Search

Background: Leptospirosis is among the major neglected zoonoses in developing countries. The prevalence of leptospirosis remains underestimated in many African countries because of limited diagnostic facilities. We studied Leptospira seropositivity prevalence in humans, sheep, goats and rodents in a semi-arid region of central Tanzania and compared findings with reports from humid tropical areas. The aims were to establish the disease burden in different settings; understand circulating Leptospira serovars and potential major reservoirs for establishing appropriate control measures., Methods: Humans, sheep, goats, rodents and shrews (insectivores) were sampled from Bahi district, a semi-arid area in central Tanzania. Samples were tested for leptospiral antibodies using microscopic agglutination test (MAT) consisting of Leptospira serovars mainly reported in Tanzania and reference strains. Findings were compared with previous data to determine the disease epidemiological patterns., Results and Conclusion: Semi-arid area showed high Leptospira seropositivity prevalence in humans and domestic animals due to intensive human-animal interactions at scarce water points and by flash flooding which occur in the area. Rodent population in the semi-arid areas was relatively low due to flooding. Leptospira seropositivity in rodents was also slightly lower, and the rodents appeared to be prolific breeders, probably as a means to compensate for the lost population during extreme drought as well as during short spells of floods. Intensive human-animal interaction in the semi-arid areas especially, in water sources in valleys where human and animals often meet, likely increased the risk of leptospirosis transmission to rice farmers in the area. Goats and sheep which are kept around homesteads had higher leptospiral antibodies prevalence (62%), nearly double of the 38% reported in same species in humid tropical regions of Tanzania. Livestock, especially goats and sheep, could be the major source of leptospirosis transmission to humans. Vaccination of livestock with vaccines against local Leptospira strains should be encouraged, and rodent control emphasized, as part of a management strategy against leptospirosis. Public awareness of leptospirosis must also be raised and supported by availability of rapid test kits in clinics for preliminary testing of leptospirosis in people with fevers of unknown origin.

Background: Leptospirosis is a bacterial zoonotic disease of worldwide importance, though relatively neglected in many African countries including sub Saharan Africa that is among areas with high burden of this disease. The disease is often mistaken for other febrile illnesses such as dengue, malaria, rickettsioses and enteric fever. Leptospirosis is an occupational disease likely to affect people working in environments prone to infestation with rodents which are the primary reservoir hosts of this disease. Some of the populations at risk include: sugarcane plantation workers, wetland farmers, fishermen and abattoir workers. In this study we investigated the prevalence of antibodies against Leptospira among sugarcane plantation and factory workers, fishing communities as well as among rodents and shrews in domestic and peridomestic environments within the study areas., Methods: The study was conducted in Kagera region, northwestern Tanzania and it involved sugarcane plantation workers (cutters and weeders), sugar factory workers and the fishing community at Kagera Sugar Company in Missenyi district and Musira island in Lake Victoria, Kagera, respectively. Blood was collected from consenting human adults, and from rodents and shrews (insectivores) captured live using Sherman traps. Serological detection of leptospiral antibodies in blood serum was carried out by the microscopic agglutination test (MAT)., Results: A total of 455 participants were recruited from the sugarcane plantation (n = 401) and fishing community (n = 54) while 31 rodents and shrews were captured. The overall prevalence of antibodies against Leptospira in human was 15.8%. Sugarcane cutters had higher seroprevalence than other sugar factory workers. Prevalent antibodies against Leptospira serovars in humans were against serovars Lora (6.8%), Sokoine (5.3%), Pomona (2.4%), Hebdomadis (1.1%) and Kenya (0.2%). Detected leptospiral serovars in reservoir hosts were Sokoine (12.5%) and Grippotyphosa (4.2%). Serovar Sokoine was detected both in humans and small mammals., Conclusion: Leptospirosis is a public health threat affecting populations at risk, such as sugarcane plantation workers and fishing communities. Public awareness targeting risk occupational groups is much needed for mitigation of leptospirosis in the study areas and other vulnerable populations in Tanzania and elsewhere., Competing Interests: Co-author (IA) is a chief medical officer at Kagera Sugar Company Ltd, that is one of the study location. However, this did not influence site selection. Employment to this company have no competing interest and did not influence the study design nor reporting of study findings.

The burden of leptospirosis in humans and animals in Africa is higher than that reported from other parts of the world. However, the disease is not routinely diagnosed in the continent. One of major factors limiting diagnosis is the poor availability of live isolates of locally circulating Leptospira serovars for inclusion in the antigen panel of the gold standard microscopic agglutination test (MAT) for detecting antibodies against leptospirosis. To gain insight in Leptospira serovars and their natural hosts occurring in Tanzania, concomitantly enabling the improvement of the MAT by inclusion of fresh local isolates, a total of 52 Leptospira isolates were obtained from fresh urine and kidney homogenates, collected between 1996 and 2006 from small mammals, cattle and pigs. Isolates were identified by serogrouping, cross agglutination absorption test (CAAT), and molecular typing. Common Leptospira serovars with their respective animal hosts were: Sokoine (cattle and rodents); Kenya (rodents and shrews); Mwogolo (rodents); Lora (rodents); Qunjian (rodent); serogroup Grippotyphosa (cattle); and an unknown serogroup from pigs. Inclusion of local serovars particularly serovar Sokoine in MAT revealed a 10-fold increase in leptospirosis prevalence in Tanzania from 1.9% to 16.9% in rodents and 0.26% to 10.75% in humans. This indicates that local serovars are useful for diagnosis of human and animal leptospirosis in Tanzania and other African countries.

Tuberculosis (TB) diagnosis in low-income countries is mainly done by microscopy. Hence, little is known about the diversity of Mycobacterium spp. in TB infections. Different genotypes or lineages of Mycobacterium tuberculosis vary in virulence and induce different inflammatory and immune responses. Trained Cricetomys rats show a potential for rapid diagnosis of TB. They detect over 28 % of smear-negative, culture-positive TB. However, it is unknown whether these rats can equally detect sputa from patients infected with different genotypes of M. tuberculosis. A 4-month prospective study on diversity of Mycobacterium spp. was conducted in Dar es Salaam, Tanzania. 252 sputa from 161 subjects were cultured on Lowenstein-Jensen medium and thereafter tested by rats. Mycobacterial isolates were subjected to molecular identification and multispacer sequence typing (MST) to determine species and genotypes. A total of 34 Mycobacterium spp. isolates consisting of 32 M. tuberculosis, 1 M. avium subsp. hominissuis and 1 M. intracellulare were obtained. MST analyses of 26 M. tuberculosis isolates yielded 10 distinct MST genotypes, including 3 new genotypes with two clusters of related patterns not grouped by geographic areas. Genotype MST-67, shared by one-third of M. tuberculosis isolates, was associated with the Mwananyamala clinic. This study shows that diverse M. tuberculosis genotypes (n = 10) occur in Dar es Salaam and trained rats detect 80 % of the genotypes. Sputa with two M. tuberculosis genotypes (20 %), M. avium hominissuis and M. intracellulare were not detected. Therefore, rats detect sputa with different M. tuberculosis genotypes and can be used to detect TB in resource-poor countries.

Tuberculosis (TB) diagnosis in regions with limited resources depends on microscopy with insufficient sensitivity. Rapid diagnostic tests of low cost but high sensitivity and specificity are needed for better point-of-care management of TB. Trained African giant pouched rats (Cricetomys sp.) can diagnose pulmonary TB in sputum but the relevant Mycobacterium tuberculosis (Mtb)-specific volatile compounds remain unknown. We investigated the odour volatiles of Mtb detected by rats in reference Mtb, nontuberculous mycobacteria, Nocardia sp., Streptomyces sp., Rhodococcus sp., and other respiratory tract microorganisms spiked into Mtb-negative sputum. Thirteen compounds were specific to Mtb and 13 were shared with other microorganisms. Rats discriminated a blend of Mtb-specific volatiles from individual, and blends of shared, compounds (P = 0.001). The rats' sensitivity for typical TB-positive sputa was 99.15% with 92.23% specificity and 93.14% accuracy. These findings underline the potential of trained Cricetomys rats for rapid TB diagnosis in resource-limited settings, particularly in Africa where Cricetomys rats occur widely and the burden of TB is high., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Trained African giant pouched rats (Cricetomys gambianus) have potential for diagnosis of tuberculosis (TB). These rats target volatile compounds of Mycobacterium tuberculosis (Mtb) that cause TB. Mtb and nontuberculous mycobacteria (NTM) species are related to Nocardia and Rhodococcus spp., which are also acid-fast bacilli and can be misdiagnosed as Mtb in smear microscopy. Diagnostic performance of C. gambianus on in vitro-cultured mycobacterial and related pulmonary microbes is unknown. This study reports on the response of TB detection rats to cultures of reference Mtb, clinical Mtb, NTM, Nocardia; Rhodococcus; Streptomyces; Bacillus; and yeasts. Trained rats significantly discriminated Mtb from other microbes (p < 0.008, Fisher's exact test). Detection of Mtb cultures was age-related, with exponential and early stationary phase detected more frequently than early log phase and late stationary phase (p < 0.001, Fisher's test) (sensitivity = 83.33%, specificity = 94.4%, accuracy = 94%). The detection of naturally TB-infected sputum exceeded that of negative sputum mixed with Mtb, indicating that C. gambianus are conditioned to detect odours of TB-positive sputum better than spiked sputum. Although further studies on volatiles from detectable growth phases of Mtb are vital for identification of Mtb-specific volatiles detected by rats, our study underline the potential of C. gambianus for TB diagnosis., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Trained African giant-pouched rats (Cricetomys gambianus) can detect Mycobacterium tuberculosis and show potential for the diagnosis of tuberculosis (TB). However, rats' ability to discriminate between clinical sputum containing other Mycobacterium spp. and nonmycobacterial species of the respiratory tract is unknown. It is also unknown whether nonmycobacterial species produce odor similar to M. tuberculosis and thereby cause the detection of smear-negative sputum. Sputum samples from 289 subjects were analyzed by smear microscopy, culture, and rats. Mycobacterium spp. were isolated on Lowenstein-Jensen medium, and nonmycobacterial species were isolated on four different media. The odor from nonmycobacterial species from smear- and M. tuberculosis culture-negative sputa detected by ≥2 rats ("rat positive") was analyzed by gas chromatography-mass spectrometry and compared to the M. tuberculosis odor. Rats detected 45 of 56 confirmed cases of TB, 4 of 5 suspected cases of TB, and 63 of 228 TB-negative subjects (sensitivity, 80.4%; specificity, 72.4%; accuracy, 73.9%; positive predictive value, 41.7%; negative predictive value, 93.8%). A total of 37 (78.7%) of 47 mycobacterial isolates were M. tuberculosis complex, with 75.7% from rat-positive sputa. Ten isolates were nontuberculous mycobacteria, one was M. intracellulare, one was M. avium subsp. hominissuis, and eight were unidentified. Rat-positive sputa with Moraxella catarrhalis, Streptococcus pneumoniae, Staphylococcus spp., and Enterococcus spp. were associated with TB. Rhodococcus, Nocardia, Streptomyces, Staphylococcus, and Candida spp. from rat-positive sputa did not produce M. tuberculosis-specific volatiles (methyl nicotinate, methyl para-anisate, and ortho-phenylanisole). Prevalence of Mycobacterium-related Nocardia and Rhodococcus in smear-negative sputa did not equal that of smear-negative mycobacteria (44.7%), of which 28.6% were rat positive. These findings and the absence of M. tuberculosis-specific volatiles in nonmycobacterial species indicate that rats can be trained to specifically detect M. tuberculosis.

Serological and microbiological studies on leptospirosis in pigs from Morogoro municipality, Tanzania were carried out between October 2007 and May 2008. Serum samples (n = 385) from apparently healthy pigs were tested by microscopic agglutination test for antibodies against live cultures of six known Leptospira interrogans serovars: Pomona, Icterohaemorrhagiae, Ballum, Tarassovi, Grippotyphosa and Hardjo. Significant positive titres were detected in 4.42% (17/385) of all the tested serum samples. Asceptically collected samples, urine (n = 236) and kidney tissues (n = 214), were cultured in enriched Fletcher's and Ellinghausen McCullough-Johnson and Harris media and assessed, at weekly intervals for 24 weeks, for growth by dark-field microscopy. Two leptospiral organisms were isolated from the urine samples. There was a statistical association between seroposivity and location that the subjects reside in (P < 0.05), whereas it was not significantly associated with sex nor age (P > 0.05). The evidence of pig exposure to different serovars and the isolation of the leptospiral organisms confirm that the infection is present in pigs although with an overall low prevalence. Apart from its economic importance on to the pig industry, this disease is a potential zoonotic public health risk in Tanzania, especially because of the lack of studies on leptospirosis among persons who handle pigs and pork products.

Setting: Resource-limited settings in sub-Saharan African countries., Objective: To utilise African giant pouched rats (Cricetomys gambianus) for the diagnosis of tuberculosis (TB) in human sputum., Design: A specially designed cage with 10 sniffing holes and cassette-carrier was used. The sputum samples were put in the sample cassette, containing 10 samples in line, placed under matching sniffing holes. Rats were trained to sniff each consecutive sample, and indicate TB positives by fixing their nose for 5 seconds at the sniffing hole. This behaviour was maintained by food reinforcement upon correct indications. A total of 3416 samples were used., Results: Of the 20 trained rats, 18 were able to discriminate positive from negative sputum samples, with average daily sensitivities ranging from 72% to 100%, and average daily false-positives ranging from 0.7% to 8.1%. The use of multiple rats significantly increased sensitivity and negative predictive value., Conclusion: Utilising trained sniffer rats for TB detection is a potentially faster screening method and is at least as sensitive as smear microscopy. This method could therefore be suitable for active case finding, especially where large numbers of samples are to be analysed in resource-limited settings, to complement existing diagnostic techniques.

Escherichia coli O157:H7 is an important agent of haemorrhagic colitis and haemolytic uraemic syndrome in children less than five years old and elderly people. The objective of this study was to investigate the prevalence of verotocytoxin producing E. coli 0157 (VTEC O157) among human patients with diarrhoea in Morogoro, Tanzania. Faecal samples originating from 275 human patients with diarrhoea were screened for presence of E. coli O157:H7. A total of 96 E. coli isolate were identified. Of these, 10 isolates were grouped into sorbitol non-fermenting and glucuronide negative and 49 isolates were sorbitol positive and glucuronide positive. The remaining 37 were sorbitol negative and glucuronide positive. Using the polymerase chain reaction techniques, a total often verotocytocin producing E. coli isolated in this study were used. The overall two (15%) and one (7%) of the isolated of E. coli possessed both attaching and effacing (eae A) and enterohemolysin (ehly) A genes respectively. Other enterobacterial agents including Pseudomonas spp, Proteus spp and coliforms were also isolated. The VTEC O157 isolates were 100% resistant to oxytetracycline, chloramphenicol, streptomycin, and amoxyclav. In conclusion, the isolation of diarrhoeaogenic E. coli O157:H7 in this region suggests that the pathogen is an important aetiology of acute gastroenteritis in Tanzania. There is therefore, need to improve sewage and refuse disposal system, the provision of safe potable water, sanitation, personal hygiene and health education in order to reduce infection with this and other enteric pathogens.

Background: Leptospira are the parasitic bacterial organisms associated with a broad range of mammalian hosts and are responsible for severe cases of human Leptospirosis. The epidemiology of leptospirosis is complex and dynamic. Multiple serovars have been identified, each adapted to one or more animal hosts. Adaptation is a dynamic process that changes the spatial and temporal distribution of serovars and clinical manifestations in different hosts. Serotyping based on repertoire of surface antigens is an ambiguous and artificial system of classification of leptospiral agents. Molecular typing methods for the identification of pathogenic leptospires up to individual genome species level have been highly sought after since the decipherment of whole genome sequences. Only a few resources exist for microbial genotypic data based on individual techniques such as Multiple Locus Sequence Typing (MLST), but unfortunately no such databases are existent for leptospires., Results: We for the first time report development of a robust MLST method for genotyping of Leptospira. Genotyping based on DNA sequence identity of 4 housekeeping genes and 2 candidate genes was analyzed in a set of 120 strains including 41 reference strains representing different geographical areas and from different sources. Of the six selected genes, adk, icdA and secY were significantly more variable whereas the LipL32 and LipL41 coding genes and the rrs2 gene were moderately variable. The phylogenetic tree clustered the isolates according to the genome-based species., Conclusion: The main advantages of MLST over other typing methods for leptospires include reproducibility, robustness, consistency and portability. The genetic relatedness of the leptospires can be better studied by the MLST approach and can be used for molecular epidemiological and evolutionary studies and population genetics.

Human plague in the Western Usambara Mountains in Tanzania has been a public health problem since the first outbreak in 1980. The wildlife reservoir is unknown and eradication measures that have proved effective elsewhere in Tanzania appear to fail in this region. We use census data from 2002 and hospital records kept since 1986 to describe the temporal, spatial and demographic variation in human plague. A seasonal peak in cases occurs from December to February with the numbers of cases during this peak varying between 0 and 1150. Variation in incidence, calculated for each village as the mean number of cases per thousand inhabitants per year, indicates that human plague is concentrated around a group of three neighbouring, relatively isolated, high-altitude villages; Nywelo, Madala and Gologolo. However, there was no evidence that these villages were acting as a source of infection for the remainder of the focus. The likelihood of becoming infected with plague is highest between the ages of 5 and 19 and lowest for adult men. This was most clear in the ward encompassing the three high-incidence villages where the risk of plague among children aged 10-14 was 2.2 times higher than for adults aged 30-34, and among adults aged 30-34, the risk was 2.4 times higher for women than men.

The prevalence of leptospirosis is generally high in domestic animals and rodents in Tanzania. Identification of Leptospira isolates from cattle was carried out to establish prevalent Leptospira serovars. Serological typing was done based on monoclonal antibodies and the standard cross-agglutination absorption test. Molecular typing involved pathogenic- and saprophytic-specific PCRs and a PCR specifically amplifying DNA from the species Leptospira kirschneri. DNA fingerprinting with primers derived from sequences of insertion elements IS1500 and IS1533 was carried out. Both serological and molecular characterization indicated that one of the Leptospira isolates, coded RM1, represents a new serovar of the species L. kirschneri of serogroup Icterohaemorrhagiae. The serovar name Sokoine is proposed for this new Leptospira isolate.

Two identical leptospiral isolates coded Sh9 and Sh25 obtained from the urine of captive African giant pouched rats (Cricetomys gambianus), destined for use as biodetector of antipersonnel landmines were typed as serovar Kenya using cross-agglutination absorption test and DNA fingerprinting with the insertion element sequences IS1533 and IS1500 derived primers. The two isolates were previously characterized using cultural and serological-microagglutination test as pathogenic leptospires of the serogroup Ballum, closely related to serovars Kenya and Peru. To our knowledge, this is the first reported in-depth characterization of leptospira isolates from Tanzania.

A localised "scalded skin syndrome" occurred in two male siblings aged five and ten years old, in Morogoro, Tanzania. This condition recurred in the same children within a period of about 12 months. A haemolytic ampicillin resistant Staphylococcus aureus was isolated from the bullous material, suggesting that the condition was staphylococcal scalded skin syndrome (SSSS). This implies that the bacterial isolate was an exfoliative toxin (ET) producing S. aureus. This infection was, however, striking because SSSS occurs mostly in newborns, and it is rarely recurrent. It is possible that a new strain with particular adaptability, or an inherent susceptibility to the S. aureus of the affected children was the cause of this recurrency.

Cholesterol oxidase (CO) and choline phosphohydrolase (CPH) exoenzymes were isolated from culture supernatants of Rhodococcus equi ATCC 33701 and their hemolytic and cytotoxic activities examined. The purifications involved differential ammonium sulphate precipitation, ion exchange and gel filtration chromatography. A purification of 32.8-fold and a yield of 0.3% of CO were determined by synergistic hemolysis of sheep red blood cells (SRBC) presensitized with Staphylococcus aureus beta toxin. The enzymatic activity of CO was also demonstrated by oxidation of aqueous cholesterol suspensions. The activity of CO was reversibly inhibited by concentration. A purification of 412.4-fold and a yield of 1.7% of CPH were determined by hydrolysis of p-nitrophenyphosphorylcholine. Purity of both exoenzymes was confirmed by immunoblotting. On sodium dodecyl sulphate polyacrylamide gel electrophoresis, the CO had a molecular mass (Mr) of 60 kd and the CPH a Mr of 65 kd. Choline phosphohydrolase did not hydrolyse sphingomyelin. Sphingomyelinase C (SMC) activity was however demonstrated in concentrated culture supernatants. This dissociation of SMC from CPH activity indicates that R. equi produces two distinct phospholipase C exoenzymes, a CPH and a SMC. Both CO and CPH combined, or individually, did not lyse native SRBC even with subsequent chilling of the cells at 4 degrees C ("hot-cold" treatment). Purified CO lysed beta toxin-sensitized SRBC. The CPH showed only minor hemolytic activity against such sensitized SRBC even at high concentrations. Combination of CO and CPH in lysis of beta toxin sensitized SRBC showed only minor additive rather than synergistic effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Rhodococcus equi produces two exoenzymes (REE), a cholesterol oxidase in large amounts and a phospholipase C, which cause lysis of sheep red blood cells (SRBC) sensitized with Staphylococcus aureus beta toxin. Two immunization studies were done in foals to determine the role of antibody to REE in protection against R. equi pneumonia. In the first study, three foals (mean age 10 days) were vaccinated four times at 2-week intervals with over 1 million units of partially purified exoenzymes (PREE). In the second study, three foals (mean age 19 days) were administered plasma from an adult horse vaccinated with PREE. Relatively low titres (16-32) of neutralizing antibody were detected in the foals of the former group, and passive transfer of neutralizing antibody (titres 32-64) occurred in the latter. Following immunization, principal foals and an equal number of similarly aged nonimmunized foals were challenged by aerosol with 1 x 10(10) live R. equi per day for 5 consecutive days. No severe clinical pneumonia developed in either group and, with one exception, only minor and resolving lung abscesses developed in these foals. These studies showed that antibody response of foals to immunization with PREE was poor, antibody to PREE did not prevent foals from developing lung abscesses following experimental infection, and that foals even as young as 3 weeks of age may be largely refractory to aerosol challenge with virulent R. equi.