitin with one of two ribosomal proteins (RPs) or of multiple ubiquitin monomers from head to tail. Here, we investigate the evolution of ubiquitin genes in mammalian malaria parasites (Plasmodium species). The ubiquitin encoded by the RPS27a fusion gene is highly divergent, as previously found in a variety of protists. However, we also find that two other forms of divergent ubiquitin sequence, each previously thought to be extremely rare, have arisen recently during the divergence of Plasmodium subgenera. On two occasions, in two distinct lineages, the ubiquitin encoded by the RPL40 fusion gene has rapidly diverged. In addition, in one of these lineages, the polyubiquitin genes have undergone a single codon insertion, previously considered a unique feature of Rhizaria. There has been disagreement whether the multiple ubiquitin coding repeats within a genome exhibit concerted evolution or undergo a birth-and-death process; the Plasmodium ubiquitin genes show clear signs of concerted evolution, including the spread of this codon insertion to multiple repeats within the polyubiquitin gene.
Topics: Animals; Ubiquitin; Polyubiquitin; Ribosomal Proteins; Magnoliopsida; Plasmodium; Mammals
PubMed: 37481258
DOI: 10.1093/gbe/evad137 -
Malaria Journal Jul 2023Malaria is a major public health problem, particularly in the tropical regions of America, Africa and Asia. Plasmodium falciparum is not only the most widespread but...
BACKGROUND
Malaria is a major public health problem, particularly in the tropical regions of America, Africa and Asia. Plasmodium falciparum is not only the most widespread but also the most deadly species. The share of Plasmodium infections caused by the other species (Plasmodium ovale and Plasmodium malariae) is clearly underestimated. The objective of the study was to determine the molecular epidemiology of plasmodial infection due to P. malariae and P. ovale in Côte d'Ivoire.
METHODS
The study was cross-sectional. The study participants were recruited from Abengourou, San Pedro and Grand-Bassam. Sample collection took place from May 2015 to April 2016. Questionnaires were administered and filter paper blood samples were collected for parasite DNA extraction. The molecular analysis was carried out from February to March 2021. A nested PCR was used for species diagnosis. The data was presented in frequencies and proportions.
RESULTS
A total of 360 patients were recruited, including 179 men (49,7%) for 181 women (50,3%). The overall Plasmodium positive rate was 72.5% (261/360). The specific index was 77.4% and 1.5% for P. falciparum and P. malariae in mono-infection, respectively. There was also 15% P. falciparum and P. malariae co-infection, 3.4% P. falciparum and P. ovale co-infection and 2.3% P. falciparum, P. malariae and P. ovale triple-infection. Typing of P. ovale subspecies showed a significant predominance of P. ovale curtisi (81.2% of cases).
CONCLUSION
Plasmodium falciparum remains the most prevalent malaria species in Côte d'Ivoire, but P. malariae and P. ovale are also endemic mostly in co-infection. Malaria elimination requires a better understanding of the specific epidemiological characteristics of P. malariae and P. ovale with a particular emphasis on the identification of asymptomatic carriers.
Topics: Male; Humans; Female; Plasmodium falciparum; Cote d'Ivoire; Molecular Epidemiology; Coinfection; Cross-Sectional Studies; Prevalence; Malaria, Falciparum; Malaria; Plasmodium ovale; Plasmodium malariae
PubMed: 37468917
DOI: 10.1186/s12936-023-04639-7 -
Mikrobiyoloji Bulteni Jul 2023Malaria is a serious, contagious infection caused by single-celled parasites. About 200 species of Plasmodium have been described that can cause infection in...
Malaria is a serious, contagious infection caused by single-celled parasites. About 200 species of Plasmodium have been described that can cause infection in vertebrates. Five different species of Plasmodium are known to cause infection in humans to date. Infection with more than one type of pathogen is called coinfection. This type of infections can be caused by different species of the same genus, as well as by different species. Malaria coinfections are mostly caused by the combination of Plasmodium vivax and Plasmodium falciparum. In this study, a case of malaria admitted to the hospital and diagnosed was presented. Thin smear blood preparations were prepared from the peripheral blood of a 54 year-old Republic of Türkiye citizen male patient who applied to the emergency department with fever and chills. The preparations were stained with Giemsa and examined under a microscope with a x 100 objective, and trophozoite and gametocyte forms belonging to Plasmodium genus were determined. As a result of probe-based quantitative real-time polymerase chain reaction (qRt-PCR) study with primers specific to Plasmodium vivax, Plasmodium malariae, Plasmodium falciparum, Plasmodium ovale and Plasmodium knowlesi for definitive species identification, co-infection of P.vivax, P.falciparum, P.ovale and P.knowlesi was detected in the patient. In addition, it was proved that our patient was infected with four different species by conventional PCR study in which five species were studied and then by DNA sequence analysis. On the fourth day of artemether-lumefantrine treatment, the patient's fever response was observed and the trophozoite forms disappeared from the third day in the daily peripheral smear follow-up. Since P.vivax and P.ovale species were also detected after species determination by molecular methods, primaquine 1 x 30 mg tablet was added to the existing drugs for the treatment of hypnozoite forms of the parasite. In recent years, there has been an increase in malaria imported cases, especially after visits to African countries. Such rare cases of malaria coinfection may be encountered during visits to geographies located at the intersection of endemic regions. According to the data of the World Health Organization, maximum attention should be paid to the prevention and prophylaxis protocols from vectors, especially in travels to countries with the highest mortality and morbidity. In co-infection cases similar to our patient, for tertian malaria and tertiary ovale malaria, hypnozoid therapy should not be overlooked. When the insecticide-resistant vectors and drug-resistant Plasmodium strains encountered in recent years are evaluated as a whole, there is a need to develop more effective strategies in the fight against malaria. In addition to microscopic examination, which is accepted as the gold standard, we believe that evaluating molecular studies together in diagnosis is extremely important for the treatment process when hypnozoite periods are considered.
Topics: Animals; Male; Humans; Middle Aged; Coinfection; Antimalarials; Cameroon; Artemether; Artemether, Lumefantrine Drug Combination; Malaria; Malaria, Vivax; Plasmodium; Plasmodium falciparum; Plasmodium vivax; Real-Time Polymerase Chain Reaction
PubMed: 37462313
DOI: 10.5578/mb.20239942 -
Frontiers in Medicine 2023Malaria remains a severe life-threatening disease caused by plasmodium parasites. Microscopy is widely used for malaria diagnosis. However, it relies heavily on the...
BACKGROUND
Malaria remains a severe life-threatening disease caused by plasmodium parasites. Microscopy is widely used for malaria diagnosis. However, it relies heavily on the skills and experience of inspectors. Due to low-level medical services and the lack of skilled inspectors, misdiagnoses are frequently made in some areas.
METHODS
In recent years, many successful applications of CNN models have been reported. Unlike images in the ImageNet, the image of plasmodium only has a tiny defect area with a large amount of information. In addition, the dataset is extremely unbalanced: the number of positive samples is much less than that of negative samples. This paper proposes a classification network by combining attention mechanism and ResNeSt for plasmodium detection and using self-supervised learning to pre-train the network. First, the positive samples were adopted to pre-train the network. Then, attention modules were taken to highlight the feature area. To support current and future research, we also constructed a plasmodium dataset with Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, and Plasmodium malaria and non-Plasmodium. Through self-supervised learning, a large amount of unlabeled data is used to mine the representational features, thus improving the feature extraction capability of the model and achieving higher accuracy, while saving the physician's labeling time and improving the classification accuracy.
RESULTS
The experiments show that our model exhibits an excellent performance and that the test accuracy, sensitivity, and specificity attain 97.8%, 96.5%, and 98.9%, respectively.
CONCLUSION
The AI classification method proposed in this paper can effectively assist clinicians in the diagnosis and provide a basis for the automatic detection of malaria parasites in the future.
PubMed: 37457573
DOI: 10.3389/fmed.2023.1117192 -
Zhongguo Xue Xi Chong Bing Fang Zhi Za... Jun 2023To analyse the epidemiological characteristics of imported malaria cases after malaria elimination in Yixing City, Jiangsu Province, so as to provide reference for...
OBJECTIVE
To analyse the epidemiological characteristics of imported malaria cases after malaria elimination in Yixing City, Jiangsu Province, so as to provide reference for malaria prevention and control in grassroots healthcare institutions.
METHODS
All data pertaining to malaria cases reported in Yixing City from 2016 to 2022 were retrieved from Chinese Disease Control and Prevention Information System, and the data pertaining to vector monitoring and human malaria parasite infections from 2016 to 2022 were collected for a descriptive statistical analysis.
RESULTS
A total of 14 imported malaria cases were reported in Yixing City from 2016 to 2022, including 12 cases with malaria, one case with malaria and one case with malaria, and all cases acquired infections in Africa and then returned to Yixing City. Malaria cases were reported across 2016 to 2022 except in 2020 and 2021. Malaria cases were predominantly reported during the period between December and February of the next year, and workers were the predominant occupation. The institutions where malaria was initially diagnosed included county-level general hospitals, county-level disease prevention and control institutions and grassroots healthcare centers, and there were 10 cases with definitive diagnosis of malaria on the day of initial diagnosis, with a 64.29% (9/14) correct rate of initial diagnosis. There were 5 cases diagnosed with severe malaria, and the standardized response rate was 100.00% following the "1-3-7" surveillance and response strategy. Of all malaria vectors, only was monitored in Yixing City from 2016 to 2022, and all humans were tested negative for blood smears exceptimportedmalariacases.
CONCLUSIONS
The correct rate of initial malaria diagnosis was not high in healthcare institutions in Yixing City from 2016 to 2022, and there are still multiple challenges for prevention of re-establishment of imported malaria.
Topics: Animals; Humans; Mosquito Vectors; Malaria; Malaria, Vivax; Malaria, Falciparum; Cities; China
PubMed: 37455103
DOI: 10.16250/j.32.1374.2023028