Leptospira Seropositivity in Humans, Livestock and Wild Animals in a Semi-Arid Area of Tanzania.

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.

Selected Endemic Zoonoses in Pigs Presenting for Slaughter in Kampala, Uganda.

Leptospirosis, brucellosis, and Q fever (coxiellosis) are bacterial zoonoses that cause acute febrile illness in people as well as reproductive losses in pigs. Pig keeping is an increasingly important livelihood to millions of smallholder farmers in Uganda because of exponential increases in demand for pork. The prevalence of leptospirosis and Q fever in pigs is unknown, and the few studies of porcine brucellosis have estimated a range of seroprevalence. Therefore, we undertook a prevalence survey of leptospirosis, brucellosis, and Q fever in pigs using quantitative real-time PCR to determine the potential importance of these zoonoses to the growing pig sector in Uganda. Six hundred forty-nine pigs were sampled in 2015-2016 at an urban pork slaughterhouse. Ten percent of pigs (n = 68) had leptospiral DNA in either their kidney or reproductive tissue. In adjusted analyses, variables predictive of leptospiral status included female sex (odds ratio [OR]: 2.37, P < 0.01) and pigs sampled in March 2016 (OR: 2.23, P = 0.02) and October 2016 (OR: 0.30, P = 0.04). DNA fingerprinting revealed circulation of at least four distinct serovars in these pigs. Brucella spp. and Coxiella burnetii DNA were not detected in any sampled pig. This is the first report of widespread circulation of pathogenic Leptospira spp. in pigs in Uganda, suggesting that leptospirosis likely has a greater impact on the health of pigs than was previously recognized. Pig farmers, pig traders, and slaughterhouse workers may be at greatest occupational risk because of their direct contact with infective leptospires in aborted fetuses, bodily fluids, and other tissues.

Seroprevalence and associated risk factors of leptospirosis in slaughter pigs; a neglected public health risk, western Kenya.

BACKGROUND: Leptospirosis is a neglected zoonosis of public health importance transmitted through contact with contaminated soil, water or urine of infected animals. In pigs the disease is characterized by abortion, still births and weak piglets. A cross-sectional study was conducted in May to July 2018 to estimate the sero-prevalence of leptospirosis and factors associated with seropositivity in slaughter pigs. A questionnaire was used to collect information on animal demographics. Serum was tested for anti-leptospiral antibodies using microscopic agglutination test (MAT) with a panel of 8 serovars. Sera were considered positive for sero-reactivity at a MAT titre ≥1:40 against at least one serovar. Chi-square tests were used to measure the strength of association between the MAT test result and exploratory variables. RESULTS: A total of 252 pig serum samples from seven slaughterhouses were tested for Leptospira antibodies by MAT. Of the 252 pigs sampled, 88.8% (244/252) were indigenous breeds; 55.6% (140/252) were female and 88.7% (220/252) were reared in extensive production systems. Eighty-three (32.9%; 83/252) sera samples tested positive on MAT against at least one serovar. Of the 8 serovars, the highest prevalence was recorded for serovar Lora 21.4% followed by Kenya 5.2%, Sokoine 3.6% and Grippotyphosa at 3.2%. Risk factors for leptospirosis seropositivity in pigs were: originating from farms with other types of livestock (OR 2.3; 95% CI 1.0-4.5) and mature pigs (OR 1.9; 95% CI 1.1-3.3). CONCLUSION: This study demonstrates that there is a high prevalence of leptospirosis positive pigs at slaughter in a small-holder livestock keeping region of the Lake Victoria basin. The potential for cross species transmission of pathogenic serovars is highlighted as well as the potential for occupational exposure to slaughterhouse personnel. Improvements in husbandry practices (confinement and rodent control) and public health education among slaughterhouse workers and other high-risk groups is recommended.

Leptospirosis in sugarcane plantation and fishing communities in Kagera northwestern Tanzania.

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.

Seropositivity of Brucella spp. and Leptospira spp. antibodies among abattoir workers and meat vendors in the city of Mwanza, Tanzania: A call for one health approach control strategies.

INTRODUCTION: Brucellosis and leptospirosis are among neglected tropical zoonotic diseases particularly in the resource limited countries. Despite being endemic in these countries, there is paucity of information on its magnitude. This study investigated seropositivity of Brucella spp. and Leptospira spp., and associated factors among abattoir workers and meat vendors in the city of Mwanza, Tanzania. METHODOLOGY: A community based cross-sectional study was conducted in Mwanza city from May to July 2017. Socio-demographic and other relevant information were collected. Detection of Brucella spp. and Leptospira spp. antibodies were done using slide agglutination test and microscopic agglutination test, respectively. Data were analyzed using STATA version 13 Software. FINDINGS: A total of 250 participants (146 abattoir workers and 104 meat vendors) were enrolled with median age of 31 (IQR: 25-38) years. The overall, seropositivity of Brucella spp. antibodies was 48.4% (95% Cl: 42-54). Seropositivity of B. abortus was significantly higher than that of B. melitensis (46.0%, 95%Cl: 39-52 vs. 23.6%, 95% Cl: 18-28, P<0.001) while seropositivity of both species was 21.2% (95%Cl: 16-26). The seropositivity of Leptospira spp. was 10.0% (95% CI: 6-13) with predominance of Leptospira kirschneri serovar Sokoine which was detected in 7.2% of the participants. Being abattoir worker (OR: 2.19, 95% CI 1.06-4.54, p = 0.035) and long work duration (OR: 1.06, 95%CI: 1.01-1.11, p = 0.014) predicted presence of both B.abortus and B. melitensis antibodies. Only being married (p = 0.041) was significantly associated with seropositivity of Leptospira spp. Primary education was the only factor independently predicted presence of Brucella spp. antibodies among abattoir workers on sub-analysis of occupational exposure. None of factors were found to be associated with presence of Brucella spp. antibodies among meat vendors on sub-analysis. CONCLUSION: Seropositivity of B.abortus antibodies among abattoir workers and meat vendors is high and seem to be a function of being abattoir worker, having worked for long duration in the abattoir and having primary education. In addition, a significant proportion of abattoir workers and meat vendors in the city was seropositive for Leptospira kirschneri serovar Sokoine. There is a need to consider 'one health approach' in devising appropriate strategies to control these diseases in the developing countries.

Pediatric tuberculosis detection using trained African giant pouched rats.

BACKGROUND: Tuberculosis (TB) diagnosis in children is a challenge with up to 94% of children with TB treated empirically in TB high-burden countries. Therefore, new diagnostic tests are needed for TB diagnosis. We determined the performance of trained rats in the diagnosis of pediatric TB and whether they can improve detection rate compared to the standard of care. METHODS: Presumptive TB patients in 24 TB clinics in Tanzania were tested. Samples indicated as TB-positive by rats underwent confirmation by concentrated smear microscopy. TB yield of bacteriologically confirmed pediatric TB patients (≤5 years) was compared with yield of standard of care. RESULTS: Sputum samples from 55,148 presumptive TB patients were tested. Nine hundred eighty-two (1.8%) were the children between 1 and 5 years. Clinics detected 34 bacteriologically positive children, whereas rats detected additional 23 children yielding 57 bacteriologically TB-positive children. Rats increased pediatric TB detection by 67.6%. Among 1-14-year-old children, clinics detected 331 bacteriologically positive TB whereas rats found the additional 208 children with TB that were missed by clinics. Relative increase in TB case detection by rats decreased with the increase in age (P<0.0001). CONCLUSION: Trained rats increase pediatric TB detection significantly and could help address the pediatric TB diagnosis challenges. Further determination of accuracy of rats involving other sample types is still needed.

Relic populations of Fukomys mole-rats in Tanzania: description of two new species F. livingstoni sp. nov. and F. hanangensis sp. nov.

Previous studies of African mole-rats of the genera Heliophobius and Fukomys (Bathyergidae) in the regions of East and south central Africa have revealed a diversity of species and vicariant populations, with patterns of distribution having been influenced by the geological process of rifting and changing patterns of drainage of major river systems. This has resulted in most of the extant members of the genus Fukomys being distributed west of the main Rift Valley. However, a small number of isolated populations are known to occur east of the African Rift Valley in Tanzania, where Heliophobius is the most common bathyergid rodent. We conducted morphological, craniometric and phylogenetic analysis of mitochondrial cytochrome b (cyt b) sequences of two allopatric populations of Tanzanian mole-rats (genus Fukomys) at Ujiji and around Mount Hanang, in comparison with both geographically adjacent and more distant populations of Fukomys. Our results reveal two distinct evolutionary lineages, forming clades that constitute previously unnamed species. Here, we formally describe and designate these new species F. livingstoni and F. hanangensis respectively. Molecular clock-based estimates of divergence times, together with maximum likelihood inference of biogeographic range evolution, offers strong support for the hypothesis that vicariance in the Western Rift Valley and the drainage patterns of major river systems has subdivided populations of mole-rats. More recent climatic changes and tectonic activity in the "Mbeya triple junction" and Rungwe volcanic province between Lakes Rukwa and Nyasa have played a role in further isolation of these extra-limital populations of Fukomys in Tanzania.

Leptospira Serovars for Diagnosis of Leptospirosis in Humans and Animals in Africa: Common Leptospira Isolates and Reservoir Hosts.

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.

Mycobacterium genotypes in pulmonary tuberculosis infections and their detection by trained African giant pouched rats.

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.

Seroprevalence of Leptospira infection in bats roosting in human settlements in Morogoro municipality in Tanzania.

Leptospirosis is a neglected emerging infectious disease with high burden in Africa. Major reservoirs of leptospirosis are rodents and other small mammals. Studies of leptospirosis in bats and the extent to which they contribute to its transmission to humans and domestic animals in Tanzania are lacking. The objective of this study was to determine the prevalence of leptospirosis and local Leptospira serovars in bat colonies found around human settlements in Morogoro, Tanzania. In this study, mist nets were used to capture bats at Sokoine University of Agriculture main campus, whereas bats roosting on trees at Nunge A, Nunge B and M were primary school were captured manually. Leptospirosis was determined using the gold standard microscopic agglutination test for determination of Leptospira antibodies. Six live Leptospira serovars including local Leptospira serovar Sokoine, Kenya and Lora reported in rodents and domestic animals in Tanzania, and reference serovars Hebdomadis, Canicola and Pomona were used in the study. Leptospirosis prevalence was high in bats (19.4%) and higher within colonies (27.3%). Leptospira serovar Sokoine was more prevalent (19.4%) compared to serovars Kenya (2.8%) and Lora (2.8%). Serovars Pomona, Canicola and Hebdomadis were not detected in bats. In conclusion, bats are potential reservoir and transmitter of Leptospira serovar Sokoine, hence should be prevented to access houses and roosting in human settlements to avoid public health risks. Routine diagnosis of human leptospirosis is needed in Tanzania as evidence show a high prevalence of Leptospira in reservoir hosts interacting with humans.

Prevalence of leptospirosis and toxoplasmosis: a study of rodents and shrews in cultivated and fallow land, Morogoro rural district, Tanzania.

Leptospirosis and toxoplasmosis are among understudied zoonotic diseases that are also not diagnosed routinely in Tanzania. Humans get leptospirosis and toxoplasmosis through contact with an environment contaminated with Leptospira bacteria and Toxoplasma protozoa from reservoir hosts, which are rodents and cats, respectively. The objective of this study was to determine the prevalence of Leptospira and Toxoplasma infections in rodents and shrews in Mikese area of Morogoro Rural District in eastern Tanzania. A total of 89 rodents and one shrew from cultivated and fallow land were tested for leptospirosis using six Leptospira serovars: Sokoine, Kenya, Canicola, Lora, Hebdomadis and Pomona. Toxoplasmosis was determined in 46 rodents brain smears. The prevalence of leptospirosis was 25.8%, and Leptospira serovar Sokoine was the most prevalent serovar (16.9%). Toxoplasma was detected in one rodent (2.17%) individual while three rodent individuals had Toxoplasma-like parasites hence were considered suspect positive. Findings suggest potential existence of human leptospirosis which needs to be further investigated. Public awareness of leptospirosis and toxoplasmosis should be promoted and their diagnosis considered in patients in health care facilities.

Seroprevalence of Leptospira Check for this Species in other Resources Infection in Bats Roosting in Human Settlements in Morogoro Municipality in Tanzania

Leptospirosis is a neglected emerging infectious disease with high burden in Africa. Major reservoirs of leptospirosis are rodents and other small mammals. Studies of leptospirosis in bats and the extent to which they contribute to its transmission to humans and domestic animals in Tanzania are lacking. The objective of this study was to determine the prevalence of leptospirosis and local Leptospira check for this species in other resources serovars in bat colonies found around human settlements in ; Tanzania. In this study; mist nets were used to capture bats at Sokoine University of Agriculture main campus; whereas bats roosting on trees at Nunge A; Nunge B and Mwere primary school were captured manually. Leptospirosis was determined using the gold standard microscopic agglutination test for determination of Leptospira antibodies. Six live Leptospira serovars including local Leptospira serovar Sokoine; Kenya and Lora reported in rodents and domestic animals in Tanzania; and reference serovars Hebdomadis; Canicola and Pomona were used in the study. Leptospirosis prevalence was high in bats (19.4) and higher within colonies (27.3). Leptospira serovar Sokoine was more prevalent (19.4) compared to serovars Kenya (2.8) and Lora (2.8). Serovars Pomona; Canicola and Hebdomadis were not detected in bats. In conclusion; bats are potential reservoir and transmitter of Leptospira serovar Sokoine; hence should be prevented to access houses and roosting in human settlements to avoid public health risks. Routine diagnosis of human leptospirosis is needed in Tanzania as evidence show a high prevalence of Leptospira in reservoir hosts interacting with humans

Mycobacterium tuberculosis volatiles for diagnosis of tuberculosis by Cricetomys rats.

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.

The volatiles of pathogenic and nonpathogenic mycobacteria and related bacteria.

Volatiles released by pathogenic and nonpathogenic mycobacteria, as well as by mycobacteria-related Nocardia spp., were analyzed. Bacteria were cultivated on solid and in liquid media, and headspace samples were collected at various times during the bacterial lifecycle to elucidate the conditions giving optimal volatile emission. Emitted volatiles were collected by using closed-loop stripping analysis (CLSA) and were analyzed by gas-chromatography-mass-spectrometry. A wide range of compounds was produced, although the absolute amount was small. Nevertheless, characteristic bouquets of compounds could be identified. Predominantly aromatic compounds and fatty-acid derivatives were released by pathogenic/nonpathogenic mycobacteria, while the two Nocardia spp. (N. asteroides and N. africana) emitted the sesquiterpene aciphyllene. Pathogenic Mycobacterium tuberculosis strains grown on agar plates produced a distinct bouquet with different volatiles, while liquid cultures produce less compounds but sometimes an earlier onset of volatile production because of their steeper growth curves under this conditions. This behavior differentiates M. tuberculosis from other mycobacteria, which generally produced fewer compounds in seemingly lower amounts. Knowledge of the production of volatiles by M. tuberculosis can facilitate the rational design of alternative and faster diagnostic measures for tuberculosis.

Ability of Cricetomys rats to detect Mycobacterium tuberculosis and discriminate it from other microorganisms.

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.

Diagnosis of tuberculosis by trained African giant pouched rats and confounding impact of pathogens and microflora of the respiratory tract.

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.

First detection of mycobacteria in African rodents and insectivores, using stratified pool screening.

With the rising number of patients with human immunodeficiency virus (HIV)/AIDS in developing countries, the control of mycobacteria is of growing importance. Previous studies have shown that rodents and insectivores are carriers of mycobacteria. However, it is not clear how widespread mycobacteria are in these animals and what their role is in spreading them. Therefore, the prevalence of mycobacteria in rodents and insectivores was studied in and around Morogoro, Tanzania. Live rodents were trapped, with three types of live traps, in three habitats. Pieces of organs were pooled per habitat, species, and organ type (stratified pooling); these sample pools were examined for the presence of mycobacteria by PCR, microscopy, and culture methods. The mycobacterial isolates were identified using phenotypic techniques and sequencing. In total, 708 small mammals were collected, 31 of which were shrews. By pool prevalence estimation, 2.65% of the animals were carriers of mycobacteria, with a higher prevalence in the urban areas and in Cricetomys gambianus and the insectivore Crocidura hirta. Nontuberculous mycobacteria (Mycobacterium chimaera, M. intracellulare, M. arupense, M. parascrofulaceum, and Mycobacterium spp.) were isolated from C. gambianus, Mastomys natalensis, and C. hirta. This study is the first to report findings of mycobacteria in African rodents and insectivores and the first in mycobacterial ecology to estimate the prevalence of mycobacteria after stratified pool screening. The fact that small mammals in urban areas carry more mycobacteria than those in the fields and that potentially pathogenic mycobacteria were isolated identifies a risk for other animals and humans, especially HIV/AIDS patients, that have a weakened immune system.