Informal Urban Settlements and Cholera Risk in Dar es Salaam,Tanzania

Background

As a result of poor economic opportunities and an increasing shortage of affordable housing, much of the spatial growth in many of the world''s fastest-growing cities is a result of the expansion of informal settlements where residents live without security of tenure and with limited access to basic infrastructure. Although inadequate water and sanitation facilities, crowding and other poor living conditions can have a significant impact on the spread of infectious diseases, analyses relating these diseases to ongoing global urbanization, especially at the neighborhood and household level in informal settlements, have been infrequent. To begin to address this deficiency, we analyzed urban environmental data and the burden of cholera in Dar es Salaam, Tanzania.

Methodology/Principal Findings

Cholera incidence was examined in relation to the percentage of a ward''s residents who were informal, the percentage of a ward''s informal residents without an improved water source, the percentage of a ward''s informal residents without improved sanitation, distance to the nearest cholera treatment facility, population density, median asset index score in informal areas, and presence or absence of major roads. We found that cholera incidence was most closely associated with informal housing, population density, and the income level of informal residents. Using data available in this study, our model would suggest nearly a one percent increase in cholera incidence for every percentage point increase in informal residents, approximately a two percent increase in cholera incidence for every increase in population density of 1000 people per km² in Dar es Salaam in 2006, and close to a fifty percent decrease in cholera incidence in wards where informal residents had minimally improved income levels, as measured by ownership of a radio or CD player on average, in comparison to wards where informal residents did not own any items about which they were asked. In this study, the range of access to improved sanitation and improved water sources was quite narrow at the ward level, limiting our ability to discern relationships between these variables and cholera incidence. Analysis at the individual household level for these variables would be of interest.

Conclusions/Significance

Our results suggest that ongoing global urbanization coupled with urban poverty will be associated with increased risks for certain infectious diseases, such as cholera, underscoring the need for improved infrastructure and planning as the world''s urban population continues to expand.     

Bioconversion of lignocellulosic waste from selected dumping sites in Dar es Salaam,Tanzania

The poor management of solid wastes in Tanzania urban centers is a chronic problem that has increasingly become a source of environmental pollution. Bioconversion offers a cheap and safe method of not only disposing these wastes, but also it has the potential to convert lignocellulosic wastes into usable forms such as reducing sugars that could be used as food. This paper reports a preliminary study on the physical characteristics, acid pretreatment, saccharification by cellulase from Trichoderma reesei and fermentation by Saccharomyces cerevisiae of the lignocellulosic component of the solid wastes collected from various dumping sites located in Kinondoni Municipality, Dar es Salaam city. The results showed that overall, the lignocellulosic component constitute about 50% of solid wastes dumped in the study areas. Maximum production of reducing sugars was obtained after 6 h of saccharification while highest concentrations of bioethanol were achieved after 48 h of fermentation. Microbial bioconversion of lignocellulose component yielded up to 21% bioethanol.     

Impact of Community-Based Larviciding on the Prevalence of Malaria Infection in Dar es Salaam,Tanzania

Background

The use of larval source management is not prioritized by contemporary malaria control programs in sub-Saharan Africa despite historical success. Larviciding, in particular, could be effective in urban areas where transmission is focal and accessibility to Anopheles breeding habitats is generally easier than in rural settings. The objective of this study is to assess the effectiveness of a community-based microbial larviciding intervention to reduce the prevalence of malaria infection in Dar es Salaam, United Republic of Tanzania.

Methods and Findings

Larviciding was implemented in 3 out of 15 targeted wards of Dar es Salaam in 2006 after two years of baseline data collection. This intervention was subsequently scaled up to 9 wards a year later, and to all 15 targeted wards in 2008. Continuous randomized cluster sampling of malaria prevalence and socio-demographic characteristics was carried out during 6 survey rounds (2004–2008), which included both cross-sectional and longitudinal data (N = 64,537). Bayesian random effects logistic regression models were used to quantify the effect of the intervention on malaria prevalence at the individual level. Effect size estimates suggest a significant protective effect of the larviciding intervention. After adjustment for confounders, the odds of individuals living in areas treated with larviciding being infected with malaria were 21% lower (Odds Ratio = 0.79; 95% Credible Intervals: 0.66–0.93) than those who lived in areas not treated. The larviciding intervention was most effective during dry seasons and had synergistic effects with other protective measures such as use of insecticide-treated bed nets and house proofing (i.e., complete ceiling or window screens).

Conclusion

A large-scale community-based larviciding intervention significantly reduced the prevalence of malaria infection in urban Dar es Salaam.     

Personal protection against mosquitoes in Dar es Salaam,Tanzania, by using a kerosene oil lamp to vaporize transfluthrin

The effectiveness of a cheap and easy method of household protection against Culex quinquefasciatus Say and other mosquitoes (Diptera: Culicidae) was investigated in Dar es Salaam, Tanzania. Kerosene-burning lamps (korobois) were modified to heat and vaporize transfluthrin, a volatile pyrethroid insecticide. When transfluthrin was added to fuel of the lamp, protection against biting was poor unless a very high concentration of insecticide was used. A modified lamp (= vaporizing koroboi) was designed to overcome this problem by mixing the insecticide with vegetable oil and heating it to 120 degrees C in a tin held just above the flame. The concentration of 0.1% transfluthrin in vegetable oil gave 50-75% reduction in biting, a similar degree of protection to that obtained from burning a mosquito coil containing a synthetic pyrethroid (0.25% d-allethrin) and significantly better protection than a locally bought coil (brand 'White Crane', probably containing DDT). Greater protection (consistently > 90%) was achieved with a higher concentration of transfluthrin (0.5%) in the vegetable oil. This modified lamp is simple, cheap and employs locally available technology. With further development, and due regard to inhalation toxicity of the vaporized materials, it may offer a more cost-effective alternative to a mosquito coil as a means of personal protection, and a useful complement to a net for the early part of the evening before bedtime.     

Prospective study of incidence of juvenile diabetes mellitus over 10 years in Dar es Salaam,Tanzania.

OBJECTIVE--To ascertain the annual incidence of diabetes requiring treatment with insulin in children and adolescents aged 0-19 years in Dar es Salaam, Tanzania, during a 10 year period from 1 January 1982 to 31 December 1991. DESIGN--Prospective registration at a major urban hospital of all patients with newly diagnosed diabetes who were resident in Dar es Salaam. SETTING--Muhimbili Medical Centre, Dar es Salaam, Tanzania. PATIENTS--86 patients: 45 male, 41 female. RESULTS--The annual incidence of juvenile diabetes for both sexes was 1.5 per 100,000 population aged 0-19 years (95% confidence interval 1.3 to 1.7). Incidence per 100,000 population per year increased with age: 0.6 (0.0 to 0.13) in the age group 0-4 years, 0.5 (0.3 to 0.7) at 5-9 years, 2.2 (1.8 to 2.6) at 10-14 years, and 3.4 (2.9 to 3.9) at 15-19 years. CONCLUSION--Juvenile diabetes mellitus is fairly rare in sub-Saharan Africa. If environmental factors such as infection and material deprivation were important determinants of insulin dependent diabetes in Africans, as they may be in Europeans, much higher rates would have been expected unless genetic factors possibly exert a protective role. The eightfold greater incidence in African Americans than in Tanzanians may be related to greater genetic admixture in African Americans with people from countries in Europe with a high incidence.     

A novel bicistronic vector for overexpressing Mycobacterium tuberculosis proteins in Escherichia coli

A putative DNA glycosylase encoded by the Rv3297 gene (MtuNei2) has been identified in Mycobacterium tuberculosis. Our efforts to express this gene in Escherichia coli either by supplementing tRNAs for rare codons or optimizing the gene with preferred codons for E. coli resulted in little or no expression. On the other hand, high-level expression was observed using a bicistronic expression vector in which the target gene was translationally coupled to an upstream leader sequence. Further comparison of the predicted mRNA secondary structures supported the hypothesis that mRNA secondary structure(s) surrounding the translation initiation region (TIR), rather than codon usage, played the dominant role in influencing translation efficiency, although manipulation of codon usage or tRNA supplementation did further enhance expression in the bicistronic vector. Addition of a cleavable N-terminal tag also facilitated gene expression in E. coli, possibly through a similar mechanism. However, since cleavage of N-terminal tags is determined by the amino acid at the P₁′ position downstream of the protease recognition sequence and results in the addition of an extra amino acid in front of the N-terminus of the protein, this strategy is not particularly amenable to Fpg/Nei family DNA glycosylases which carry the catalytic proline residue at the P₁′ position and require a free N-terminus. On the other hand, the bicistronic vector constructed here is potentially valuable particularly when expressing proteins from G/C rich organisms and when the proteins carry proline residues at the N-terminus in their native form. Thus the bicistronic expression system can be used to improve translation efficiency of mRNAs and achieve high-level expression of mycobacterial genes in E. coli.     

结核分枝杆菌sRNA Mpr5的功能验证

【目的】探究结核分枝杆菌(Mycobacterium tuberculosis, M. tb)中sRNA Mpr5对分枝杆菌的抗逆性及宿主细胞生理的影响。【方法】构建结核分枝杆菌sRNA Mpr5过表达的重组耻垢分枝杆菌(Mycolicibacterium smegmatis, M. smeg) M3-Atc,以转入空载质粒(pSI)的野生型耻垢分枝杆菌T1-Atc作为对照,观察细菌体外生长能力和菌落形态变化。通过非生物胁迫处理(低氧、饥饿、0.02%十二烷基硫酸钠)探究重组菌株的抗逆能力。用重组耻垢分枝杆菌侵染人非小细胞肺癌上皮细胞系A549,活菌涂板计算细菌的胞内增殖能力,利用免疫荧光染色观察感染后细胞的生理结构变化。【结果】sRNA Mpr5过表达菌株的体外生长与菌落形态均与野生型相似。抗逆性实验表明Mpr5过表达菌株(M3-Atc)的抗表面活性剂能力在4 h时显著提高(P<0.05);在饥饿模型中M3-Atc早期(2–12 h)就表现出生长劣势(P<0.05);低氧模型中M3-Atc菌株0–3 d均处于生长优势状态,增长均高于对照组(P<0.05),3d后增长...     

Three-step purification of lipopolysaccharide-free, polyhistidine-tagged recombinant antigens of Mycobacterium tuberculosis

Previous work has shown that the study of host immune responses against Mycobacterium tuberculosis, the causative agent of tuberculosis, requires the availability of multiple mycobacterial antigens. Since purification of protein from M. tuberculosis cells is extremely cumbersome, we developed a protocol for purifying milligram amounts of ten recombinant antigens of M. tuberculosis from E. coli cells. Purified proteins were immunologically active and free of contaminants that confound interpretation of cell-based immunological assays. The method utilizes a three-step purification protocol consisting of immobilized metal-chelate affinity chromatography, size exclusion chromatography and anion-exchange chromatography. The first two chromatographic steps yielded recombinant protein free of protein contaminants, while the third step (anion-exchange chromatography) efficiently removed E. coli lipopolysaccharide, a potent polyclonal activator of lymphoid cells. The recombinant proteins were immunologically indistinguishable from their native (i.e., purified from M. tuberculosis) counterparts. Thus the method provides a way to utilize recombinant proteins for immunological analyses that require highly purified antigens.     

Prevalence of Cryptosporidium parvum/hominis,Entamoeba histolytica and Giardia lamblia among Young Children with and without Diarrhea in Dar es Salaam,Tanzania

Background

Although enteroparasites are common causes of diarrheal illness, few studies have been performed among children in Tanzania. This study aimed to investigate the prevalence of Cryptosporidium parvum/hominis, Entamoeba histolytica and Giardia lamblia among young children in Dar es Salaam, Tanzania, and identify risk factors for infection.

Methodology/Principal Findings

We performed an unmatched case-control study among children < 2 years of age in Dar es Salaam, recruited from August 2010 to July 2011. Detection and identification of protozoans were done by PCR techniques on DNA from stool specimens from 701 cases of children admitted due to diarrhea at the three study hospitals, and 558 controls of children with no history of diarrhea during the last month prior to enrollment. The prevalence of C. parvum/hominis was 10.4% (84.7% C. hominis), and that of G. lamblia 4.6%. E. histolytica was not detected. The prevalence of Cryptosporidium was significantly higher in cases (16.3%) than in controls (3.1%; P < 0.001; OR = 6.2; 95% CI: 3.7–10.4). G. lamblia was significantly more prevalent in controls (6.1%) than in cases (3.4%; P = 0.027; OR = 1.8; 95% CI: 1.1–3.1). Cryptosporidium infection was found more often in HIV-positive (24.2%) than in HIV-negative children (3.9%; P < 0.001; OR = 7.9; 95% CI: 3.1–20.5), and was also associated with rainfall (P < 0.001; OR = 2.41; 95% CI: 1.5–3.8). Among cases, stunted children had significantly higher risk of being infected with Cryptosporidium (P = 0.011; OR = 2.12; 95% CI: 1.2–3.8). G. lamblia infection was more prevalent in the cool season (P = 0.004; OR = 2.2; 95% CI: 1.3–3.8), and more frequent among cases aged > 12 months (P = 0.003; OR = 3.5; 95% CI: 1.5–7.8). Among children aged 7–12 months, those who were breastfed had lower prevalence of G. lamblia infection than those who had been weaned (P = 0.012).

Conclusions

Cryptosporidium infection is common among young Tanzanian children with diarrhea, particularly those living with HIV, and infection is more frequent during the rainy season. G. lamblia is frequently implicated in asymptomatic infections, but rarely causes overt diarrheal illness, and its prevalence increases with age.     

用小鼠模型分析可溶性表达结核分枝杆菌Ag85A的免疫原性

【目的】可溶性表达结核分枝杆菌Ag85A蛋白,并评价其免疫原性。【方法】利用冷休克表达质粒和含有伴侣质粒的大肠杆菌对Ag85A蛋白进行可溶性原核表达,并进行纯化与鉴定,通过C57BL/6小鼠模型对Ag85A蛋白的免疫原性,包括诱导机体特异性体液免疫应答和细胞免疫应答水平进行分析。【结果】重组菌诱导后裂解上清中检测到可溶性Ag85A蛋白的表达,经过亲和层析纯化收获了纯度在90%以上的Ag85A蛋白,Western blot鉴定显示其具有较好的免疫反应性。Ag85A蛋白免疫小鼠后,血清中可以检测到高水平的Ig G抗体效价,其中Ig G2b水平要高于Ig G1。通过特异性多肽、蛋白刺激脾脏和腹股沟淋巴结细胞可分泌高水平的IFN-γ、TNF-α等Th1型细胞因子。【结论】实现了Ag85A蛋白的可溶性表达,免疫特性评价显示Ag85A蛋白可诱导机体产生强烈的特异性体液免疫应答及Th1型的细胞免疫应答,从而为其进一步免疫学功能的研究奠定了重要基础。     

Identifying Programmatic Gaps: Inequities in Harm Reduction Service Utilization among Male and Female Drug Users in Dar es Salaam,Tanzania

Introduction

Current estimates suggest an HIV prevalence of 42% among people who inject drugs (PWIDs) in Dar es Salaam, while HIV prevalence is estimated to be 8.8% among the general population in the city. To address the HIV epidemic in this population, the government of Tanzania began establishing HIV prevention, treatment and care services including outreach and medication assisted treatment (MAT) for PWIDs in 2010. We assessed gender inequities in utilization of outreach and MAT services and evaluated differences in HIV risk behaviors between female and male PWIDs.

Materials and Methods

Routine outreach data between December 2010 to mid-August 2012 and baseline data on clients enrolling in methadone from February 2011 to August 2012 were utilized. Binomial regression was used to estimate adjusted relative risk estimates comparing females to males.

Results

From December 2010 to August 2012, 8,578 contacts were made to drug users; among them 1,898 were injectors. A total of 453 injectors were eligible and referred to MAT, of which, 443 enrolled in treatment. However, regarding total outreach contacts, outreach to PWID, referral to MAT and enrollment in MAT, 8% or less of drug users accessing services were women. In contrast, weighted estimations from surveys suggest that 34% of PWIDs are female, and this approximation is similar to recent population size estimations. Overall, 43% of traditional outreach workers conducting outreach with drug users were female. Though reporting higher levels of condom usage, female PWID were more likely to report multiple sex partners, anal sex, commercial sex work and struggle under a higher burden of addiction, mental disorders and abuse.

Conclusions

Services have not been mobilized adequately to address the clear needs of females who inject drugs. A clear and urgent need exists for women-centered strategies that effectively engage female PWID into HIV prevention services.     

结核分枝杆菌Rv2626c诱导的细胞免疫应答特性

全球有近1/4的人感染结核分枝杆菌(Mycobacterium tuberculosis,M.tb)并长期处于潜伏感染状态。Rv2626c是结核分枝杆菌受DosR调控的重要潜伏感染相关蛋白。本研究对Rv2626c蛋白进行了原核表达和纯化,并以RAW264.7细胞和小鼠为感染模型,对其免疫生物学特性进行了分析。SDS-PAGE及Western blotting鉴定结果表明,Rv2626c-His融合蛋白主要以可溶形式表达,能与兔抗H37Rv多抗血清发生特异性免疫反应。此外,本研究发现Rv2626c蛋白能结合到巨噬细胞RAW 264.7表面并上调细胞NO的生成;显著诱导促炎细胞因子IFN-γ、TNF-α、IL-6和MCP-1的产生;并能诱导小鼠产生更强的Th1免疫应答。上述研究有利于揭示结核分枝杆菌的致病机制,为新型结核病疫苗的研制奠定了理论基础。     

The diet of the edible cockle Anadara antiquata L. (Bivalvia,Arcidae) in Dar es Salaam,Tanzania, during the northeast monsoons

The diet of the cockle Anadara antiquata L. was studied along Dar es Salaam coast during the N-E Monsoon season. No zooplankton were found in the 80 cockle guts examined. 27 phytoplankton genera were identified, an indication of wide phytophagy. Members of classes Bacillariophyceae, Chlorophyceae, Pyrophyceae and Prasnophyceae were very prevalent in the diet. Scenedesmus, Chlamydomonas, Palmeria, Exuviella, and Coscinodiscus showed the highest recurrence frequencies per gut.     

A systems biology framework for modeling metabolic enzyme inhibition of Mycobacterium tuberculosis

Background  

Because metabolism is fundamental in sustaining microbial life, drugs that target pathogen-specific metabolic enzymes and pathways can be very effective. In particular, the metabolic challenges faced by intracellular pathogens, such as Mycobacterium tuberculosis, residing in the infected host provide novel opportunities for therapeutic intervention.     

扩增子测序法筛选结核分枝杆菌的耐药突变

结核病是严重的公共健康问题之一,而耐药结核病的增加是控制结核病流行的难点之一。快速、准确的诊断是提高结核患者治愈率和降低死亡率的关键因素。本研究建立了基于二代测序技术的扩增子测序方法,对5种一线抗结核药物的17个耐药基因进行检测。在26个临床耐药结核菌株中共鉴定出65个突变,包括33个热点突变,9个稀有突变和23个新突变。对18个新发现的错义突变进行了蛋白质序列保守性和蛋白质局部结构的分析。结果表明,14个新的错义突变在9种分枝杆菌中显示出高度保守性,并且导致了该蛋白质局部结构的改变。根据本研究检测和分析结果,推测这些新发现的突变可能是潜在的耐药突变。在本研究中,构建了扩增子测序的检测方法,可同时检测10株临床结核菌株的17个耐药基因,是一种快速、准确并且全面的检测耐药结核分枝杆菌一线治疗药物耐药突变的方法,该方法不仅能检测热点突变和稀有突变,还能发现一些未报道过的新突变。该检测方法或可用于临床诊断和基础研究。