Decreased prevalence of the Plasmodium falciparum Pfcrt K76T and Pfmdr1 and N86Y mutations post-chloroquine treatment withdrawal in Katete District, Eastern Zambia.

BACKGROUND: In 2002, Zambia withdrew chloroquine as first-line treatment for Plasmodium falciparum malaria due to increased treatment failure and worldwide spread of chloroquine resistance. The artemisinin combination regimen, artemether-lumefantrine, replaced chloroquine (CQ) as first choice malaria treatment. The present study determined the prevalence of CQ resistance molecular markers in the Pfcrt and Pfmdr1 genes in Eastern Zambia at 9 and 13 years after the removal of drug pressure. METHODS: Samples collected from Katete District during the drug therapeutic efficacy assessments conducted in 2012 and 2016 were assayed by polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLP) to determine the prevalence of genetic mutations, K76T on the Pfcrt gene and N86Y on the Pfmdr1 gene. A total of 204 P. falciparum-positive DBS samples collected at these two time points were further analysed. RESULTS: Among the samples analysed for Pfcrt K76T and Pfmdr1 N86Y in the present study, 112 (82.4%) P. falciparum-infected samples collected in 2012 were successfully amplified for Pfcrt and 94 (69.1%) for Pfmdr1, while 69 (65.7%) and 72 (68.6%) samples from 2016 were successfully amplified for Pfcrt and Pfmdr1, respectively. In 2012, the prevalence of Pfcrt 76K (sensitive) was 97.3%, 76T (resistant) was 1.8%, and 0.8% had both 76K and 76T codons (mixed). Similarly in 2012, the prevalence of Pfmdr1 86N (sensitive) was 97.9% and 86Y (resistant) was 2.1%. In the 2016 samples, the prevalence of the respective samples was 100% Pfcrt 76K and Pfmdr1 86N. CONCLUSION: This study shows that there was a complete recovery of chloroquine-sensitive parasites by 2016 in Katete District, Eastern Zambia, 13 years following the withdrawal of CQ in the country. These findings add to the body of evidence for a fitness cost in CQ-resistant P. falciparum in Zambia and elsewhere. Further studies are recommended to monitor resistance countrywide and explore the feasibility of integration of the former best anti-malarial in combination therapy in the future.

Data on selected antimalarial drug resistance markers in Zambia.

This article describes data on selected resistance markers for antimalarial drugs used in Zambia. Antimalarial drug resistance has hindered the progress in the control and elimination of malaria. Blood samples were collected during a cross-sectional household survey, conducted during the peak malaria transmission, April to May of 2017. Dried blood spots were collected during the survey and transported to a laboratory for analysis. The analysed included polymerase chain reaction (PCR) followed by high resolution melt (HRM) for mutations associated with Sulfadoxine-pyrimethamine resistance in the Plasmodium falciparum dihydrofolate reductase (Pfdhfr) and P. falciparum dihydropteroate synthase (Pfdhps) genes. Mutations associated with artemether-lumefantrine resistance in falciparum multi-drug resistance gene 1 (Pfmdr1) were also assessed using PCR and HRM analysis, whereas the P. falciparum Kelch 13 (PfK13) gene was assessed using nested PCR followed by amplicon sequencing.

Surveillance of molecular markers for antimalarial resistance in Zambia: Polymorphism of Pfkelch 13, Pfmdr1 and Pfdhfr/Pfdhps genes.

Antimalarial resistance is an inevitable feature of control efforts and a key threat to achieving malaria elimination. Plasmodium falciparum, the deadliest of several species causing human malaria, has developed resistance to essentially all antimalarials. This study sought to investigate the prevalence of molecular markers associated with resistance to sulfadoxine-pyrimethamine (SP) and artemether-lumefantrine (AL) in Southern and Western provinces in Zambia. SP is used primarily for intermittent preventive treatment during pregnancy, while AL is the first-line antimalarial for uncomplicated malaria in Zambia. Blood samples were collected from household members of all ages in a cross-sectional survey conducted during peak malaria transmission, April to May of 2017, and amplified by polymerase chain reaction (PCR). Amplicons were then analysed by high-resolution melt following PCR to identify mutations associated with SP resistance in the P. falciparum dihydrofolate reductase (Pfdhfr) and P. falciparum dihydropteroate synthase (Pfdhps) genes and lumefantrine resistance in the P. falciparum multi-drug resistance 1 (Pfmdr1) gene. Finally, artemether resistance was assessed in the P. falciparum Kelch 13 (PfK13) gene using nested PCR followed by amplicon sequencing. The results showed a high frequency of genotypic-resistant Pfdhps A437G (93.2%) and Pfdhfr C59R (86.7%), N51I (80.9%), and S108N (80.8%) of which a high proportion (82.4%) were quadruple mutants (Pfdhfr N51I, C59R, S108N +Pfdhps A437G). Pfmrd1 N86Y, Y186F, and D1246Y - NFD mutant haplotypes were observed in 41.9% of isolates. The high prevalence of quadruple dhps/dhfr mutants indicates strong antifolate drug pressure from SP or other drugs (e.g., co-trimoxazole). Three samples contained PfK13 mutations, two synonymous (T478 and V666) and one non-synonymous (A578S), none of which have been associated with delayed clearance. This suggests that artemisinin remains efficacious in Zambia, however, the moderately high prevalence of approximately 40% Pfmdr1 NFD mutations calls for close monitoring of AL.

Prevalence and genotypic characterization of Giardia duodenalis isolates from asymptomatic school-going children in Lusaka, Zambia.

Giardia duodenalis is one of the most common causes of diarrhea in humans with about 250-300 million cases per year. It is considered to be a species complex comprising of eight genetic assemblages (A to H), with assemblages A and B being the major causes of human infections. In this study we carried out genotypic characterization of G. duodenalis isolates detected in asymptomatic school-going children aged 3-16 years. Between May and September 2017, a total of 329 fecal samples were collected from school-going children from Chawama compound of Lusaka City and were screened for Giardia by microscopic examination. All microscopically positive fecal samples were analyzed by semi-nested polymerase chain reaction (PCR) targeting the glutamate dehydrogenase (gdh) gene. Genotyping of amplified PCR products was conducted by restriction fragment length polymorphism (RFLP) and DNA sequence analysis. Microscopically, Giardia was found in 10% (33/329) of fecal samples. The PCR-RFLP analysis of the gdh gene revealed assemblages A and B in 27.3% (9/33) and 72.7% (24/33), respectively. Furthermore, analysis with restriction enzymes identified sub-assemblages AII (27.3%, 9/33), BIII (12.1%, 4/33), BIV (51.5%, 17/33) and mixed infections of BIII and BIV (9.1%, 3/33). Phylogenetic analysis showed the clustering of 27.6% (8/29) and 72.4% (21/29) of Zambian Giardia gdh gene sequences into assemblages A and B, respectively. This study has revealed the presence of both assemblage A and B and that spread of G. duodenalis in school-going children appears to be mostly through anthroponotic transmission. To our knowledge, this is the first report of genotypic characterization of G. duodenalis identified in Zambia.

En-route to the 'elimination' of genotypic chloroquine resistance in Western and Southern Zambia, 14 years after chloroquine withdrawal.

BACKGROUND: Anti-malarial resistance is, and continues to be a significant challenge in the fight against malaria and a threat to achieving malaria elimination. In Zambia, chloroquine (CQ), a safe, affordable and well-tolerated drug, was removed from use in 2003 due to high levels of resistance evidenced with treatment failure. This study sought to investigate the prevalence of chloroquine resistance markers in Southern and Western Provinces of Zambia 14 years after the withdrawal of CQ. METHODS: Data from a cross-sectional, all-age household survey, conducted during the peak malaria transmission season (April-May 2017) was analysed. During the all-age survey, socio-demographic information and coverage of malaria interventions were collected. Consenting individuals were tested for malaria with a rapid diagnostic test and a spot of blood collected on filter paper to create a dried blood spot (DBS). Photo-induced electronic transfer-polymerase chain reaction (PET-PCR) was used to analyse the DBS for the presence of all four malaria species. Plasmodium falciparum positive samples were analysed by high resolution melt (HRM) PCR to detect the presence of genotypic markers of drug resistance in the P. falciparum chloroquine resistance transporter (Pfcrt) and P. falciparum multi-drug resistance (Pfmdr) genes. RESULTS: A total of 181 P. falciparum positive samples were examined for pfcrt K76T and MDR N86. Of the 181 samples 155 successfully amplified for Pfcrt and 145 for Pfmdr N86. The overall prevalence of CQ drug-resistant parasites was 1.9% (3/155), with no significant difference between the two provinces. No N86Y/F mutations in the Pfmdr gene were observed in any of the sample. CONCLUSION: This study reveals the return of CQ sensitive parasites in Southern and Western Provinces of Zambia 14 years after its withdrawal. Surveillance of molecular resistant markers for anti-malarials should be included in the Malaria Elimination Programme so that resistance is monitored country wide.

Distribution of Plasmodium species and assessment of performance of diagnostic tools used during a malaria survey in Southern and Western Provinces of Zambia.

BACKGROUND: Zambia continues to make strides in reducing malaria burden through the use of proven malaria interventions and has recently pledged to eliminate malaria by 2021. Case management services have been scaled up at community level with rapid diagnostic tests (RDTs) providing antigen-based detection of falciparum malaria only. Key to national malaria elimination goals is the ability to identify, treat and eliminate all Plasmodium species. This study sought to determine the distribution of non-falciparum malaria and assess the performance of diagnostic tests for Plasmodium falciparum in Western and Southern Provinces of Zambia, two provinces planned for early malaria elimination. METHODS: A sub-set of individuals' data and samples from a cross-sectional household survey, conducted during peak malaria transmission season in April and May 2017, was used. The survey collected socio-demographic information on household members and coverage of malaria interventions. Malaria testing was done on respondents of all ages using blood smears and RDTs while dried blood spots were collected on filter papers for analysis using photo-induced electron transfer polymerase chain reaction (PET-PCR). Slides were stained using Giemsa stain and examined by microscopy for malaria parasites. RESULTS: From the 1567 individuals included, the overall prevalence of malaria was 19.4% (CI 17.5-21.4) by PCR, 19.3% (CI 17.4-21.4) by RDT and 12.9% (CI 11.3-14.7) by microscopy. Using PET-PCR as the gold standard, RDTs showed a sensitivity of 75.7% (CI 70.4-80.4) and specificity of 94.2% (CI 92.8-95.4). The positive predictive value (PPV) was 75.9% (CI 70.7-80.6) and negative predictive value (NPV) was 94.1% (CI 92.1-95.4). In contrast, microscopy for sensitivity, specificity, PPV, and NPV values were 56.9% (CI 51.1-62.5), 97.7% (CI 96.7-98.5), 85.6% (CI 80.0-90.2), 90.4% (CI 88.7-91.9), respectively. Non-falciparum infections were found only in Western Province, where 11.6% of P. falciparum infections were co-infections with Plasmodium ovale or Plasmodium malariae. CONCLUSION: From the sub-set of survey data analysed, non-falciparum species are present and occurred as mixed infections. As expected, PET-PCR was slightly more sensitive than both malaria RDTs and microscopy to detecting malaria infections.

Human mobility and factors associated with malaria importation in Lusaka district, Zambia: a descriptive cross sectional study.

BACKGROUND: Malaria is a major public health problem in Zambia with an estimated 4 million confirmed cases and 2389 deaths reported in 2015. Efforts to reduce the incidence of malaria are often undermined by a number of factors such as human mobility which may lead to introduction of imported infections. The aim of this study was to establish the burden of malaria attributed to human mobility in Lusaka district and identify factors associated with malaria importation among residents of Lusaka district. METHODS: A cross sectional study was conducted in five randomly selected health facilities in Lusaka district from November 2015 to February 2016. Data was collected from 260 patients who presented with malaria and whose status was confirmed by rapid diagnostic test or microscopy. Each confirmed malaria case was interviewed using a structured questionnaire to establish their demographic characteristics, travel history and preventive measures. Travel history was used as a proxy to classify cases as either imported or local. Residency was also used as a secondary proxy for importation to compare characteristics of residents vs non-residents in relation to malaria importation. Logistic regression was used to determine factors associated with malaria importation among residents of Lusaka district. RESULTS: Out of 260 cases, 94.2% were classified as imported cases based on participants' travel history. There were 131 (50.4%) males and 129 (49.6%) females. Age distribution ranged from 0 to 68 years with a median age of 15 years (IQR 8-27). Imported cases came from all the ten provinces of Zambia with the Copperbelt Province being the highest contributor (41%). Of all imported cases, use of prophylaxis was found to be highly protective [AOR = 0.22 (95% CI 0.06-0.82); p-value = 0.024]. Other factors that significantly influence malaria transmission and importation by residents include duration of stay in a highly endemic region [AOR = 1.25 (95% CI 1.09-1.44); p-value = 0.001] and frequency of travel [AOR = 3.71 (95% CI 1.26-10.84); p-value = 0.017]. CONCLUSION: Human mobility has influenced malaria transmission in Lusaka district through a number of factors by importing infections. This leads to onward transmission and poses a challenge to malaria elimination and control. However, taking of prophylaxis is highly protective and must be highly recommended.

Signal functions for emergency obstetric care as an intervention for reducing maternal mortality: a survey of public and private health facilities in Lusaka District, Zambia.

BACKGROUND: Zambia's maternal mortality ratio was estimated at 398/100,000 live births in 2014. Successful aversion of deaths is dependent on availability and usability of signal functions for emergency obstetric and neonatal care. Evidence of availability, usability and quality of signal functions in urban settings in Zambia is minimal as previous research has evaluated their distribution in rural settings. This survey evaluated the availability and usability of signal functions in private and public health facilities in Lusaka District of Zambia. METHODS: A descriptive cross sectional study was conducted between November 2014 and February 2015 at 35 public and private health facilities. The Service Availability and Readiness Assessment tool was adapted and administered to overall in-charges, hospital administrators or maternity ward supervisors at health facilities providing maternal and newborn health services. The survey quantified infrastructure, human resources, equipment, essential drugs and supplies and used the UN process indicators to determine availability, accessibility and quality of signal functions. Data on deliveries and complications were collected from registers for periods between June 2013 and May 2014. RESULTS: Of the 35 (25.7% private and 74.2% public) health facilities assessed, only 22 (62.8%) were staffed 24 h a day, 7 days a week and had provided obstetric care 3 months prior to the survey. Pre-eclampsia/ eclampsia and obstructed labor accounted for most direct complications while postpartum hemorrhage was the leading cause of maternal deaths. Overall, 3 (8.6%) and 5 (14.3%) of the health facilities had provided Basic and Comprehensive EmONC services, respectively. All facilities obtained blood products from the only blood bank at a government referral hospital. CONCLUSION: The UN process indicators can be adequately used to monitor progress towards maternal mortality reduction. Lusaka district had an unmet need for BEmONC as health facilities fell below the minimum UN standard. Public health facilities with capacity to perform signal functions should be upgraded to Basic EmONC status. Efforts must focus on enhancing human resource capacity in EmONC and improving infrastructure and supply chain. Obstetric health needs and international trends must drive policy change.

Seroprevalence and determinants of toxoplasmosis in pregnant women attending antenatal clinic at the university teaching hospital, Lusaka, Zambia.

BACKGROUND: Toxoplasmosis is a neglected zoonotic disease which is prevalent among pregnant women especially in Africa. This study aimed to determine the seroprevalence and determinants of the disease among pregnant women attending antenatal clinic at the University Teaching Hospital (UTH). METHOD: A cross-sectional study was employed where 411 pregnant women attending antenatal clinic at UTH were interviewed using closed ended questionnaires. Their blood was also tested for Toxoplasma gondii IgG and IgM antibodies using the OnSite Toxo IgG/IgM Combo Rapid test cassettes by CTK Biotech, Inc, USA. RESULT: The overall seroprevalence of the infection (IgG) was 5.87%. There was no seropositive IgM result. Contact with cats showed 7.81 times the risk of contracting the infection in the pregnant women and being a farmer/being involved in construction work showed 15.5 times likelihood of contracting the infection. Socio-economic status of the pregnant women also presented an inverse relationship (showed association) with the infection graphically. However, though there were indications of association between contact with cats, employment type as well as socioeconomic status of the pregnant women with the infection, there was not enough evidence to suggest these factors as significant determining factors of Toxoplasma gondii infection in our study population. CONCLUSION: There is a low prevalence of Toxoplasma gondii infection among pregnant women in Lusaka, Zambia. Screening for the infection among pregnant women can be done once or twice during pregnancy to help protect both mother and child from the disease. Health promotion among women of child bearing age on the subject is of immense importance in order to help curb the situation. Further studies especially that of case-control and cohort studies should be carried out in the country in order to better ascertain the extent of the condition nationwide.

Factors associated with late antenatal care booking: population based observations from the 2007 Zambia demographic and health survey.

INTRODUCTION: In spite of the extreme importance of an early antenatal care visit, more than 50% of Zambian pregnant women book for antenatal care late. We aimed to determine factors associated with late antenatal care booking in Zambia. METHODS: Data stem from the 2007 Zambia Demographic and Health Survey where information on socio-demographic, social-economic, obstetrical characteristics and timing of the first antenatal visit were extracted on all women aged 15 to 49 years. A weighted survey analysis using STATA version 12 was applied. Firstly, we explored proportions of ANC booking at 0-3 months, 4-5 month and 6-9 months. Secondly, we investigated the association between predictor variables and late antenatal care booking using univariate and multivariate logistic regression. RESULTS: Overall (n= 3979), the proportion of late ANC booking (booking between 4th to 9th month) was 81% disaggregated as 56% and 19% at 4 to 5 months and 6 to 9 months respectively. Women who wanted their last child later were more likely to book late than those with wanted pregnancies then (AOR: 1.35 95% CI 1.10-1.66). Women with higher education were 55% less likely to book for ANC late compared to women with no education (AOR: 0.45 95%CI: 0.27-0.74). Women aged 20-34 years were 30% more likely to book earlier than women younger than 20 years (AOR: 0.69 95% CI 0.50-0.97). CONCLUSION: We found high proportion of late ANC booking associated with presence of unplanned or unwanted pregnancies in this population. The concentration of this problem in lower or no education groups may be an illustration of existing inequalities which might further explain limitations in health promotion messages meant to mitigate this challenge. There is thus urgent need to re-pack health promotion message to specifically target this and related poor groups.

Reported incidence of fever for under-5 children in Zambia: a longitudinal study.

BACKGROUND: Childhood fever is the most common clinical sign of Plasmodium falciparum infection. It is used as a measure of burden of the disease and the effectiveness of control programs for malaria. This study aimed to determine the incidence of fever in under-5 children of Magoye and Chivuna rural areas of Mazabuka district, Zambia. METHODS: Incidence of fever was evaluated longitudinally over a period of 16 months (July 2006 and November 2007) among children aged 12-59 months in Magoye and Chivuna rural communities. The data was collected for a study on community directed treatment of soil-transmitted helminth infections in under-five children. Data from caretakers of 1221 children were collected using a structured interviewer-administered questionnaire. Cox proportion hazard regression was used to determine predictors of multiple episodes of fever and Kaplan-Meier survival curves was used to compare survival between two groups. RESULTS: A total of 1221 under-5 children [median age 32 months; IQR 12-58] participated in the study and 696 (57 %) were from Magoye and 525 (43 %) from Chivuna. The incidence rate of fever was 162.4 per month per 1000 children for the 16 months period. The proportion of fever was not statistically related to children' age [p = 0.779] and sex [p = 0.546]. Predictors of multiple episodes of fever were: age (37-48 vs. 12-24 months) [HR 0.81; 95 % CI 0.67, 0.98; p = 0.030]; location (Chivuna vs. Magoye) [HR 1.35; 95 % CI 1.17, 1.56; p < 0.001]; and season (dry vs. rainy) [HR 0.17; 95 % CI 0.12, 0.23; p < 0.001]. CONCLUSION: The study has shown that the incidence of fever was high in the study areas. Febrile illnesses like malaria still have a significant effect on the health of under-5 children in the study population. There still exists the need for interventions aimed at reducing the incidence of fever in under five children, more especially in rural areas.

Prevalence of pathogenic trypanosomes in anaemic cattle from trypanosomosis challenged areas of Itezhi-tezhi district in central Zambia.

BACKGROUND: The measure of anaemia status using packed cell volume (PCV) can be a reliable indicator of African trypanosomosis (AT) in the absence of other anaemia-causing conditions. However, studies that have estimated prevalence of anaemia in cattle from AT endemic areas have rarely reported the prevalence of the disease in the anaemic cattle. Therefore we investigated the prevalence of AT in anaemic cattle at sites that had recently reported the disease in Itezhi tezhi district of central Zambia. METHODS: During a survey, blood samples were collected from 564 randomly selected cattle for anaemia determination from seven crush pens (Mutenda, Kapulwe, Banachoongo, Itumbi, Iyanda, New Ngoma and Shinampamba). At a PCV- value cut off of 26 %, all samples positive for anaemia were subjected to both parasitological examination on thick and thin blood smears and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for detection of trypanosome DNA. Fisher's exact test and a mixed effect logistic regression analyses were used to determine and measures associations, respectively. RESULTS: Of 564 cattle screened, 58 (10.3 %; 95 % CI: 7.8-12.8 %) had anaemia. PCR-RFLP results showed that 17 (29.3 %; 95 % CI; 17.2-41.4 %) anaemic cattle were positive for pathogenic trypanosomes compared to 1 (1.7 %; 95 % CI: 0.0-5.2 %) on parasitological examination using thick smears. The infections were caused by Trypanosoma congolense and Trypanosoma vivax. Fisher's exact test showed a strong association between PCV and pathogenic trypanosome infection (P = 0.004). A mixed effect multivariate logistic regression showed that a one unit increase in PCV reduced the likelihood of detecting AT with PCR-RFLP by 24.7 % (95 % CI: 4.6-40.6 %; P = 0.019) in anaemic cattle, taking into account their age and sex, with random effects for crush pen. CONCLUSION: These results suggest that T. congolense and T. vivax could be important causes of anaemia in cattle reared in AT endemic areas of Itezhi tezhi in Central Zambia. This also suggests that even though pathogenic trypanosomal infection was strongly associated with PCV, it could only account for up to 41 % of the anaemia in cattle. Therefore further investigation to ascertain other factors responsible for anaemia in AT endemic areas of Itezhi tezhi in Central Zambia is needed.

Mortality and morbidity patterns in under-five children with severe acute malnutrition (SAM) in Zambia: a five-year retrospective review of hospital-based records (2009-2013).

BACKGROUND: Severe acute malnutrition has continued to be growing problem in Sub Saharan Africa. We investigated the factors associated with morbidity and mortality of under-five children admitted and managed in hospital for severe acute malnutrition. METHODS: It was a retrospective quantitative review of hospital based records using patient files, ward death and discharge registers. It was conducted focussing on demographic, clinical and mortality data which was extracted on all children aged 0-60 months admitted to the University Teaching Hospital in Zambia from 2009 to 2013. Cox proportional Hazards regression was used to identify predictors of mortality and Kaplan Meier curves where used to predict the length of stay on the ward. RESULTS: Overall (n = 9540) under-five children with severe acute malnutrition were admitted during the period under review, comprising 5148 (54%) males and 4386 (46%) females. Kwashiorkor was the most common type of severe acute malnutrition (62%) while diarrhoea and pneumonia were the most common co-morbidities. Overall mortality was at 46% with children with marasmus having the lowest survival rates on Kaplan Meier graphs. HIV infected children were 80% more likely to die compared to HIV uninfected children (HR = 1.8; 95%CI: 1.6-1.2). However, over time (2009-2013), admissions and mortality rates declined significantly (mortality 51% vs. 35%, P < 0.0001). CONCLUSIONS: We find evidence of declining mortality among the core morbid nutritional conditions, namely kwashiorkor, marasmus and marasmic-kwashiorkor among under-five children admitted at this hospital. The reasons for this are unclear or could be beyond the scope of this study. This decline in numbers could be either be associated with declining admissions or due to the interventions that have been implemented at community level to combat malnutrition such as provision of "Ready to Use therapeutic food" and prevention of mother to child transmission of HIV at health centre level. Strategies that enhance and expand growth monitoring interventions at community level to detect malnutrition early to reduce incidence of severe cases and mortality need to be strengthened.

Patterns of mixed Plasmodium species infections among children six years and under in selected malaria hyper-endemic communities of Zambia: population-based survey observations.

BACKGROUND: Although malaria is preventable and treatable, it still claims 660,000 lives every year globally with children under five years of age having the highest burden. In Zambia, malaria rapid diagnostic tests (RDTs) that only detect Plasmodium falciparum are the main confirmatory means for malaria diagnosis in most health facilities without microscopy services. As a consequence of this P. falciparum species diagnostic approach, non-falciparum malaria is not only under-diagnosed but entirely missed, thereby making the exact disease burden unknown. We thus investigated the prevalence of various Plasmodium spp. and associated burden of infection in selected communities in Zambia. METHODS: Data from two malaria hyper-endemic provinces (Eastern and Luapula) of the 2012 National Malaria Indicator Survey (MIS), conducted between April and May 2012, were used. The MIS is a nationally representative, two-stage cluster survey conducted to coincide with the end of the malaria transmission season. Social, behavioural and background information were collected from households as part of the survey. Thick blood smears, RDTs and dried blood spots (DBS) were collected from children below six years of age. Slides were stained using Giemsa and examined by microscopy while polymerase chain reaction (PCR) was used to analyse the DBS for malaria Plasmodium spp. Multivariate logistic regression was employed to examine the association between background factors and malaria. RESULTS: Overall, 873 children younger than six years of age were surveyed. The overall prevalence of Plasmodium spp. by PCR was 54.3% (95% CI 51-57.6%). Of the total Plasmodium isolates, 88% were P. falciparum, 10.6% were mixed infections and 1.4% were non-falciparum mono infections. Among the mixed infections, the majority were a combination of P. falciparum and P. malariae (6.5% of all mixed infections). Children two years and older (2-5 years) had three-fold higher risk of mixed malaria infections (aOR 2.8 CI 1.31-5.69) than children younger than two years of age. CONCLUSION: The high prevalence of mixed Plasmodium spp. infections in this population stresses review of the current malaria RDT diagnostic approaches. The observed less incidence of mixed infections in children under two years of age compared to their older two-to-five-year-old counterparts is probably due to the protective maternal passive immunity, among other factors, in that age group.

Adherence to the Food and Agricultural Organization guidelines on trypanocide usage among cattle farmers in Itezhi tezhi, Central Zambia.

Trypanocides will continue to play an important role in the control of tsetse fly transmitted trypanosomosis now and in the near future. The drugs are mostly administered by farmers without any veterinary supervision leading to misuse and under dosing of medication, and these could be factors that promote trypanocidal drug resistance (TDR) development. In order to delay or prevent TDR, the Food and Agriculture Organization (FAO) recommended guidelines on trypanocide use. It is not known if these recommended guidelines are adhered to in Itezhi tezhi district of Zambia. A survey was undertaken to examine how socio-economic and environmental factors were associated with adherence to the recommended guidelines on trypanocide use in Itezhi tezhi, Central Zambia. Ninety farmers who use trypanocides were interviewed using a questionnaire to collect their socio-economic characteristics (age, education in years, cattle herd size, competence on trypanocide use and their access to extension on trypanocide use) and trypanocide usage practices while crush pens which they use were stratified according to location, whether in the Game Management Area (GMA) (Mutenda, Itumbi, Kapulwe and Banachoongo) or non-GMA (Iyanda, New Ngoma and Shinampamba) as an environmental factor. Associations and measures of associations to adherence of FAO guidelines were determined. The results showed that 25.6% of the farmers adhered to guidelines by FAO on trypanocide use and that none of the socio-economic factors under investigation were significantly associated with it. Further the farmers that used crush pens that were in the GMA had an 80% reduction in the likelihood of adhering to the FAO guidelines on trypanocide use than those that used crush pens in the non-GMA (AOR 0.20, 95% CI: 0.05-0.81, P=0.02). There was low adherence to the recommended FAO guidelines on trypanocide use and it was associated with the location of the crush pen whether in the GMA or not, as an environmental factor. With farmers in the GMA less likely to adhere to FAO guidelines than those in the non-GMA, we recommend an integrated approach of measures to control trypanosomosis in the GMA of Itezhi tezhi to lessen overuse of trypanocides by the farmers.