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Degefa Bekala,1 Dawit Yihdego Reda,2 Musa Mohammed Ali2 1Bulchana Health Center, Oromia, Ethiopia; 2School of Medical Laboratory, College of Medicine and Health Sciences, Hawassa University, Hawassa, EthiopiaCorrespondence: Musa Mohammed Ali Email ysnmss@yahoo.comIntroduction: Leprosy is one of the neglected tropical diseases that affect skin and peripheral nervous system often results in severe, lifelong disabilities and deformities. Even though multidrug therapy was in place for more than 30 years to treat and prevent leprosy worldwide including Ethiopia, its epidemiology is not well studied in the West Arsi zone.Objective: The aim of this study was to determine the prevalence of acid-fast bacilli (AFB) positivity rate and associated factors among leprosy suspected cases.Methods: A health facility-based cross-sectional study was conducted among 422 leprosy suspected cases from August 2020 to December 2020. To detect AFB, skin slit specimens were collected and examined using the Ziehl-Neelsen staining technique. Socio-demographic and clinical data were collected using a structured questionnaire. Data were analyzed using Statistical Package for Social Sciences (SPSS) version 24. Logistic regression was employed to determine predictors of AFB positivity rate.Results: Acid-fast bacilli were detected among 46 leprosy suspected cases which gives a prevalence of 10.9% with 95% CI (8.2‒15.6). Suspected leprosy cases with multibacillary type were 4 times more likely to be AFB positive (p=0.021) than their counterparts. Study participants who had contact with known leprosy cases were 2 times more likely to be AFB positive (p = 0.032) and those with no formal education were 2 times more likely to be AFB positive (p = 0.03). Participants who had close contact with leprosy patients for ≥ 3 years were 8 times more likely to be AFB positive (p = 0.02).Conclusion: This study revealed a high prevalence of AFB positivity rate in the era of multidrug therapy. Types of leprosy, close contact with known leprosy cases, educational status, and duration of closer contact with leprosy cases were significantly associated with AFB positivity rate.Keywords: acid-fast bacilli, Ethiopia, Mycobacterium leprae, prevalence, West Arsi zone, Ziehl-Neelsen staining

Tuberculosis is one of the major causes of death worldwide, millions of people have been infected with pulmonary tuberculosis and over 95% of tuberculosis deaths occur in low and middle income countries as per World Health Organization report. The study assessed the performance of Ziehl Neelsen technique and GeneXpert used for diagnosis of Pulmonary tuberculosis in Uasin Gishu county health facilities, Kenya, between November 2019 and February 2020. A cross-sectional study was conducted in three selected health facilities and a total of 104 participants were enrolled into the study. Sputum samples from suspected tuberculosis patients were tested using Ziehl Neelsen technique, GeneXpert and culture method was used as gold standard. Performance of the tests was done by considering their sensitivity, specificity, predictive values. The results indicated that among the 104 sputum samples tested Ziehl Neelseen technique used detected 23(22.1%) bacilli from spot samples and 33(31.7%) bacilli from morning sputum samples. GeneXpert test detected 49 (47.1%) bacilli. The results also revealed that there was significant difference between Ziehl Neelsen technique using spot samples and GeneXpert and culture test (p-value< 0.05) and Ziehl Neelsen technique using morning test done with culture test had no significant difference (p-value>0.05). Performance of tests should be done accurately for proper diagnosis of TB, GeneXpert should always be used since its sensitivity is high and also detects Rifampicin resistance. Also, Performance of tests should be done using standard procedures to avoid errors in the test results.

Objective: To assess the performance of real-time polymerase chain reaction (RT-PCR) to diagnosis pulmonary tuberculosis in patients with scanty positive acid-fast bacilli sputum smears, in a single hospital. Materials and Methods: All patients, who had scanty positive AFB sputum smears in Songklanagarind Hospital; between 2015 and 2019 were included. Demographic data, clinical data, radiographic findings, RT-PCR and mycobacterial culture results were reviewed. Results: From a total of 269 patients reporting scanty AFB smears, 116 patients (43.1%) had cultures confirmed as M. tuberculosis. From overall, samples from 92 patients with scanty AFB smear were processed for RT-PCR. There were 26 (28.3%) isolates having positive RT-PCR test results. Of these 26 isolates that RT-PCR positive, 25 (96.2%) were culture positive, while only 1 (3.8%) were culture negative. A remaining 66 samples that RT-PCR negative, 15 (22.7%) were culture positive for tuberculosis. Using mycobacterial cultures as the gold standard, the sensitivity, specificity, positive predictive value and negative predictive value of RT-PCR were 62.5%, 98.1%, 96.2%, and 77.3%, respectively. Pulmonary consolidation and cavity on chest radiograph were associated with the growth of M. tuberculosis, with an OR of 2.3 (95% C.I. 0.26-0.73) and 3.4 (95% C.I. 1.2-9.9), respectively. Conclusion: Less than half of the patients with scanty smears had culture-confirmed tuberculosis; RT-PCR also has low sensitivity. Consequently, a negative RT-PCR does not exclude tuberculosis; especially in cases of a high index for clinical suspicion. Radiographic findings; including pulmonary consolidation and cavities, are helpful predictors for supporting this diagnosis.

Objective: To evaluate the cytospin slide microscopy method for detection of acid fast bacilli (AFB) in bronchoalveolar lavage (BAL) fluid comparing it with concentrated smear microscopy. Study Design: Cross-sectional validation study. Place and Duration of Study: Department of Microbiology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, from Dec 2016 to Sep 2018. Methodology: BAL specimens from suspected tuberculosis patients who were sputum smear negative, submitted to AFIP for diagnosis were included in the study. Smears for microscopy were prepared with the modified cytospin method as well as the standard concentrated technique. The prepared smears from both methods were stained with Ziehl–Neelsen (ZN) staining method and examined under 100 × oil immersion lens. TB culture performed on BACTEC MGIT 960 automated culture system (Becton Dickinson, USA) was taken as gold standard for TB diagnosis. The two methods were compared in terms of sensitivity, specificity, positive predictive value and negative predictive value. Results: Out of the 130 samples tested, 62 (47.7%) were positive on culture using MGIT 960 system. The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of the modified cytospin method for pulmonary TB diagnosis was found to be 68.3%, 100%, 100%, 77% and 84.6%, respectively. Conclusion: The sensitivity of the modified cytopsin smear method was significantly higher than that of the concentrated method. The study concludes that this is a simpler and more accurate method for BAL fluid microscopy.

Tuberculosis (TB) is a communicable disease caused by M. tuberculosis and known as Acid Fast Bacilli (AFB). The purpose of this study was to analyze the correlation between AFB at the end of the intensive phase and continuation phase and to analyze the variables that associated with sputum conversion at the end of continuation phase in pulmonary TB patient’s category I in several primary public health in Surabaya. This was an observational analytic with a retrospective cohort design. Data were collected by medical records of TB patients from January 2017 to December 2019. One hundred twenty-four of TB patients met the inclusion criteria with 69 male and 55 females, and mean of age was 44.09 ± 12.05 years old. The initial AFB was mostly 1+ (41%). There was a significant correlation between AFB at the end of intensive phase and continuation phase (p-value=0.000; r=0.657) by Kendal-Tau test. Age was significantly associated with sputum conversion at the end of continuation phase (p-value=0.022). The conclusion of this study that there was a significant correlation between AFB at the end of intensive phase and continuation phase

Canada is a low-incidence country for tuberculosis (TB). The BC Public Health Laboratory diagnostic algorithm for pulmonary TB includes acid fast bacilli (AFB) smear and mycobacterial culture of all submitted sputa. TB nucleic acid amplification testing (NAT) is routinely performed on AFB-smear-positive (AFB+) sputa only. We assessed the laboratory-associated costs of implementing the international recommendations for TB NAT on AFB-smear-negative (AFB-) sputa.Two data sets were obtained: (1) all AFB- samples for a 3-year period (October 1, 2014-September 30, 2017) and (2) all AFB-, TB-culture-positive samples for the same period. One AFB- sample/patient from each defined diagnostic set of sputa was deemed eligible for TB NAT. To stratify patients by ordering location, a 1-year subset of data (October 1, 2016-September 30, 2017) was examined.In the 3-year period, 0.7% of all diagnostic sets were AFB- and culture-positive. In the 1-year period, the provincial TB Services clinics submitted 26% of all AFB- samples received, but these constituted 78% of AFB-, culture-positive samples.The annual cost of TB NAT on one AFB- sputum sample from each eligible diagnostic set would total approximately $247,000. Targeting only TB Services clinic patients would reduce this cost to approximately $64,000/year while capturing more than 75% of AFB-, culture-positive patients. On the basis of our provincial positivity rate, it would cost approximately $6,000 to provide an early TB diagnosis for an AFB-, culture-positive patient. The cost-effectiveness to public health of this approach in a TB low-incidence setting needs to be carefully evaluated.Le Canada est un pays à faible incidence de tuberculose. L’algorithme diagnostique de tuberculose pulmonaire duLes chercheurs ont obtenu deux groupes de données : 1) tous les échantillons BAAR– sur une période de trois ans (du 1Pendant la période de trois ans, 0,7 % de tous les groupes diagnostiques étaient BAAR– et positifs à la culture. Pendant la période d’un an, les cliniques provinciales de services pour la tuberculose ont soumis 26 % de tous les échantillons BAAR– reçus, mais ils représentaient 78 % des échantillons BAAR– positifs aux cultures.Le coût annuel du TAN TB sur un échantillon d’expectoration BAAR– de chaque groupe diagnostique admissible totaliserait environ 247 000 $. Si on ciblait seulement les patients ayant consulté des services pour la tuberculose, ce coût fléchirait à environ 64 000 $ par année, mais saisirait plus de 75 % des patients BAAR– dont la culture est positive. D’après le taux de positivité provinciale, un diagnostic précoce de tuberculose chez un patient BAAR– positif à la culture coûterait environ 6 000 $. Le rapport coût-efficacité de cette approche pour la santé publique dans les milieux à faible incidence de tuberculose devra faire l’objet d’une évaluation attentive.

Obwohl die histologischen Charakteristika der Lungentuberkulose seit langem bekannt sind, ist die Diagnose einer nicht mikrobiologisch gesicherten Tuberkulose mit den der Pathologie zur Verfugung stehenden Mitteln eine Herausforderung. Eine nekrotisierende epitheliodzellige Granulomatose ist zwar fur eine Tuberkulose typisch, kommt jedoch auch bei einer Reihe anderer infektios, aber auch nichtinfektios bedingter Erkrankungen vor. Mikroskopische und molekularpathologische Verfahren konnen eine Tuberkulose bestatigen und ggf. eine andere Diagnose bahnen. Die Molekularpathologie findet jedoch am formalinfixierten und in Paraffin eingebetteten Material ihre methodischen Grenzen. Dies sollte den einsendenden KlinikerInnen gegenuber offen kommuniziert werden. Nach interdisziplinarer Reevaluation der Befunde muss daher bei negativem Ergebnis der Zusatzuntersuchungen entsprechend eine alternative Losung zur Diagnosesicherung gefunden werden.

Background: Microbiological confirmation of tuberculosis disease in children remains difficult due to paucibacillary disease and inability to obtain optimal samples. Recently introduced Cartridge based nucleic acid amplification test (CBNAAT) has improved microbiological diagnosis in pediatric tuberculosis. Objectives: We aimed to study association of CBNAAT grading based on cycle threshold value with conventional microbiological diagnosis. Methodology: This prospective study was conducted over a period from November 2016 to October 2017 in the Departments of Microbiology and Pediatrics, University College of Medical Sciences and Guru Teg Bahadur Hospital, Delhi. CBNAAT positive pediatric TB cases ≤12 years were recruited and subjected to Ziehl-Neelsen staining for acid fast bacilli (AFB) & culture on Lowenstein Jensen medium. CBNAAT positivity was graded based on cycle threshold value: very low, low, medium and high. Results: Smear and culture positivity was highest (100%) among specimens with high positive CBNAAT result based on CT value. Time to culture positivity was inversely related to CBNAAT grading (p=0.000). Conclusion: CBNAAT grading has significant positive association with smear and culture positivity. Bangladesh Journal of Infectious Diseases 2020;7(2):72-77

Mycobacterium paratuberculosis causes Johne's disease in cattle, a chronic granulomatous enteritis for which a killed vaccine is commercially available. We present a case of a veterinarian who had an unintended needle-stick with injection of vaccine material. He developed a granulomatous dermatitis with acid-fast bacilli identified on pathology of affected tissue. It is important for health-care providers who care for patients at risk for unintended vaccine exposure to be aware of the possible sequelae after exposure and provide appropriate treatment based on the severity of the injury and subsequent tissue response.

Background Active case finding (ACF) of tuberculosis (TB) has been recommended by the WHO for early detection of pulmonary TB. Nonetheless, there is little evidence about the cost-effectiveness of different ACF strategies. Objective The aim was to assess the cost-effectiveness of ACF using sputum Xpert MTB/RIF and acid-fast bacilli tests. Materials and methods Economic modeling was employed to assess the effectiveness and costs of three ACF strategies, based on applying Xpert MTB/RIF (‘Xpert only’), acid-fast bacilli (‘AFB only’), or both AFB and Xpert (‘AFB+Xpert’) in the screening protocols. Outcomes included TB case detection rate, lost to follow-up rate, costs, and incremental cost-effectiveness ratios of the strategies. Costs were estimated in US dollars from the societal perspective, with base year 2018. Results The ‘AFB only’ strategy showed a low case detection rate with higher costs, in relation to other strategies. Compared with the ‘Xpert only’ strategy, a rising rate of lost to follow-up was observed from the ‘AFB+Xpert.’ The case detection rate in the ‘Xpert only’ strategy is approximately twice the rate compared with the ‘AFB+Xpert’ strategy. The costs per TB case detected of the ‘AFB+Xpert’ and the ‘Xpert only’ strategies were $17 778.33 and $8334.47, respectively. The incremental cost-effectiveness ratio of the ‘Xpert only’ strategy was $257.28 per case detected as compared with the ‘AFB+Xpert’ strategy. Conclusion This study showed that the ‘AFB only’ strategy was dominated, in comparison with other strategies. It should not be recommended in the settings where application of Xpert MTB/RIF is feasible.

This study investigates the association of the AFB stain with the cycle threshold (Ct) value of the Cobas TaqMan MTB test (CTM test, Roche Diagnostics, Basel, Switzerland), and it establishes the base data for semi-quantitative identification of M. tuberculosis by the Ct value. CTM test were simultaneously conducted on 8,389 specimens submitted to the Samsung Medical Center from January 2015 to December 2015, and the results were analyzed and compared retrospectively investigates the association of the AFB stain with the Ct value of the CTM test, and it establishes the base data for semi-quantitative identification of M. tuberculosis by the Ct value. The Ct values for 135 positive specimens of the CTM were inversely correlated with the AFB stain (rs=−0.545, P＜0.01). When the Ct value of the CTM test and the time to positivity (TTP) of the mycobacteria cultures were verified based on the AFB stain, they were found to have a positive correlation (rs=0.136, P＜0.01). The negative correlation between the CTM test and the AFB stain grade was demonstrated. The clinical significance was verified by applying these criteria to the clinical results. The semi- quantitative criteria of this study can be used to facilitate the rapid isolation of patients with active tuberculosis and infection control in the hospital.

Cryptosporidium is recognized globally as a major etiology of persistent and chronic diarrheaespecially in immunocompromised patients and children with significant morbidity and mortalityrates. Moreover, it is one of the most important pathogens causing waterborne outbreaks.Hence, rapid diagnosis is crucial. In this study, we tried to determine the diagnostic performanceof Rida®Quick Cryptosporidium cassettes versus microscopy of the modified acid-faststained smear. Forty fecal samples were collected from immunocompromised patients complainingof chronic diarrhea. All samples were examined by Rida®Quick Cryptosporidium cassettesfor copro-antigen detection, and microscopically using concentration techniques andmodified acid-fast staining. Cryptosporidium was detected in (45%) of immunocompromisedpatients. The sensitivity, specificity, PPV, and NPV of Rida®Quick Cryptosporidium cassetteswere 88.9%, 95.5%, 94.1% & 91.3% respectively with an accuracy of 92.5%. There was nocross-reactivity with other intestinal parasites. Rida®Quick Cryptosporidium cassettes provideadequate sensitivity and specificity and give rapid results. Our results recommend its use as analternative test in certain situations especially when screening large populations is needed urgentlyin a short time, as in outbreaks and epidemiological surveys.

Drug-resistant tuberculosis (DR-TB) is a worldwide global burden and related to poor treatment outcomes. Monitoring the progress of DR-TB treatment can be carried out microscopic with Acid-Fast Bacilli (AFB) sputum smear and assessed from the beginning or the first time the patient is diagnosed and monthly to determine the sequential conversion of AFB baseline to the next month twice until it becomes negative. The prolonged conversion has been associated with infectiousness and treatment outcomes. This study was aimed to determine the correlation between positivity of AFB sputum and time to conversion in DR-TB patients with short-term treatment (STR) regimen. An analytic retrospective at hospital collected from medical records of DR-TB patients, from September 2017 to July 2018. Spearman technique was used to analyze the data with p < 0.05. From the total of 151 DR-TB patients on STR regimen, 51 patients were enrolled consisting of 30 (58.8%) males and 21 (41.2%) females with ages average were 51 ± 12.9 years old. Overall, 39 (76.5%) patients had time of AFB conversion in the first month, 9 (17.6%) patients in the second months, 2 (3.9%) patients in the third months, and 1 (2%) patient in the fourth months. Among those patients, 26 (51%) patients had completed the treatment outcomes, 22 (43.1%) were loss to follow-up, 1 (2%) patient had the treatment failure, and 2 (3.9%) were died. There was no significant correlation between AFB sputum baseline (Scanty, 1+, 2+, 3+) with the time to sputum conversion (p > 0.05). AFB sputum have significant correlation with time of culture conversion (p < 0.05), and treatment compliance was not associated with time of AFB sputum conversion (p > 0.05). There was no significant relation between positivity of AFB baseline and time to sputum conversion in DR-TB patients on short-term treatment (STR) regimen. AFB sputum have significant correlation with time of culture conversion in DR-TB patients on short-term treatment (STR) regimen.

Background The spectrum of morphological pattern in tubercular lymphandenopathies was observed to study the various cytomorphological patterns and their correlation with acid fast bacilli. Methods FNAC smears of 210 cases of granulomatous lymphadenitis stained with Giemsa, Pap and haematoxylin and eosin were used to analyze cytomorphological pattern and Zeihl Neelsen stained smears for acid fast bacilli (AFB) detection. Results 193 cases with necrotising granulomatous inflammation or positive acid fast bacilli were included. Age group 21–30 years was most common (38.3%) followed by age group 11–20 years (30.05%). Females constituted 66.3% of patients and 33.7% were male. Overall the most common pattern in present study was pattern A (Epitheloid granuloma with caseous necrosis 33.7% followed by pattern B (caseous necrosis with few scattered epitheloid histiocytes and lymphocytes) 31.1% and pattern C (caseous necrosis with suppurative inflammation) 30.6%, followed by pattern D (Caseous necrosis only) (3.6%) and pattern E (non necrotising epitheloid granuloma with positive acid fast bacilli) (1.03%). Acid fast bacilli were demonstrable in 175 cases (90.7%). Amongst the acid fast bacilli positive cases highest bacillary load 3+ grade was seen in pattern C in 6/59 (10.16%) cases. Conclusion FNAC is a simple useful tool and should be attempted in all cases of lymphandenopathies. It helps in establishing a diagnosis of tubercular etiology based on its morphological patterns however demonstration of acid fast bacilli on aspirated material confirms the diagnosis.

We studied 31 US healthcare facilities to characterize trends in mycobacterial testing. During 2009-2015, testing for acid-fast bacilli increased 3.2% annually, and prevalence of pathogenic nontuberculous mycobacteria increased 4.5% annually. These increases were highest for subpopulations at high risk of infection, including older women, Asians, and patients with concurrent conditions.

Tuberculosis (TB) basically originates due to bacteria, influences the developing nations and disrupts their economy severely. TB like diseases are with a high mortality rate worldwide, but early detection highly increases the chances of survival. This paper presents a novel method for TB bacteria segmentation and classification using microscopic images (MI). Manual identification of the bacterial cell is a very difficult process. The automation in TB bacteria detection is the objective of this article using MI processing. The proposed segmentation method first performs the image enhancement followed by bacteria region masking. Further, the marking of bacteria points is performed by the marked point process model. Finally, the complete bacteria are identified by the superellipse and supervised variational contour models. The features are extracted using bag of visual words and handcrafted work for the image classification. Simulation results confirm the superiority of the proposed method as compared with the state-of-the-art methods.

Background: Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis which enters the droplet nuclei into the mouth or nasal passages to reach the pulmonary alveoli. The sputum smear examination for smear-positive and negative smear pulmonary tuberculosis is challenging due to requiring a longer time to get the results. Therefore, a specific marker is needed that is able to detect bacterial infections immediately, such as procalcitonin (PCT). This study aims to determine the levels of PCT in pulmonary tuberculosis patients with acid-fast bacilli positive and negative smear at Haji Adam Malik General Hospital, Medan, Indonesia.Method: A cross sectional was conducted among 40 pulmonary tuberculosis patients who met the inclusion and exclusion criteria at Haji Adam Malik General Hospital, Medan, Indonesia. The subjects were 20 positive smears and 20 negative smears. PCT examination with mini VIDAS BRAHMS was carried out using the Sandwich principle using the ELFA method (Enzyme-Linked Fluorescent Assay). Sputum examination was carried out by smear of Zhiel Neelsen smear, which was analyzed by the Mann-Whitney (Non-Parametric) test using SPSS version 17 for Windows. Results: Most of the respondents were males in both positive and negative smears group (65.0% and 70.0%, respectively) and not significantly different (P>0.05). The age of subjects was slightly older in positive smear (49.75 ± 17.993 years) compare with negative smear group (42.50 ± 14.816 years) but not statistically significant (P=0.172). The PCT levels in pulmonary tuberculosis patients with positive smear (0.1550; 21.65 ng/mL) differ significantly from the PCT levels in pulmonary tuberculosis patients with negative smear (0.05; 3.14 ng/mL) (p=0.0001). The cut off value using ROC found 0.06 ng/mL (AUC: 0.842) with a sensitivity of 80% and a specificity of 80%.Conclusion: There was a significant difference in the level of procalcitonin in patients with pulmonary tuberculosis who smear-positive and smear-negative.

Cryptosporidium intestinal coccidian is a preeminent account for infectious diarrhea with reference to children. Oocyst may escape ordinary microscopic analysis or immune detection; molecular techniques are of convenient use. This study intent to detect cryptosporidiosis infection among children, and debates risks for infection. A total of 189 children with age range from under one to five complaining of gastrointestinal manifestations were examined coproscopically, by acid fast stain, ELISA and nested PCR (nPCR) for Cryptosporidium from June to September. Infection was proved by nPCR among 13 (6.8%) samples. ELISA sensitivity merit stool staining (77%, 53.8%, respectively), however all positive samples by microscopy were settled by nPCR. Age groups (P = 0.02) were statistically associated with infection. Source ofwater (OR=0.069, 95% CI= 0.004/1.165, P = 0.01) and presence of undigested food particles on microscopy (OR= 1.058, 95% CI= 0.521/6.154, P = 0.02) were risks for infections. No other variables were predicated risks. Infection was potent among preschool children (46.2%), toddlers (38.5%) and then infants (15.4%). Cryptosporidiosis should be doubtful among children with diarrhea during preschool age. Source of water for ordinary drinking activities or for infantswith non-exclusive breast or complementary feeding should be considered as hazards aspects for acquiring Cryptosporidium.

Introduction: Indonesia is a high incidence country of multidrug-resistant tuberculosis. There are approximately 11,000 MDR TB cases, 2.8% of them are new cases and 16% of them are relapse cases. Although guidelines for MDR-TB are frequently designed, medication freely provided, and centers for treatment duly expanded, studies on time to sputum culture conversion have been very limited in Indonesia. Therefore the aim of the study is to identify risk factors that effect on sputum and culture conversion.Methods: Data on short-term regimen for MDR TB in Dr. Soetomo General Hospital from January 1st – December 31st, 2018 were collected with a total sampling approach, and fulfill the inclusion and exclusion. Data were analyzed by computer software IBM SPSS Statistic 24 for windows. Results: Male is more likely to have delayed sputum conversion and culture conversion but no statistical difference is observed (p>0.05). Smoking history is more likely to have delayed sputum and culture conversion but only in sputum smear test shows a significant difference (p≤ 0.05). Alcohol consumption has delay effect on sputum and culture conversion but there is statistically difference in only culture conversion (p

— Bovine nodular thelitis in dairy cows is a chronic and enzootic infection. Four stages were distinguished: early stage, evolutive stage, ulcerative stage and final stage. Lesions were located mainly in the lower part of the udder or on the teat and recognized as solid papules and nodules. The histopathological study revealed a tuberculoid granulomatous pattern characterized by a subacute focal dermatitis with lymphoid cells, macrophages, epitheloid cells and a variable number of multinucleate giant cells (Langhans type) but no necrosis. Acid-fast bacilli were demonstrated in two of 14 cases. Three strains of acid fast bacilli were isolated but the only one identified was Mycobacterium terrae. M. terrae is a non-pathogenic slowly growing, non-photochromogenic mycobacterium. Its role in tuberculosis-like skin lesion is the bovine udder is uncertain. Resume— La thelite nodulaire est une infection chronique et enzootique chez les vaches laitieres. On distingue 4 phases: une phase precoce, une phase evolutive, une phase ulcerative et une phase finale. Vingt echantillons histologiques, mammelles et trayons, ont ete examines. Les lesions etaient localisees essentiellement a la partie basse de la mammelle ou sur le trayon et se presentaient sous l'aspect de papules solides ou de nodules. Les echantillons etaient divises en deux parties: l'une destinee a l'isolement de mycobacteries, l'autre a l'examen histologique. Les lesions histologiques des trayons etaient des granulomes tuberculoides, caracterises par une dermatite subaigue focale avec presence de lymphocytes, macrophages, cellules epithelioides et une nombre variable de cellules multinucleees geantes (type Langhans). Les infiltrats inflammatoires etaient principalement perivasculaires. Parfois, ils infiltraient toute la paroi du trayon. Aucune necrose n'a ete observee. Des bacilles acidoalcooloresistants furent mis en evidence de maniere inconstante (2 cas sur 14). Trois types de bacilles acidoalcooloresistants ont ete isoles raais, un seul a ete identifie comme etant Mycobacterium terrae. M. terrae est une mycobacterie non pathogene, de croissance lente et non photochromogene. L'implication de cet organisme dans les lesions de type tuberculeux dans la mammelle de la vache est discutee. Zusammenfassung— Bei Milchkuhen stellt die bovine nodulare Thelitis eine chronische und enzootische Infektion dar. Vier Stadien lassen sich unterscheiden: Frustadium evolutorisches Stadium, ulzeratives Stadium und Endstadium. 20 pathologische Proben von Euter und Zitzen wurden untersucht. Die Veranderungen waren hauptsachlich am unteren Teil des Euters oder an den Zitzen lokalisiert und liesen sich als solide Papeln oder Knoten ansprechen. Das Untersuchungsgul wurde fur die Isolierung von Mykobakterien und fur eine histopathologische Untersuchung geteilt. Die histopathologischen Veranderungen der Zitzen zeigten ein tuberkuloides granulomatoses Bild, das durch subakute fokale Dermatitis mit lymphyoiden Zellen, Makrophagen, Epitheloidzellen und einer variablen Anzahl von vielkernigen Riesenzellen (Langhans-Typ) gekennzeichnet war. Die entzundlichen Infiltrate befanden sich hauptsachlich perivaskular. Manchmal liesen sie sich aber auch in der gesamten Zitzenwand nachweisen. Nekrosen bestaznden nicht. Saurefeste Bazillus-Keime waren inkonstant vorhanden (2 von 14 Fallen). Drei Arten von saurefesten Bazillus-Keimen wurden isoliert, aber nur eine wurde speziell identifiziert (Mycobacterium terrae). M. terrae ist ein nicht pathogenes, langsam wachsendes und nich photochromogenes Mykobakterium. Die Beteiligung dieses Organismus bei den tuberkulose-ahnlichen Hautveranderungen am bovinen Euter wird diskutiert. Resumen La Telitis Nodular Bovina es una infeccion enzootica cronica de las vacas lecheras. Se distinguen cuatro etapas: etapa inicial, etapa evolutiva, etapa ulcerativa y etapa final. En el presente trabajo se examinaron 20 muestras de mama y de pezon. Las lesiones se localizaban principalmente en la parte inferior de la glandula mamaria o en el pezon y consistian en papulas solidas y en nodulos. Las muestras se dividian en dos partes, una para el aislamiento de micobacterias y la otra para los estudios histopatologicos. Las lesiones histopatologicas consistian en una reaccion granulomatosa de tipo tuberculoide, caracterizada por una dermatitis focal subaguda con celulas linfoides, macrofagos, celulas epitelioides y un numero variable de celulas gigantes multinucleadas (tipo Langhans). Los infiltrados inflamatorios estaban localizados predominantemente en la zone perivascular. En ocasiones, aparecian a lo largo de toda la pared de la glandula mamaria. No se observo necrosis. Se han aislado tres cepas de bacilos acido-alcohol resistentes, pero solo una fue identificada especificamente como Mycobacterium terrae. M. Terrae es una micobacteria no fotocromogenica de crecimiento lento y no patogena. Se discute la posible participacion de este microorganismo en la genesis de las lesiones pseudotuberculosas observadas en estos animales.

Extrapulmonary Tuberculosis [EPTB] affects mainly lymph nodes, pleura, central nervous system, bones, gastrointestinal tract and genitourinary tract. Most commonly it presents as isolated lymphadenopathy and also as effusion in the body cavities. Tests to diagnose EPTB include Fine Needle Aspiration Cytology [FNAC], Acid Fast Bacilli [AFB] demonstration by Zeihl-Neelson [Z-N] stain and Cartridge Based Nucleic Acid Amplification Test [CBNAAT]. A retrospective record based study was conducted by taking the data of 100 clinically suspected EPTB patients who had undergone FNAC, CBNAAT and Z-N stain in Mandya Institute of Medical Sciences, Mandya, Karnataka. The study was conducted for a period of four months from March 2018 to June 2018 after taking approval from the Institutional Ethics Committee. The study showed 47 cases with FNAC findings favouring EPTB. Out of these 47 cases, 30 cases were CBNAAT positive. CBNAAT was positive in 5 cases out of 53 cases with negative FNAC findings. Among 35 CBNAAT positive cases, only 13 cases were AFB positive. To conclude, CBNAAT is rapid, accurate and simple test for early diagnosis of EPTB because of its high specificity and positive predictive value. CBNAAT can also detect the cases missed by FNAC and Z-N stain.

Objective To compare the performance of the Fluorescein diacetate (FDA) vital staining method with Ziehl-Neelsen staining method in detecting the viability of acid-fast bacilli using MGIT culture as “reference standard”. Methods This was a single centre prospective observational study conducted from October 2015 to November 2016. Microbiologically confirmed ZN-Smear positive (3+) sputum specimens were obtained from 30 pulmonary tuberculosis patients taking anti-tuberculosis treatment at DOTS centre of NITRD, New Delhi. Patients were made available to collect the first baseline sputum sample before commencing treatment, and an early morning sputum sample was collected as per RNTCP guidelines. After starting treatment, sputum specimens were collected weekly in the first month and thereafter twice-weekly until 18th week. All sputum specimens from patients receiving anti-tuberculosis treatment were examined using Ziehl-Neelsen (ZN) smear microscopy, FDA vital staining, and MGIT culture. Result Out of 360 follow up sputum specimens collected from 30 adult microbiologically confirmed ZN- Smear (3+) pulmonary tuberculosis patients, 146 were ZN-positive and 130 FDA-positive. Of 130 FDA-positive sputum samples, mycobacteria tuberculosis (MTB) growth was found in 116 sputum samples, of which 116 sputum specimens were positive for FDA. Additionally, 14 culture-negative specimens were FDA positive. No FDA-negative sputum samples were positive for MGIT culture. Among ZN positive specimens, FDA had 100% sensitivity and 85.3% specificity with an accuracy of 96.58% for the detection of viable mycobacteria. Among ZN negative sputum specimens, FDA had comparatively high specificity (95.7%). Using positive MGIT culture as a reference for viability, negative predictive value (NPV) and positive predictive value (PPV) from FDA vital staining method were found to be 100 and 89% respectively. Conclusion FDA staining is a simple and rapid tool for identifying viable MTB bacilli. Because of its excellent NPV and encouraging specificity, FDA staining is useful to identify patients with non-viable bacilli (FDA negative) among retreatment cases at diagnosis and patients on anti-tuberculosis treatment for both drug-sensitive and drug-resistant tuberculosis for follow up for the response of treatment.

Mycobacterium genavense, a nontuberculous Mycobacterium, is found in immunosuppressed patients, particularly in those with HIV. Mycobacterium genavense incubation under standard culture conditions is difficult, and its identification is challenging using routine culture methods. Herein, we report the case of a 40-year-old Japanese man with HIV presenting with disseminated M. genavense infection. An analysis using an automated blood culture system did not show positive signals during 6 weeks of incubation. However, an acid-fast bacilli smear of his blood sample was positive for the bacterium. Mycobacterium genavense was identified using sequencing analysis, targeting the heat shock protein 65 gene. The patient recovered from the infection, following antibiotic therapy for 18 months. Under suspicion of disseminated M. genavense infection and the absence of bacterial growth in blood culture samples, an acid-fast bacilli smear test of the sample may be useful for timely diagnosis.

Osteoarticular tuberculosis (TB) is an uncommon form of extrapulmonary TB, comprising approximately 5% of all TB and 10%-15% of extrapulmonary TB cases. Multifocal skeletal TB is rare and accounts for 10% of all osteoarticular TB cases. Sometimes, the diagnosis is difficult. The potential delay in the clinical diagnosis may be critical for patients since it can cause the spread of the infection from the bone to the adjacent joints and surrounding tissues. We present a rare case of military TB with multiarticular involvement in a patient with chronic tophaceous gout. The initial diagnosis was confirmed throughout the positive analysis for Ziehl-Nielsen acid-fast staining in synovial fluid of two different joints, which is unusual. The patient was treated with antituberculosis drugs and presented good recovery signs.

Introduction: Tuberculosis (TB) is one of the world’s deadliest disease. Presently, one fourth of the world’s population is thought to be infected with TB. Early diagnosis of tuberculosis is very important for therapeutic reasons and to control the spread of infection. The purpose of this study was to compare the efficacy of fluorescence (FL) microscopy in comparison to Ziehl- Neelsen (ZN) staining Methods: A cross-sectional study of 310 TB suspects attending Dhaka Medical College hospital subjected to four sputum examinations by ZN and FM. Löwenstein- Jensen culture was used as the gold standard. Of 310 suspects enrolled, 304 (98%) had a complete diagnostic work-up involving three sputum specimens for ZN and FM, culture and chest X-ray (CXR). Result: Out of 320 suspected case, 10 persons were out of our study. Among 310, 204 suspected case sputum culture was positive. Though we took sputum culture as gold standard, so there were 204 tuberculosis patient present in our study. Out of 204 TB patient 125 (61.2%) cases were positive in ZN staining. Negative ZN methods were also high 79 (39.7%) which indicate less sensitivity. On the other hand, FM methods showed 193 (94.6%) cases positive out of 204 sputum culture positive patient. It also presence of 11 cases which showed negative result in this methods. From this data we can assumed higher sensitivity of FL methods from ZN methods. Here 106 cases were culture negative. Out of these 106 people only 05 persons were positive in ZN staining whereas 10 persons were find to positive in FL methods. This message demonstrates us that specificity of ZN methods are slightly higher over FM methods. Overall sensitivity of ZN microscopic (61%)) methods are much lower than FL microscopic methods (95%). On the other hand, specificity was slightly reverse, which showed 95.5% in case ZN methods and 91% in FL methods. It also demonstrates that positive predictive value of ZN microscopy (96%) is slightly higher than FL microscopic (95%) methods but negative predictive value showed gross difference ZN microscopic (56%) and FL microscopic (89%) methods. Conclusions: The FL methods is more sensitive and shortens the diagnostic process of tuberculosis. Consequently, more patients can be put on a regimen for smear-positive TB, contributing to improved treatment and reducing transmission. For that Front-loaded (FL) microscopic approach may apply for early diagnosis and detection of tuberculosis. J MEDICINE JAN 2020; 21 (1) : 36-40

Infectious agents and chemical substances can trigger the production and the release of cytokines. To determine the pattern of infectious agents in herbal homes patients with elevated TNFα (≥ 5.0 pg/ml) in Saki – West-Nigeria. The subjects include patients with elevated TNFα (≥ 5.0 pg/ml) in 15 herbal homes(n=23 ; aged 5-69 years; males – 13 ; females – 10) and patients with elevated TNFα in 3 major hospitals (n = 32; aged 6 -67years; males – 12 ; females - 20). Plasma TNFα, anti-HCV, HBsAg and HIV1-p24 antigen were determined by ELISA, Plasmodium spp., by Giemsa thick film staining and Acid Fast Bacilli by Ziehl-Nelson staining. The frequency of infectious agents obtained in the patients include 4.3 %(1) anti-HCV; 17.4% (4) HBsAg; 4.3 %(1) HIV1-p24 antigen; 17.4% (4) Plasmodium spp.; 8.7 %(2) Acid Fast Bacilli and 4.3 %(1) HBsAg + Plasmodium spp., in patients of herbal homes and 6.3 %(2) anti-HCV; 9.4% (3) HBsAg; 3.1 %(1) HIV1-p24 antigen; 15.6% (6)Plasmodium spp., 0%(0) Acid Fast Bacilli and 3.1 %(1) HBsAg + Plasmodium spp., in patients of the three major hospitals. The frequency of those with 5.0 – 6,0 pg/ml TNFα (65.2% (15)) was more than those whose TNFα was between 6.1 – 6.8 pg/ml (34.8% (8) in herbal home patients while the frequency of those with 5.0 – 6,0 pg/ml TNFα (40.6%(13)) was lower than those whose TNFα was between 6.1 – 6.8 pg/ml (59.4% (19)) in hospital patients. The pattern of the infectious agents was more in herbal home patients with 5.0 – 6,0 pg/ ml TNFα than those with 6.1 – 6, 8 pg/ml TNFα while in hospital patients it was more in those with 6.1 – 6,8 pg/ml TNFα than those with 5.0 – 6,0 pg/ml TNFα. The overall frequency of the infectious agents was more in herbal homes patients (58.5%(13)) than 37.5%(12) obtained in hospital patients. The frequency of viral immunochemical biomarkers, Acid Fast Bacilli and Plasmodium spp., was more in herbal home patients than the patients recruited from the hospital which varies with the TNFα range.

Microscopic examination of the specimen smear for acid fast bacilli (AFB) provides a simple and rapid means of detecting AFB using fluorescent stain methods and remains a valuable diagnostic test used worldwide to identify and manage suspect cases of tuberculosis (TB). Methods to improve AFB smear staining protocols could provide better detection of suspect TB cases. In particular, decreasing background debris may improve the detection of smears with low numbers of bacilli. We assessed staining by the standard rack method compared to bulk container staining using an acetone rinse step to decrease background debris. No cross-contamination was observed in the bulk container staining, and higher accuracy with less reading time was achieved with the acetone rinse. Most importantly, more bacilli were detected per positive smear using the acetone rinse method.

Aim: We determine the prevalence of TB and MDRTB within the study population. Methodology: The sampling consists of 192 (54.4%) males and 161 (45.6%) females. Ziehel Neelsen staining method was used for the detection of Acid Fast Bacilli (AFB) in the sputum samples collected. Rifampicin resistant (MDRTB) isolates were detected by GeneXpert and the conventional PCR technique. Results: In this study, 6 (9.5%) out of the 63 positive samples were rifampicin resistant Mycobacterium tuberculosis in which 4(6.3%) were found among females and 2 (3.2%) were found among males. Infection with Mycobacterium tuberculosis was not dependent on gender statistically in this study P>0.05. Secondary institution among the health care centers had highest cases of tuberculosis18 (33.3%) while tertiary institute had higher rifampicin resistant tuberculosis 5(2.5%). Statistically there was significant relationship between occurrence of tuberculosis and health care centers p

Diagnostic procedures in pediatric dermatology are different and significantly more challenging than those in adult patients, especially on how to approach the patients. We need to acknowledge that pediatric patients have unique anatomical, physiological, and psychological aspects. Compared to adults, children have smaller procedure area, are less cooperative, and more difficult to understand. For these reasons, we should perform diagnostic procedures on children cautiously. Having extensive knowledge in this field can facilitate us to carry out an ethical, efficient, targeted procedure with less risk. This article reviewed the most appropriate, most comfortable, and least invasive diagnostic procedures for children. Some of the most commonly performed pediatric dermatology procedures highlighted in this article are potassium hydroxide examination, Gram staining examination, acid fast bacilli examination, skin test for allergies, and skin biopsy for children. We also discuss the process of obtaining a written informed consent from the parents who were involved in decision making process and play a crucial role in assisting physicians to calm their children during the diagnostic procedures. Keywords: children, dermatology, diagnostic procedures

It was established that when stored for many years (10–13 years) in low-temperature conditions (3°C), without sub-culture on a nutrient medium, Mycobacterium bovis grew as visible colonies along the line of inoculation. However, due to long-term storage in conditions of low temperature (3°C) morphology of mycobacteria differed significantly from initial cultures formed by rod-shaped bacteria. Some of them became pigment-forming and smooth on the surface. Unlike the initial strain of mycobacteria, a perennial bacteria stored under hard conditions did not cause the death of guinea pigs or their sensitization to a purified protein derivative for mammals. Morphological forms of the perennial mycobacteria had the following changes: pigment forming, L-forms of the vesicular type, non-acid-fast thread-like (filamentous) bacillary forms, and elementary bodies when compared to the initial strain. There were also some genetic changes in the target DNA due to the long-term storage of M. bovis. It may indicate a mutation in the pathogen’s DNA. These mycobacteria had altered biochemical activity during storage. The number of passages on the solid nutrient medium did not affect their fermentative activity. However, the low cultivation temperature increases mycobacterial catalase activity and the ability to hydrolyze Tween-80.

Study Design: Retrospective study.Purpose: To identify the rate of positive acid-fast bacillus (AFB) and fungal cultures during spine debridement, determine whether these infections are more common in certain spine segments, identify comorbidities associated with these infections, and determine whether the universal performance of fungal and AFB cultures during spine debridement is cost effective.Overview of Literature: Spine infections are associated with significant morbidity and costs. Spine fungal and AFB infections are rare, but their incidence has not been well documented. As such, guidance regarding sample procurement for AFB and fungal cultures is lacking.Methods: A retrospective review of medical record data from patients undergoing spine irrigation and debridement (I&D) at the University of Missouri over a 10-year period was performed.Results: For patients undergoing spine I&D, there was a 4% incidence of fungal infection and 0.49% rate of AFB infection. Steroid use was associated with a higher likelihood (odds ratio, 5.62; 95% confidence interval, 1.33–23.75) of positive fungal or AFB cultures. Although not significant, patients undergoing multiple I&D procedures had higher rates of positive fungal cultures during each subsequent I&D. Over a 10-year period, if fungal cultures are obtained for each patient, it would cost our healthcare system $12,151.58. This is compared to an average cost of $177,297.64 per missed fungal infection requiring subsequent treatment.Conclusions: Spine fungal infections occur infrequently at a rate of 4%. Physicians should strongly consider obtaining samples for fungal cultures in patients undergoing spine I&D, especially those using steroids and those undergoing multiple I&Ds. Our AFB culture rates mirror the false positive rates seen in previous orthopedic literature. It is unlikely to be cost effective to send for AFB cultures in areas with low endemic rates of AFB.

Two patients whom we discuss below were 47 and 70y had shared common symptoms such as dyspnea, intermittent but profuse productive cough and newly onset wheeze whereas both were long term diabetics...