Acta Microbiologica Hellenica

The excessive use of antimicrobials drives the emergence of multidrug resistance (MDR) in bacterial strains, which harbor resistance genes to survive under diverse drug pressures. Such resistance can result in life-threatening infections. The predominance of MDR Pseudomonas spp. poses significant challenges to public health and environmental sustainability, particularly in ecosystems affected by human activities. Characterizing MDR Pseudomonas spp. is crucial for developing effective diagnostic tools and biosecurity protocols, with broader implications for managing other pathogenic bacteria. Strains were diagnosed through 16S rRNA PCR and sequencing, complemented by phylogenetic analysis to evaluate local and global evolutionary connections. Antibiotic susceptibility tests revealed extensive resistance across multiple classes, with MIC values surpassing clinical breakpoints. This study examined the genetic diversity, resistance potential, and phylogenetic relationships among Pseudomonas aeruginosa strain DG2 and Pseudomonas fluorescens strain FM3, which were isolated from solid waste dump sites (n = 30) and dairy farms (n = 22) in West Bengal, India. Phylogenetic analysis reveals distinct clusters that highlight significant geographic linkages and genetic variability among the strains. Significant biofilm production under antibiotic exposure markedly increased resistance levels. RAPD-PCR profiling revealed substantial genetic diversity among the isolates, indicating variations in their genetic makeup. In contrast, SDS-PAGE analysis provided insights into the protein expression patterns that are activated by stress, which are closely linked to MDR. This dual approach offers a clearer perspective on their adaptive responses to environmental stressors. This study underscores the need for vigilant monitoring of MDR Pseudomonas spp. in anthropogenically impacted environments to mitigate risks to human and animal health. Surveillance strategies combining phenotypic and molecular approaches are essential to assess the risks posed by resilient pathogens. Solid waste and dairy farm ecosystems emerge as critical reservoirs for the evolution and dissemination of MDR Pseudomonas spp. Full article

Trichomonas vaginalis infection (TVI) is the most common curable sexually transmitted infection (STI). In this study, we aimed to assess the performances of different tests for TVI diagnosis in symptomatic Greek women, evaluating the TVI prevalence rate (PR) in Greece and comparing the latter with TVI-PR estimates from Europe. A laboratory-based cross-sectional analysis and a meta-analysis were conducted. Of 399 symptomatic Greek women, 17 had TVI, corresponding to a TVI-PR of 4.3%. The commercial nucleic acid amplification test (NAAT) achieved a sensitivity of 94.1%, which was 6% higher than the sensitivity of the culture method, 35% higher than that of the wet mount test, and 59% higher than that of the Giemsa stain test. The wet mount test achieved the lowest positive predictive value of 76.9%. All the tests had high specificity levels and negative predictive values. Data from 34 European TVI-PR studies in symptomatic women were pooled. The TVI-PR established in our study was similar to the TVI-PR estimates of 4.8% in Europe and 4.5% in Greece, with the second being higher than those of 2.1% in Northwestern Europe and 1.5% in Southern Europe but closer to that of 6.7% in Türkiye. In Greece, a European country with a relatively high TVI-PR among symptomatic women, the highly sensitive and specific, automated, point-of-care NAAT would facilitate rapid, accurate TVI diagnosis and the treatment of this target population to meet the WHO’s goal of ending STI epidemics by 2030. Full article

Beta-thalassemia major is an inherited disorder that requires lifelong blood transfusions, with the risk of complications including poor oral health and dental caries. The objective of this study was to compare the oral microbiome diversity and composition in transfusion-dependent thalassemia patients and relate it to oral hygiene and dental caries. A cross-sectional analysis of 35 patients of beta-thalassemia major aged 6–18 years was performed. The status of oral hygiene was examined through the Oral Hygiene Index—Simplified (OHI-S) and Decayed, Missing, and Filled Teeth (DMFT) index. Saliva was taken for DNA extraction, followed by the 16S rRNA sequencing of V3-V4 hypervariable regions. The bioinformatics pipeline in QIIME2 was utilized for analyzing the comparison of microbial composition and diversity in groups of varying oral hygiene status and severity of caries. Metagenomic analysis revealed 3334 Amplicon Sequence Variants (ASVs), of which the most prevalent genera were Streptococcus, Haemophilus, Veillonella, Rothia, and Prevotella. High-oral-hygiene groups presented increased levels of cariogenic bacteria, while moderate-oral-hygiene groups presented an equilibrated microbiome. No statistically significant differences in microbial diversity were found between the study groups (p > 0.05). This study sheds light on the critical importance of oral hygiene in microbiome diversity in patients with beta-thalassemia major. Full article

Dengue virus infection (DVI), a mosquito-borne arboviral infection, is prevalent in tropical and subtropical regions, including Bangladesh, where incidence has surged over the past three decades—particularly in urban and peri-urban areas. This study investigates the factors influencing DVI seropositivity among clinically suspected patients admitted to the selected hospitals of Savar, Dhaka, and Chattogram. Data were collected from 850 clinically suspected patients admitted to two hospitals in Savar, Dhaka, and two in Chattogram during 2019. Questionnaire responses and laboratory test results (NS1, IgM, and IgG) were analyzed using descriptive statistics, chi-square tests, and logistic regression. Out of 450 admissions in Savar, 330 tested positive, while Chattogram reported 145 positives from 400 cases. No significant differences were observed between regions in relation to hospital type, season, gender, or household preventive measures. In Savar, DVI status was significantly associated with season, mosquito net use, and patient contact. In Chattogram, household repellent use and patient contact were key factors. Diagnostic tests varied in detection capability. These findings can inform targeted intervention strategies and public health messaging, such as promoting personal protection measures and community awareness campaigns, particularly in high-incidence urban settings. However, further research across diverse geographic and socio-ecological contexts is needed to enhance the generalizability and policy relevance of these results. Full article

Vaginitis is a major health concern among women, with inadequate treatment potentially leading to reproductive complications. This study aimed to assess vaginitis prevalence, identify predominant pathogens, and evaluate associated risk factors among female patients at Da Nang Dermato-Venereology Hospital. A prospective study of 796 female patients undergoing physical examinations was conducted, with demographic, clinical, and microbiological data collected. Vaginitis was diagnosed in 180 (22.6%) of 796 female patients, predominantly caused by Candida spp. (66.1%) and bacterial pathogens (31.7%), with no Trichomonas vaginalis detected, and was most prevalent in women aged 20–30 years. Poor hygiene practices, including infrequent sanitary pad changes (OR = 5.01, p < 0.001) and routine vaginal douching (OR = 6.77, p < 0.001), were significantly associated with vaginitis. The Amsel criteria showed high specificity (99.1%) for bacterial vaginosis diagnosis. The absence of T. vaginalis suggests a potential shift in the epidemiology of vaginal infections. The Amsel criteria are a practical diagnostic tool in resource-limited settings. Our findings highlight the need for targeted hygiene education to reduce vaginitis prevalence in Vietnam. Full article

Talaromyces marneffei is a zoonotic dimorphic pathogen endemic to Southeast Asia and reported in 33 countries, with an estimated 17,300 human cases and 4900 deaths annually. We aimed to identify the best available evidence regarding the epidemiological and clinical features and the prevalence of T. marneffei reported in companion animals, wildlife, and humans in Europe. A systematic literature review was conducted by searching three databases under PRISMA guidelines for “Talaromyces marneffei” or “talaromycosis” in Europe or the equivalent. References from the obtained publications were also checked to identify additional papers that met the inclusion criteria. The search was not limited by language or year. Studies published until 30 April 2025 were included. Due to the limited number of publications on animals, the geographic scope was expanded to a global level. Of the 915 studies identified, 33 were eligible and categorised according to the subject they addressed: talaromycosis in humans (n = 26), talaromycosis in companion animals (n = 4), and talaromycosis in wildlife (n = 3). Talaromycosis has been reported 28 times in 11 different European countries among humans. Additionally, one case of T. marneffei in wildlife has been documented in Europe. There is a potential liaison host between bamboo rats and humans. Talaromycosis is an emerging planetary neglected disease. Confusion with other diseases and potential misdiagnosis leads to delayed diagnosis and unnecessary risk to lives. Immunocompromised and HIV-positive patients should be screened for talaromycosis. The unexplained worldwide reports in atypical species and locations prompt a call to action for a more proactive search for T. marneffei in other domestic and wild animals, as well as in soil, to fully understand its hosts and transmission, which must incorporate the Stockholm Paradigm and Planetary Health perspectives. Full article

Acute diarrhoeal disease caused by antibiotic-resistant diarrhoeagenic bacteria is a significant global public health issue, particularly in low- and middle-income countries. This study provides the first molecular characterisation of antimicrobial resistance profiles, including the detection of CTX-M-15 and CTX-M-55 extended-spectrum beta-lactamases (ESBLs), among diarrhoeagenic Enterobacterales in Bioko Island, Equatorial Guinea, offering novel epidemiological insights into an understudied region. This study investigated the antibiotic resistance profiles of pathogenic bacteria isolated from diarrhoeal samples on Bioko Island. A total of 153 clinical isolates were collected between 1 February and 30 May 2014, and antimicrobial susceptibility testing was performed at Loeri Comba Polyclinic (Malabo) using the Kirby–Bauer method. The molecular characterisation of β-lactamase-associated genes was performed on different isolates of diarrhoeagenic pathotypes—144 Escherichia coli, 7 Salmonella enterica, and 2 Shigella flexneri—at the National Centre for Microbiology (Majadahonda, Spain). High resistance rates were detected against ampicillin (98%), tetracycline (93.5%), sulfonamides (94.8%), sulfamethoxazole–trimethoprim (88.2%), and cefotaxime (78.8%), while moderate rates of resistance were noted for ciprofloxacin (26.7%), and all isolates remained susceptible to imipenem. Of the isolates, 107 (69.9%) produced either single or multiple β-lactamases. Among these, 73 (68.2%) harbored classical β-lactamases, specifically TEM and OXA-1 types, representing 47.7% of the total sample. Additionally, 34 (31.8%) of the isolates were identified as producers of extended-spectrum β-lactamases (ESBLs), specifically CTX-M enzymes. Sequencing identified CTX-M-15 and CTX-M-55 variants. The predominant ESBL-producing bacteria were enteroaggregative Escherichia coli (56.2%), followed by enteropathogenic and enterotoxigenic E. coli. These findings confirm the circulation of multidrug-resistant diarrhoeagenic Enterobacterales in Equatorial Guinea, raising concerns about limited treatment options due to widespread resistance to multiple antibiotic classes, including third-generation cephalosporins and quinolones. The most important conclusion drawn from this study is that a high percentage of diarrhoeagenic bacteria have an antibiotic resistance and multi-resistance profile, especially to beta-lactams and other groups of antibiotics such as tetracyclines and sulphonamides. There is also a moderate prevalence of isolates carrying ESBLs on Bioko Island, Equatorial Guinea, which could indicate the inappropriate use of antimicrobials. Full article

Sexually transmitted infections (STIs) remain a global issue, causing health problems and financial burdens. This study aimed to provide an update on the invasive pathogens and analyze any associated factors in patients visiting Da Nang Dermato-Venereology Hospital who were diagnosed with genital tract infections in males and lower genital tract infections in females; 535 participants underwent clinical examinations and microbiological tests to identify the invasive microorganisms, before we analyzed previously gathered laboratory results and associated risk factors. The rate of infection was 37.6% amongst 535 participants. Treponema pallidum infection accounted for the highest rate of 21.3%, followed by HSV, standing at 6%. The prevalence of syphilis infection was highest in the 20–29 age group, standing at 51.7%. There was a positive correlation between age under 20 and infected conditions (OR = 3.78, 95% CI: 1.41–10.11, p = 0.008). Having multiple sexual partners was identified as a risk factor for infection, as those with three or more sexual partners showed a high correlation (OR = 3.19, 95% CI: 1.44–7.05, p = 0.004). The high syphilis prevalence among young adults and teenagers underscores the need for improved STI education and screening programs in Vietnam. Full article

Hepatitis B and C (HBV and HCV) infections persist as significant public health concerns. Specific occupational groups, such as waste collectors, continue to face elevated risk due to exposure to contaminated materials. Research studies have underscored heightened infection rates, notably from needle prick injuries. The present study aspires to re-evaluate the prevalence of HBV and HCV among waste collectors and to scrutinize associated risk factors, thereby contributing to the development of effective public health and occupational safety strategies. The study analyzed data from 116 Italian waste collectors who were undergoing regular occupational visits, examining demographics, health conditions, HBV immunization, and annual blood tests for the HBsAg, HBsAb, and HCVAb. The HBsAb was detected in 66 individuals (56.9%), while the HCVAb was found in 4 (3.4%). Logistic regression showed HBV immunization and longer job experience to be significant factors associated with HBsAb presence. Waste collection may increase HBV infection risk due to occupational exposure, unlike HCV, which requires blood-to-blood contact. Preventive strategies, including education, protective equipment, and HBV vaccination, are essential. Ensuring vaccination coverage among waste collectors could reduce infection risk. Further research should assess the occupational risks and the effectiveness of preventive measures. Full article

Antimicrobial resistance (AMR) is a growing concern in laboratory animals, where antibiotic use can disrupt the gut microbiome and promote resistant strains. Rabbits, commonly used in biomedical research, are particularly susceptible to gut dysbiosis, increasing the risk of infection and subsequent antibiotic use. This study used 16S rRNA amplicon sequencing and PICRUSt2 to predict resistance-related functions in rabbits under standard laboratory conditions. Among 2427 amplicon sequence variants, 42 functional classes were identified, with AMR comprising 1.55% of the predicted functions, including beta-lactam, vancomycin, and cationic antimicrobial peptide resistance. These AMR functions were associated with commensal genera such as Bacteroides and Ruminococcus, while no associations were observed with pathogens such as Escherichia or Salmonella. The study provides functional evidence of intrinsic AMR in the rabbit gut microbiome and underscores the importance of antimicrobial stewardship in laboratory animal research. Full article

Surgical site infections (SSIs) are one of the most common causes of hospital-acquired infections worldwide, with significant clinical and economic implications. The aim of this review was to summarize the latest body of evidence on associations between microbial air load and SSIs. The systematic review was conducted using the revised Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA, 2020) method. Pubmed and Scopus databases were searched for the period 2014–2024. English language articles were searched for their reports on the microbial burden of operating room air and its association with surgical site infections. The present review includes a total of 36 articles related to microbial air load as an aggravating factor to air quality in the operating room and its association with SSIs. A direct correlation between microbial air load and the occurrence of SSIs was established through sampling methods and genetic analysis. A lack of consensus on the effectiveness of laminar air flow (LAF) systems was underlined, while temperature-controlled air flow seemed a promising alternative. One study found that each additional person in the operating room increases the number of bacterial colonies by 4.93 CFU/m³ while another did not find significant changes in air quality. More than 20 air changes per hour (ACH) appeared to have better results in improving the quality of the air in the operation room. Airborne microbial contamination is multifactorial, and for some of those factors, a revision of the guidelines seems necessary. Artificial Intelligence (AI) and Next-Generation Sequencing methods show great promise for improving air quality in the future. This review calls for the implementation of international guidelines regarding air contamination limits in operating rooms and standardized air sampling methods, as well as further research for the efficacy assessment of air flow systems and emerging technologies based on AI in order to reduce the burden of SSIs and improve patient outcomes. Full article

The vaginal microbiome can be perturbed by various intrinsic and extrinsic factors, resulting in a state of dysbiosis that decreases the abundance of commensal lactobacilli and often leads to pathological conditions such as bacterial vaginosis, yeast infections, sexually transmitted infections, and other vaginal disorders. This narrative review explores the molecular and pathophysiological mechanisms of several microbial diseases associated with the dysbiosis of the vaginal microbiome, as well as the efficacy of therapeutic tools for these conditions, such as antibiotic treatment and the use of live biotherapeutic products. A non-systematic, narrative approach was employed. Searches and data extraction were performed using the PubMed and Scopus databases between January and February 2025. All reviewed studies reported vaginal microbiome dysbiosis, with microbial pathogens inducing a specific immune response in the host. Current treatments for vaginal microbiota dysbiosis-related pathologies often result in high relapse and recurrence rates, suggesting microbial resistance and the need for alternative therapeutic strategies. In turn, live biotherapeutic products have demonstrated beneficial effects, restoring microbial balance in dysbiotic conditions. While these findings suggest promising potential for live biotherapeutic products, further rigorous clinical studies are necessary to gain a deeper understanding of the female genital tract ecosystem and to identify novel biomarkers along with their associated health implications. Moreover, the development of new diagnostic and management strategies will facilitate personalized therapeutic approaches. Ultimately, a comprehensive perspective on vaginal care is pivotal, taking into account both microbial and immune dynamics to enhance women’s health outcomes. Full article

The human microbiota plays a crucial role in maintaining host health through its considerable influence on immune function, nutrient metabolism, and overall homeostasis. While trace metals such as cobalt, lithium, and nickel are essential micronutrients at low concentrations, their increasing environmental accumulation presents emerging risks for microbial dysbiosis and related systemic health effects. This review examines the dual role of these trace metals as both beneficial nutrients and potential disruptors of microbial balance. Specifically, cobalt supports microbial diversity through its role in vitamin B12 synthesis, but excessive exposure can lead to dysbiosis. Lithium, beneficial at therapeutic concentrations by enhancing beneficial microbial populations, adversely affects gut barrier integrity by promoting inflammation and epithelial damage at higher concentrations. Similarly, nickel participates in essential enzymatic activities but promotes dysbiosis and inflammatory responses at elevated exposures. Furthermore, the growing environmental contamination by these metals poses risks to food systems and various microbial communities in the environment. Highlighting these environmental concerns, this review calls for sustainable management and multidisciplinary research to mitigate health risks to mitigate health risks associated with trace metal exposure. Full article

In developing countries like Bangladesh, livestock is one of the main sources of income. Among several infectious diseases, the Gram-positive bacterium Bacillus anthracis causes a zoonotic disease named anthrax. Animal anthrax outbreaks are a frequently occurring problem in Bangladesh. Our present study aims to molecularly identify and characterize B. anthracis from three districts of Bangladesh by 16S rRNA gene sequencing. B. anthracis was confirmed in soil, meat, and blood samples using PCR. Anthrax-affected soil (n = 128), blood (n = 1), and meat (n = 2) samples were analyzed using PCR. One of the positive samples was randomly chosen for sequencing, and MEGA5 software was used to generate the phylogenetic tree from the sequencing result. A total of 21 (16.40%) soil samples and all of the blood and meat samples were positive for the presence of bacteria, confirmed by PCR. The 16S rRNA gene of B. anthracis Sirajganj-1 was identical to that of other strains. To fulfill the Sustainable Development Goals, it is important to control zoonotic diseases. Our results may help discover the virulent genes of B. anthracis for future investigation and control this zoonotic disease. Also, a proper awareness of vaccination and effective surveillance system is important to eradicate any kind of zoonotic disease in developing nations. Full article

The present study investigates the prevalence and diversity of Plasmodium and Trypanosoma infections in Allochrocebus solatus, a vulnerable primate species native to Gabon. Using molecular techniques like nested PCR and phylogenetic analysis, we found 34.0% infection rate for malaria parasites infection, 21.3% for Trypanosoma spp., and 12.8% co-infections. Additionally, Hepatocystis was exclusively detected among malaria parasites, while Trypanosoma brucei brucei, T. vivax, and T. congolense were identified. These results underscore the complex host–parasite interactions influenced by captivity and the ecological and immunological consequences of such infections, particularly the increased susceptibility associated with captivity-induced stress. This preliminary study highlights the need for ongoing surveillance to mitigate health risks in primates and prevent potential zoonotic spillovers, providing critical data for conservation efforts. Full article

Device-associated infections (DAIs) are a significant public health concern because of their attributable mortality, along with the extra length of stay and cost. This two- year prospective surveillance study aimed to assess the incidence of DAIs and their clinical impact on four Greek adult medical-surgical Intensive Care Units (ICUs). Centers for Disease Control and Prevention (CDC) definitions were used to diagnose DAIs. Of the 500 patients hospitalized for 12,624 days, 254 (50.8%) experienced 346 episodes of DAIs. The incidence of DAIs was 27.4 episodes per 1000 bed-days. The incidence of ventilator-associated events (VAEs), central line-associated bloodstream infections (CLABSIs), and catheter-associated urinary tract infections (CAUTIs) was 20.5 episodes per 1000 ventilator-days, 8.6 episodes per 1000 central line-days, and 2.5 episodes per 1000 catheter-days, respectively. The most common pathogens isolated were Acinetobacter baumannii (35.7%) and Klebsiella pneumoniae (29.9%). All gram-negative pathogens were carbapenem-resistant. The ICU’s mortality was 44.9% for patients with DAIs and 24.8% for patients without a DAI (attributable mortality 20.1%, p < 0.001), while the mean ICU length of stay was 34.5 days for patients with DAIs and 15.6 days for patients without a DAI (attributable length of stay 18.9 days, p < 0.001). The high incidence of multidrug-resistant pathogens and the attributable length of stay and mortality of DAIs emphasize the need to establish an organized antimicrobial surveillance program and implement a care bundle for DAI prevention in ICUs with personnel educational training, monitoring, and feedback. Full article

Antibiotic resistance is a global health crisis exacerbated by the misuse of antibiotics in healthcare, agriculture, and the environment. In an intensive care unit (ICU), where high antibiotic usage, invasive procedures, and immunocompromised patients converge, resistance risks are amplified, leading to multidrug-resistant organisms (MDROs) and poor patient outcomes. The human microbiome plays a crucial role in the development and dissemination of antibiotic resistance genes (ARGs) through mechanisms like horizontal gene transfer, biofilm formation, and quorum sensing. Disruptions to the microbiome balance, or dysbiosis, further exacerbate resistance, particularly in high-risk ICU environments. This study explores microbiome interactions and antibiotic resistance in the ICU, highlighting machine learning (ML) as a transformative tool. Machine learning algorithms analyze high-dimensional microbiome data, predict resistance patterns, and identify novel therapeutic targets. By integrating genomic, microbiome, and clinical data, these models support personalized treatment strategies and enhance infection control measures. The results demonstrate the potential of machine learning to improve antibiotic stewardship and predict patient outcomes, emphasizing its utility in ICU-specific interventions. In conclusion, addressing antibiotic resistance in the ICU requires a multidisciplinary approach combining advanced computational methods, microbiome research, and clinical expertise. Enhanced surveillance, targeted interventions, and global collaboration are essential to mitigate antibiotic resistance and improve patient care. Full article

Cervical lymphadenitis in children, often caused by bacterial infections, may require surgical drainage if initial antibiotic treatment fails. Identifying factors associated with surgical intervention may aid in treatment decision-making and improve patient outcomes. We assessed the demographic, clinical, and laboratory factors associated with the need for surgical drainage in children diagnosed with cervical lymphadenitis. We conducted a retrospective cohort study of children diagnosed with cervical lymphadenitis or abscesses between 2015 and 2021. Data collected included demographics, clinical presentation, laboratory results, imaging findings, and treatment outcomes. Surgical intervention was compared to the conservative treatment group to identify predictors for drainage. Overall, 201 children were included; 87 (43%) underwent surgical drainage, while 114 (57%) received conservative treatment. In univariate analysis, young age (<3 years), leukocytosis (>15,000/mm³), and longer hospitalization duration (5.6 vs. 3.2 days, p < 0.001) were associated with surgical intervention. Methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) were the most common pathogens isolated from abscess cultures. In multivariate analysis, factors associated with surgical drainage included large lymph node size (>3 cm), erythema, and fluctuance in the physical examination and imaging (ultrasound) findings of abscess or phlegmon. Surgical intervention in pediatric cervical lymphadenitis was found to be associated with young age, large lymph nodes, erythema, and abscesses on imaging. Early assessment and recognition of these factors can guide the timely initiation of appropriate treatment, including surgery, when indicated. Full article

Tuberculosis (TB) is an infectious disease caused by bacteria that primarily target the lungs. The transmission of this disease occurs through the air in the form of droplet nuclei. Unfortunately, there has been an emergence of resistance to strains of such infections, such as multidrug- as well as extensively drug-resistant strains. Healthcare workers (HCWs) are particularly vulnerable to contracting TB due to their direct contact with patients. This study aims to evaluate the knowledge, attitude, and practices among Lebanese healthcare workers towards TB and its resistant forms, particularly multidrug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB). This study is a cross-sectional, descriptive, questionnaire-based research study that was conducted on HCWs in Lebanon. A total of 517 HCWs were included in this study. The findings displayed that 48.52% of HCWs had good knowledge of TB and MDR-TB/XDR-TB, 49.52% had average knowledge, and 2.13% had bad knowledge. Moreover, only 16.25% had a good attitude, 71.92% had an average attitude, and 11.8% had a bad attitude. Furthermore, only 14.7% had good practices, 54.74% had average practices, and 30.56% had bad practices. Having a history of testing for TB was found to be related to increased attitude. Also, the history of having TB-infected family members was found to be linked to both increased attitude and practice scores. Moreover, this study highlights the idea that high knowledge scores do not mean high attitude or high practices scores. On the same note, acceptable attitude scores do not inflict acceptable practice scores. The findings of this study showed that there is an overall good knowledge regarding TB, MDR-TB, and XDR-TB, average attitude, and average-to-bad practices. Gaps are seen in all sectors, even knowledge, especially with matters related to the diagnosis of MDR-TB/XDR-TB and its treatment duration. Also, the attitude section revealed a gap in the understanding of the modes of transmission of such an infection. Full article