Tuberculin Research Articles

Use of commercial ELISA for the diagnosis of bovine tuberculosis in animals negative in the intradermal tuberculin test

Tuberculosis is a globally distributed infectious disease of great importance to public health due to its zoonotic nature. In addition to its major impact on public health, bovine tuberculosis causes major economic losses due to reduced herd productivity and carcass losses in slaughterhouses. Given the critical importance of the disease, it is of utmost importance to enhance the accuracy of diagnostic methods in programs aimed at controlling and eradicating bovine tuberculosis in Brazil. The objective of this research project is to diagnose bovine tuberculosis in dairy herds that previously tested negative in the intradermal tuberculin test using commercial ELISA (enzyme-linked immunosorbent assay). Samples from 17,917 cattle from dairy herds distributed across five farms in the Campos Gerais region of the state of Paraná were analyzed. For serological diagnosis, the commercial ELISA/IDEXX® kit was used to detect anti-Mycobacterium bovis antibodies in cattle herds. Of the 17,917 animals tested, the ELISA Mycobacterium bovis Antibody Test (IDEXX®) identified 399 (2.3%) reactive animals and 17,518 (97.7%) non-reactive animals. The highest number of positive animals in the ELISA was observed on farm B, with 375 (2.5%) reactive animals. Given the challenges faced in Brazil regarding the diagnostic confirmation of tuberculosis and the absence of validated reagents and methods, the importance of implementing new validated diagnostic methods is evident. The use of ELIST could serve as a complementary alternative to the official intradermal tuberculin test, potentially detecting false-negative animals and aiding in the control of the disease.

Endoscopic Monitoring of Treatment of Indeterminate Intestinal Lesions in a Prospective "Real-Life" Cohort in Eastern India Where Tuberculosis Remains Endemic: Distinguishing Intestinal Tuberculosis From Crohn's Disease.

Introduction It is sometimes difficult to differentiate between intestinal tuberculosis (ITB) and Crohn's disease (CD) in India, as both conditions may mimic each other. The aim was to differentiate ITBfrom CDin indeterminate intestinal lesions with a therapeutic trial of anti-tubercular therapy (ATT) and follow-up to find out the clinical, endoscopic, radiological, and histological predictors for differentiation between ITB and CD. Methods A prospective observational cohort study of patients diagnosed with ITB and CD according to the Asia-Pacific Guidelines in a "real-life" clinical setting was conducted. ITB was diagnosed by Paustian criteria with Logan's modification. CD was diagnosed according to European Crohn's and Colitis Organization (ECCO) guidelines. We put the patients with a definite diagnosis of ITB and those with an indeterminate diagnosis on ATT and followed them up clinically, endoscopically, and radiologically. Patients were reassessed clinically, endoscopically, and histologically eight weeks after the start of therapy. They were again evaluated endoscopically and radiologically after completion of six months of ATT. The CD patients continued anti-inflammatory, immunomodulator, biological, and/or steroid treatments. Results We conducted this prospective study on consecutive Indian patients who had 21 definite diagnoses of ITB, 26 definite diagnoses of CD, and 42 indeterminate diagnoses. We diagnosed 49 with ITB and 28 (57%) after a therapeutic trial. Ultimately, 40 patients received a CD diagnosis, with 14 (35%) not responding to the ATT therapeutic trial. In patients with ITB, symptomatic improvement after eight weeks of ATT is correlated with endoscopic healing, especially for ulcers but not necessarily for nodularity or strictures. In 50% of these patients, minimal nodularity/pseudopolypii as well as residual scarring was seen on endoscopy even after completion of therapy. Strictures in ITB patients persisted on endoscopy in 40% despite six months of ATT. GI bleeding (64% vs. 10%; p < 0.0001), chronic diarrhea (71% vs. 35%; P = 0.02), fistula or sinuses (21% vs. 0%;p < 0.01), and multiple site involvement of the intestine (73% vs. 6%; p < 0.0001) were significantly more common in CD than in patients with ITB. Fever (82% vs. 50%; p < 0.01) and positive tuberculin tests were more common in ITB patients. PCR positivity and the presence of AFB in smear and culture could be demonstrated in only a small percentage of ITB patients. Conclusion Therapeutic trials in indeterminate intestinal lesions can distinguish ITB from CD without significant adverse effects. Strictures in patients with ITB do not resolve in all patients. GI bleeding, chronic diarrhea, fistulas or sinuses, multiple sites of involvement, and fever have the highest accuracy in differentiating ITB from CD.

A high titer antibody response against P22 protein immunocomplex is not correlated with protection in naturally tuberculosis-infected goats

Caprine livestock are significant reservoirs of the Mycobacterium tuberculosis complex (MTBC), contributing to tuberculosis (TB) transmission among animals and humans. The P22 protein immunocomplex (P22PI), derived from bovine tuberculin, shows immunostimulating capacity and is used for TB diagnosis. This study assessed the immunogenicity and protective efficacy of P22PI in two groups of goats: 24 naïve goats (12 immunised, 12 controls) from a TB-free herd, and 24 infected goats (12 immunised, 12 controls), referred to as pre-infected animals, from a M. bovis-infected herd. Both were exposed for 5 months to M. bovis-naturally infected goats. Reactors to single and comparative intradermal tuberculin (SIT and SCIT, respectively) tests and interferon-gamma release assay (IGRA) significantly increased (p < 0.05) in both groups 5 months’ post-exposure, with no significant differences between immunised and control animals. However, immunised animals exhibited a significantly higher (p < 0.05) antibody response against P22PI. Most naïve animals (83.3%) and all pre-infected animals developed TB-compatible lesions, with extensive necrosis in the lungs and associated lymph nodes, compared to 50% and 83.3% of control animals, respectively. These findings suggest that while P22PI stimulates an intense antibody response under the conditions of the present study, it does not confer protection against TB and may exacerbate disease severity.

Cost-effectiveness of diagnostic technologies for mycobacterium tuberculosis infection in India and Brazil.

The economic value of new skin-based tests and blood-based interferon-γ release assays (IGRAs) for tuberculosis (TB) infection is not yet well-established. This study evaluates the cost and cost-effectiveness in two high-burden countries by comparing:(a) new skin-based tests(Diaskintest and Cy-Tb) with the purified protein derivative (PPD)-tuberculin test (TST);(b) IGRAs (Standard E TB-Feron ELISA (TBF))with approved IGRAs (QuantiFERON-TB Gold Plus (QFT-GP)and TSPOT.TB); and (c) the best performing skin-based test with the best performing IGRA) based on cost effectiveness. In this paper, we developed a decision tree model for India and Brazil from a health system perspective. To quantify the effect of parameter variability and uncertainty, we performed both univariate and probabilistic sensitivity analysis. The study findings reveal that among skin-based tests, the Diaskintest is more cost-effective compared to TST-PPD at 22.6 USD and 41.0 USD per correctly diagnosed case of TB infection for Brazil and India, respectively. For blood-based assays, TSPOT.TB outperforms QFT-GP and TBF due to its lower cost and higher effectiveness. When compared with Diaskintest, TSPOT.TB has an incremental cost of approximately 8 USD and 6 USD for India and Brazil respectively but is more effective. The incremental cost-effectiveness ratio (ICER) was 74 USD and 55 USD for India and Brazil, respectively. In summary, while Diaskintest is potentially cost-saving when compared to TSPOT.TB in these two high-burden TB countries but the TSPOT.TB demonstrates higher effectiveness.

A conservative management approach for spinal tuberculosis causing cauda equine syndrome in pregnancy

Background: Spinal tuberculosis in pregnancy leading to neurological sequelae is rare and a challenging condition to manage. The usual approach is to perform urgent decompression surgery, but patients with a more stable course of the disease may benefit from a conservative approach with anti-tuberculosis (ATT) therapy.Case presentation: A pregnant mother in her second trimester presented with a history of chronic backache exacerbated by right-sided foot drop, L5 root sensory loss and intermittent urinary retention. MRI revealed L5 vertebral collapse with para-vertebral collection causing cauda equine compression at the L5-S1 level. High ESR, strongly positive tuberculin test, and the clinical and radiological findings were compatible with a diagnosis of spinal tuberculosis. Based on the multi-disciplinary team’s decision, a conservative management approach with ATT and supportive therapy was initiated with close observation. The patient showed a remarkable response to ATT with clinical and radiological improvement. She underwent an elective caesarean section at 37 weeks and delivered a healthy baby. On the fourth week postpartum, she underwent decompressive surgery with complete neurological recovery,Discussion and conclusion: Spinal TB in pregnancy causing neurological impairment is usually managed with urgent surgical interventions. This is one of the few reported cases where a conservative approach was undertaken, with close observation and a positive outcome.Therefore, it is important to carefully select patients needing surgical interventions and those who would benefit better from a conservative approach. High suspicion, early diagnosis, prompt treatment initiation and close monitoring are vital components in achieving the best outcome.

A case report: Reactivation of tubercular infection presenting as bilateral frosted branch angiitis

: Frosted branch angiitis is a rare form of angiitis having multifactorial associations. Tubercular retinitis is a common association with frosted branch angiitis. Aim of this study is to show that frosted branch angiitis can be the only presenting symptom of reactivated tubercular infection.: A female patient presented with severe diminution of vision in both eyes and bilateral severe form of retinal vasculitis. She had a positive skin tuberculin test. Radiological investigation (HRCT Chest) revealed an old healed infection in the form of fibrotic streaks. She had previous history of Tuberculosis 17 years back and she didn’t complete the ATT course at that time.: Reactivation of Tubercular infection can present as frost branch angiitis. Prompt diagnosis and effective management helps in preventing as well as reversing the loss of vision.

A disposable, portable electrochemical immunosensor for rapid in situ detection of bovine tuberculosis

This contribution describes the development of a simple, fast, cost-effective, and sensitive impedimetric immunosensor for quantifying bovine tuberculosis (TB) in bovine serum samples. The construction of the immunosensor involved immobilizing the purified protein derivative (PPD) of M. bovis onto a screen-printed electrode that was modified with gold nanoparticles (AuNPs) and a polypyrrole (pPy) film synthesized electrochemically. The immunosensor exhibited a linear range from 0.5 μg mL−1 to 100 μg mL−1 and achieved a limit of detection (LD) of 100 ng mL−1 for the detection of anti-M. bovis antibody. The recovery percentages obtained in bovine serum samples were excellent, ranging between 98 % and 103 %.This device presents several advantages over alternative methods for determining TB in bovine serum samples. These include direct, in situ measurement without the need for pre-treatment, utilization of small volumes, thus avoiding harmful solvents and expensive reagents, and portability. In addition, the immunosensor exhibits both physical and chemical stability, retaining effectiveness even after 30 days of modification. This allows simultaneous incubations and facilitates large-scale detection.Hence, this immunosensor presents itself as a promising diagnostic tool for detecting anti-M. bovis antibodies in bovine serum. It serves as a viable alternative to tuberculin and ELISA tests.

Evaluating the ability of non-tuberculous mycobacteria to induce non-specific reactions in bovine tuberculosis diagnostic tests in guinea pigs and cattle

Limitations in diagnostic test performance are one of the major challenges hampering the eradication of bovine tuberculosis (bTB). Non-tuberculous mycobacteria (NTM) are considered one of the main causes of non-specific reactions in the intradermal tuberculin test, the most widely used bTB diagnostic test. To determine the role of NTMs in bTB misdiagnosis in Spain, an experimental study including the NTM species most commonly found in bTB-positive animals from bTB-free farms in the country (M. avium subsp. avium (Maa), “Mycobacterium avium subsp. hominissuis” (Mah), M. bourgelatii, M. intermedium, M. kansasii and M. nonchromogenicum) was carried out on guinea pigs and cattle. First, guinea pigs were sensitized with the selected NTMs, and six weeks post-sensitization four antigen mixtures (bovine-PPD, avian-PPD, P22 and ESAT6-CFP10) were inoculated intradermally and their effect was measured 24- and 48-h post-inoculation. Larger erythematous reactions were observed in guinea pigs sensitized with Mah, M. kansasii, and Maa, with significant differences in the reactions measured at the bovine-PPD inoculation site for the two first bacteria compared with other NTMs. The sensitization process was repeated in cattle, and five months post-sensitization the same antigen mixtures were inoculated in the cervical region and responses were measured at 48- and 72-h post-inoculation. A significantly higher increase in the skinfold thickness measured at the bovine-PPD inoculation site was observed in calves sensitized with Mah, Maa, M. intermedium and M. kansasii. These results demonstrate that certain NTM species may play a more significant role in bTB diagnostic interferences and show that results obtained in guinea pig and bovine models do not always coincide.

Extensive differential DNA methylation between tuberculosis skin test positive and skin test negative cattle

Bovine tuberculosis (bTB), caused by Mycobacterium bovis (M. bovis), represents a significant problem for the agriculture industry as well as posing a risk for human health. Current diagnostic tests for bTB target the cell-mediated immune (CMI) response to infection with M. bovis, primarily through screening of animals with the tuberculin skin test. Epigenetic modifications have been shown to alter the course of the immune response and differentially methylated regions (DMRs) might also influence the outcome of the skin test in cattle. Whole Genome Bisulphite Sequencing (WGBS) was used to profile DNA methylation levels from peripheral blood of a group of cattle identified as test positive for M. bovis (positive for the single intradermal comparative tuberculin test (SICTT) and/or the interferon-γ release assay compared to a test negative control group [n = 8/group, total of 16 WGBS libraries]. Although global methylation profiles were similar for both groups across the genome, 223 DMRs and 159 Differentially Promoter Methylated Genes (DPMGs) were identified between groups with an excess of hypermethylated sites in SICTT positive cattle (threshold > 15% differential methylation). Genes located within these DMRs included the Interleukin 1 receptor (IL1R1) and MHC related genes (BOLA and BOLA-DQB). KEGG pathway analysis identified enrichment of genes involved in Calcium and MAPK signalling, as well as metabolism pathways. Analysis of DMRs in a subset of SICTT negative cattle that were IFN-γ positive showed differential methylation of genes including Interleukin 10 Receptor, alpha (IL10RA), Interleukin 17 F (IL17F) and host defence peptides (DEFB and BDEF109). This study has identified a number of immune gene loci at which differential methylation is associated with SICTT test results and the degree of methylation could influence effective host immune responses.

Differentiation of Bovine Tuberculosis and Paratuberculosis Infections with Antemortem Diagnostic Methods

In this study, based on the results of tuberculin skin tests (Bovine and Avian PPD) used in the antemortem diagnosis and differentiation of Bovine Tuberculosis, the animals in the farms with suspected Tuberculosis were serologically examined to diagnose Paratuberculosis infection and fecal bacterioscopy was performed. In addition, it was aimed to obtain data that will contribute to the eradication studies of Bovine Tuberculosis disease by comparing the antemortem diagnostic methods of Bovine Tuberculosis disease, which is endemic in Türkiye and by determining the sensitivity and specificity values of the interferon gamma (IFN-γ) test. In this context, intradermal tuberculin test was applied to 423 cattle with suspected Tuberculosis in a total of 5 dairy cattle farms, one each from Çankırı, Çorum, Ankara, Eskişehir and Konya regions, and this test was determined as the gold standard method and the sensitivity and specificity of the IFN-γ test were determined as 86% and 97%, respectively. For the diagnosis of Paratuberculosis infection, antibody ELISA, fecal bacterioscopy and IFN-γ ELISA were performed on these animals and the prevalence of these tests were 10.4%, 5.44% and 4.96% respectively and 4 (0.95%) of the cattle were positive for each of the diagnostic methods for Map infection. As a result, it was concluded that IFN-γ test, which gives similar results to intradermal tuberculin test results, can also be used in the antemortem diagnosis of Bovine Tuberculosis. Also, in the comparative intradermal tuberculin test for the diagnosis of Tuberculosis infection, avian PPD positive animals were found to play a decisive role in the detection of nonspecific reactions or Paratuberculosis infected animals, supported by other tests used for the diagnosis of Paratuberculosis.

How to plan and provide general anesthesia for a troop of 98 hamadryas baboons (Papio hamadryas) for contraceptive and preventative health interventions.

Present an approach to the safe and efficient provision of anesthesia and birth control measures to a large group of primates. 98 hamadryas baboons (Papio hamadryas) held in a German zoological institution. A group of 12 veterinarians, 2 zookeepers, and 6 volunteers anesthetized all animals within 2 days. The baboons were orally premedicated with midazolam (0.1 to 0.5 mg/kg) and anesthetized with medetomidine (40 to 60 µg/kg, IM) and ketamine (2 to 4 mg/kg, IM); isoflurane at rates of 1.5% to 2% was used for maintaining anesthesia if necessary. All animals received a physical examination, prophylactic medication, and tuberculin testing. For population management, the animals received a contraceptive implant (adult females), orchiectomy (young males), or vasectomy (breeding males). Young males received intratesticular blocks with lidocaine. All animals received atipamezole (125 to 150 µg/kg) before recovery. Premedication resulted in anxiolysis, which facilitated separating and darting. Median time from darting to access to the animal was 10 minutes. Mean anesthetic times were 25 minutes for females and 55 minutes for males. The depth of anesthesia was appropriate for the procedures. No fatalities were recorded. One animal was injured by other baboons but recovered after treatment. Health management and birth control measures are necessary in baboon troops under human care. Anesthesia and/or contraception of individual animals often leads to intraspecific aggression. This case series describes how to provide anesthesia and contraception to an entire troop as an alternative approach that can be adopted to future similar interventions.

PREVALENCE OF BOVINE TUBERCULOSIS IN NILI RAVI BUFFALOES IN DISTRICT KASUR

This study aimed to determine the prevalence of Bovine Tuberculosis (BTB) in Nili Ravi buffaloes in Kasur district, Punjab, Pakistan. A total of 350 buffaloes were sampled from August 2023 to April 2024, comprising 200 buffaloes from the Progeny Testing Program (PTP) and 150 random field samples. The Skin Tuberculin Test (TST) and Enzyme-Linked Immunosorbent Assay (ELISA) were employed to diagnose BTB. The overall prevalence of BTB was found to be 3.5%, with TST and ELISA indicating prevalence rates of 2.6% and 2.9%, respectively. The study also examined risk factors such as age and management practices. Although older buffaloes (&gt;8 years) showed a higher prevalence compared to younger animals, the difference was not statistically significant (P &gt; 0.05). No significant difference in prevalence was observed between the PTP group and random field samples. The findings suggest that while BTB prevalence in Kasur district is relatively low, there is a need for ongoing surveillance and improved management practices. Variations in historical prevalence rates highlight the importance of continuous monitoring and targeted control measures. The study underscores the necessity of using multiple diagnostic methods and considering various risk factors to develop effective BTB control strategies.

Expression and field evaluation of new Mycobacterium bovis antigens

Bovine tuberculosis (bTB) represents a threat to livestock production. Mycobacterium bovis is the main causative agent of bTB and a pathogen capable of infecting wildlife and humans. Eradication programs based on surveillance in slaughterhouses with mandatory testing and culling of reactive cattle have failed to eradicate bTB in many regions worldwide. Therefore, developing effective tools to control this disease is crucial. Using a computational tool, we identified proteins in the M. bovis proteome that carry predictive binding peptides to BoLADRB3.2 and selected Mb0309, Mb1090, Mb1810 and Mb3810 from all the identified proteins. The expression of these proteins in a baculovirus-insect cell expression system was successful only for Mb0309 and Mb3810. In parallel, we expressed the ESAT-6 family proteins EsxG and EsxH in this system. Among the recombinant proteins, Mb0309 and EsxG exhibited moderate performance in distinguishing between cattle that test positive and negative to bTB using the official test, the intradermal tuberculin test (IDT), when used to stimulate interferon-gamma production in blood samples from cattle. However, when combined as a protein cocktail, Mb0309 and EsxG were reactive in 50 % of positive cattle. Further assessments in cattle that evade the IDT (false negative) and cattle infected with Mycobacterium avium paratuberculosis are necessary to determine the potential utility of this cocktail as an additional tool to assist the accurate diagnosis of bTB.

Serological and epidemiological investigation of Mycobacterium avium subspecies paratuberculosis in bovines in Pakistan.

This study aimed to investigate the prevalence of paratuberculosis in cattle and buffaloes at twelve public dairy farms in Punjab, Pakistan. A total of 2,181 more than two-year-old animals (1,242 cattle and 939 buffaloes) were tested by avian tuberculin, i.e., killed purified protein derivative of Mycobacterium avium paratuberculosis and indirect enzyme linked immunosorbent assay (ELISA). Blood and fecal samples were collected from tuberculin positive animals. These samples were further processed by indirect ELISA. The data were analyzed using frequency analysis and logistic analysis procedures. The prevalence of paratuberculosis at public dairy farms was 3.8%, as determined by tuberculin+ELISA test. It varied from 0.71% to 13.5% with a 100% herd prevalence. Multivariate logistic regression analysis revealed that species, milk production, total animals, total small ruminants, and total buffaloes were significantly associated with the occurrence of paratuberculosis. Odd ratio analysis revealed that with a one-kilogram increase in body weight, there will be a 0.006% increase in disease occurrence. With the increase in one animal in small ruminants and buffaloes, there will be 0.008% and 0.42% greater chances of developing paratuberculosis, respectively. Bivariate logistic regression analysis of cattle and buffaloes revealed that farm number, age, and total number of cattle were significantly associated with the occurrence of paratuberculosis. A one-month increase in lactation length increases the chance of tuberculosis by 0.004%; similarly, a one-liter increase in milk production increases the chance of disease by 10%. With each additional buffalo in the herd, there will be a 0.007% greater chance for the occurrence of paratuberculosis. This study concluded that tuberculin testing can be used in conjunction with ELISA to screen animals for paratuberculosis in countries with scarce resources, such as Pakistan. The efficacy of disease diagnosis can be improved by combining multiple tests.

Occurence and risk factors of tuberculosis infection in orphanage children in Bali

Background Tuberculosis (TB) is an infectious disease that is still a common threat worldwide, especially in pediatric populations. TB transmission occurs particularly when the transmitter has no obvious manifestation of the disease. There is a higher incidence of TB infection in children than in the general population, especially in high risk populations such as children in orphanages. However, the incidence of TB infection in orphaned children in Indonesia, including Bali, is unknown. Objective To describe the incidence and risk factors for TB infection in children in orphanages in Bali. Methods This case-control study was conducted in 12 orphanages in Bali. Subjects were divided into a case group comprised of children with TB infection, and a control group comprised of those without TB infection. TB infection was diagnosed by positive tuberculin test without clinically confirmed TB. Results A total of 175 children were recruited as subjects. There were 49 (28.0%) children with TB infection. Bivariate analysis revealed significant associations between density, humidity, ventilation/room area ratio, and area of ??origin with TB infection. Multivariate analysis showed that ventilation/room area ratio and room humidity of ?73% were independently positive correlated with TB infection. Natural lighting and BCG scar were not significantly different between groups. Conclusion The occurence of TB infection in children residing in orphanages in Bali is high (28%). The risk factors identified independently are lower ventilation and higher room humidity.

Prevalence of pulmonary tuberculosis and HIV infections and risk factors associated to tuberculosis in detained persons in Antananarivo, Madagascar

The incidence rate of tuberculosis in prisons is estimated to be 8 times greater than that in the general population in Madagascar. Our objectives were to estimate the prevalence of pulmonary tuberculosis and HIV infection among prisoners and to identify risk factors associated with tuberculosis. We conducted a cross-sectional study at the central prison of Antananarivo from March to July 2021. Individual male and female inmates aged ≥ 13 years who had lived in the prison for at least three months prior to the study period were included as participants. Acid-fast bacilli detection by microscopy and/or culture, an intradermal tuberculin test, a chest X-ray, and a rapid diagnostic orientation test for HIV were performed. Among 748 participants, 4 (0.5%) were confirmed to have pulmonary tuberculosis. Overall, 14 (1.9%) patients had “confirmed” or “probable” tuberculosis [0.90–2.84, 95% CI]. The proportion of participants with latent tuberculosis infection was 69.6% (517/743) based on a positive tuberculin test without clinical symptoms or radiography images indicating tuberculosis. Out of 745 HIV screening tests, three showed reactive results (0.4%). Age (OR = 4.4, 95% CI [1.4–14.0]) and prior tuberculosis treatment (or episodes) were found to be associated with confirmed and probable tuberculosis.

GAMBARAN PEMERIKSAAN TES CEPAT MOLEKULER (TCM) GENEXPERT PADA PASIEN DI RUMAH SAKIT UMUM DAERAH PALEMBANG BARI

Background: Tuberculosis is an infectious disease caused by the bacterium Mycobacterium tuberculosis. The diagnosis of pulmonary TB is made based on the clinical picture, physical examination, radiological picture, laboratory examination, and tuberculin test. The pulmonary TB diagnosis currently used routinely in laboratories including hospitals and health centers is bacteriological diagnosis using the rapid molecular test (TCM) technique, while microscopic examination is used as a follow-up examination (monitoring) in patients receiving treatment. TCM examination is a molecular detection method based on nested real-time PCR. The use of TCM is a priority for TB testing because it has several advantages, including: it can diagnose TB and resistance to rifampicin quickly, accurately with high sensitivity, and results can be known in approximately 2 hours.&#x0D; Objectives: The aim of this study is to see the description of TCM examinations in patients at Palembang BARI Regional Hospital based on gender and age groups. Method: This research was conducted at the Palembang BARI Regional General Hospital Laboratory Installation. The samples taken were from patients at the Palembang BARI Regional General Hospital and not referrals from other health service facilities. 50 samples were taken and grouped based on TCM results, gender and age. Result: In grouping based on TCM results, the highest result was negative. In gender grouping, more male patients underwent TCM examinations. In the age grouping, the elderly are more likely to undergo TCM examinations. Conclusion: There are several factors that cause a person to become infected with TB, namely social status, smoking, and having the habit of drinking alcohol.

Bovine Tuberculosis Testing in Colombia: Comparative Histopathological, Microbiological, and Molecular Biology Findings

Introduction: Bovine tuberculosis (bTB) is a zoonotic infectious disease present in Colombia, caused by Mycobacterium bovis, and causes tuberculosis in water buffalo (Bubalus bubalis). Diagnosis of bovine tuberculosis through the intradermal test is difficult; evaluating and understanding the behavior of other diagnostic tests is necessary.&#x0D; Objective: To describe the behavior and results of different diagnostic methods for bovine tuberculosis in water buffalo positive for the Purifed Proteic Derivate (DPP) intradermal test.&#x0D; Methodology: In water buffaloes positive for comparative cervical tuberculin test, different diagnostic methods were applied, described, and compared: Ziehl-Neelsen staining, microbiological culture, histopathological analysis, and PCR-HRM.&#x0D; Results: Histopathological tests showed that 26 water buffalo positive for DPP (52%) had histological lesions compatible with bovine tuberculosis. 37% of the evaluated samples from tuberculin-positive Buffalo's lungs and secondary lymph nodes showed acid-alcohol-resistant bacillus with Ziehl-Neelsen staining. Four samples of Mycobacterium bovis from tuberculin-positive buffalo were isolated and identified, with two of these isolates confirmed from tissues with PCR-HRM, and three buffalo with microbiological isolates presented granulomatous lesions through histological analysis. Seventeen tuberculin-positive buffalo (34%) tested positive for real-time PCR HRM, and nine of these buffalo did not have histological lesions compatible with bTB and were confirmed with the molecular test.&#x0D; Conclusion: Our results provide positive evidence of histological findings, microbiological isolation, and molecular diagnosis of tuberculin-positive water buffalo in the lowlands of Colombia. None of the complementary tests performed showed 100% concordance with the comparative cervical tuberculin test results for bTB.

Fluvastatin Converts Human Macrophages into Foam Cells with Increased Inflammatory Response to Inactivated Mycobacterium tuberculosis H37Ra.

Cholesterol biosynthesis inhibitors (statins) protect hypercholesterolemic patients against developing active tuberculosis, suggesting that these drugs could help the host to control the pathogen at the initial stages of the disease. This work studies the effect of fluvastatin on the early response of healthy peripheral blood mononuclear cells (PBMCs) to inactivated Mycobacterium tuberculosis (Mtb) H37Ra. We found that in fluvastatin-treated PBMCs, most monocytes/macrophages became foamy cells that overproduced NLRP3 inflammasome components in the absence of immune stimulation, evidencing important cholesterol metabolism/immunity connections. When both fluvastatin-treated and untreated PBMCs were exposed to Mtb H37Ra, a small subset of macrophages captured large amounts of bacilli and died, concentrating the bacteria in necrotic areas. In fluvastatin-untreated cultures, most of the remaining macrophages became epithelioid cells that isolated these areas of cell death in granulomatous structures that barely produced IFNγ. By contrast, in fluvastatin-treated cultures, foamy macrophages surrounded the accumulated bacteria, degraded them, markedly activated caspase-1 and elicited a potent IFNγ/cytotoxic response. In rabbits immunized with the same bacteria, fluvastatin increased the tuberculin test response. We conclude that statins may enhance macrophage efficacy to control Mtb, with the help of adaptive immunity, offering a promising tool in the design of alternative therapies to fight tuberculosis.

Prevalence and risk factors of tuberculosis infection in orphanage children in Bali, Indonesia

Background: Tuberculosis (TB) is an infectious disease that causes high morbidity and mortality worldwide. Indonesia ranks third in the most TB cases in 2020 globally. Orphanages are densely populated places with a high risk of transmission, but routine TB screening programs have yet to be carried out. This study aims to find the prevalence of tuberculosis infection and its risk factors in orphanage children. Methods: This study used a cross-sectional approach. The data collected included age, gender, nutritional status, stunting, BCG scars, and abnormalities in physical examination. History, physical examination, and tuberculin test are the screening programs performed. All children with a positive tuberculin test as an induration of 10 mm 72 hours after the tuberculin test underwent a chest X-ray. Results: We conducted tuberculosis screening for 262 children in 15 orphanages in Denpasar and Badung from July to October 2022. There is no known history of contact with active TB sufferers. No active tuberculosis infection was detected. Latent tuberculosis infection (LTBI) occurred in 57 children (21.7%), 48 (84.2%) aged 10-17 years, 28 (49.1%) were male, 54 (94.7%) had normal nutritional status, 13 (22.8%) were stunted, and no BCG scars in 28 (49.1%). No BCG scar was a risk factor for LTBI with OR 2.29 (95% CI 1.06 - 4.97; p 0.03). Conclusion: One-fifth of orphanage children suffer from LTBI, and no BCG scar was a risk factor for LTBI. TB screening should be done routinely and actively in the orphanage.