Ziehl-Neelsen Research Articles

Occurrence of Cryptosporidium Oocysts in Faeces, Milk and Water Sources in Sedentary Fulani Herds in Selected Local Government Areas of Kaduna State, Nigeria

Abstract Cryptosporidiosis is a neglected tropical zoonotic disease, commonly associated with a diarrhoea, caused by a protozoan parasite of the genus Cryptosporidium. The aim of this study was to determine the occurrence of Cryptosporidium oocysts in faeces, milk, and water sources, and its associated risk factors in sedentary Fulani herds. Seven hundred and fifty faecal samples (240, 180, 240 and 90 from cattle, sheep, goats and dogs, respectively), and 120 samples each, of milk and water were collected for this study. Faecal samples were concentrated by formol ether concentration technique, then stained using Modified Ziehl Neelsen staining method, to detect Cryptosporidium oocysts. Cryptosporidium oocysts in milk and water samples were also concentrated and stained with same stain. The prevalence of Cryptosporidium oocysts in (cattle, dog, sheep and goat faeces were 18.3 %, 15.6 %, 13.9 % and 4.2 %, respectively.), Significantly higher prevalence was observed in calves ≤ 1 year (P = 0.002), female cattle (P = 0.007) and animals with poor body condition (P < 0.001) than in the other groups. Goats with poor body condition scores had significantly higher prevalence (P = 0.008) than other groups while dogs ≤ 6 months of age also had significantly higher prevalence (P = 0.03) than older dogs. The prevalence of Cryptosporidium oocysts in bulk cow milk was 11.7 % while a prevalence of 13.3 % was observed in the water bodies sampled. Hence the findings of this study are of great public health significance, therefore, inhabitants of these LGAs should be informed and educated on the need for improvement of sanitary measures during the course of milking these animals, and the need for adequate pasteurization, of milk before consumption.

Prevalence of opportunistic intestinal coccidian parasites and associated factors in HIV/AIDS patients attending anti-retroviral therapy (ART) clinic at Debre Tabor Comprehensive Specialized Hospital, Northwest Ethiopia: A cross-sectional study.

A growing number of acquired immunodeficiency syndrome (AIDS) patients suffer from opportunistic intestinal coccidian infections. Instead of human immuno deficiency virus (HIV) infection itself, opportunistic infections like intestinal coccidian parasites cause death of over 80% AIDS patients. Factors like exposed drinking water sources and poverty aid the prevalence of opportunistic intestinal coccidian parasitic infections in HIV/AIDS patients. The goal of this study was to determine the prevalence of intestinal coccidian parasites and associated factors in HIV/AIDS patients. A health facility based cross sectional study was conducted from 140 HIV/AIDS patients attending ART clinic in Debre Tabor Comprehensive Specialized Hospital, Northwest Ethiopia from September to December 2023. The sociodemographic characteristics were collected through face-to-face interviews. Stool samples were processed with Modified Acid Fast staining technique. Statistical Package for Social Sciences software version 20 was used to analyze the data. Logistic regression was used to assess factors associated with dependent variable and p < 0.05 was considered significantly associated. The total prevalence of opportunistic intestinal coccidian parasites (OICPs) in HIV/AIDS patients was 16.4% (23/140). Drinking surface water [p = 0.015, COR = 3.4] compared to tape water, drinking alcohol [p = 0.001, COR = 18] compared to not drinking alcohol, diarrhea [p = 0.005, COR = 1] compared to non-diarrheic, drug dropout [p = 0.01, COR = 11] compared to regular drug intake and low CD4 count [p = 0.042, COR = 9] compared to CD4 > 500/µL showed significant association with increased prevalence of OICPs in HIV/AIDS patients. OICPs are still the common causes of morbidity and mortality in HIV/AIDS patients. Surface water consumption, alcoholism, interruption of treatment drugs, diarrhea, and reduced CD4+ T-cells significantly contribute to acquisition and prevalence of OICPs in HIV/AIDS patients. Routine screening of OICPs with sensitive diagnostic techniques in HIV/AIDS patients regardless of symptoms is crucial and has to be practiced in health settings.

Prevalence and associated risk factors of Cryptosporidium infection in calves and hospitalized humans in Libo Kemkem, North Western Ethiopia.

Cryptosporidium infection is one of the major causes of acute gastroenteritis and diarrhoea caused by a protozoan parasite affecting vertebrates and humans. The disease is prevalent in cases of immunocompromised individuals. Despite the impact of the diseases in calf and hospitalized humans, well-documented studies are not available in the study area. The objectives of this study were to determine the prevalence of Cryptosporidium infection in calves and hospitalized humans and assess the major associated risk factors associated with Cryptosporidium infection in calves and hospitalized humans. A cross-sectional study was conducted from November 2020 to March 2021 on calf and human Cryptosporidium infection in Libo Kemkem District, North West Ethiopia. A total of 193 calves and 122 human stool samples admitted to the hospital were used for this study. Three kebeles were selected purposely, and individual calves were selected using a simple random sampling method. A number of sampled calves were allocated proportionally to the selected kebeles. Human samples were collected using a systematic random sampling method. Faecal and stool samples were examined using a modified Ziehl-Neelsen staining method. The overall prevalence of calf and human Cryptosporidium infection found in this study was 15.5% and 11.5%, respectively. Age of calf, breed, body condition, water source, faecal consistency and hygienic condition were found significantly (p<0.05) associated with Cryptosporidium infection in the calf. Similarly, the source of potable water, immunocompromisation and contact with domestic animals were found to be significantly (p<0.05) associated with Cryptosporidium infection in humans. There was a higher prevalence of Cryptosporidium infection in calves and humans in Libo Kemkem District. Therefore, the implementation of proper prevention methods of zoonotic Cryptosporidium infection between calf and human beings through significant risk factors is mandatory. Furthermore, additional studies to investigate the levels of economic importance of the disease should be conducted.

Microsporidial stromal keratitis: treatment outcomes, clinical manifestations, confocal microscopy and histopathology findings: a retrospective observational study

ObjectiveMicrosporidial stromal keratitis (MSK) is an uncommon disease. Only several case series have been reported. We aimed to describe the clinical manifestations, histopathology and treatment outcomes of MSK.Methods and analysisRetrospective...

Utility of GeneXpert in Diagnosis of Pediatric Tuberculosis and Detection of Rifampicin Resistance

Abstract Background: Tuberculosis (TB) remains a significant public health challenge in developing countries, including India, where children under 15 years account for 12% of all TB cases. The National Tuberculosis Elimination Programme (NTEP) recommends the use of GeneXpert/Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) for TB diagnosis, followed by Line Probe Assay (LPA) for detecting drug resistance. This study aimed to determine the prevalence of TB and rifampicin resistance in pediatric patients and to evaluate the utility of CBNAAT in comparison with LPA and Ziehl-Neelsen staining. Methods: This retrospective study was conducted over one year, from 1st January 2022 to 31st December 2022, at a tertiary care hospital in western Maharashtra. A total of 528 samples from pediatric patients (aged &lt;18 years) suspected of having TB were included. Samples underwent CBNAAT testing and Ziehl-Neelsen staining. Positive samples were further analyzed using LPA or Mycobacterial Growth Indicator Tube (MGIT) culture. Results: Among the 528 samples analyzed, gastric lavage accounted for 81.25% of the specimens. CBNAAT detected TB in 61 (11.55%) of the samples, with 15 (24.59%) of these showing rifampicin resistance. Samples with very low or low bacterial loads were often missed by Ziehl-Neelsen staining. Conclusion: Pediatric tuberculosis continues to be a public health concern in developing countries. The study found an 11.55% positivity rate in children under 18 years of age, with 24.59% of positive cases exhibiting rifampicin resistance. The findings underscore the importance of using advanced diagnostic techniques like CBNAAT and highlight the limitations of Ziehl-Neelsen staining in detecting low bacterial loads.

Diagnóstico de &lt;i&gt;Mycobacterium leprae&lt;/i&gt; con Ziehl Neelsen en frotis de piel

The diagnosis of leprosy or Hansen's disease is based on a multifaceted approach that includes clinical evidence and laboratory methods that allow the demonstration of Mycobacterium spp to establish the treatment and prognosis of patients. Ziehl Neelsen staining is an alternative to detect the bacillus in skin smears, determine the classification of leprosy based on its bacillary load and control treatment in confirmed patients.

Integrating computational and experimental approaches in discovery and validation of MmpL3 pore domain inhibitors for specific labelling of Mycobacterium tuberculosis

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), continues to pose a significant global health threat. Identifying new druggable targets is crucial for the advancement of drug development. Equally critical is the development of precise methods for monitoring Mtb to effectively combat this disease. Addressing these needs, our study pinpointed the pore domain (PD) of MtbMmpL3 as a new binding site for virtual screening, which led to the discovery of the small molecule ZY27. To confirm the binding site and action mode of ZY27, we employed cosolvent molecular dynamics (CMD), steered molecular dynamics (SMD), and long timescale molecular dynamics (MD) simulations of 5 μs. These in silico studies verified that ZY27 binds to the PD of MtbMmpL3. In antimicrobial activity tests, ZY27 exhibited potent anti-Mtb activity and high selectivity among mycobacterial species. Whole-genome sequencing of spontaneous ZY27-resistant Mtb variants, complemented by acid-fast staining experiments, confirmed that ZY27 specifically targets MtbMmpL3. Utilizing the ligand-protein binding data, we designed and synthesized two solvatochromic fluorescent probes, 27FP1 and 27FP2, based on ZY27. Further investigations through flow cytometry and confocal microscopy confirmed that these probes specifically label Mtb cells via the MtbMmpL3 binding mechanism.

Mycobacterium marseillense bloodstream infection combined with skin fungal infection: a case report and literature review

BackgroundNon-tuberculous mycobacteria (NTM) are present widely in the natural environment and can invade the human body through the respiratory tract, gastrointestinal tract, and skin. Immunocompromised patients are particularly prone to infection, which primarily affects multiple organs, including the lungs, lymph nodes, and skin. However, cases of NTM bloodstream infections are rare. Here, we report a rare case of Mycobacterium marseillense bloodstream infection with concurrent skin fungal infection in a patient after kidney transplantation. Related literature was reviewed to enhance the understanding of this rare condition.Case presentationA 58-year-old male with a history of long-term steroid and immunosuppressant use after kidney transplantation presented with limb swelling that worsened over the past two months. Physical examination revealed redness and swelling of the skin in all four limbs, with a non-healing wound on the lower left limb. Skin tissue analysis by metagenomic next-generation sequencing (mNGS) and fungal culture indicated infection with Trichophyton rubrum. Blood culture results suggested infection with Mycobacterium marseillense. After receiving anti-NTM treatment, the patient’s symptoms significantly improved, and he is currently undergoing treatment.ConclusionMycobacterium marseillense is a NTM. Gram staining suffered from misdetection, and the acid-fast staining result was positive. This bacterium was identified by mass spectrometry and mNGS analyses. Antimicrobial susceptibility tests for NTM were performed using the broth microdilution method. The results of the susceptibility test showed that Mycobacterium marseillense was sensitive to clarithromycin, an intermediary between moxifloxacin and linezolid. Bacterial clearance requires a combination of drugs and an adequate course of treatment. NTM bloodstream infections are relatively rare, and early identification and proactive intervention are key to their successful management.

Comparison of sampling techniques and diagnostic tests for Cryptosporidium serpentis in eastern indigo snakes (Drymarchon couperi).

The purpose of this study was to compare various sampling techniques and commercially available diagnostic tests for Cryptosporidium serpentis. A colony of 80 eastern indigo snakes (Drymarchon couperi) in human care was screened for the presence of C serpentis using endoscopic gastric mucosal biopsies for histologic and molecular analyses. At the time of endoscopic examination and biopsy, a cloacal swab, gastric swab, and gastric lavage sample were also collected. A C serpentis-specific probe hybridization quantitative PCR (qPCR) was performed on each sample. The gastric lavage sample was divided equally for direct microscopy, acid-fast stain, rapid qualitative immunochromatographic assay, direct fluorescent antibody, and 5 different PCR analyses. If a fecal sample was available at the time of endoscopic evaluation, it was also evaluated for Cryptosporidium oocysts by direct microscopy and acid-fast staining. When comparing test results to histologic analyses, the sensitivity of the probe hybridization qPCR of gastric biopsy, gastric lavage, and gastric swab was 100% while the cloacal swab was 72%. When gastric lavage tests were compared, qPCRs outperformed the other tests. Endoscopic biopsy for histologic and qPCR analyses is recommended for disease diagnosis, while gastric lavage or gastric swab samples for qPCR analysis are as sensitive as endoscopic biopsy for screening for the pathogen but cannot diagnose disease. The results from this study allow the veterinary practitioner to select the most appropriate sample and testing methodology when evaluating an ophidian patient for gastric cryptosporidiosis.

Bridging dermatology and hematology: a case of lepromatous leprosy with bone marrow involvement and pancytopenia.

Leprosy, caused by Mycobacterium leprae (M. leprae), primarily manifestswith cutaneous and peripheral nerve involvement. Systemic involvement, particularly in the bone marrow, is exceedingly rare. This report presents a case of lepromatous leprosy with bone marrow involvement, emphasizing the systemic nature of the disease and the importance of comprehensive diagnostic and management approaches. We aim to present a case of lepromatous leprosy with bone marrow involvement, detailing the clinical presentation, diagnostic evaluation, and management approach. A 65-year-old male with lepromatous leprosy and severe erythema nodosum leprosum developed pancytopenia. After undergoing comprehensive clinical evaluation, including history taking, physical examination, and laboratory investigations, bone marrow examination and molecular diagnostics using polymerase chain reaction (PCR) were performed to confirm the presence of M. leprae as an etiology for his pancytopenia. The bone marrow aspirate revealed hypercellularity with erythropoiesis and thrombopoiesis within normal limits. Foamy histiocytes with erythrophagocytosis were observed, along with the presence of M. leprae on Modified Ziehl-Neelsen stain. Molecular analysis confirmed M. leprae DNA in the bone marrow aspirate. Treatment with multi-drug therapy (MDT) and thalidomide resulted in normalization of blood counts and healing of skin lesions. This case underscores the systemic nature of leprosy and the rarity of bone marrow involvement, highlighting the importance of thorough evaluation in cases of persistent symptoms. Comprehensive diagnostic approaches, including bone marrow examination and molecular diagnostics, are essential for accurate diagnosis and timely initiation of appropriate treatment, ultimately improving patient outcomes and minimizing disease complications.

Peritoneal dialysis-associated peritonitis caused by Tsukamurella inchonensis: a case report and literature review

BackgroundPeritoneal dialysis (PD)-associated peritonitis is a serious complication that can lead to PD discontinuation and mortality. Tsukamurella species are uncommon opportunistic pathogens that can cause peritonitis, often necessitating catheter removal.Case presentationA 46-year-old woman with a 1.5-year history of PD presented with fever and lower abdominal pain, along with cloudy peritoneal effluent showing an elevated cell count. Empiric antibiotic therapy with ciprofloxacin, cefazolin, and ceftazidime was started for PD-associated peritonitis. The peritoneal effluent culture yielded a gram-positive rod with weak acid-fast staining. A rebound in the cell count necessitated a switch of antibiotics to meropenem and vancomycin. This led to improvement, and the treatment was switched to oral levofloxacin on day 30 and the patient was discharged on day 35. Subsequently, 16S rRNA gene sequencing confirmed the isolate as Tsukamurella inchonensis.ConclusionThis case highlights the challenges in identifying and treating Tsukamurella peritonitis. Successful treatment without catheter removal in this case suggests that early detection and appropriate antibiotics may enable catheter salvage in this rare infection. However, further research is needed to establish optimal treatment strategies.

An immunocompetent lady with invasive aspergillosis presenting as disseminated lesions: a case report

BackgroundInvasive Aspergillosis is a fungal infection caused by Aspergillus species, typically posing life-threatening risks to immunocompromised individuals. While occurrences in immunocompetent hosts are rare, a recent case report documented fulminant pulmonary aspergillosis in an immunocompetent patient during autopsy. Here, we present a case of invasive aspergillosis in an immunocompetent woman, manifesting with disseminated lesions.Case presentationA 29-year-old Asian woman presented to our hospital in March 2022, reporting chest pain and shortness of breath persisting for two months. Upon examination, she appeared thin and unwell, with no notable abnormalities otherwise. Radiographic imaging revealed an ill-defined lesion in her left lung. Subsequent bronchoscopy and lavage were performed, followed by initiation of empirical antibiotic therapy. Lavage results were negative for gram staining, culture, and ZN staining for AFB, but revealed numerous septate hyphae on fungal smear. Histopathological examination indicated chronic granulomatous inflammation with septal fungal hyphae, indicative of aspergillosis. Subsequent culture confirmed Aspergillus species, prompting initiation of voriconazole therapy. Remarkably, the patient exhibited significant improvement, with weight gain and restored appetite observed within a short period. Within 2 months of treatment, her symptoms resolved, and she resumed near-normal daily activities.ConclusionThis case highlights the diagnosis of aspergillosis in an immunocompetent individual presenting with disseminated nodular lesions across the lungs, mediastinum, and abdomen. Clinicians should maintain a high index of suspicion for aspergillosis in cases of non-resolving pneumonia and disseminated nodular lesions, even in patients lacking traditional predisposing factors.

Mycobacterium genavense detection in a blood smear from a domestic ferret

AbstractThe domestic ferret is sensitive to mycobacterial infections, which are likely underdiagnosed. There are limited reports of therapy for mycobacteriosis in ferrets and most have failed. A ferret in poor body condition revealed marked organomegaly on diagnostic imaging. The complete blood count revealed a mild pseudoeosinophilia, which might be attributed to negatively stained bacilli phagocytised by monocytes observed in the blood smear. Ziehl–Neelsen stain was performed and showed acid‐fast bacilli within monocytes. Fine‐needle aspirates of the spleen and liver revealed the same finding. Mycobacterium genavense was identified by PCR and DNA sequencing from peripheral blood. Treatment with rifampicin, clarithromycin and enrofloxacin was initiated. The disseminated mycobacteriosis was suspected from the observation of bacilli on the blood smear. After 16 months, the ferret is still undergoing treatment. To prevent recurrence, continued monitoring of drug toxicity and effective therapy will be maintained even after the resolution of clinical signs (mostly organomegaly).

Granulomatous mastitis and other inflammatory mastitis simulating breast neoplasms – A retrospective study in the fine needle aspiration cytology clinic of a tertiary care center in eastern India

Background: Fine needle aspiration cytology (FNAC) is an important diagnostic modality in palpable breast lesions. This together with imaging modality is important in distinguishing benign from malignant lesions of the breast. Granulomatous/inflammatory mastitis forms a small subset of breast lesions that may present as a breast lump. Clinical and radiological features in these cases may mimic malignant breast pathology. Aims and Objectives: The aim of this study was to study the frequency of granulomatous mastitis and other inflammatory mastitis with respect to other neoplastic lesions of the breast, presented in FNAC clinics and to correlate the cytological findings with the clinical presentation of granulomatous and other inflammatory mastitis. Materials and Methods: A retrospective study was conducted in the Department of Pathology in a tertiary care center in Eastern India for duration of 4 years 5 months. Details of all patients who underwent FNAC for breast lesions during the study period were recorded in terms of their clinical presentation, cytological findings, imaging, and ancillary investigations. The findings were then tabulated in a master sheet and analysis done by descriptive statistics using SPSS 25. Results: A total of 154 cases were included with 5 cases (3.2%) of granulomatous and 12 cases (7.8%) of inflammatory mastitis. Age range for granulomatous/inflammatory mastitis was 20–70 years. Lump and pain were the most common complaints followed by inflammatory skin changes and nipple discharge. Very few cases also showed nipple retractions and very occasional systemic symptoms like fever. History of lactation was seen among five cases. Three cases showed positive acid-fast bacilli in Ziehl–Neelsen stain. Radiology was discordant with cytological findings in three cases, where the former suggested a breast neoplastic process. Conclusion: Granulomatous/inflammatory mastitis is a rarely diagnosed entity in breast cytopathology with no standard guidelines for management. Antibiotics antitubercular therapy and corticosteroids are some proposed lines of medical management. In treatment-resistant cases, wide local excision may be done depending on available resources and surveillance opportunities.

Relative Value of Immunohistochemistry in Detection of Mycobacterial Antigen in Suspected Cases of Tuberculosis in Tissue Section.

Due to its infectious nature, complex immunological response, chronic progression, and the necessity for long-term treatment, tuberculosis has always been a major health burden. Immunohistochemistry (IHC) has the capacity to highlight the occurrence of mycobacterial antigens for tissue diagnosis. This study was conducted to understand the advantage of immunostaining over culture of Mycobacterium tuberculosis. A cross-sectional study was conducted on 30 samples of suspected cases of tuberculosis. Specimens received were fixed in 10% formalin and processed; 3-5 µm thick sections were made from paraffin block, stained with hematoxylin and eosin, Ziehl-Neelsen stain, and immunohistochemistry. Culture was done using Lowenstein-Jensen medium. Immunohistochemistry was interpreted as fine granular brownish cytoplasmic, coarse granular brownish cytoplasmic, and bacillus staining. Out of the 30 samples studied, 12 (40.0%) were culture positive while 20 (66.7%) of them were IHC positive. Immunohistochemistry showed 17 granulomatous lesions of which 11 (55.0%) were well-formed granulomas. The sensitivity and negative predictive value were found to be high with immunohistochemistry, while specificity and positive predictive value were found to be on the lower side. Among the 20 positive IHC cases, the degree of staining was fine granular cytoplasmic staining in 13 cases (65.0%) and coarse granular staining in 7 cases (35.0%). Immunohistochemistry is a reliable test with high sensitivity as well as high negative predictive value which can be done rapidly for establishing an etiological diagnosis of tuberculosis in histologic specimens.

Survey of gastrointestinal coccidian parasites of pet birds in Tehran

ABSTRACT Birds of various families and genera comprise a notable proportion of veterinary clients in veterinary practice. Regarding the scarcity of information on avian coccidia in pet birds, this study was designed to investigate and identify the diversity of gastrointestinal coccidian protozoa in caged pet birds. Accordingly, droppings samples were collected in 2.5% potassium dichromate and examined for the presence of oocysts by wet smear and by flotation with the saturated salt solution. Detected oocysts were further morphologically specified. Special Modified Ziehl-Neelsen stain was used for the detection of Cryptosporidium spp. oocysts. Out of the 145 droppings that were collected for parasitological investigations, it was discovered that a Japanese quail (Coturnix japonica) had a Cryptosporidium sp. infection (1/145: 0.7%). Additionally, Isospora spp. was found in a common mynah and a canary (2/145: 1.4%), while Eimeria sp. was detected in a canary (1/145: 0.7%). Studied birds were clinically normal and no signs of intestinal infection were observed. The oocysts are described morphologically in detail and compared with the available data. Periodic faecal examination should be recommended for monitoring intestinal parasites in pet birds both from a public health perspective for Cryptosporidium and for avian host health in the case of non-zoonotic parasites. Additionally, the identification of parasite/host species composition will enhance our understanding of species diversity and the variety of hosts that coccidian parasites are capable of infecting.

Evaluation of Diagnostic Methods and Rifampicin Resistance in Pulmonary Tuberculosis: A Hospital-Based Study.

Background Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis, predominantly affecting the lungs (pulmonary TB) and is a significant public health challenge in India. The study aims to analyze demographic, radiological, and clinical subgroups of pulmonary TB cases, examine the relationship between smear acid-fast bacillus (AFB examination) and cartridge-based nucleic acid amplification test (CBNAAT), evaluate CBNAAT sensitivity for Mycobacterium tuberculosis (MTB) in new and previously treated patients, and determine the proportion of rifampicin resistance. Methods This hospital-based prospective study was conducted among patients diagnosed with pulmonary TB at the Respiratory Medicine Department of a Government Hospital over 16 months (August 2019 to December 2020). The study included 150 diagnosed TB cases (new and previously treated). Data collection encompassed demographic details, clinical symptoms, comorbidities, radiological findings (chest X-ray), and microbiological results (smear AFB examination, CBNAAT). Sputum samples were subjected to Ziehl-Neelsen staining and CBNAAT for MTB detection and rifampicin resistance testing. Statistical analysis was performed using IBM SPSS Statistics version 21.0 (IBM Corp., Armonk, NY, USA). Results Of the 150 patients, 69.3% were male, and 48% were aged 21-40 years. The majority had a BMI of 18.5-24.9 kg/m² (50%) and resided in urban areas (63.3%). Common symptoms included cough (95.3%), fever (80%), and weight loss (74%). Cavitary lesions on chest X-ray were observed in 84% of patients. Smear microscopy detected MTB in 72.7% of cases, while CBNAAT detected MTB in 94% of cases. CBNAAT sensitivity for smear-positive and smear-negative samples was 93.97% and 94.12%, respectively. Rifampicin resistance was found in 3% of new cases and 6% of previously treated cases. The sensitivity of smear microscopy was 77.33%, and the sensitivity of CBNAAT was 94%. Conclusion The study underscores the high burden of pulmonary TB and the utility of CBNAAT in detecting MTB and rifampicin resistance, particularly in smear-negative samples. The findings highlight the necessity of universal drug susceptibility testing (DST) for effective TB management and the importance of addressing drug resistance to improve treatment outcomes.

Detection of caprine paratuberculosis (Johne's disease) in pre- and post-vaccinated herds: morphological diagnosis, lesion grading, and bacterial identification.

Samples from the mesenteric lymph nodes (MS LNs) and ileocecal valves (ICV) of 105 goats, comprising 61 non-vaccinated and 44 vaccinated against Mycobacterium avium subspecies paratuberculosis (MAP), were collected at slaughter from a farm with a confirmed history of paratuberculosis (PTB). These goats had subclinical infections. PTB-compatible lesions in the MS LNs, ICV lamina propria (LP), and Peyer's patches (PPs) were graded separately. Furthermore, the load of acid-fast bacilli was quantified using Ziehl-Neelsen staining (ZN), MAP antigens by immunohistochemistry (IHC), and MAP DNA by PCR targeting the IS900 sequence. Gross PTB-compatible lesions were found in 39% of the goats, with 31.72% vaccinated (V) and 68.29% non-vaccinated (nV). Histopathological lesions induced MAP were observed in 58% of the animals, with 36.07% vaccinated and 63.93% non-vaccinated. The inclusion of histopathology as a diagnostic tool led to a 28% increase in diagnosed cases in MS LNs and 86.05% in ICV. Grade IV granulomas with central mineralization and necrosis were the most common lesions in MS LNs. In the ICV, mild granulomatous enteritis with multifocal foci of epithelioid macrophages was predominant, occurring more frequently in the PPs than in the LP. Furthermore, statistical differences in the presence of histopathological lesions between vaccinated and non-vaccinated goats were noted in MS LNs, ICV LPs, and ICV PPs. Non-vaccinated animals showed higher positivity rates in ZN, IHC, and PCR tests, underscoring the benefits of anti-MAP vaccination in reducing PTB lesions and bacterial load in target organs. Our findings emphasize the necessity of integrating gross and histopathological assessments with various laboratory techniques for accurate morphological and etiological diagnosis of PTB in both vaccinated and non-vaccinated goats with subclinical disease. However, further studies are required to refine sampling protocols for subclinical PTB in goats to enhance the consistency of diagnostic tools.

A comparative study of diagnostic yield of Ziehl-Neelsen staining and fluorescent staining in diagnosis of primary pulmonary tuberculosis in a resource limited country like India

Background: Tuberculosis (TB) is a major public health disease. Early diagnosis and treatment are essential for effective TB control program. In developing countries, diagnosis of TB is mainly by microscopy because of the simplicity of the procedure and cost-effectiveness. In the present study the two methods were compared to derive a conclusion to adopt the better method to provide more effective services for early diagnosis of TB. Methods: This study was conducted to compare the Ziehl-Neelsen (ZN) staining with Fluorescent staining in the diagnosis of primary pulmonary TB patients. A total of 514 pulmonary tuberculosis suspected patients were included for a period of 2 years. Sputum samples were taken and subjected to both ZN staining and fluorescent staining and results were compared. All samples were subjected to solid culture and taking it as gold standard sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of both the staining methods were defined. Results: Out of total 514 samples, 265 (51.56%) were ZN positive and 326 (63.43%) samples were positive for FM. Positive samples detected by ZN staining were compared with positive samples by FM. Sensitivity, specificity, PPV, NPV for ZN microscopy were 80.76%, 97.27%, 98.05% and 74.79% respectively. Sensitivity, specificity, PPV, NPV for FM were 98.71%, 95.63%, 97.47% and 97.76% respectively. Conclusions: In developing country like India where there are large number of cases and gross financial constraints microscopy can be adopted as the main method of diagnosis of pulmonary TB and preferably FM because of higher sensitivity and lesser time taking character of the method.

Severe Bilateral Sialadenitis of the Mandibular and Parotid Salivary Glands with Severe Panniculitis in a 2-Year-old Standard Poodle.

A 2-year-old male neutered Standard Poodle weighing 17.9 kg was presented to their primary care veterinarian for enlarged bilateral submandibular swellings following an interdog altercation sustained in the previous weeks. Cytology performed following fine-needle aspirates of the regions of swelling was inconclusive, and the patient was treated empirically with Clavaseptin. Despite treatment, the submandibular swellings continued to enlarge, and right-sided intermittent epistaxis was reported. On biochemical profile, there was mild hypercalcemia and mild hyperglobulinemia. The computed tomography (CT) findings were indicative of severe multifocal sialadenitis with severe regional cellulitis and inflammatory lymphadenopathy. Histopathology and cytology results described mixed inflammation of the salivary gland. Methenamine silver staining and Fite's acid-fast staining were negative. Aerobic and anaerobic cultures were negative. Targeted, next-generation DNA sequencing detected no known fungi or bacterial pathogens. These findings were consistent with the diagnosis of severe bilateral mandibular sialadenitis, panniculitis, and lymphadenopathy. The patient was prescribed enrofloxacin, clindamycin, phenobarbital, and prednisolone for 1 month. One week after initiating treatment, the patient had a significant reduction in size of the salivary glands. CT imaging was helpful in the diagnosis of this patient and allowed the clinician to identify which submandibular anatomical structures were abnormal, guiding subsequent diagnostic decisions to provide medical management to resolve the condition.