Genetic Polymorphisms of Pfcrt K76T and Pfmdr1 N86Y among Asymptomatic School Children in Forest Communities of Ekondo Titi Subdivision along the Cameroon-Nigeria Border Area

Aims: The study sought to quantify Plasmodium infection and molecular markers for chloroquine resistance among asymptomatic school children. Study Design: The study was cross-sectional. Place and Duration of Study: The study was carried out in Ekondo Titi Subdivision near Cameroon's south-western border with Nigeria from March to May and from September to October 2014. Methodology: The prevalence of human Plasmodium species was determined by nested PCR (Polymerase Chain Reaction) using DNA from dried blood spot in six primary schools. A PCR/RFLP analysis (Restriction Fragment Length Polymorphism) was used to determine the prevalence of chloroquine resistance (CQR) associated pfcrt 76T and pfmdr 1 56Y point mutations in Plasmodium falciparum asymptomatic school children. Results: A nested PCR amplifying the 18S small-subunit ribosomal RNA (SSU rRNA) gene of Plasmodium in 205 samples confirmed 76.1% of the isolates as asymptomatic P. falciparum infections, with a substantial proportion 22% of P. malariae infection. Among these, 3.6% were single P. malariae infections and 15.1% were P. falciparum and P. malariae mixed infections. Mixed P. falciparum and P. ovale infections were 2.0%. Of the 156 Plasmodium falciparum, positive samples by species-specific PCR, 107 samples with P. falciparum mono-infection were analyzed for the presence of drug resistant alleles pfcrt 76T and pfmdr1- Y 86. The prevalence of pfcrt 76T mutation (74.6%) was higher than that of the pfmdr1-Y86 mutation (25.4%). Logistic regression analysis of socio-demographic factors predicted no significant association between pfcrt 76T mutation with gender and communities. Conclusions: The results indicated a high prevalence of P. malariae and mixed infection in the area under study. The high-level distribution of the pfcrtT76 observed in the study could be possibly attributed to the fact that CQ remained widely used at the community level more than 14 years after withdrawal.

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Fixation of pfcrt chloroquine resistance alleles in Plasmodium falciparum clinical isolates collected from unrest tribal agencies of Pakistan

Brazilian Journal of Biology ◽

10.1590/1519-6984.247422 ◽

2023 ◽

Vol 83 ◽

Author(s):

M. F. Nadeem ◽

N. Zeeshan ◽

A. A. Khattak ◽

U. A. Awan ◽

A. Yaqoob

Keyword(s):

Plasmodium Falciparum ◽

Nested Pcr ◽

Resistant Mutant ◽

Small Subunit ◽

Clinical Isolates ◽

Chloroquine Resistance ◽

Mortality And Morbidity ◽

Blood Smears ◽

Tribal Areas ◽

Pcr Targeting

Abstract Plasmodium falciparum resistance to Chloroquine (CQ) is a significant cause of mortality and morbidity worldwide. There is a paucity of documented data on the prevalence of CQ-resistant mutant haplotypes of Pfcrt and Pfmdr1 genes from malaria-endemic war effected Federally Administered Tribal Areas of Pakistan. The objective of this study was to investigate the prevalence of P. falciparum CQ-resistance in this area. Clinical isolates were collected between May 2017 and May 2018 from North Waziristan and South Waziristan agencies of Federally Administrated Trial Area. Subsequently, Giemsa-stained blood smears were examined to detect Plasmodium falciparum. Extraction of malarial DNA was done from microscopy positive P. falciparum samples, and P. falciparum infections were confirmed by nested PCR (targeting Plasmodium small subunit ribosomal ribonucleic acid (ssrRNA) genes). All PCR confirmed P. falciparum samples were sequenced by pyrosequencing to find out mutation in Pfcrt gene at codon K76T and in pfmdr1 at codons N86Y, Y184F, N1042D, and D1246Y. Out of 121 microscopies positive P. falciparum cases, 109 samples were positive for P. falciparum by nested PCR. Pfcrt K76T mutation was found in 96% of isolates, Pfmdr1 N86Y mutation was observed in 20%, and 11% harboured Y184F mutation. All samples were wild type for Pfmdr1 codon N1042D and D1246Y. In the FATA, Pakistan, the frequency of resistant allele 76T remained high despite the removal of CQ. However, current findings of the study suggest complete fixation of P. falciparum CQ-resistant genotype in the study area.

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Molecular Pattern of Anti-malarial Drug Resistance of Plasmodium falciparum in Bangladeshi Troops Working in Endemic Areas of Bangladesh and Africa

Bangladesh Journal of Microbiology ◽

10.3329/bjm.v37i1.51202 ◽

2020 ◽

Vol 37 (1) ◽

pp. 1-6

Author(s):

Md Nurul Amin ◽

Mahmuda Yasmin ◽

Marufa Zerin Akhtar ◽

Chowdhury Rafiqul Ahsan

Keyword(s):

Plasmodium Falciparum ◽

Nested Pcr ◽

Plasmodium Species ◽

Armed Forces ◽

Chloroquine Resistance ◽

Chittagong Hill Tracts ◽

Resistance Marker ◽

African Countries ◽

Endemic Areas ◽

Sub Saharan

Members of Bangladesh Armed Forces work in two different malaria endemic area, Chittagong Hill Tracts (CHT) in Bangladesh and Sub-Saharan countries in Africa. This under-recognized group remained unexplored for long in respect to drug resistant falciparum malaria they usually suffer from. In this study, a total of 252 ‘dried blood samples on filter paper’ were collected between November 2014 and February 2016, from Plasmodium falciparum positive Bangladeshi troops working in Chittagong Hill Tracts (CHT), Bangladesh and five Sub Saharan African Countries namely, Central African Republic (CAR), Democratic Republic of Congo (DRC), Liberia, Mali and Ivory Coast. After DNA extraction from all these samples (94 from Bangladesh and 138 from African countries), plasmodium species was confirmed by a nested PCR following standard protocol with minor modifications. Thereafter, a multiplex nested PCR followed by restriction fragment length polymorphism (RFLP) method was employed to investigate the presence of chloroquine resistance marker ‘K76T mutation’ in P. falciparum chloroquine resistance transporters (pfcrt) gene and lumifantrine and mefloquine resistance marker ‘N86Y mutation’ in P. falciparum multidrug resistance1 (pfmdr1) gene. The P. falciparum DNA was confirmed in 35 (37.23%) Bangladeshi and 45 (28.48%) African samples. The ‘pfcrt (K76T) mutation’ that confers resistance to chloroquine, was detected in 93.10% Bangladeshi and 29.27% African samples. The ‘pfmdr1 (N86Y) mutation’ that confers resistance to lumifantrine and mefloquine, was detected in 20.69% Bangladeshi and only 2.44% African samples. The higher prevalence of chloroquine resistance of P. falciparum in Bangladesh than in African countries revealed that possible withdrawal of chloroquine from endemic areas and also periodic molecular survey to monitor pf resistance to chloroquine, mefloquine, lumefantrine and artemisinin among these troops working in both endemic areas. Bangladesh J Microbiol, Volume 37 Number 1 June 2020, pp 1-6

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Deletions of pfhrp2 and pfhrp3 genes in rapid diagnostic test-negative Plasmodium falciparum isolates from Uganda

10.21203/rs.3.rs-42855/v1 ◽

2020 ◽

Author(s):

Sam L Nsobya ◽

Andrew Walakira ◽

Elizabeth Namirembe ◽

Moses Kiggundu ◽

Joaniter I Nankabirwa ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Protein Expression ◽

False Negative ◽

Plasmodium Species ◽

Pcr Amplification ◽

Rapid Diagnostic Tests ◽

False Negatives ◽

Cross Sectional ◽

True Prevalence ◽

Blood Smears

Abstract Background: Rapid diagnostic tests (RDTs) play a key role in malaria case management. The most widely used RDT identifies Plasmodium falciparum based on immunochromatographic recognition of P. falciparum histidine-rich protein 2 (PfHRP2). Deletion of the homologous pfhrp2 and pfhrp3 genes leads to false-negative PfHRP2-based RDTs, and has been reported in P. falciparum from South America and Africa. However, identification of pfhrp2/pfhrp3 deletions has usually been based only on failure to amplify these genes using PCR, without confirmation based on protein expression, and our understanding of the true prevalence of deletions is incomplete. Methods: We investigated pfhrp2 / pfhrp3 deletions in blood samples from cross-sectional surveys in 2012-13 in three regions of varied malaria transmission intensity in Uganda. We evaluated samples with positive Giemsa-stained thick blood smears and negative PfHRP2-based RDTs by PCR amplification of conserved subunit ribosomal DNA for Plasmodium species, PCR amplification of pfhrp2 and pfhrp3 genes to identify deletions, and bead-based immunoassays for expression ofPfHRP2. Results: Of 3516 samples collected in cross-sectional surveys, 1493 (42.5%) had positive blood smears, of which 96 (6.4%) were RDT-negative. Of these 96 RDT-negative samples, P. falciparum was identified in 56 (58%) and only non-falciparum plasmodial DNA in 40 (42%). In all 56 P. falciparum -positive samples there was a failure to amplify pfhrp2 or pfhrp3 :in 25 (45%) pfhrp2 was not amplified, in 39 (70%) pfhrp3 was not amplified, and in 19 (34%) neither gene was amplified. For the 39 P. falciparum -positive, RDT-negative samples available for analysis of protein expression, PfHRP2 was not identified by immunoassay in only four samples (10.3%); these four samples all had failure to amplify both pfhrp2 and pfhrp3 by PCR. Thus, only four of 96 (4.2%) smear-positive, RDT-negative samples had P. falciparum infections with deletion of pfhrp2 and pfhrp3 confirmed by failure to amplify the genes by PCR and lack of expression of PfHRP2 demonstrated by immunoassay. Conclusion: False negative RDTs were uncommon, and deletions in pfhrp2 and pfhrp3 explained some of these findings, although most false negatives were not due to deletion of the pfhrp2 and pfhrp3 genes.

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Soil Transmitted Helminths and Plasmodium falciparum Co-infections among School Children in Bugesera District, Rwanda: Implications for National Control Programs

International Journal of TROPICAL DISEASE & Health ◽

10.9734/ijtdh/2021/v42i1630527 ◽

2021 ◽

pp. 30-36

Author(s):

Umwanankundi Marcelline ◽

Mazigo D. Humphrey ◽

Tumusiime David ◽

Mucumbitsi Joseph ◽

Arpita Sharma ◽

...

Keyword(s):

School Children ◽

Primary Schools ◽

Spatial Scales ◽

Cross Sectional Survey ◽

Cross Sectional ◽

Soil Transmitted Helminths ◽

Control Programs ◽

Stool Samples ◽

The Impact ◽

National Control

Soil Transmitted Helminths and Plasmodium infections are ubiquitous with morbidity and mortality within the tropical and subtropical regions. However, the extent and consequences of STH-Plasmodium co-infection at different spatial scales are poorly understood. This study aimed at determining the prevalence of Soil Transmitted Helminths and Plasmodium coinfection, and, the impact thereof among school children in Bugesera District of Eastern Rwanda. Study Design: Cross Sectional Survey Place and Duration of Study: The survey was conducted in Bugesera district, Eastern Rwanda, between May and December 2020. Methodology: The survey was conducted among children between ages 5 and 18 years, across 21 randomly selected primary schools. Stool samples were collected and screened for soil transmitted helminths using Kato-Katz, while finger-prick blood samples were examined under the microscope to determine Plasmodium infection. Results: Overall the prevalence of A. lumbricoides, T. trichiura, Hookworm and P. falciparum was 4.43%, 0.76%, 0.16% and 3.15% respectively. On the other hand, 36.15% were co-infected with A. lumbricoides and P. falciparum. Surprisingly, the coinfection was higher 41.79%, (P<0.001) in girls than the overall coinfection prevalence. Conclusion: Helminthiasis and malaria remain dominant, in spite of the continuing and tremendous national control programs. The strikingly surprising higher prevalence of A. lumbricoides-P. falciparum co-infection in girls calls for additional investigations.

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Prevalence of pfcrt point mutations and level of chloroquine resistance in Plasmodium falciparum isolates from Africa

Infection Genetics and Evolution ◽

10.1016/j.meegid.2005.07.002 ◽

2006 ◽

Vol 6 (4) ◽

pp. 262-268 ◽

Cited By ~ 18

Author(s):

Carlo Severini ◽

Michela Menegon ◽

Anna Rosa Sannella ◽

Maria Grazia Paglia ◽

Pasquale Narciso ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Point Mutations ◽

Chloroquine Resistance

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Drug sensitivity of Plasmodium falciparum field isolates to selected antimalarial drugs in Ghana using the in-vitro DAPI assay

10.21203/rs.3.rs-16372/v1 ◽

2020 ◽

Author(s):

Michael Fokuo Ofori ◽

Emma E. Kploanyi ◽

Benedicta A. Mensah ◽

Emmanuel K. Dickson ◽

Eric Kyei Baafour ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Drug Sensitivity ◽

Geometric Mean ◽

Antimalarial Drugs ◽

Chloroquine Resistance ◽

Cape Coast ◽

African Region ◽

Cross Sectional ◽

Ecological Zones

Abstract Background: Malaria continues to be a major health issue globally with nine out of ten cases reported in Africa. Although the current artemisinin derived combination therapies in Ghana are still efficacious against the Plasmodium falciparum parasite, compounding evidence of artemisinin and amodiaquine resistance in the African region establish the need for a full, up-to-date understanding and monitoring of antimalarial resistance to provide evidence for planning control strategies.Methods: The study was cross-sectional and was conducted during the peak transmission seasons of 2015, 2016, and 2017 in two study sites located in different ecological zones of Ghana involving children aged 0.5-14 years presenting with symptomatic uncomplicated Plasmodium falciparum (Pf) malaria with parasitaemia greater than 1000 parasites/µl of blood. Using in vitro 4-,6-diamidino-2-phenylindole (DAPI) drug sensitivity assays, 328 Pf parasites collected were used to investigate susceptibility to five selected antimalarial drugs: chloroquine, amodiaquine, dihydroartemisinin, artesunate and mefloquine.Results: The geometric mean B (GMIC50) of five drugs against the parasites collected from Cape Coast were 9.6, 23.6, 9.1, 3.5 and 8.1nM for chloroquine, amodiaquine, artemisinin, artesunate, and mefloquine respectively in 2015. There was a 2 fold increase in the GMIC50 levels of all the drugs against the isolates collected in 2016 as compared to the 2015 data from Cape Coast .The a of the five drugs against the parasites collected from Cape Coast were significantly higher than those isolates collected from Begoro in 2016 and 2017 (P<0.001) . The chloroquine resistance ranged between 1.9% and 9.1% among isolates collected from Cape Coast but remained 0% in Begoro over the period. High amodiaquine resistance levels were recorded at both sites whilst that of artesunate resistance ranged between 4 and 10% over the study period.Conclusions: The study has assessed the antimalarial drug sensitivities of Ghanaian Pf isolates collected over 3 consecutive years. The parasites showed variable resistance levels to all the drugs used over the period. The study has demonstrated the continual return of chloroquine-sensitive parasites. The in vitro DAPI assay is a useful method for monitoring individual drugs used in combinations in Ghana for the generation of data on their sensitivities over time.

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Anaemia and Its Relation to Demographic, Socio-economic and Anthropometric Factors in Rural Primary School Children in Hai Phong City, Vietnam

Nutrients ◽

10.3390/nu11071478 ◽

2019 ◽

Vol 11 (7) ◽

pp. 1478 ◽

Cited By ~ 2

Author(s):

Hoang ◽

Orellana ◽

Le ◽

Gibson ◽

Worsley ◽

...

Keyword(s):

Primary School ◽

School Children ◽

Rural Areas ◽

Primary Schools ◽

Economic Factor ◽

Primary School Children ◽

Public Health Issue ◽

Microcytic Anaemia ◽

Cross Sectional ◽

Normocytic Anaemia

Little is known about the prevalence of anaemia and associated factors in school children in Vietnam. In this cross-sectional study, we aimed to determine the prevalence of anaemia and its subtypes, and the associations of types of anaemia with demographic, socio-economic and anthropometric factors among 6–9-year-old primary school children in rural areas of Hai Phong City, Vietnam. Haemoglobin (Hb) and mean corpuscular volume (MCV) were measured, and demographic, socio-economic and anthropometric data were collected in 893 children from eight primary schools. The prevalence of anaemia (Hb < 115 g/L) was 12.9% (95% CI: 8.1%, 19.9%), microcytic anaemia (Hb < 115 g/L and MCV < 80 fL) was 7.9% (95% CI: 5.3%, 11.6%) and normocytic anaemia (Hb < 115 g/L and MCV 80–90 fL) was 5.3% (95% CI: 2.9%, 9.5%). No child presented with macrocytic anaemia (Hb < 115 g/L and MCV > 90 fL). Children who were underweight, wasted, or in anthropometric failure (either underweight, stunted or wasted) were more likely to be anaemic (all p ≤ 0.004), and specifically, to have normocytic anaemia (all p ≤ 0.006), than those who were not underweight, wasted or in anthropometric failure. Stunted children were more likely to be anaemic (p = 0.018) than those who were not stunted. Overweight/obese children were less likely to be anaemic (p = 0.026) or have normocytic anaemia (p = 0.038) compared with children who were not overweight/obese. No anthropometric status indicator was associated with the risk of microcytic anaemia. No demographic or socio-economic factor was associated with any type of anaemia. Anaemia remains a public health issue in rural areas in Hai Phong City, Vietnam, and future approaches for its prevention and control should target undernourished primary school children.

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School bag weight and the occurrence of back pain among elementary school children

Journal of Public Health Research ◽

10.4081/jphr.2020.1841 ◽

2020 ◽

Author(s):

Semuel Layuk ◽

Tri Martiana ◽

Bongakaraeng Bongakaraeng

Keyword(s):

Elementary School ◽

Elementary Schools ◽

Back Pain ◽

School Children ◽

Primary Schools ◽

Elementary School Children ◽

Total Sample ◽

The Body ◽

Cross Sectional ◽

Suburban Areas

Background: Children in primary schools use school bags to carry study material, although the heavy bags are often associated with musculoskeletal problems, especially back pain. This practice requires strength, which significantly changes the body posture and walking pattern, subsequently leading to the incidence of back pain as a side effect. The aim of this study, therefore, is to analyze the differences in bag loads between elementary school children in urban and suburban area, and also analyze the relationship with the occurrence of back pain. Design and methods: This was a cross sectional study, which used stratified random sampling to choose 2 elementary schools each representing the urban and suburban areas. Furthermore, a total sample of 164 students were selected, whose school bag weight were measured for 5 consecutive days, particularly in the morning on respondents arrival at school. Moreover, the occurrence of back pain was evaluated using a modified Nordic map, and data analysis required the use of independent sample analysis t test and χ2 test.Results: The results showed the presence of highly significant differences in the school bags weight of respondents in the urban and suburban areas, as well as between schools using the 2013 curriculum and otherwise. Conclusions: In conclusion, there is a correlation between the weight measured and the occurrence of back pain, hence the 2013 curriculum is recommended to be adopted by all elementary schools. Also, the use of lockers to store items used at school repeatedly by children is also highly encouraged.

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Detection of Pneumocystis in lungs of bats from Brazil by PCR amplification

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2009000600005 ◽

2009 ◽

Vol 29 (6) ◽

pp. 469-473 ◽

Cited By ~ 2

Author(s):

Edna Maria Cavallini Sanches ◽

Susi M. Pacheco ◽

Alison S. Cericatto ◽

Rosane M. Melo ◽

Edson Molleta Colodel ◽

...

Keyword(s):

Nested Pcr ◽

Urban Areas ◽

Pcr Amplification ◽

Small Subunit ◽

Rrna Gene ◽

Wild Animals ◽

Tadarida Brasiliensis ◽

Desmodus Rotundus ◽

Mato Grosso ◽

Wide Range

Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3% = 5/19), Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis (2/3, 66.7%)and Diphylla ecaudata (1/2, 50%). PCR products which could indicate the presence of Pneumocystis (21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the first report of detection of Pneumocystis in bats from Brazil.

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Prevalence of Overweight and Obesity among Primary School Children Aged 8–13 Years in Dar es Salaam City, Tanzania

Advances in Preventive Medicine ◽

10.1155/2016/1345017 ◽

2016 ◽

Vol 2016 ◽

pp. 1-5 ◽

Cited By ~ 14

Author(s):

Ismail N. Pangani ◽

Festus K. Kiplamai ◽

Jane W. Kamau ◽

Vincent O. Onywera

Keyword(s):

Public Schools ◽

Primary School ◽

School Children ◽

Primary Schools ◽

Weight Status ◽

Age Groups ◽

Dar Es Salaam ◽

Overweight And Obesity ◽

Primary School Children ◽

Cross Sectional

Background. The understanding of obesity as a growing health problem in Africa and Tanzania in particular is hampered by lack of data as well as sociocultural beliefs in which overweight and obesity are revered. This study sought to determine the prevalence of overweight and obesity among primary school children aged 8–13 years in Dar es Salaam, Tanzania.Method. A cross-sectional analytical research design was used to study overweight and obesity in primary schools in Dar es Salaam, Tanzania. The target population was 150,000 children aged 8–13 years. Stratified random sampling was used to select 1781 children. Weight and height were taken and WHO standards for children were used to determine weight status.Results. Findings showed that the prevalence of overweight and obesity was 15.9% and 6.7%, respectively (N=1781). However, 6.2% of the children were underweight. There were significant differences in mean BMI between children in private and public schools (p=0.021), between male and female (p<0.001), and across age groups of 8–10 and 11–13 years (p<0.001).Conclusion. The prevalence of overweight and obesity among primary school children is significant and requires management and prevention strategies.

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