Serosurvey of Treponema pallidum infection among children with skin ulcers in the Tarangire-Manyara ecosystem, northern Tanzania.

BACKGROUND: The first yaws eradication campaign reduced the prevalence of yaws by 95%. In recent years, however, yaws has reemerged and is currently subject to a second, ongoing eradication campaign. Yet, the epidemiological status of Tanzania and 75 other countries with a known history of human yaws is currently unknown. Contrary to the situation in humans in Tanzania, recent infection of nonhuman primates (NHPs) with the yaws bacterium Treponema pallidum subsp. pertenue (TPE) have been reported. In this study, we consider a One Health approach to investigate yaws and describe skin ulcers and corresponding T. pallidum serology results among children living in the Tarangire-Manyara ecosystem, an area with increasing wildlife-human interaction in northern Tanzania. METHODS: To investigate human yaws in Tanzania, we conducted a cross-sectional study to screen and interview skin-ulcerated children aged 6 to 15 years, who live in close proximity to two national parks with high numbers of naturally TPE-infected monkeys. Serum samples from children with skin ulcers were tested for antibodies against the bacterium using a treponemal (Treponema pallidum Particle Agglutination assay) and a non-treponemal (Rapid Plasma Reagin) test. RESULTS: A total of 186 children aged between 6 and 15 years (boys: 10.7 ± 2.1 (mean ± SD), N = 132; girls: 10.9 ± 2.0 (mean ± SD), N = 54) were enrolled. Seven children were sampled at health care facilities and 179 at primary schools. 38 children (20.4%) reported active participation in bushmeat hunting and consumption and 26 (13.9%) reported at least one physical contact with a NHP. None of the lesions seen were pathognomonic for yaws. Two children tested positive for treponemal antibodies (1.2%) in the treponemal test, but remained negative in the non-treponemal test. CONCLUSIONS: We found no serological evidence of yaws among children in the Tarangire-Manyara ecosystem. Nevertheless, the close genetic relationship of human and NHPs infecting TPE strains should lead to contact prevention with infected NHPs. Further research investigations are warranted to study the causes and possible prevention measures of spontaneous chronic ulcers among children in rural Tanzania and to certify that the country is free from human yaws.

Father absence but not fosterage predicts food insecurity, relative poverty, and poor child health in northern Tanzania.

OBJECTIVES: The importance of fathers in ensuring child health in rural developing populations is questioned by anthropologists and population health scientists. Existing literature focuses on paternal death and child mortality. A relative lack of studies consider alternative forms of father absence and/or more subtle health outcomes. Here we determine the frequency and form of father absence in northern Tanzania, and its relationship to household food security, wealth, and child anthropometric status. METHODS: We conducted a cross-sectional survey of 3136 children under 5 years of age from 56 villages. Using multilevel regression we contrast children residing with both parents to those that (i) have experienced paternal death, (ii) reside with their mother but not their living father and (iii) are fostered apart from both living parents. RESULTS: Of the total, 3.5% of children had experienced paternal death. Thirteen percent resided with their mother but away from their living father. Supporting data indicate such cases primarily reflect parental divorce/separation, extra-marital birth, or polygynous fathers residing with an alternative cowife. Paternal death and residing apart from one's living father was associated with lower food security and/or relative poverty and there is suggestive evidence that children in such circumstances achieve lower height-for-age. Six percent of children were fostered, usually with grandparents, and were comparable to children residing with both parents in terms of household food security, wealth, and anthropometric status. CONCLUSION: Our results highlight diversity in the form and consequences of father absence. We discuss limitations of the current study and wider literature on fatherhood and make suggestions for future research.

Increased isolation of nontuberculous mycobacteria among TB suspects in Northeastern, Tanzania: public health and diagnostic implications for control programmes.

BACKGROUND: Non-tuberculous mycobacteria (NTM) are increasingly reported worldwide associated with human disease. Defining the significance of NTM in settings with endemic tuberculosis (TB) requires the discrimination of NTM from TB in suspect patients. Correct and timely identification of NTM will impact both therapy and epidemiology of TB and TB-like diseases. The present study aimed at determining the frequency and diversity of NTM among TB suspects in northeastern Tanzania. METHODS: A cross-sectional study was conducted between November 2012 through January 2013. Seven hundred and forty-four sputum samples were collected from 372 TB suspects. Detection was done by using phenotypic, GenoType(®) Mycobacterium CM/AS kits, 16S rRNA and hsp65 gene sequencing for identification of isolates not identified by Hain kits. Binary regression model was used to analyse the predictors of NTM detection. RESULTS: The prevalence of NTM was 9.7% of the mycobacterial isolates. Out of 36 patients with confirmed NTM infection, 12 were HIV infected with HIV being a significant predictor of NTM detection (P < 0.001). Co-infection with Mycobacterium tuberculosis (M. tb) was found in five patients. Twenty-eight NTM isolates were identified using GenoType(®) Mycobacterium CM/AS and eight isolates could not be identified. Identified species included M. gordonae and M. interjectum 6 (16.7%), M. intracelullare 4 (11.1%), M. avium spp. and M. fortuitum 2 (5.5%), M. kansasii, M. lentiflavum, M. simiae, M. celatum, M. marinum 1 (2.8%) each. Of isolates not identified to subspecies level, we identified M. kumamotonense (2), M. intracellulare/kansasii, M. intermedium/triplex, M. acapulcensis/flavescens, M. stomatepiae, M. colombiense and M. terrae complex (1) each using 16S rRNA sequencing. Additionally, hsp65 gene sequencing identified M. kumamotonense, M. scrofulaceum/M. avium, M. avium, M. flavescens/novocastrense, M. kumamotonense/hiberniae, M. lentiflavum, M. colombiense/M. avium and M. kumamotonense/terrae/hiberniae (1) each. Results of the 16S rRNA and hsp65 gene sequencing were concordant in three and discordant in five isolates not identified by GenoType(®) Mycobacterium CM/AS. CONCLUSION: NTM infections may play a vital role in causing lung disease and impact management of TB in endemic settings. GenoType(®) Mycobacterium CM/AS represents a useful tool to identify clinical NTM infections. However, 16S rRNA gene sequencing should be thought for confirmatory diagnosis of the clinical isolates. Due to the complexity and inconsistence of NTM identification, we recommend diagnosis of NTM infections be centralized by strengthening and setting up quality national and regional infrastructure.

Anti-TB drug resistance in Tanga, Tanzania: A cross sectional facility-base prevalence among pulmonary TB patients.

OBJECTIVE: To determine the prevalence and risk factors associated with drug resistance tuberculosis (TB) at facility-base level in Tanga, Tanzania. METHODS: A total of 79 Mycobacterium tuberculosis (MTB) isolates included in the study were collected from among 372 (312 new and 60 previously treated) TB suspects self-referred to four TB clinics during a prospective study conducted from November 2012 to January 2013. Culture and drug susceptibility test of the isolates was performed at the institute of medical microbiology and epidemiology of infectious diseases, University hospital, Leipzig, Germany. Data on the patient's characteristics were obtained from structured questionnaire administered to the patients who gave informed verbal consent. Unadjusted bivariate logistic regression analysis was performed to assess the risk factors for drug resistant-TB. The significance level was determined at P < 0.05. RESULTS: The overall proportions of any drug resistance and MDR-TB were 12.7% and 6.3% respectively. The prevalence of any drug resistance and MDR-TB among new cases were 11.4% and 4.3% respectively, whereas among previously treated cases was 22.2% respectively. Previously treated patients were more likely to develop anti-TB drug resistance. There was no association between anti-TB drug resistances (including MDR-TB) with the risk factors analysed. CONCLUSIONS: High proportions of anti-TB drug resistance among new and previously treated cases observed in this study suggest that, additional efforts still need to be done in identifying individual cases at facility-base level for improved TB control programmes and drug resistance survey should continuously be monitored in the country.

No evidence that polygynous marriage is a harmful cultural practice in northern Tanzania.

Polygyny is cross-culturally common and a topic of considerable academic and policy interest, often deemed a harmful cultural practice serving the interests of men contrary to those of women and children. Supporting this view, large-scale studies of national African demographic surveys consistently demonstrate that poor child health outcomes are concentrated in polygynous households. Negative population-level associations between polygyny and well-being have also been reported, consistent with the hypothesis that modern transitions to socially imposed monogamy are driven by cultural group selection. We challenge the consensus view that polygyny is harmful, drawing on multilevel data from 56 ethnically diverse Tanzanian villages. We first demonstrate the vulnerability of aggregated data to confounding between ecological and individual determinants of health; while across villages polygyny is associated with poor child health and low food security, such relationships are absent or reversed within villages, particularly when children and fathers are coresident. We then provide data indicating that the costs of sharing a husband are offset by greater wealth (land and livestock) of polygynous households. These results are consistent with models of polygyny based on female choice. Finally, we show that village-level negative associations between polygyny prevalence, food security, and child health are fully accounted for by underlying differences in ecological vulnerability (rainfall) and socioeconomic marginalization (access to education). We highlight the need for improved, culturally sensitive measurement tools and appropriate scales of analysis in studies of polygyny and other purportedly harmful practices and discuss the relevance of our results to theoretical accounts of marriage and contemporary population policy.

Ethnicity and child health in northern Tanzania: Maasai pastoralists are disadvantaged compared to neighbouring ethnic groups.

The Maasai of northern Tanzania, a semi-nomadic ethnic group predominantly reliant on pastoralism, face a number of challenges anticipated to have negative impacts on child health, including marginalisation, vulnerabilities to drought, substandard service provision and on-going land grabbing conflicts. Yet, stemming from a lack of appropriate national survey data, no large-scale comparative study of Maasai child health has been conducted. Savannas Forever Tanzania surveyed the health of over 3500 children from 56 villages in northern Tanzania between 2009 and 2011. The major ethnic groups sampled were the Maasai, Sukuma, Rangi, and the Meru. Using multilevel regression we compare each ethnic group on the basis of (i) measurements of child health, including anthropometric indicators of nutritional status and self-reported incidence of disease; and (ii) important proximate determinants of child health, including food insecurity, diet, breastfeeding behaviour and vaccination coverage. We then (iii) contrast households among the Maasai by the extent to which subsistence is reliant on livestock herding. Measures of both child nutritional status and disease confirm that the Maasai are substantially disadvantaged compared to neighbouring ethnic groups, Meru are relatively advantaged, and Rangi and Sukuma intermediate in most comparisons. However, Maasai children were less likely to report malaria and worm infections. Food insecurity was high throughout the study site, but particularly severe for the Maasai, and reflected in lower dietary intake of carbohydrate-rich staple foods, and fruits and vegetables. Breastfeeding was extended in the Maasai, despite higher reported consumption of cow's milk, a potential weaning food. Vaccination coverage was lowest in Maasai and Sukuma. Maasai who rely primarily on livestock herding showed signs of further disadvantage compared to Maasai relying primarily on agriculture. We discuss the potential ecological, socioeconomic, demographic and cultural factors responsible for these differences and the implications for population health research and policy.

Genetic profile of Mycobacterium tuberculosis and treatment outcomes in human pulmonary tuberculosis in Tanzania.

Information on the different spoligotype families of Mycobacterium tuberculosis in Tanzania is limited, and where available, restricted to small geographical areas. This article describes the genetic profile of M tuberculosis across Tanzania and suggests how spoligotype families might affect drug resistance and treatment outcomes for smear positive pulmonary tuberculosis patients in Tanzania. We conducted the study from 2006 to 2008, and the isolates were obtained from samples collected under the routine drug resistance surveillance system. The isolates were from specimens collected from 2001 to 2007, and stored at the Central and Reference Tuberculosis Laboratory. A total of 487 isolates from 23 regions in the country were spoligotyped. We were able to retrieve clinical information for 446 isolates only. Out of the 487 isolates spoligotyped, 195(40.0%) belonged to the Central Asian (CAS) family, 84 (17.5%) to the Latin American Mediterranean (LAM) family, 49 (10.1%) to the East-African Indian (EAI) family, and 33 (6.8%) to the Beijing family. Other isolates included 1 (0.2%) for H37Rv, 10 (2.1%) for Haarlem, 4 (0.8%) for S family, 58 (11.9%) for T family and 52 (10.7%) for unclassified. No spoligotype patterns were consistent with M bovis. Regarding treatment outcomes, the cure rate was 80% with no significant variation among the spoligotype families. The overall level of MDR TB was 2.5% (3/12 1), with no significant difference among the spoligotype families. All Beijing strains (11.8%, 30/254) originated from the Eastern and Southern zones of the country, of which 80% were from Dar es Salaam. Isolates from the CAS and T families were reported disproportionately from the Eastern-Southern zone, and EAI and LAM families from the Northern-Lake zones but the difference was not statistically significant. Five isolates were identified as non-tuberculous Mycobacteria. In conclusion, M. tuberculosis isolates from pulmonary tuberculosis cases in Tanzania were classified mostly within the CAS, LAM, and EAI and T families, while the Beijing family comprised about 7% isolates only. Consistently good treatment outcomes were recorded across these spoligotype families. The proportion of drug resistance strains was low. The findings also suggest variation of spoligotype families with varying geographical localities within the country, and identify this area for further research to confirm this finding.

Genetic profile of Mycobacterium Tuberculosis and Treatment Outcomes in Human Pulmonary Tuberculosis in Tanzania

Information on the different spoligotype families of Mycobacterium tuberculosis in Tanzania is limited; and where available; restricted to small geographical areas. This article describes the genetic profile of M. tuberculosis across Tanzania and suggests how spoligotype families might affect drug resistance and treatment outcomes for smear positive pulmonary tuberculosis patients in Tanzania. We conducted the study from 2006 to 2008; and the isolates were obtained from samples collected under the routine drug resistance surveillance system. The isolates were from specimens collected from 2001 to 2007; and stored at the Central and Reference Tuberculosis Laboratory. A total of 487 isolates from 23 regions in the country were spoligotyped. We were able to retrieve clinical information for 446 isolates only. Out of the 487 isolates spoligotyped; 195(40.0) belonged to the Central Asian (CAS) family; 84 (17.5) to the Latin American Mediterranean (LAM) family; 49 (10.1) to the Latin American Mediterranean (LAM) family; 49 (10.1) to the East-African Indian (EAI) family; and 33 (6.8) to the Beijing family. Other isolates included 1 (0.2) for H37Rv; 10 (2.1) for Haarlem; 4 (0.8) for S family; 58 (11.9) for T family and 52 (10.7) for unclassified. No spoligotype patterns were consistent with M. bovis. Regarding treatment outcomes; the cure rate was 80 with no significant variation among the spoligotype families. The overall level of MDR TB was 2.5 (3/121); with no significant difference among the spoligotype families. All Beijing strains (11.8; 30/254) originated from the Eastern and Southern zones of the country; of which 80 were from Dar es Salaam. Isolates from the CAS and T families were reported disproportionately from the Eastern-Southern zone; and EAI and LAM families from the Northern-Lake zones but the difference was not statistically significant. Five isolates were identified as non-tuberculous Mycobacteria. In conclusion; M. tuberculosis isolates from pulmonary tuberculosis cases in Tanzania were classified mostly within the CAS; LAM; and EAI and T families; while the Beijing family comprised about 7 isolates only. Consistently good treatment outcomes were recorded across these spoligotype families. The proportion of drug resistance strains was low. The findings also suggest variation of spoligotype families with varying geographical localities within the country; and identify this area for further research to confirm this finding

Implementation of a national anti-tuberculosis drug resistance survey in Tanzania.

BACKGROUND: A drug resistance survey is an essential public health management tool for evaluating and improving the performance of National Tuberculosis control programmes. The current manuscript describes the implementation of the first national drug resistance survey in Tanzania. METHODS: Description of the implementation process of a national anti-tuberculosis drug resistance survey in Tanzania, in relation to the study protocol and Standard Operating Procedures. RESULTS: Factors contributing positively to the implementation of the survey were a continuous commitment of the key stakeholders, the existence of a well organized National Tuberculosis Programme, and a detailed design of cluster-specific arrangements for rapid sputum transportation. Factors contributing negatively to the implementation were a long delay between training and actual survey activities, limited monitoring of activities, and an unclear design of the data capture forms leading to difficulties in form-filling. CONCLUSION: Careful preparation of the survey, timing of planned activities, a strong emphasis on data capture tools and data management, and timely supervision are essential for a proper implementation of a national drug resistance survey.

Immunohistochemistry using a Mycobacterium tuberculosis complex specific antibody for improved diagnosis of tuberculous lymphadenitis.

The clinical and histological criteria used to diagnose lymphadenitis caused by Mycobacterium tuberculosis complex organisms have poor specificity. Acid-fast staining and culture has low sensitivity and specificity. We report a novel method for diagnosis of tuberculosis that uses immunohistochemistry to detect the secreted mycobacterial antigen MPT64 on formalin-fixed tissue biopsies. This antigen has not been detected in non-tuberculous mycobacteria. Polymerase chain reaction (PCR) for amplification of IS6110 from DNA obtained from the biopsies was used as a gold standard. Fifty-five cases of granulomatous lymphadenitis with histologically suspected tuberculosis obtained from Norway and Tanzania were evaluated. Four known tuberculosis cases were used as positive controls, and 16 biopsies (12 foreign body granulomas and four other non-granulomatous cases) as negative controls. With immunohistochemistry, 64% (35/55) and with PCR, 60% (33/55) of granulomatous lymphadenitis cases were positive. Using PCR as the gold standard, the classical tuberculosis histology had sensitivity, specificity, positive and negative predictive values of 92, 37, 60, and 81%, respectively, and immunohistochemistry had sensitivity, specificity, positive and negative predictive values of 90, 83, 86, and 88%, respectively. The observed agreement between PCR and immunohistochemistry was 87% (kappa = 0.73). Immunohistochemistry with anti-MPT64 antiserum is a rapid, sensitive, and specific method for establishing an etiological diagnosis of tuberculosis in histologic specimens. Immunohistochemistry has the advantages over PCR of being robust and cheap, and it can easily be used in a routine laboratory.

A first insight into the genetic diversity of Mycobacterium tuberculosis in Dar es Salaam, Tanzania, assessed by spoligotyping.

BACKGROUND: Tanzania has a high tuberculosis incidence, and genotyping studies of Mycobacterium tuberculosis in the country are necessary in order to improve our understanding of the epidemic. Spoligotyping is a potentially powerful genotyping method due to fast generation of genotyping results, high reproducibility and low operation costs. The recently constructed SpolDB4 database and the model-based program 'Spotclust' can be used to assign isolates to families, subfamilies and variants. The results of a study can thus be analyzed in a global context. RESULTS: One hundred forty-seven pulmonary isolates from consecutive tuberculosis patients in Dar es Salaam were spoligotyped. SpolDB4 and 'Spotclust' were used to assign isolates to families, subfamilies and variants. The CAS (37%), LAM (22%) and EAI (17%) families were the most abundant. Despite the dominance of these three families, diversity was high due to variation within M. tuberculosis families. Of the obtained spoligopatterns, 64% were previously unrecorded. CONCLUSION: Spoligotyping is useful to gain an overall understanding of the local TB epidemic. This study demonstrates that the extensive TB epidemic in Dar es Salaam, Tanzania is caused by a few successful M. tuberculosis families, dominated by the CAS family. Import of strains was a minor problem.

Immunohistochemical analysis of cytokines and apoptosis in tuberculous lymphadenitis.

Relatively little is known about the effector mechanisms whereby the human immune system controls Mycobacterium tuberculosis infection. In this study we elaborate on the immune response and mechanisms of persistence of mycobacteria in lesions by analysing, using immunohistochemistry, the expression of cytokines [tumour necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), transforming growth factor-beta (TGF-beta) and interferon-gamma (IFN-gamma)], apoptotic cells and apoptosis-related proteins [Bcl2, Bax, Fas ligand (FasL) and Fas] in the human tuberculous lymphadenitis lesions. The expression of apoptosis-related proteins has been shown to be exploited by mycobacteria to evade the immune response and persist in the host. Foreign body (FB) granulomas were used as controls. In tuberculosis (TB) granulomas, epithelioid cells and multinucleated giant cells expressed cytokines differently. In epithelioid cells, the numbers of TNF-alpha-, IL-10- and TGF-beta-stained cells were higher than IFN-gamma-stained cells (P < 0.01). TGF-beta and FasL were strongly expressed in the necrotic centres as compared with other cytokines. More giant cells expressed IL-10 and TGF-beta than expressed TNF-alpha and IFN-gamma (P < 0.01). Staining of consecutive sections revealed that some giant cells expressed IL-10 but not TNF-alpha. Apoptotic TB giant cells correlated positively with the expression of TNF-alpha, IFN-gamma and TGF-beta, but not with the expression of IL-10. The percentage of giant cells expressing Bax was lower than those expressing Fas, unlike the epithelioid cells, suggesting that TB giant cells are less susceptible to apoptosis. Compared with FB giant cells, there were fewer TB giant cells showing TNF-alpha, IFN-gamma, FasL, Fas expression or undergoing apoptosis (P < 0.05). Taken together, these observations show that the cellular microenvironment of TB granulomas down-regulates microbicidal functions, favouring bacillary survival and persistence. TGF-beta and FasL may be responsible for tissue destruction. The giant cells, being less susceptible to apoptosis, may remain a continuous source of pro-inflammatory cytokines, causing immune pathology.

Significance of Fas and Fas ligand in tuberculous lymphadenitis.

The Fas/Fas-ligand (FasL) system plays an important role in regulation of apoptosis and the immune response, and is exploited by mycobacteria to evade the immune response. This study was performed to investigate the distribution and levels of FasL and Fas in lymph node granulomas and sera of tuberculous lymphadenitis patients by immunohistochemistry and enzyme-linked immunosorbent assay. The validity of soluble Fas (sFas) or soluble FasL (sFasL) as a diagnostic tool was also examined. Levels of sFasL in serum were elevated among patients. The numbers of FasL stained cells in lymph node granulomas were higher than Fas. Children had significantly higher levels of sFasL as compared to adults. The human immunodeficiency virus (HIV)-tuberculosis (TB)-coinfected patients displayed no differences in the levels of sFasL or sFas compared with HIV-negative patients. The healthy controls from a high endemic tuberculosis country (having latent TB) had significantly higher levels of sFasL than from a country with no TB transmission. The sensitivity and specificity of the FasL and Fas test were low when compared with the culture results as the gold standard. However, by using histology as the gold standard, the sensitivity and specificity of the FasL test were increased to 66.7% and 100%, respectively, but for the Fas test remained low. In conclusion, sFasL and sFas cannot be used as diagnostic tests for tuberculous lymphadenitis. However, its utility in detecting latent TB and childhood tuberculous lymphadenitis remains to be evaluated. FasL seems to play a role in immune modulation and pathogenesis of TB. Modulators of Fas/FasL-mediated apoptosis may therefore be clinically useful.