Cryptosporidium Infections in Neonatal Calves on a Dairy Farm.

This study was conducted with the aim of the molecular identification of the protozoan parasite Cryptosporidium spp. in calves in the early stage of their development on a dairy farm in Eastern Slovakia. Twenty-five Holstein and Holstein cross calves were included in the study and monitored from their birth to the fifth week of life (1-5 weeks). Fresh fecal samples were collected from the same group of calves each week, except during the fourth week, and with the exception of Sample 8. All samples were analyzed using the Ziehl-Neelsen staining method and coproantigen was tested using the ELISA test as the screening method. Using the ELISA method, the highest incidence of cryptosporidiosis was observed in the second week of life of the calves, while the antigen was detected in 21 (91.6%) calves. Using the Ziehl-Neelsen staining method, the highest incidence was also observed in the second week, with an incidence rate of 62.5%. Positive isolates confirmed by the ELISA test were molecularly characterized. The species and subtypes of Cryptosporidium in the positive isolates were identified using PCR and the sequence analysis of the small subunit of the ribosomal 18S RNA (ssu rRNA) and the 60 kDa glycoprotein (gp60) genes of the parasite. The sequence analysis of 29 isolates at the 18S rRNA loci confirmed the presence of two species-Cryptosporidium parvum and Cryptosporidium ryanae. Out of 29 isolates, 25 were assigned to the species C. parvum, with the gp60 locus identified as genotype IIaA17G1R1. Among the individual animal groups, calves are the most common reservoirs of the C. parvum zoonotic species. This disease has significant public health implications as contact with livestock and their feces and working with barn manure are major sources of infection, not only for other animals but also for humans.

Nocardia otitidiscaviarum causing pulmonary nocardiosis: a case report and its review of the literature.

Background: Infections caused by Nocardia spp. can occur in immunocompromised as well as immunocompetent individuals. Although nocardiosis is rare, it is being increasingly recognized owing to the rise in occurrence rate over the years. The documentation of pleural involvement in nocardiosis is rare in India. Case: We report a case of pulmonary nocardiosis in an immunocompromised individual caused by Nocardia otitidiscaviarum. Discussion: Pulmonary nocardiosis caused by Nocardia otitidiscaviarum may go unnoticed without clinical suspicion. Correct and timely identification is the key to proper patient management. Conclusion: Coordination between clinicians and microbiologists is necessary for early diagnosis and appropriate management of nocardiosis.

The Use of Xpert MTB/RIF Ultra Testing for Early Diagnosis of Tuberculosis: A Retrospective Study from a Single-Center Database.

Tuberculosis (TB) is a multisystemic contagious disease produced by Mycobacterium tuberculosis complex bacteria (MTBC), with a prevalence of 65:100,000 inhabitants in Romania (six times higher than the European average). The diagnosis usually relies on the detection of MTBC in culture. Although this is a sensitive method of detection and remains the "gold standard", the results are obtained after several weeks. Nucleic acid amplification tests (NAATs), being a quick and sensitive method, represent progress in the diagnosis of TB. The aim of this study is to assess the assumption that NAAT using Xpert MTB/RIF is an efficient method of TB diagnosis and has the capacity to reduce false-positive results. Pathological samples from 862 patients with TB suspicion were tested using microscopic examination, molecular testing and bacterial culture. The results show that the Xpert MTB/RIF Ultra test has a sensitivity of 95% and a specificity of 96.4% compared with 54.8% sensitivity and 99.5% specificity for Ziehl-Neelsen stain microscopy, and an average of 30 days gained in the diagnosis of TB compared with bacterial culture. The implementation of molecular testing in TB laboratories leads to an important increase in early diagnostics of the disease and the prompter isolation and treatment of infected patients.

Pathological and molecular identification of Mycobacterium avium infection in a loft of domestic pigeons (Columba livia var. domestica) from India.

Mycobacterium avium is a zoonotic pathogen associated with a wide range of pulmonary and extrapulmonary manifestations in a range of host species like humans, animals, and birds. The disease is more common in the avian population, and opportunistic infections have been reported in immune-compromised or debilitated animals and humans. This study reports the pathological and molecular identification of Mycobacterium avium causing avian mycobacteriosis in a loft of domestic pigeons (Columba livia var. domestica). Out of 30 pigeons aged 2-3 years, ten adult racing pigeons revealed a severe chronic and debilitating disease followed by death. The clinical signs included chronic emaciation, dullness, ruffled feathers, lameness, and greenish, watery diarrhea. Post-mortem examination of birds revealed multifocal gray- to yellow-colored raised nodules in the liver parenchyma, spleen, lungs, intestines, bone marrow, and joints. Avian mycobacteriosis was suspected based on the tissue impression smears stained by Ziehl-Neelsen staining. Histopathological examination also revealed multifocal granulomatous lesions in affected organs, which is characteristic of avian mycobacteriosis. The PCR analysis based on 16S rRNA, IS1245, and IS901 regions suggested the presence of Mycobacterium avium infection belonging to either subspecies avium or sylvaticum. This is the first detailed report of avian mycobacteriosis in pigeons from India, warranting a strict surveillance program to identify the carrier status of these microorganisms in the pigeons, which may prove a fatal zoonotic infection in humans.

Comparison of conventional diagnostic methods with molecular method for the diagnosis of pulmonary tuberculosis.

BACKGROUND: Tuberculosis remains one of the deadliest communicable diseases. Prompt diagnosis of active tuberculosis cases facilitates timely therapeutic intervention and minimizes the community transmission. Although conventional microscopy has low sensitivity, still it remains the corner stone for the diagnosis of pulmonary tuberculosis in high burden countries like India. On the other hand, Nucleic acid amplification techniques due to their rapidity and sensitivity, not only help in early diagnosis and management of tuberculosis but also curtail the transmission of the disease. This study therefore was aimed at assessing the diagnostic performance of Microscopy by Ziehl Neelsen (ZN) and Auramine Staining (AO) with Gene Xpert/CBNAAT (Cartridge based nucleic acid amplification test) in the diagnosis of Pulmonary Tuberculosis. METHODS: A prospective comparative study was done on the sputum samples of 1583 adult patients from November 2018 to May 2020 suspected of having pulmonary tuberculosis as per NTEP criteria visiting the Designated Microscopic Centre of SGT Medical College, Budhera, Gurugram. Each sample was subjected to ZN staining, AO staining, and was run on CBNAAT as per National Tuberculosis Elimination Program (NTEP) guidelines. The sensitivity, specificity, PPV and NPV and Area under the curve of ZN microscopy and Fluorescent Microscopy were calculated taking CBNAAT as reference in absence of culture. RESULTS: Out of the 1583 samples studied, 145 (9.15%) and 197 (12.44%) were positive by ZN and AO staining methods respectively. By CBNAAT 246 (15.54%) samples were positive for M. tuberculosis. AO was also able to detect more pauci-bacillary cases than ZN. While CBNAAT detected M. tuberculosis in 49 sputum samples which were missed by both methods of microscopy. On the other hand there were 9 samples which were positive for AFB by both the smear microscopy techniques but M. tuberculosis was not detected by CBNAAT, these were considered as Non-Tuberculous Mycobacteria. Seventeen samples were resistant to rifampicin. CONCLUSION: Auramine Staining technique is more sensitive and less time consuming for the diagnosis of pulmonary tuberculosis as compared to the conventional ZN Staining. CBNAAT can be a useful tool for early diagnosis of patients with high clinical suspicion of pulmonary tuberculosis and detecting rifampicin resistance.

Effect of 80% ethanol or 10% formalin fixation, freezing at -20 °C and staining on Myxobolus (Myxosporea) spores to be deposited in parasitological collections.

The preparation of myxosporeans for the description of myxospores and their preservation as type material in parasitological collections show great variations. Most frequently, formalin and ethanol are used for fixation and Giemsa solution for staining spores. In this work, authors studied the effect of 80% ethanol and 10% formalin fixation, freezing at -20 °C and staining on the size and transparency of two Myxobolus species of cyprinid fishes, M. bramae and M. bliccae spore, and recommended a new method for the deposition of type material to parasitological collections in museums. The studies have commended that fresh spores from mature plasmodia are the best material for measuring the size and studying the inner structures, the number of polar tubules in polar capsules and the morphological characters of the intercapsular appendix. The obtained quantitative data suggest that cryo- and chemical preservation do not have a notable negative effect on spores compared to fresh samples but they decrease the transparency of spores. Staining the spores with Ziehl-Neelsen has proved to be a useful method for studying the fine structure without size reduction, while Giemsa staining induced a shrinkage of spores so it seems to be not ideal for description of a new species. When treating spores of Myxobolus spp. with Lugol's solution, iodinophilous vacuoles in the sporoplasm were not recognised but visualisation of the coils of polar tubules was enhanced. As a type material for newly described species, authors suggest phototypes and spores fixed in 80% ethanol to be deposited into collections, as this preservation method is suitable for subsequent research, such as re-measurements and molecular analysis.

Pulmonary and Disseminated Mycobacterium avium Complex Cases Confirmed by Tissue-Direct Polymerase Chain Reaction-Based Nucleic Acid Lateral Flow Immunoassay of Formalin-Fixed Paraffin-Embedded Tissues.

Background: Detection of Mycobacterium avium complex (MAC) in tissue is essential for the diagnosis of MAC infections when the Mycobacterium is not isolated from sputum. However, detection of MAC in paraffin-embedded sections has not been established. Methods: We encountered two patients with suspected MAC infections after surgery: patient 1 had a pulmonary nodule that was initially suspected to be lung cancer and was excised under video-assisted thoracoscopic surgery (VATS). Patient 2, who was under treatment with steroids and anti-IL-6 inhibitors for rheumatoid arthritis, was suspected to have disseminated ileocecal cancer with metastasis to the lung and skin. In both cases, we postoperatively detected MAC genes in paraffin-embedded tissue sections using the novel mycobacterial nucleic acid identification test, ie tissue-direct polymerase chain reaction (tdPCR)-based nucleic acid lateral flow immunoassay (NALFIA). Both patients showed granulomatous lesions with hematoxylin-eosin staining, and mycobacteria by Ziehl-Neelsen staining in tissue sections from the lung and skin, respectively, although MAC were not isolated from the sections. MAC genes were finally detected by tdPCR-NALFIA in both cases. Conclusion: Although Ziehl-Neelsen staining and culture tests are the gold standard in identifying causative mycobacteria, the rapid results of tdPCR-NALFIA performed simultaneously with sputum and/or tissue culture may make it an important auxiliary diagnostic tool for identifying mycobacterial infection, leading to improvement in the management of MAC patients.

'Primary Thyroid Tuberculosis'-The Masquerader: A Case Report.

Tuberculosis of thyroid is a rare entity even among highly prevalent regions of tuberculosis. Primary tuberculosis of thyroid is even more rarer. The reason is attributed to the inherent relative immunity of the thyroid gland. Clinical manifestation is unpredictable accounting to both asymptomatic and variable benign and malignant mimicking symptoms. Clinical course may too vary depending on the thyroid dysfunction and complications. Aspiration cytology is diagnostic, though the yield is low. Histological diagnosis, depicting caseating granuloma added with acid fast staining confirms the diagnosis. High clinical suspicion is to be maintained to prevent total thyroidectomy.

Acid-Fast Bacilli Positivity Rate and Associated Factors among Leprosy Suspected Cases attending Selected Health Facilities located in West Arsi Zone, Oromia, Ethiopia.

INTRODUCTION: 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.

Molecular prevalence of Cryptosporidium isolates among Egyptian children with cancer.

Immunocompromised individuals especially children with cancer are at risk for acquiring cryptosporidiosis, which can result in severe morbidity and mortality. This work was conducted to evaluate the prevalence of Cryptosporidium parasite and its genotypes in children with cancer. Stool specimens were collected from 145 children in the Oncology unit of Pediatric Department, Zagazig University Hospital, Sharqiyah province, Egypt. Cryptosporidium infection was evaluated using modified Ziehl-Neelsen (MZN) staining and nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The prevalence of Cryptosporidium infection in oncological children was 29.7% using microscopy and 25.5% using nested PCR. Genotypic characterization showed that 23 (62.2%) had C. hominis, 11 (29.7%) C. parvum, and 3 specimens (8.1%) were mixed infection of both genotypes. Cryptosporidiosis was significantly associated with diarrhea. However, no statistically significant difference was detected between the age, gender, residency, animal contact and malignancy type concerning to Cryptosporidium infection. This study concluded that Cryptosporidium is a prevailing opportunistic parasite among children with cancer. It should be considered in oncological patients especially those suffering from diarrhea which requires proper management to reduce its complications.

Molecular detection of Cystoisospora belli by single-run polymerase chain reaction in stool samples.

INTRODUCTION: Cystoisospora belli (C. belli) is the only pathogenic species of the Cystoisospora genus responsible for severe diarrhea in immunocompromised patients. Most common microscopic method of diagnosis is less sensitive due to intermittent shedding of oocysts. We developed a new single-run polymerase chain reaction (PCR)-based diagnostic assay for C. belli. METHODS: A new single-run PCR-based diagnostic assay was standardized for the detection of C. belli. Diagnostic reproducibility and repeatability of the PCR assay were evaluated. A cross-sectional analytical study was done on a total of 354 stool samples collected from 331 immunocompromised patients with diarrhea. All the stool samples were tested for the presence of oocysts of C. belli and were also tested by our new PCR assay for C. belli. Three of the representative PCR products were confirmed by sequencing. Fisher's exact test was used to compare the two proportions. RESULTS: Microscopy detected C. belli in 11/354 (3.1%) of stool samples, and the new PCR-based assay detected C. belli in 16/354 (4.5%). The new single-run PCR-based assay detected C. belli in all the stool samples which were tested positive by microscopy and additionally detected C. belli in five stool samples. The developed PCR assay detected statistically significant proportion of C. belli (p < 0.001) as compared to microscopy. The 795 base pair PCR product from one microscopy positive stool sample and two microscopy negative stool samples were confirmed by sequencing. CONCLUSION: Our newly developed single-run PCR-based detection assay for C. belli is robust and reproducible. It may be used for molecular diagnosis of cystoisosporiasis especially in transplant, pediatrics, and human immunodeficiency virus (HIV) positive patients.

Identification of suspected paragonimiasis-endemic foci using a questionnaire and detection of Paragonimus ova using the Ziehl-Neelsen technique in Zamboanga Region, the Philippines.

Improving paragonimiasis surveillance, which is crucial for disease control, requires adopting new tools and techniques useful in mapping endemic areas. This study aimed to (1) develop a questionnaire to identify suspected paragonimiasis-endemic foci, (2) describe the epidemiology of paragonimiasis, and (3) evaluate Ziehl-Nielsen Staining technique (ZNS) in detecting Paragonimus ova. The questionnaire, which municipal health officers filled out, was based on proposed site inclusion criteria utilized in the integrated tuberculosis (TB)-paragonimiasis surveillance and control project. Newly deployed medical technologists in Zamboanga Region underwent training, which included laboratory diagnosis of paragonimiasis using preserved and fresh specimens and an integrated tuberculosis-paragonimiasis survey in nine selected barangays (villages). Paragonimiasis cases were found in seven out of the nine barangays identified by the questionnaire. Of the 373 patients, three (0.80%) were TB-positive, and 29 (7.77%) were paragonimiasis-positive. The highest paragonimiasis prevalence (27%) was found in Barangay Libato. Ziehl-Neelsen Staining technique (ZNS) correctly detected 8 out of the 29 samples positive (sensitivity - 27.59%; 95% CI: 12.73-47.24%) and all the 334 samples negative (specificity - 100%; 95% CI: 98.90-100%) for Paragonimus ova. The questionnaire may be improved by refining the inclusion criteria. In paragonimiasis-endemic areas, the ZNS and the NaOH concentration technique may be used for detecting Paragonimus ova. Modifying the ZNS, for instance by including a concentration step, may improve its sensitivity. The model for the integrated capacity building of health workers and surveillance and research demonstrated in this project may contribute to improving surveillance and control of paragonimiasis and other neglected tropical diseases.

Comparison of immunohistochemistry and Ziehl-Neelsen staining for detecting the distribution of Mycobacterium avium subsp avium in naturally infected domestic Pekin ducks (Anas platyrhynchos domestica).

In order to detect the distribution of Mycobacterium avium subsp avium (MAA) in naturally infected domestic Pekin ducks, immunohistochemistry (IHC) and Ziehl-Neelsen (ZN) staining were used and compared. Six organs, the liver, spleen, lung, kidney, duodenum and pectoralis muscle, were collected from naturally infected Pekin ducks. Paraffin embedded tissues were examined, and the results were compared. Statistical analysis was performed using Chi-Square test. The results showed that the detection rates by IHC were similar with ZN staining in liver, lung, spleen and pectoralis muscle, but the detection rates by IHC were much higher than ZN staining in kidney and duodenum (p = .013, p = .0044). The liver (87.5%) and lung (81.3%) had the highest detection rates. Acid-fast bacilli (AFB) were primarily found intracellularly in six organs using ZN staining. Similarly, the MAA antigens in those selected organs were also detected in the cytoplasm with different cell types. Specifically, MAA antigen was distributed in epithelioid macrophages and necrotic centres within the liver, lung, spleen and kidney, while they were observed in macrophages of the lamina propria and duodenal glands and degenerative myocytes in the pectoralis muscle. This comparative study provides an important insight into the distribution of MAA in infected domestic ducks and indicates that the detection rate by IHC was higher than that of ZN staining.

Prevalence and associated risk factors of Cystoisospora belli and Cyclospora cayetanensis infection among Iranian patients with colorectal cancer.

From among intestinal parasites, coccidian intestinal parasites such as Cystoisospora belli (previously known as Isospora belli), and Cyclospora cayetanensis are well-known as opportunist parasites, particularly in patients with cancer. This study assessed the prevalence of C. belli and C. cayetanensis in patients with cancer in Lorestan Province, Southwest of Iran. This cross-sectional descriptive study was conducted on 87 patients with colorectal cancers, referred to the general hospitals of Lorestan from October 2017 to August 2018. A fresh stool specimen was collected from each subject in a sterile labeled container. The collected stool samples were concentrated through sucrose flotation method and then prepared for Ziehl-Neelsen staining for microscopic examination. Demographic and possible risk factors such as age, sex, education, residence, and unwashed vegetable/fruit consumption were collected by an applied questionnaire. Out of the 87 patients with colorectal cancer, eight (9.2%) were found positive for C. belli and C. cayetanensis infections, with five (5.74%) and three (3.44%) patients positive for C. belli and C. cayetanensis infections, respectively. Results also showed that sex and unwashed vegetable/fruit consumption were significantly associated with the prevalence of these parasites (p < 0.01). The findings revealed the considerable prevalence of C. belli and C. cayetanensis in patients with colorectal cancers. It is therefore essential for gastrointestinal specialists to pay special attention to the prevalence of coccidian parasites in patients with colorectal cancer.

Application of Laser Scanning Confocal Microscopy for the Visualization of M. tuberculosis in Lung Tissue Samples with Weak Ziehl-Neelsen Staining.

One of the key requirements for the diagnosis of pulmonary tuberculosis is the identification of M. tuberculosis in tissue. In this paper, we present the advantages of specific fluorescent antibody labelling, combined with laser scanning confocal microscopy (LSCM), for the detection of M. tuberculosis in histological specimens of lung tissues. We demonstrate that the application of LSCM allows: (i) The automatic acquisition of images of the whole slice and, hence, the determination of regions for subsequent analysis; (ii) the acquisition of images of thick (20-40 µm) slices at high resolution; (iii) single bacteria identification; and (iv) 3D reconstruction, in order to obtain additional information about the distribution, size, and morphology of solitary M. tuberculosis; as well as their aggregates and colonies, in various regions of tuberculosis inflammation. LSCM allows for the discrimination of the non-specific fluorescence of bacteria-like particles and their aggregates presented in histological lung samples, from the specific fluorescence of labelled M. tuberculosis, using spectrum emission analysis. The applied method was effective in the identification of M. tuberculosis in lung histological samples with weak Ziehl-Neelsen staining. Altogether, combining immunofluorescent labelling with the application of LSCM visualization significantly increases the effectiveness of M. tuberculosis detection.

Preliminary analysis of the value of Fite staining in the diagnosis of leprosy / 中华皮肤科杂志

Objective To evaluate the diagnostic value of Fite staining in leprosy histopathology.Methods Between 2013 and 2017,13 patients diagnosed with leprosy or suspected leprosy (high suspicion of leprosy based on clinical manifestations and hematoxylin-eosin staining,but negative acid-fast staining) in our department,were enrolled into this study.The histopathological sections were subjected to Fite staining,and the results were compared with those of acid-fast staining,so as to assess the value of Fite staining in the diagnosis of leprosy.Results Six patients with positive acid-fast staining still showed positive Fite staining.Among 7 patients with suspected leprosy and negative acid-fast staining,6 patients showed positive Fite staining with varying numbers of Mycobacterium leprae,and 1 showed negative Fite staining.Conclusion Fite staining can increase the detection rate of Mycobacterium leprae.

Mycobacterium tuberculosis found at both skin lesions and Mantoux testing site in a patient with erythema induratum of Bazin.

Mycobacterium tuberculosis is very rarely found in erythema induratum of Bazin; recently, we found an unusual case with positive acid-fast bacilli and polymerase chain reaction for detecting M. tuberculosis in both skin lesions of the extremities and the site of Mantoux test.

Performance of nested multiplex PCR assay targeting MTP40 and IS6110 gene sequences for the diagnosis of tubercular lymphadenitis.

The conventional methods for diagnosis of tubercular lymphadenitis (TBLN) such as - fine needle aspiration cytology, Ziehl-Neelsen staining and culture have limitations of low sensitivity and/or specificity. So, it becomes essential to develop a rapid, sensitive, and specific method for an early diagnosis of TBLN. Therefore, the present study was conducted to evaluate nested multiplex polymerase chain reaction (nMPCR) targeting MTP40 and IS6110 gene sequences of Mycobacterium tuberculosis and Mycobacterium tuberculosis complex, respectively in 48 successive patients of TBLN and 20 random patients with non-tubercular lymph node lesions. Out of the 48 cases of TBLN, 14 (29.2%) were found to be positive by Ziehl-Neelsen staining, 15 (31.2%) were positive by culture and 43 (89.6%) cases were positive after first round of PCR while 48 (100%) cases were positive by nMPCR assay. The sensitivity and specificity of nMPCR was found to be 100% for the diagnosis of TBLN. The results thus obtained indicate that nMPCR assay is a highly sensitive and specific tool for the diagnosis of TBLN.

Outbreak of Avian Tuberculosis in Commercial Domestic Pekin Ducks ( Anas platyrhynchos domestica).

Avian tuberculosis is a contagious disease affecting various domestic and wild bird species, and is caused by Mycobacterium avium . It is reported extremely rarely in commercial poultry flocks and has not been reported in commercial domestic ducks to date, with domestic ducks reported to be moderately resistant to M. avium infection. Here, we report the outbreak of avian tuberculosis in commercial Pekin duck ( Anas platyrhynchos domestica) flocks. Postmortem and histopathologic findings included nodules presenting in the visceral organs of ducks, and granulomas with central caseous necrosis surrounded by infiltrating lymphocytes. The M. avium pathogen was isolated and further identified by Ziehl-Neelsen staining and PCR based on insert sequence IS901 and the 16S rRNA gene. We highlight that avian tuberculosis not only has economic significance for the duck industry, but also presents a potential zoonotic hazard to humans.

Staining with two observational methods for the diagnosis of tuberculous meningitis.

Ziehl-Neelsen (Z-N) staining of cerebrospinal fluid (CSF) for acid-fast bacilli (AFB) is the cornerstone of the laboratory diagnosis of tuberculous meningitis (TBM). However, the sensitivity of conventional Z-N staining for the detection of AFB in CSF specimens is suboptimal. The present study aimed to compare the practicality of modified Z-N staining with light microscopy and fluorescence microscopy in the same smear without auramine O. A total of 155 patients with 223 CSF specimens were enrolled and grouped according to the uniform case definition. The smears of each CSF specimen were subjected to modified Z-N staining and then observed using a light microscope under transmitted light and under fluorescence with a green-excitation wavelength in the same microscopic field. The results for different groups, inspection times, and prior to and following treatment were compared. Results indicated that the fuchsin-stained AFB were visible as bright orange-red fluorescing rods under fluorescence, or as red, lightly curved rods under transmitted light. The sensitivity of fluorescence microscopy was 96.2% while that of light microscopy was 84.6%. The positive rate of fluorescence microscopy was 79.2% prior to treatment compared with 61.7% post-treatment. In the same microscopic field, a greater number of AFB were observed using fluorescence compared with transmitted light, and AFB that were not visible under transmitted light were clearly observed under fluorescence. Furthermore, transmitted light and fluorescence could be interchanged directly when equivocal smears were encountered. The combination of modified Z-N staining and fluorescence microscopy without auramine O is sensitive and convenient for the diagnosis of TBM.