First Report of Fusarium equiseti Causing Damping-Off Disease on Aleppo Pine in Algeria.

The Aleppo pine (Pinus halepensis Mill.) is a conifer native to the Mediterranean region. In 2008 and 2009, a survey of Aleppo pine seedling diseases was performed in three forest nurseries from Relizane, Sidi Bel Abbes, and Tlemcen provinces in northwestern Algeria. Aleppo pine seedlings showed symptoms of pre- and post-emergence damping-off disease, with an incidence of 64 to 77%. Four composite samples were taken from each location. Disinfested root and root collar segments, approximately 5 mm in length, were cultured on potato dextrose agar (PDA) and incubated at 25°C, and hyphal tips were transferred to PDA. Fusarium equiseti (Corda) Sacc. (teleomorph: Gibberella intricans Wollenw.) was identified from roots of two seedlings from the Sidi Bel Abbes nursery. Morphological identification was done according to Fusarium keys (2). PDA colonies with abundant, loosely floccose, whitish aerial mycelium and beige pigmentation were observed. Macroconidia with usually 5 to 6 septa, 31 to 45 µm long. A pronounced dorsiventral curvature, tapered and elongated apical cell, and prominent foot shape were observed. Microconidia were absent. Chlamydospores were produced in hyphae, most often intercalary, solitary, in pairs, frequently forming chains or clusters, globose (7 to 13 µm). To confirm the identity of this fungus, the internal transcribed spacer of F3RS1 and F19RS1 isolates of F. equiseti were amplified and sequenced using ITS1 and ITS4 primers (4), GenBank accession nos. JX114784 and JX114791, respectively. Those sequences bore 100% (HQ671182) similarity with sequences of F. equiseti in GenBank. Pathogenicity tests were performed to fulfill Koch's postulates. Inoculum was produced by adding a 5-mm-diameter plug from a 7-day-old CMA petri dish culture to a previously sterilized 500 ml flask (237.5 g sand, 12.5 g cornmeal, 80 ml sterile distilled water), shaken over 9 days at 25°C, and mixed with sterile sandy clay soil at 1:3 (v:v). Infested soil was then transferred to 500 ml pots, and 10 Aleppo pine seeds were planted per pot. A completely randomized design was used with three replicates per isolate and three control pots with a similar non-infested soil. After 1 month at 25°C the two tested isolates caused typical damping-off symptoms (collar rot) on seedlings and were re-isolated from recently infected tissues. The percentages of the inoculated plants that became infected were 59 to 65% among isolates (0% in control pots). To our knowledge, infection by F. equiseti is a first report on Aleppo pine in northwestern Algeria, Northern Africa, and globally, and on conifers in the Mediterranean region (1,3). In Algeria, F. equiseti is associated with black pepper (Piper nigrum L.) (3). These findings highlight the moderate impact of F. equiseti on the production of Aleppo seedling stock for reforestation activities in northwestern Algeria. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory. ARS, USDA, Beltsville, MD. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , February 20, 2013. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) D. W. Minter. Cybertruffle's Robigalia, Observations of Fungi and their Associated Organisms. Retrieved from http://www.cybertruffle.org.uk/robigalia/eng/ , February 20, 2013. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

First Report of Globisporangium ultimum Causing Pythium Damping-Off on Aleppo Pine in Algeria, Africa, and the Mediterranean Region.

Globisporangium ultimum (Trow) Uzuhashi, Tojo & Kakish. (syn. Pythium ultimum Trow, syn. P. ultimum Trow var. ultimum) is a known oomycetal species from Pythium s.l. causing damping-off and/or root rot on a great variety of plants throughout the world, including some pine species (Pinus L.) and conifers (2,3). Aleppo pine (Pinus halepensis Mill.) is a common native forest tree in the Mediterranean region. Pre- and post-emergence damping-off disease symptoms were observed during 2008 and 2009 in four forest nurseries from northwestern Algeria (Relizane, Sidi Belabes, and Tlemcen departments). This disease occurred under cool conditions, and Aleppo pines were significantly affected, reducing seedling emergence. Disinfected segments, about 5 mm in length, from decayed root and collar, were cultured on CMA at 25°C. This oomycetal species was identified based on the species description in Pythium keys (3,4). For the molecular identification, PCR was used to amplify the ITS region of Pythium isolates. It was amplified with the flanking primers ITS1 and ITS4, and these products were directly sequenced. Sequence data were compared to known sequences deposited in the NCBI non redundant database to confirm morphological identification. A BLAST search identified U3CR, U7CR, U1RT, U2CR, U4CR, U14CR, U7RT, and U17RT isolates (GenBank Accession Nos. JX191921, 22, 27, 29, 31, and 33 to 35, respectively) as G. ultimum based on 100% similarity with corresponding sequence of the reference isolate no. UZ056 MAFF240024 (AB468781) (3). Phytopathogenicity testing was conducted in a petri dish and pot experiment. In the petri dish experiment, a 3 mm diameter plug was transferred from a 7-day-old CMA colony to the center of a CMA petri dish, with three replicates per isolate, and three control plates were inoculated with sterile agar plugs. After 72 h, 10 Aleppo pine seeds were placed equally spaced to 1 cm from the edge of each plug. After 7 days at 22°C in the dark, germination inhibition (46.1 to 87.6%) and root growth inhibition (62.3 to 92.2%) were calculated. In the control plates, germination failure (13.4%) and root length (27.7 cm) were observed. For the pot experiment, inocula were produced by adding a 5 mm diameter plug from a 7-day-old CMA culture to a previously sterilized 500 ml flask containing 237.5 g sand, 12.5 g cornmeal, and 80 ml SDW. Nine-day-old inoculum was mixed with sterile soil at a rate of 1:3 (v:v). Inoculum was transferred to 500 ml pot, and 10 Aleppo pine seeds were planted, with three replicates per isolate, and three control pots were used. After 2 weeks, all of the isolates tested caused typical symptoms of Aleppo pine Pythium damping-off, the percentage of inoculated plants that became infected was 36.6 to 83.3%. In the control pots, no infected plants were observed. To our knowledge, this is the first report of G. ultimum causing damping-off on Aleppo pine in Algeria, Africa, and the Mediterranean Region. Before, Aleppo pine damping-off caused by G. ultimum was reported in Australia (1). References: (1) R. P. Cook and A. J. Dubé. Host-pathogen index of plant diseases in South Australia. SADA, Melbourne, Australia, 1989. (2) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory. ARS, USDA, Bestville, MD. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , June 24, 2012. (3) S. Uzuhashi et al. Mycoscience 51:337, 2010. (4) A. J. van der Plaats-Niterink. Stud. Mycol. 21:1, 1981.

First Report of Fusarium redolens as a Causal Agent of Aleppo Pine Damping-Off in Algeria.

Aleppo pine (Pinus halepensis Mill.) is a common native coniferous tree in the natural forests of the Mediterranean region. In 2008 and 2009, a survey of Aleppo pine seedling diseases was performed in three forest nurseries from Relizane, Sidi Bel Abbes, and Tlemcen departments in northwestern Algeria. Aleppo pine seedlings showed symptoms of pre- and post-emergence damping-off, resulting in severe crop losses. The problem was widespread with a disease incidence of 64 to 77%. Fusarium redolens Wollenw. was isolated from Relizane and Sidi Bel Abbes forest nurseries. Disinfested root and root collar segments, approximately 5 mm in length, were cultured on potato dextrose agar (PDA) and incubated at 25°C. Morphological identification was done according to Fusarium keys (2). PDA colonies consisted of flat mycelium with sparse white aerial hyphae. Macroconidia with three to five septa, 24 to 43.8 µm long, widest upper third, hooked apical cell, and foot shaped basal cell were observed. Microconidia with zero to one septa, 6.8 to 10.4 µm long, oval to cylindrical, and produced on monophialides were also observed. Chlamydospores were produced abundantly in terminal and intercalary chains, in 3- to 4-week-old cultures. To confirm the identity of the fungus, the internal transcribed spacer (ITS) of F5RS3, F91SR, F55RS1, F8RS3, and F09SS1 isolates of F. redolens were amplified and sequenced using ITS 1 and ITS 4 primers (3). GenBank Accession Nos. are JX051323 to 26, and JX114783, respectively. Those sequences bore 99% (JF311916) and 100% (U34565) similarity with sequences of F. redolens in GenBank. A Fusarium pathogenicity assay was used to complete Koch's postulates. Inoculum was produced by adding a 5 mm diam. agar disc from a 7-day-old CMA petri dish culture to a previously sterilized 500 ml flask (237.5 g of sand, 12.5 g of cornmeal, and 80 ml of deionized H2O). Isolates were allowed to colonize the medium for 9 days, and flasks were shaken every day. The inoculum was mixed with sterile soil at a rate of 3:1 (v:v). Infested soil was then transferred to 500 ml pots, and 10 Aleppo pine seeds were planted. A completely randomized design was used with three replicates. After 1 month, all tested isolates caused typical damping-off symptoms on seedlings. The percentage of the inoculated plants that became infected was 53 to 91%. To our knowledge, this is the first report of F. redolens being pathogenic on Aleppo pine in northwestern Algeria and throughout the world. In Algeria, F. redolens has been reported on tomato (Solanum lycopersicum L.) (1), suggesting that it is adapted to the conditions of this area and could become a major threat to regional plant production. The annual economic impact of this disease was estimated at approximately US$50,000 per forest nursery. References: (1) N. Hamini-Kadar et al. New Dis. Rep. 22:3, 2010. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, Iowa, 2006. (3) T. J. White et al. Pages 315-322 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

First Report of Fusarium chlamydosporum Causing Damping-Off Disease on Aleppo Pine in Algeria.

The Aleppo pine (Pinus halepensis Mill.) is a conifer native to the Mediterranean region. In 2008 and 2009, a survey of Aleppo pine seedling diseases was performed in three forest nurseries from the Relizane, Sidi Bel Abbes, and Tlemcen departments in northwestern Algeria. One- to two-month-old Aleppo pine seedlings showed symptoms of damping-off in pre- and post-emergence (typical seedling collar rot). The problem was widespread with a disease incidence of 64 to 77% and an annual impact of US$50,000. Disinfested root and root collar segments (from four composite samples per location), approximately 5 mm in length, were cultured on PDA and incubated at 25°C and day/night light. Two (from 21) isolates were identified morphologically (2) as the anamorph Fusarium chlamydosporum Wollenw. & Reinking and isolated from collar rots of Relizane forest nursery seedlings. Colony development on PDA media was fast; 32 mm diameter colonies developed after 3 days. Colonies were white. Mycelia were floccose, fairly dense, off-white, and turned a lilac color in older portions of the colony. Macroconidia were thick-walled and moderately curved with unequal dorsiventral curvature (the lower wall is almost straight), short, curved and pointed apical cell, usually notched, but occasionally foot shaped basal cell, 3- to 5-septate, and 2 × 8 to 21 µm. Microconidia were abundant, 0-septate, and 2 × 6 to 9 µm. Chlamydospores were abundant, formed rapidly in single chains or clusters, and 8 to 15 µm diameter. To confirm the identity of this fungus, the internal transcribed spacer of F12RR and F4SR isolates of F. chlamydosporum were amplified and sequenced using ITS1 and ITS4 primers (4). Sequences were deposited in GenBank under accessions JX114795 and JX114789, respectively. Those sequences bore 99% similarity with reference sequence AY213655 (2) and 100% with HQ671187, also found 99 to 100% similarity with F. equiseti (Corda) Sacc. but with different conidia. Pathogenicity tests were performed to fulfill Koch's postulates. Inoculum was produced by adding a 5 mm diam. plug from a 7-day-old CMA petri dish culture to a previously sterilized 500 ml flask (237.5 g sand, 12.5 g cornmeal, 80 ml SDW), shaken over 9 days, and mixed with sterile soil at 1:3 (v:v). Infested soil was then transferred to 500 ml pots, and 10 seeds were planted. A completely randomized design was used with three replicates per isolate and three control pots. After 1 month, two tested isolates caused typical damping-off symptoms on seedlings. The percentage of the plants that became infected was 65 to 77%. To our knowledge (1,3), this is the first report of F. chlamydosporum on Aleppo pine in northwestern Algeria. It is also the first report of this fungal species affecting the Aleppo pine throughout the world, and on conifers in Africa and the Mediterranean region (1,3). References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases, Syst. Mycol. Microbiol. Lab. ARS, USDA, Beltsville, MD. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , February 20, 2013. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) D. W. Minter. Cybertruffle's Robigalia, Observations of Fungi and their Associated Organisms. Retrieved from http://www.cybertruffle.org.uk/robigalia/eng/ , February 20, 2013. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

First Report of Fusarium acuminatum Causing Damping-Off Disease on Aleppo Pine in Algeria.

The Aleppo pine (Pinus halepensis Mill.) is a conifer native to the Mediterranean region. In autumn and spring of 2008 to 2009, a survey of Aleppo pine seedling diseases was carried out in three forest nurseries from the Relizane, Sidi Bel Abbes, and Tlemcen departments in northwestern Algeria. Aleppo pine seedlings were potted from the soil. In all three nurseries, 1- to 2-month old seedlings showed symptoms of damping-off disease in pre- and post-emergence (collar rot) with a disease incidence of 64, 77, and 72%, respectively. Disinfected collar segments, about 5 mm in length, were plated on PDA and petri dishes incubated at 25°C. A Fusarium sp. was consistently isolated from tissues and all isolates were morphologically identified as Fusarium acuminatum Ellis & Everh. (teleomorph: Gibberella acuminata Wollenw.) according to Fusarium keys (2). Colony growth was 43 mm after 3 days on PDA; the aerial mycelium was white, developing a brownish tinge in the center on PDA; macroconidia were formed in orange sporodochia, broadly falcate, strongly septate, 3 to 5 septa, the apical cell with an incurved elongation, distinct foot shape, 3 to 4 × 20 to 50 µm; microconidia were usually absent for isolates other than F12SS1, reniform, septate, 5 to 6 × 6 to 10 µm, in monophialides; chlamydospores were formed in chains, 6 to 13 µm. For the molecular identification, ITS regions of Fusarium isolates were amplified with the primers ITS1 and ITS4, and products were directly sequenced in both strands using the same primers ITS 1 and ITS4. Sequences were compared to known sequences deposited in the NCBI non redundant database to confirm morphological identification. An NCBI BLAST search identified isolates F12SS1, F14SS3, F30SS3, and F25SR as F. acuminatum based on 100% similarity with corresponding sequences. GenBank Accession Nos. were JX114788, JX114785, JX114782, and JX114790, respectively. Pathogenicity tests were performed to fulfill Koch's postulates. Inocula were produced by adding a 5-mm diameter plug from a 7-day-old CMA petri dish culture to a previously sterilized 500-ml flask (237.5 g sand, 12.5 g cornmeal, 80 ml SDW), shaken over 9 days, and mixed with sterile soil at 1:3 (v:v). The inocula were transferred to a 500-ml pot, and 10 Aleppo pine seeds were planted with three replicates. After 1 month, all tested isolates caused typical symptoms on seedlings and the proportion of infected seedlings per each isolate was 50, 53.33, 56.66, 60, and 63.33%, respectively. There are many reports of F. acuminatum associated to conifer seedlings in nurseries (1,3) and most of them are conflicting because in some reports this species is considered non-pathogenic or only a seed contaminant and others consider it as a pathogen. To our knowledge, F. acuminatum is a first report on the Aleppo pine in northwestern Algeria, northern Africa. It is also the first report of this fungal species affecting the Aleppo pine throughout the world, and on conifers in Africa and the Mediterranean region. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory. ARS, USDA., Bestville, Maryland, USA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , June 18, 2012. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, Iowa, USA, 2006. (3) D. W. Minter. Cybertruffle's Robigalia, Observations of Fungi and their Associated Organisms. Retrieved from http://www.cybertruffle.org.uk/robigalia/eng/ , June 18, 2012.

Rituximab induction immunotherapy for first-line low-tumor-burden follicular lymphoma: survival analyses with 7-year follow-up.

BACKGROUND: The purpose of this study was to report long-term results of rituximab induction monotherapy in patients with low-tumor-burden follicular lymphoma (LTBFL). PATIENTS AND METHODS: Of 49 first-line LTBFL patients who received weekly doses of rituximab (375 mg/m(2)), 46 have been followed with a long-term analysis of clinical and molecular responses. RESULTS: Best clinical response (at any staging within a year following treatment) was 80%, 24 (52%) patients had complete or unconfirmed complete response, 13 (28%) had partial response and 9 (20%) had stable or progressive disease. Of 31 patients having a positive bcl2-JH rearrangement, 15 (48%) became negative following treatment. After 83.9 months of follow-up (95% confidence interval 6.4-92.8 months), the median progression-free survival is 23.5 months and overall survival (OS) is 91.7%. Five patients died (one progression, one myelodysplasia, one diffuse large B-cell lymphoma and two solid tumors). Seven patients (15%) are progression-free including five who are bcl2 informative. No unexpected long-term adverse event has been observed. CONCLUSION: A significant proportion of patients remain progression-free 7 years after a single 4-dose rituximab treatment in first-line LTBFL. The 7-year overall survivalOS is very high in this selected population of patients.

A 4-weekly course of rituximab is safe and improves tumor control for patients with minimal residual disease persisting 3 months after autologous hematopoietic stem-cell transplantation: results of a prospective multicenter phase II study in patients with follicular lymphoma.

BACKGROUND: This study explored the efficacy and safety of rituximab as treatment of clinical or molecular residual disease after autologous stem-cell transplantation (ASCT) in follicular lymphoma (FL). PATIENTS AND METHODS: Forty patients with CD20+ FL and clinically (group A, n = 14) or clono-specific PCR-detectable (group B, n = 25) residual disease persisting 3 months after ASCT received rituximab 375 mg/m² once weekly for 4 weeks. RESULTS: Response rate at day 50 was 36% [90% confidence interval (CI) 15-61] in group A (World Health Organization criteria) and 52% (90% CI 34-70) in group B (conversion PCR-undetectable status to undetectable status). The best response rate was 71% [nine complete responses (CRs) and one partial response] in group A and 76% in group B. At 36 months, all 10 responses persisted in group A, whereas 46% of patients in group B still had PCR-undetectable disease. Furthermore, 68% of patients in group B were still in clinical CR. Rituximab after ASCT was safe with few grade 3-4 toxic effects (15% patients), mainly acute reactions and infections. CONCLUSION: Rituximab induced a high rate of durable CRs in patients with clinically detectable disease, as well as durable eradication of PCR-detectable disease in patients with FL after ASCT. Continued molecular responses assessed with a highly sensitive and clono-specific PCR technique were correlated with an excellent disease control.

[Study of overweight and obesity in a population of urban school children in Sousse, Tunisia]. / Etude de la surcharge pondérale et de l'obésité dans une population d'enfants et d'adolescents scolarisés en milieu urbain à Sousse en Tunisie.

AIM: The aim of this study was to assess the prevalence of obesity and overweight and their relationship with cardiovascular disease risk factors. METHODS: Epidemiological survey based on a representative sample of 1569 urban school children of Sousse, Tunisia. RESULTS: Overweight (BMI > or = 25) was significantly higher in girls (16.1%) than in boys (11.6%); (chi 2 = 8.2; p = 0.004). Obesity (BMI > or = 30) was slightly higher in girls (3.7%) than in boys (2.7%); (chi 2 = 0.89; p = 0.34). Girls had significantly higher BMI, diastolic blood pressure, cholesterol and HDL cholesterol levels than boys who had however significantly higher levels of systolic blood pressure. Overweight was significantly higher in children who did not practice sport at school: 22 versus 13.1% (p < 0.002), in groups of youngsters who were not affiliated to school sport or city associations. Overweight children had a significantly higher levels of cholesterol, HDL cholesterol and means of systolic and diastolic blood pressures. CONCLUSION: These results will serve to set up a regional program of health promotion at schools.

[Smoking among school children in Sousse, Tunisia]. / Le tabagisme en milieu scolaire à Sousse, Tunisie.

In order to study smoking habits of children and adolescents from Sousse in Tunisia, we undertook an epidemiological survey on a representative sample of 1569 pupils aged between 13 and 19 years with a global answer rate of 95,4%. The objective of the study was both to describe Tunisian adolescent smoking behaviour and also to evaluate the influence of the home environment, friends and the different socio-demographic factors on acquiring or maintaining the habit. Students were surveyed in schools using a self-administered, anonymous questionnaire. Overall 7,6% of our sample smoked tobacco with prevalence amongst boys being much higher than in girls: 14,7% versus 1,1%; X(2)=103,4, p=0,00001. The smoking prevalence rose with age: in boys it increased from 3,4% at 13 years to 32,3% at 19 years; X(2)=40,9, p=0,0001. 60,6% of youngsters interrogated were exposed to passive smoking at home. Peer smoking behaviour has a clear effect on the tobacco habits of boys. These findings suggest school and medical authorities should design specific programs to limit the spread of the tobacco phenomenon in youngsters.

[Profile of tobacco use among the teachers of Sousse, Tunisia]. / Les facteurs déterminants le comportement tabagique chez les enseignants de la ville de Sousse, Tunisie.

OBJECTIVE: To examine the prevalence of tobacco use among the teachers in the region of Sousse (Tunisia) and to identify the factors, which determine this behavior. PATIENTS AND METHODS: It is a transactional study; using a self-administered and pre-tested questionnaire to 800 teachers. RESULTS: The population being studied was made up of 739 teachers including 50.6% of professors. The sample was 35.4% male and the average age was of 45.3±8.1 years. The total prevalence of tobacco use was 17.8% (41.4% among men and 4.7% among women). Half of these teachers smoked in their school establishments and in the presence of their pupils. The multivariate analysis had made it possible to identify four factors, which determine the profile of tobacco use in our teachers; these factors were: the sex, the age, knowledge and attitudes. CONCLUSION: It's necessary to intervene with the teachers of the town of Sousse with specific trainings on the topic of the tobacco use and dispose their adapted tools which will be used in the educational programmes of tobacco use prevention in schools.

Rapid detection of immunoglobulin G against Mycobacterium tuberculosis antigens by two commercial ELISA kits.

SETTING: Tunisia. OBJECTIVE: To assess the clinical usefulness of the commercial Pathozyme-Myco G (Myco G) and Pathozyme TB complex plus (Patho) enzyme-linked immunosorbent assay (ELISA) kits for the rapid diagnosis of active tuberculosis (TB) and to distinguish between active TB and non-TB pulmonary diseases in Tunisian patients. DESIGN: Immunoglobulin G mediated humoral immune response against mycobacterial antigens (38 kDa and lipoarabinomannan, Myco G; 16 and 38 kDa, Patho) was evaluated in a group of active TB patients (128 smear-positive pulmonary and 33 extra-pulmonary samples) and in a control group (107 patients with non-tuberculous lung disease and two with leprosy). Active TB cases were confirmed by Mycobacterium tuberculosis culture from clinical samples. RESULTS: The sensitivity of the Myco G test was 71% in active TB (pulmonary and extra-pulmonary), while the specificity was 100%. The Patho test showed a sensitivity of 43.5% with a specificity of 96.3%. A combination of both tests showed a sensitivity of 81% and a specificity of 96.3%. CONCLUSIONS: Both ELISA tests were simple and easy to perform. Their combined use led to an increase in the diagnostic accuracy of active TB and its discrimination from non-TB pulmonary diseases. They could therefore be used as screening tools in poorly equipped laboratories in TB-endemic regions.