Assessment of MTR Rs1805087 SNP as Possible Modifier of Sickle Cell Disease Severity in a Nigerian Population.

BACKGROUND: Sickle cell disease is the commonest genetic disorder in Nigeria, affecting 2-3% of an estimated population of 160 million people. The role of genetic mutations in folate cycle genes, and the variable phenotypic expressions constituting disease severity, needs to be critically examined. OBJECTIVE: This study was carried out to establish the pattern of methionine synthase gene mutations (rs1805087 SNP), and its possible association with disease severity in adults with sickle cell anaemia in Lagos, Nigeria. METHODOLOGY: This is a cross-sectional study of seventy (70) subjects with sickle cell disease (HbSS) matched for age and gender with known apparently healthy haemoglobin genotype AA (HbAA) subjects, as cases and controls respectively. Structured questionnaires were used to obtain demographic, clinical and other phenotypic data needed to compute disease severity. Pattern of MTR A2756G gene mutation and homocysteine assay (Hcy) were assessed by Polymerase Chain Reaction and Enzyme- linked Immunosorbent Assay respectively. Full blood count analysis of participants was done using the KX-21 Automated Analyzer (Sysmex Corporation, Japan). RESULTS: The mutant genotypes MTR 2756 AG/GG were recorded in 46.4% (n =55) of subjects with disease severity score >7. Elevated plasma homocysteine (HHcy) was significantly associated with disease severity among HbSS subjects (OR=17.2, CI: 3.490-86.079; p=0.0001). Conversely, no significant association was observed with the mutant genotypes MTR 2756 AG/GG and disease severity (p>0.05). CONCLUSION: While HHcy is significantly associated with phenotypic expression of HbSS, the MTR 2756 SNPs did not appear to independently influence homocysteine level or disease severity in HbSS subjects.

CONTEXTE: La drépanocytose est la maladie génétique la plus répandue au Nigeria, affectant 2 à 3 % d'une population estimée à 160 millions de personnes. Le rôle des mutations génétiques dans les gènes du cycle du folate, et les expressions phénotypiques variables constituant la gravité de la maladie, doivent être examinés de façon critique. OBJECTIF: Cette étude a été menée pour établir le schéma des mutations du gène de la méthionine synthase (rs1805087 SNP), et son association possible avec la gravité de la maladie chez les adultes atteints de drépanocytose à Lagos, au Nigeria. MÉTHODOLOGIE: Il s'agit d'une étude transversale de soixantedix (70) sujets atteints de drépanocytose (HbSS) appariés pour l'âge et le sexe avec des sujets connus apparemment sains de génotype d'hémoglobine AA (HbAA), comme cas et contrôles respectivement. Des questionnaires structurés ont été utilisés pour obtenir des données démographiques, cliniques et autres données phénotypiques nécessaires au calcul de la gravité de la maladie. Le profil de la mutation du gène MTR A2756G et le dosage de l'homocystéine (Hcy) ont été évalués respectivement par réaction en chaîne par polymérase et par test immunologique enzymatique. L'analyse de la formule sanguine complète des participants a été effectuée à l'aide de l'analyseur automatisé KX-21 (Sysmex Corporation, Japon). RÉSULTATS: Les génotypes mutants MTR 2756 AG/GG ont été enregistrés chez 46,4 % (n =55) des sujets présentant un score de gravité de la maladie > 7. L'homocystéine plasmatique élevée (HHcy) était significativement associée à la gravité de la maladie chez les sujets HbSS (OR=17,2, CI : 3,490­86,079 ; p=0,0001). À l'inverse, aucune association significative n'a été observée entre les génotypes mutants MTR 2756 AG/GG et la gravité de la maladie (p>0,05). CONCLUSION: Alors que l'HHcy est significativement associée à l'expression phénotypique de l'HbSS, les SNP MTR 2756 ne semblent pas influencer indépendamment le niveau d'homocystéine ou la gravité de la maladie chez les sujets HbSS.

Assessment of MTR Rs1805087 SNP as Possible Modifier ofSickle Cell Disease Severity in a Nigerian Population

BACKGROUND: Sickle cell disease is the commonest geneticdisorder in Nigeria, affecting 2­3% of an estimated population of 160million people. The role of genetic mutations in folate cycle genes,and the variable phenotypic expressions constituting disease severity,needs to be critically examined.OBJECTIVE: This study was carried out to establish the pattern ofmethionine synthase gene mutations (rs1805087 SNP), and its possibleassociation with disease severity in adults with sickle cell anaemia inLagos, Nigeria.METHODOLOGY: This is a cross-sectional study of seventy (70)subjects with sickle cell disease (HbSS) matched for age and genderwith known apparently healthy haemoglobin genotype AA (HbAA)subjects, as cases and controls respectively. Structured questionnaireswere used to obtain demographic, clinical and other phenotypic dataneeded to compute disease severity. Pattern of MTR A2756G genemutation and homocysteine assay (Hcy) were assessed by PolymeraseCh ain Reaction and Enzyme- linked Immun osorbent Assayrespectively. Full blood count analysis of participants was done usingthe KX-21 Automated Analyzer (Sysmex Corporation, Japan).RESULTS: The mutant genotypes MTR 2756 AG/GG were recordedin 46.4% (n =55) of subjects with disease severity score >7. Elevatedplasma homocysteine (HHcy) was significantly associated withdisease severity among HbSS subjects (OR=17.2, CI: 3.490-86.079;p=0.0001). Conversely, no significant association was observed withthe mutant genotypes MTR 2756 AG/GG and disease severity(p>0.05).CONCLUSION: While HHcy is significantly associated withphenotypic expression of HbSS, the MTR 2756 SNPs did not appearto independently influence homocysteine level or disease severity inHbSS subjects

Epigenetic Modification in Methylene Tetrahydrofolate Reductase (MTHFR) Gene of Women with Pre-eclampsia.

BACKGROUND: Genetic and epigenetic factors play significant roles in the aetio-pathogenesis of pre-eclampsia (PE). The effects may vary across racial and geographical boundaries. The role of epigenetic modification in pre-eclampsia was studied among African populations in Lagos, Nigeria. AIM AND OBJECTIVES: This study aimed to determine the pattern of Methylene tetrahydrofolate reductase gene (MTHFR) CpG island methylation in pre-eclampsia, and evaluate associated covariates. METHODOLOGY: This study was an observational, cross-sectional, study conducted at the Lagos University Teaching Hospital and the Lagos State Island Maternity Hospital. A total of 400 pregnant women consisting of 200 pregnant women diagnosed with pre-eclampsia (study group) and 200 pregnant normotensive and apparently healthy women (control group) were recruited for the study. Demographic and clinical histories were obtained through questionnaires. The DNA Methylation status of the CpG Island in promoter region of the MTHFR gene was assessed using bisulphite conversion and methylation specific PCR method. The biochemical parameters measured in the study were: red cell folate, vitamin B12, plasma homocysteine (Hcy) and methylene tetrahydrofolate reductase enzyme level. RESULTS: Homozygous MTHFR CpG island hypomethylation pattern was significantly associated with pre-eclampsia (χ 2 = 22.96; p = 0.000), Mean values of plasma homocysteine in PE women with homozygous hypomethylation (26.1 ± 9.1 umol/L) were significantly higher than (20.1 ± 4.2 umol/L) observed in PE subjects with homozygous hypermethylation (p = 0.008). Homozygous CpG island hypomethylated pattern of the MTHFR promoter region, was associated with the lowest median MTHFR enzyme level (72.8 ± 39.8 pmol/L) compared with heterozygous methylated pattern (91.3 ± 60.9 pmol/L; p = 0.047) and homozygous methylated pattern (82.3 ± 31.0 pmol/L; 0.047). Red cell folate and Vitamin B12 levels were not significantly associated with CpG island methylation status. CONCLUSION: Epigenetic modification plays significant role in the pathogenesis of pre-eclampsia.

Methylene tetrahydrofolate reductase and methionine synthase gene polymorphisms as genetic determinants of pre-eclampsia.

BACKGROUND: Pre-eclampsia (PE) is a leading cause of maternal and neonatal mortality in Africa; and has been associated with the interplay of genetic, metabolic and environmental factors. Polymorphisms of methylene tetrahydrofolate reductase (MTHFR) and methionine synthase (MTR) folate cycle genes, have been controversially associated with pre-eclampsia in studies from different human populations. OBJECTIVES: To determine the distribution of MTHFR C677T and MTR A2756G polymorphisms in a Nigerian population and evaluate possible associations with the occurrence of pre-eclampsia and homocysteine metabolic derangement. MATERIALS AND METHODS: This study was a hospital based study carried out in Lagos, South-western Nigeria. Two hundred pregnant women clinically diagnosed with pre-eclampsia (study group) and 200 apparently healthy non-pre-eclamptic pregnant women (control group) were recruited for the study after written informed consent. Pre-eclampsia was diagnosed based on the International Society for the Study of Hypertension in Pregnancy re-classification of 2013. MTHFR C677T and MTR A2756G polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Statistical analyzes were performed using SPSS version 23. Hardy-Weinberg distribution were tested with χ2 test. Logistic regression model was used to evaluate the relationship of variables with pre-eclampsia. A value of p < 0.05 was considered statistically significant. RESULTS: MTHFR genotype frequencies of CC, CT and TT were 59.8%; 31.2% and 9.0% in study group and 76.6%; 22.3% and 1.0% in the control group respectively. MTR A2756G genotype frequencies of AA, AG and GG genotypes were 71.9%; 20.1% and 8.0% for the study group and 81.5%; 16.4% and 2.1% for the control group. Occurrence of pre-eclampsia was significantly associated with presence of T allele of MTHFR (OR = 1.855; p < 0.05) and G allele of MTR genes (OR = 1.269; p < 0.05), Homozygosity of TG haplotype significantly increased the occurrence of pre-eclampsia among Nigerian women (OR = 2.252; p < 0.05). Population attributable risk fraction percent for the T and G alleles were 16.4% and 11.5% respectively. Mean plasma Hcy level was not, however, significantly affected by MTHFR/MTR haplotypes (F = 1.54; p = 0.157). CONCLUSION: MTHFR C677T and MTR A2756G polymorphisms were associated with pre-eclampsia in a population of pregnant women in Lagos, Nigeria.

Methylenetetrahydrofolate Reductase Enzyme Level and Antioxidant Activity in Women with Gestational Hypertension and Pre-eclampsia in Lagos, Nigeria.

BACKGROUND: Deficiencies of enzymes in the folate cycle may lead to the generation of homocysteine, a toxic metabolic intermediate with pro-oxidant effect and ability to induce oxidant stress and lipid peroxidation as part of the pathophysiological process in gestational hypertension (GH) and pre-eclampsia (PE). AIM: The aim of this study is to assess the reliability of plasma homocysteine (hcy) 5, 10 methylenetetrahydrofolate reductase (MTHFR) enzyme and oxidative stress parameters as indicators of aetio-pathogenesis and severity of gestational hypertension and pre-eclampsia. SUBJECTS AND METHODS: This was a comparative cross-sectional study conducted over 6 months. Two hundred pregnant women were recruited from two sites. They were divided into gestation hypertension (n = 40), pre-eclampsia (n = 60) and control groups (n = 100). Parameters evaluated for statistical analysis were MTHFR enzyme level, plasma homocysteine and malondialdehyde (MDA) levels, with glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) activities. RESULTS: Mean plasma hcy level and MDA were significantly higher in pre-eclampsia and gestational hypertension when compared to control group (p < 0.05). However, MTHFR enzyme level, GSH, SOD and CAT were significantly higher in normotensive females when compared to PE and GH subgroups (p < 0.05). Pre-eclampsia was significantly associated with an increased risk of lipid peroxidation (OR = 4.923; p = 0.007). CONCLUSION: Pre-eclampsia and gestational hypertension are associated with marked homocysteine metabolic derangement and increased lipid peroxidation induced by oxidative stress and reduced MTHFR enzyme activity which may be the significant risk factors in the aetio-pathogenesis of GH and PE.

Plasma D-dimer reference ranges in pregnant Nigerians.

BACKGROUND: The use of D-dimer as a screening test for thromboembolic disorders has not been validated in pregnancy thus necessitating further studies. This cross sectional study was carried out among Pregnant Nigerians at the Nigerian Air Force Hospital Antenatal Clinic, Ikeja, Lagos. OBJECTIVE: This study was to determine local reference ranges for plasma D-dimer in both pregnant and non-pregnant females and compare differences in both populations. METHODS: Structured questionnaires were administered on a total of 365 participants which included 71 apparently healthy non-pregnant females, 64 women in the first trimester of pregnancy (≤13 weeks gestation); 65 women at the second trimester of pregnancy (14-26 weeks), and 82 pregnant females at the third trimester of pregnancy (> = 27 weeks). Citrated blood was collected for estimation of prothrombin time (PT), activated partial thromboplastin time (aPTT), and D-dimer estimation by ELISA method. Aspartate aminotransferase (AST) and alanine aminotransferase enzymes (ALT) were estimated using the Hitachi chemistry analyzer. Reference ranges were estimated non-parametrically using the Reference Value Advisor V 2.1. RESULTS: Median D-dimer level for non-pregnant females was 190 ng/mL, while the median D-dimer levels for pregnant female in 1st, 2nd, and 3rd trimester were 485 ng/ml; 620 ng/mL; and 1185 ng/mL respectively.Reference ranges were calculated to be 86-494 ng/mL; 338-624 ng/mL; 451-799 ng/mL and 665-1262 ng/mL for non-pregnant females, 1st, 2nd, and 3rd trimester of pregnancy respectively. CONCLUSION: A diagnostic algorithm for venous thromboembolism (VTE) in pregnant women which combines clinical suspicion with elevated plasma D-dimers levels above estimated reference range for each trimester should precede definitive formal imaging.

Haematotoxic and reproductive toxicity of fixed dose combined anti-tuberculous agents: protective role of antioxidants in rats.

BACKGROUND: Tuberculosis (TB) is the world's greatest infectious killer of women of reproductive age and the leading cause of death among people with HIV/AIDS. The major problem militating against the management of tuberculosis is the lack of compliance to medication by the infected patients as a result of multidrug needed to be taking daily leading to resistance. Occurrences of hepatic toxicity, teratogenicity, sperm quality damage, haematotoxicity and meningeal congestion of individual anti-tuberculous agents have been reported. OBJECTIVE: The study is aimed to determine the reproductive and haematological toxicity of combined antituberculous agents and the modulatory role of antioxidants using animal model. METHODS: Fifty rats (10 per group) were randomly allotted to five groups, consisting of the control, the fixed dose combined anti TB agents treated group, the fixed dose combined anti TB agents plus vitamin C treated group, the fixed dose combined anti TB agents plus vitamin E treated group and the fixed dose combined anti TB agents plus vitamin C plus vitamin E treated group. Therapeutic doses of the fixed dose combined anti TB agents (25 mg/kg/day), vitamin E (5 mg/kg) and vitamin C (8 mg/kg) were administered to the animals via oral gavage, daily over 28 days. After 28days, rats were sacrificed for internal macroscopic and histological examination of the organs, sperm analysis and haematological investigations were carried out. RESULTS: The results showed a significant increase (p < or = 0.05) in the levels of white blood cells (WBC), red blood cell (RBC) and haemoglobin (HB) of the combined anti-TB plus vitamins C or E treated groups compared with combined anti-TB treated group alone (56.34 +/- 0.11) that decreased the haematological parameters. A significant decrease (p < or = 0.05) in the sperm counts (22.26 +/- 0.02; 35.40 +/- 0.02) and motility (77.03 +/- 0.02; 94.50 +/- 0.01) of the combined anti-TB treated rats as compared with the control group were observed. The combined anti-TB plus vitamin C treated rats demonstrated a significant increase (p < or = 0.05) in the sperm motility (90.23 +/- 0.01) as compared with the control group. There was also a remarkable decrease in the abnormal morphology of the sperm in the combined anti-TB plus vitamins E and C treated rats (0.05 +/- 0.02) as compared with the combined anti-TB group alone (1.10 +/- 0.02). CONCLUSION: Vitamins C and E positively modulated the sperm quality and haematological damage produced by the Fixed Dose Combined Anti-Tuberculous agents.

Pattern of lymph node pathology in Lagos.

BACKGROUND: Enlarge lymph node is a common finding in clinical practice, which can be caused by several factors, such as infections, drugs etc. OBJECTIVE: To review the pathology of lymph node diseases in children and adults with both deep seated and peripheral lymphadenopathy in Lagos. METHODS: A 12- year retrospective study of lymph node biopsies at Histopathology Department of LUTH, and two main histopathology private laboratories in Lagos; The specialist Laboratory (TSL) and Histolab was carried out. RESULTS: 733 samples of lymph nodes were seen. Of these, 229 (31.37%) were reactive, 109 (14.93%) were chronic granulomatuos lesions while lymphoma was seen in 123 cases (16.85%) and metastatic lesions in 269 cases (36.50%). The male to female ratio was 1:1.5. (286:436). In 670 cases, the lymph nodes were peripheral in location with the distribution of 249, 140 and 42 from the axilla, cervical and inguinal regions respectively. Only 60 of the samples were deeply seated lymph nodes; 55 from the intra-abdominal cavity and 5 from the mediastenium. About 239 samples did not have their sites indicated and majority of these (106) were diagnosed to be reactive lymphadenitis. CONCLUSION: The categories of lymphoid disease are similar to that of other third world countries but there is slight disparity in the frequency distribution of these diseases compared with findings in literatures within the countries.

Accuracy of fine needle aspiration in the diagnosis of peripheral lymph node enlargements. Lagos University Teaching Hospital, Nigeria.

BACKGROUND: The usefulness of fine needle aspiration technique for cytology diagnosis of lymph node lesions has been a subject of controversy over years. OBJECTIVE: This study aims to determine the accuracy of this technique in the diagnosis of peripheral lymph node lesions, in our center, using histology diagnosis as a gold standard. METHODS: The cytopathological diagnosis of 49 cases of peripheral lymph node lesions seen in the Morbid Anatomy Department of the Lagos University Teaching Hospital, Lagos from Jan. 2000 to December 2009 were compared with their corresponding histological diagnosis. Lymph node lesions were categorized into inflammatory, Hodgkin's lymphoma, Non Hodgkin's lymphoma and metastatic. The sensitivity, specificity and positive as well as the negative predictive values of the technique were determined. RESULTS: The overall sensitivity, specificity and positive and negative predictive values were 79.6%, 95.9.0%, 79.6% and 95.9% respectively. For non neoplastic lesions, FNAC has a high false negative report. For primary lymphoid neoplasm, the sensitivity, specificity and both predictive values were above 80% while the procedure produced sensitivity and specificity values of 100 and 97.1 percent respectively in the diagnosis of metastatic lesions. CONCLUSION: Despite the pitfalls and limitations of the procedure, the study demonstrated that fine-needle aspiration is a valuable tool for the evaluation of peripheral adenopathy. But FNAC technique for cytological diagnosis of lymphoid lesions using H and E and Giemsa stains alone cannot be used as the sole diagnosis of lymphoid enlargements. Its accuracy can be improved with the availability immunocytochemistry and flow cytometry.

Haemoglobin and ferritin concentrations in cord blood in a tertiary health centre in Nigeria.

BACKGROUND: Haemoglobin(Hb) and serum ferritin (SF) concentrations of cord blood of babies born at term at the Lagos State University Teaching Hospital, Maternity Centre (Ayinke House), Ikeja in the South-Western part of Nigeria were determined to establish mean values for these substances in our locality. OBJECTIVES: To establish the mean values for haemoglobin and serum ferritin concentrations of cord blood of babies born at term in our environment and to determine the prevalence of foetal anaemia and low iron store in cord blood in our locality. METHODS: Haemoglobin and ferritin levels in cord blood of 142 newborns were determined. Two millilitres of blood was collected from the cord of each newborn into EDTA bottle for complete blood count analysis and another 2mls into a plain bottle for serum ferritin assay. Cut-off values for cord blood Hb and serum ferritin concentrations were 12.5g/dL and 60 microg/L respectively. RESULTS: The mean Hb and ferritin values were 13.024 +/- 2.41 g/dL and 70.85 +/- 97.07 microg/dL respectively. The prevalence of foetal anaemia is 32.4 %. About 59.2% of full term newborns had low iron store. Birth weight was significantly associated with Hb concentration (p=0.039) and apga sscore (p=0.002). CONCLUSION: The prevalence of foetal anaemia was 32.4%. More than half (59.2%) of the newborns had low cord blood serum ferritin.

Pattern of demographic risk factors in the seroprevalence of anti-Toxoplasma gondii antibodies in HIV infected patients at the Lagos University Teaching Hospital.

BACKGROUND: The Apicomplexan protozoan, Toxoplasma gondii, is a human parasite, with an ubiquitous distribution. Prevalence of the infection varies widely, depending on cultural, geographic and climatic factors. More often asymptomatic, T. gondii infection may be a severe and life-threatening disease. OBJECTIVES: This study was conducted to determine the seroprevalence of Toxoplasma gondii IgG antibody (TIgG) among HIV infected persons at the Lagos University Teaching Hospital PEPFAR site, and the demographic characteristics of the study group. METHODS: 460 plasma specimens were tested for TIgG antibodies by enzyme immuno assay technique and close ended questionnaires were applied on all respondents to obtain relevant data on demographics. Plasma was obtained from two study groups comprising of 380 HIV positive patients and 80 HIV negative adults who served as the control group. RESULTS: The overall seroprevalence was 30% in the immunocompetent control group (37.5% of the males tested and 16.7% among the females). The over all seroprevalence of TIgG antibody among the HIV positive respondents was 54% (206 Of 380). The study therefore showed statistically significant difference between the seroprevalence of TIgG antibody among the immunocompetent control group and HIV positive study group (p = 0.00356). Seroprevalence of TIgG antibody was lowest among the educated subjects (19% of subjects with tertiary education). T. gondii antibody seroprevalence for males, married respondents and rural dwellers were 70.4%, 72.3% and 69% respectively, and were all statistically significant at p < 0.000. Moreso, a significant association was observed between the seroprevalence of anti toxoplasma gondii IgG antibody and the consumption of beef among the study groups. (P < 0.0001). CONCLUSION: Socio-cultural and nutritional habits,contribute significantly to the prevalence of Toxoplasmosis and thus any effective control must be centred around these issues.

Preliminary study on the induction of sperm head abnormalities in mice, Mus musculus, exposed to radiofrequency radiations from global system for mobile communication base stations.

The exposure of male mice to radiofrequency radiations from mobile phone (GSM) base stations at a workplace complex and residential quarters caused 39.78 and 46.03%, respectively, in sperm head abnormalities compared to 2.13% in control group. Statistical analysis of sperm head abnormality score showed that there was a significant (p < 0.05) difference in occurrence of sperm head abnormalities in test animals. The major abnormalities observed were knobbed hook, pin-head and banana-shaped sperm head. The occurrence of the sperm head abnormalities was also found to be dose dependent. The implications of the observed increase occurrence of sperm head abnormalities on the reproductive health of humans living in close proximity to GSM base stations were discussed.

Pattern of Demographic Risk Factors in the Seroprevalence of Anti-Toxoplasma Gondii Antibodies in HIV Infected Patients at the Lagos University Teaching Hospital

BACKGROUND:The Apicomplexan protozoan; Toxoplasma gondii; is a human parasite; with an ubiquitous distribution. Prevalence of the infection varies widely; depending on cultural; geographic and climatic factors. More often asymptomatic; T. gondii infection may be a severe and life-threatening disease. OBJECTIVES: This study was conducted to determine the seroprevalence of Toxoplasma gondii IgG antibody (TIgG) among HIV infected persons at the Lagos University Teaching Hospital PEPFAR site; and the demographic characteristics of the study group. METHODS: 460 plasma specimens were tested for TIgG antibodies by enzyme immuno assay technique and close ended questionnaires were applied on all respondents to obtain relevant data on demographics. Plasma was obtained from two study groups comprising of 380 HIV positive patients and 80 HIV negative adults who served as the control group. RESULTS: The overall seroprevalence was 30in the immunocompetent control group (37.5of the males tested and 16.7among the females). The over all seroprevalence of TIgG antibody among the HIV positive respondents was 54(206 Of 380). The study therefore showed statistically significant difference between the seroprevalence of TIgG antibody among the immunocompetent control group and HIV positive study group (p = 0.00356). Seroprevalence of TIgG antibody was lowest among the educated subjects (19of subjects with tertiary education). T. gondii antibody seroprevalence for males; married respondents and rural dwellers were 70.4; 72.3and 69respectively; and were all statistically significant at p 0.000. Moreso; a significant association was observed between the seroprevalence of anti toxoplasma gondii IgG antibody and the consumption of beef among the study groups.(P 0.0001). CONCLUSION: Socio-cultural and nutritional habits;contribute significantly to the prevalence of Toxoplasmosis and thus any effective control must be centred around these issues