Ziehl-Neelsen Research Articles

A Review on Paragonimiasis and its Differential Diagnosis Technique

This review highlights the existence of Paragonimus (PRG) and how they are mistaken for Tuberculosis (TB) during diagnosis. PRG is a parasitic lung fluke (flatworm) that infect the lungs, human serves as the first hosts, crabs, crayfish, and snails as the second intermediate hosts. The north-eastern states of India are endemic to PRG and infection is acquired by consuming uncooked or partially cooked crustaceans. Due to their similar clinical presentation to TB, PRGs are usually misdiagnosed, leading to delayed treatment. Ziehl-Neelsen (ZN) stain, conventional wet film for staining the sputum, pleural fluid, and stool, and lung biopsy are different laboratory tests to determine PRG infection. In addition, efficient technique that can aid diagnosis are immunological assays, like Enzyme-linked immunosorbent assay (ELISA), dot-ELISA, complement fixation test (CFT), intradermal test (ID), Western blot, immunodiffusion and indirect haemagglutination test (IHA). In India, PRG infection is most likely to persist until medical professionals and governing bodies raise awareness to implement adequate management measures.

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

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

Identification of Cytomegalo Virus Infection Among Mycobacterium Tuberculosis in Baghdad City. Correlation between TB and CMV infection

Objectives: Human cytomegalovirus (HCMV) infection and immunological activation are risk factors for tuberculosis disease, according to recent research. Materials and Methods: The best procedure for the detection (TB) was investigated by Ziehl Neelsen and Auramine O (AO) technology. 315 sputum samples were collected randomly. The ZN approach produced a smear positivity rate for AFB of 49/315 (15.55%) while the LED Auramine O staining method produced a positivity rate of 69/315 (21.9%), One hundred serum samples were subjected to HCV IgG and IgM detection using Cassette method and ELISA techniques. Results: The results of the cassette method documented there were 62/100 (62%) cases of M. tuberculosis patients who had HCMV IgG versus 16/100 (16%) cases who had HCMV having IgM (P-value= 0.001). While the results of ELISA techniques observed there were 65 (65%) cases of M. tuberculosis patients had HCMV IgG versus 17/100 (17%) cases had HCMV had IgM these differences statistically were highly significant. These results observed the sensitivity of the cassette method and ELISA procedure for diagnosis of CMV among M.tuberculosis were 92% for IgG and 94% for IgM antibodies. Conclusions: According to recent findings from CMV infection significantly increases the probability of active TB disease.

Retracted: Application Value of SAT-TB Combined with Acid-Fast Staining in the Diagnosis and Treatment of Pulmonary Tuberculosis.

[This retracts the article DOI: 10.1155/2020/3620425.].

The Enhancing Accuracy in Mycobacterium Tuberculosis Counting on Acid-Fast Bacteria Sputum through Microscope Camera and Image Processing with Thresholding Method

Tuberculosis (TB) is a chronic infectious illness. Pulmonary TB is still a global threat and concern. Currently, TB is among the top 10 causes of death in the world in 2015 and it is estimated that there are around 10.4 million new cases of TB in the world. 6 countries account for around 60% of new cases in the world: India, Indonesia, China, Nigeria, Pakistan, and South Africa. Globally, the TB death rate was around 22% of total world deaths from 2000 to 2015. This study introduces an innovative tool, the "Microscope Camera for Enhanced Mycobacterium Tuberculosis Counting in Acid-Fast Bacteria Sputum through Image Processing with Thresholding Method," designed to revolutionize the examination of tuberculosis (TB) samples. In response to the global threat of Pulmonary Tuberculosis, the tool employs a state-of-the-art CMOS-HD (Complementary metal-oxide-semiconductor High Definition) camera module and leverages the Python application for efficient image processing. The primary objective is to address the limitations faced by conventional diagnostic methods, particularly the challenges associated with the Rapid Molecular Test Method due to its dependence on expensive equipment, thereby making it less feasible for implementation in resource-constrained public health centers. The study underscores the persistent threat of TB as a global health concern, emphasizing its prevalence, with approximately 10.4 million new cases worldwide. Focusing on Ziehl-Neelsen stain sputum samples, the research seeks to elevate the accuracy of TB diagnosis by proposing a streamlined approach. This involves integrating a digital microscope system with a CMOS-HD camera module and utilizing advanced image processing techniques, specifically the Python Thresholding method. In the methodology, the research outlines a comprehensive data collection and analysis process. Mechanical and block diagrams illustrate the design of the module, while the system flow diagram elucidates the sequence of steps involved in TB sample analysis. The data analysis section employs metrics such as average, percentage error, and success percentage, providing a quantitative assessment of the system's performance. The results reveal an encouraging average system accuracy of 85.30%, demonstrating the potential of the developed tool to enhance the diagnostic process. However, the study transparently acknowledges areas for improvement, particularly in addressing discrepancies between manual and system calculations. The conclusion emphasizes the tool's promising role in assisting healthcare workers in detecting and counting TB bacteria in sputum, underscoring the need for ongoing refinements to achieve a definitive 100% accuracy and eliminate the possibility of misdiagnoses.

Identification of major protozoal enteropathogens causing calf diarrhea in dairy farms in and around Holeta Town, Oromia Special Zone, Ethiopia

Calf diarrhea can be caused by a variety of pathogens, including bacteria, viruses, protozoa, and intestinal parasites. Giardia, Eimeria, and Cryptosporidium are the most significant protozoan parasites and are all individually and ollectively infectious. A case-series study was carried out in and around Holeta dairy farms, Oromia Special Zone, Ethiopia, from November 2017 to April 2018 to identify the main protozoan enteropathogens from diarrheic calves. Samples were purposely collected from three dairy farms: Serkalem Dairy Farm (SDF), Holeta Agricultural Research Centre (HARC), and Ada Berga Agricultural Research Centre (ABARC). A total of 93 fecal samples were taken from diarrheic calves of up to 4 months of age. Protozoan enteropathogens were identified using flotation and modified Ziehl Neelsen staining methods. Cryptosporidium, Eimeria, and Giardia were detected in 44 (47.3%), 50 (53.8%), and 34 (36.6%) of the diarrheic fecal samples examined, respectively. The findings indicated that there were 17 (18.27%) cases of Cryptosporidium, Eimeria, and Giardia as mixed infections, 16 (17.2%) cases of Cryptosporidium and Eimeria, 5 (5.4%) cases of Cryptosporidium and Giardia, and 4 (4.3%) cases of Eimeria and Giardia, compared to 13 (13.97%), 6 (6.45%), and 5 (5.4%) cases of Cryptosporidium, Eimeria and Giardia as single infections, respectively. The selected farms had significant prevalence levels of three common protozoa: Eimeria, Giardia, and Cryptosporidium. Further molecular research is required to identify the species and genotype levels of protozoal enteropathogens and related risk factors.

Prevalence of enteroparasites in rural and urban children in the municipalities of Bocaiúva do Sul and Colombo, Paraná, Brazil

The aim of this study was to diagnose enteroparasites and to relate them to socioeconomic factors in schoolchildren from the public system in the municipalities of Colombo and Bocaiúva do Sul, located in the Metropolitan Region from the city of Curitiba, Brazil. Fecal samples were processed using the modified Ritchie and Ziehl-Neelsen staining methods. Analysis of the risk factors, using Tukey test and Odds ratio (OR), was obtained by connecting questionnaires with parasitological results. Of the 414 schoolchildren analyzed, 32.4% (134) were positive for at least one enteroparasite species. The most prevalent were Blastocystis spp. (21.7%), Entamoeba coli (5.5%), Endolimax nana (5.5%), Giardia duodenalis (3.9%), Trichuris trichiura (3.7%) and Ascaris lumbricoides (2.9%). Schoolchildren from rural areas had a higher occurrence of enteric parasites (p<0.05), showing that is necessary to continue with health education, being careful with the food, basic sanitation and parasitological examination of feces before treatment. KEY WORDS: Enteroparasites; monoparasitism; multiparasitism; children; socio-echonomic factors.

Concomitant Tuberculous and Lepromatous Lymphadenitis: Clues and Pitfalls of Leprosy Concealed by Tuberculosis in Lymph Nodes.

Comorbidities between tuberculosis and leprosy are expected in endemic regions. Pulmonary tuberculosis and cutaneous leprosy are the most prevalent coinfections. One of the common manifestations of tuberculosis is generalized lymphadenopathy. In contrast, leprosy is clinically less suspected to manifest as a generalized lymphadenopathy, and it is pathologically unusual to diagnose leprosy primarily in lymph nodes. Concomitant tuberculous and lepromatous lymphadenitis are unprecedented and clinically unexpected, particularly in nonendemic countries. This imposes diagnostic challenges. We report concurrent tuberculosis and leprosy that were diagnosed in a lymph node in 45-year-old man with generalized lymphadenopathy. The effaced lymph node was predominantly replaced by caseating epithelioid granulomas alternating with foamy histiocytes. Ziehl-Neelsen stain showed positive acid-fast bacilli in the necrotizing granulomas only. The initial differential diagnosis of the nodal foamy macrophages included fungal infections, leishmaniasis, and Whipple disease, for which the special stains were negative. The vacuolated macrophages were disregarded as nonspecific lipogranuloma. A modified acid-fast stain was not considered. The histopathologic clues to nodal lepromatous leprosy included the presence of intracytoplasmic globi, intermixed microabscesses, and lymphoplasmacytic infiltrate and involved pericapsular nerves. Wade-Fite stain was subsequently performed. It revealed numerous lepra bacilli within the foamy histiocytes. The final diagnosis was concurrent disseminated tuberculosis and leprosy. Nodal lepromatous leprosy could be missed when compounded by concurrent nodal tuberculosis, particularly in developed countries. The clinicians and pathologists should have a high index of suspicion, particularly in patients from or with history of travel from endemic regions. Certain histopathologic features are helpful clues to avoid pitfalls.

Intestinal Parasitic Infections in People Referring to the Central Laboratory of Meshkin Shahr County, Ardabil Province, Iran.

Intestinal parasitic infections are still a considerable global public health problem. We aimed to determine the frequency of intestinal parasitic infections among people referring to the central laboratory of Meshkin Shahr City, Ardabil Province, Iran. In this cross-sectional survey, 460 fecal samples were collected randomly from persons referred to the central laboratory of Meshkin Shahr City, from January to June 2022. The samples were examined by direct wet-mount, Trichrome and modified Ziehl-Neelsen staining, formalin ethyl acetate sedimentation, and agar plate culture. The frequency of intestinal parasites was 15.7% (72 out of 460 cases), with some people with numerous intestinal parasites. The frequency of protozoan infections (13.9%) was higher than the helminthic infections (2.6%). Blastocystis spp. (8.1%) was the most prevalent detected intestinal protozoan. Entamoeba coli (5.7%), Dicrocoelium dendriticum (2.2%), Giardia lamblia (1.5%), Fasciola spp. (0.2%), and Hymenolepis nana (0.2%) were other detected parasites. In- spite of betterment of the health condition in Iran and reduction of parasitic infection, intestinal parasitic infections are still a considerable public health issue in some parts of Iran.

Absence of Tuberculosis-Causing Mycobacteria from Slaughtered Livestock Tissues and Environmental Samples, Gauteng Province, South Africa.

Mycobacterium tuberculosis complex (MTBC) is a group of bacteria responsible for causing tuberculosis in animals and humans. In South Africa (S.A), slaughterhouses are registered by the government and closely inspected and audited for hygienic slaughter practices. Meat inspection to detect lesions has been used for passive surveillance, monitoring, and diagnosis of the disease status. Information on the current status of bovine tuberculosis (bTB) in livestock in the country is limited. Hence, we investigated the occurrence of Mycobacterium spp. in the tissues of slaughtered livestock and environmental samples in abattoirs in Gauteng province of South Africa (S.A). The cross-sectional study employing random sampling from cattle, pigs, and sheep (with the collection of liver, lung, spleen, and different lymph nodes) irrespective of lesions was carried out in 19 red meat abattoirs. Five hundred animals were sampled, comprising cattle (n = 369), pigs (n = 90), and sheep (n = 41). Additionally, 19 environmental samples were collected from feedlots, or where animals drink water while awaiting slaughter, to identify mycobacterial species using culture, acid-fast bacteria staining, and polymerase chain reaction (PCR). The Chi-square and Fisher's Exact tests were used to detect statistically significant differences in the frequency of detection of Mycobacterium spp. according to the variables investigated (types of tissues, livestock, abattoirs, etc.). The PCR assays detected no MTBC complex species DNA in the bacterial isolates from cattle (n = 32). Sequence analysis (16S rDNA) of the isolates from eight cattle confirmed only two species, namely Mycobacterium colombiense (99.81% identity) and Mycobacterium simiae (99.42% identity). The remaining isolates were identified as members of the Actinomadura species. From the environmental samples, bacterial isolation was made from three samples, and two could only be identified up to the genus level (Mycobacterium species) while the remaining isolate was identified as Mycobacterium senuense (99.22% identity). The study revealed the absence of bovine tuberculosis-causing pathogens in red meat abattoirs of the Gauteng province. Although non-tuberculous Mycobacteria have been implicated as potentially causing tuberculosis-like diseases in livestock, their occurrence in the current study was found to be low, but the potential to cause disease cannot be ignored.

Granulomas co-occurring in malignancies – tale of etiological relationship

Objectives To determine the possible aetiopathogenesis of co-occurrence of granulomas with different malignancies in different body sites. Material and methods All cases with granuloma formation observed in draining lymph nodes or in the primary site of malignant tumors were included in the present study. After routine histopathology examination, modified Ziehl-Neelsen (ZN) staining for Mycobacteria was carried out in all the cases. Detailed history, especially of Tuberculosis, sarcoidosis, neoadjuvant chemotherapy, radiation, or previous procedure, was recorded. Results 11 out of 35 cases (31.4%) had granulomas within the primary tumor, while 24 cases out of 35 (68.6%) showed nodal granulomas. Of the 24 cases, 5 cases had nodal metastatic tumor deposits. Also, necrotizing granulomas with AFB were significantly more as compared to AFB in non-necrotizing granulomas (p value of 0.05). Of the total cases, 20% had an attributed risk factor. Three cases received neoadjuvant chemotherapy, and three had a history of systemic tuberculosis, while one case had associated Crohn’s disease. Conclusion We recommend to characterize granulomas as necrotizing/nonnecrotizing, confluent/discrete, tumor deposit present/absent; prior history of systemic/local illnesses (like SS, TB, fungal infections, IBD, etc.); prior history of CT/RT; and to follow routine ZN staining in all cases of granulomas with malignancy.

Diagnosis of multiple tuberculous muscle abscesses in a patient with systemic lupus erythematosus by metagenomic next-generation sequencing- a case report and literature review

BackgroundEarly diagnosis of muscular tuberculosis (TB) without coexistent active skeletal involvement is often challenging because the disease is very rare and its clinical manifestation is nonspecific and misleading. To raise the awareness and emphasize early diagnosis of muscular TB, we present a case of multiple tuberculous muscle abscesses in a systemic lupus erythematosus (SLE) female, but without pulmonary tuberculosis (PTB), in order to increase awareness of and stress the need of early detection of muscular TB.Case presentationA 44-year-old woman with a 6-year history of SLE who had been treated with methylprednisolone for a long time complained of erythema on her trunk and extremities for five months, along with edema and myalgia for two months, and fever for one month. The patient was first misdiagnosed as SLE overlap dermatomyositis. However, an ultrasound-guided drainage of muscle abscesses revealed positive acid-fast staining combined with positive deoxyribonucleic acid fragment of Mycobacterium tuberculosis using metagenomic next-generation sequencing (mNGS). The patient was cured and released following standard anti-tuberculosis medication, local puncture drainage, and an intravitreal injection of streptomycin. Literature search found only 19 cases of tuberculous muscle abscesses occurring in the extremities reported from 1999 to 2023.ConclusionsExtrapulmonary TB with predominantly muscle involvement is rare and with no specific clinical presentation. Muscular tuberculosis may be disdiagnosed for dermatomyositis due to the high muscle enzyme levels, delaying diagnosis and treatment. mNGS technology is helpful in the early and rapid diagnosis of muscular TB. On the basis of traditional anti-tuberculosis treatment, an ultrasound-guided percutaneous puncture drainage and intracavitary injection of streptomycin for the treatment of tuberculous muscle abscess is easy to operate, safe and effective, which is worthy of clinical popularization and application.

First detection of bovine tuberculosis by Ziehl-Neelsen staining and polymerase chain reaction at dairy farms in the Lekok Sub-District, Pasuruan Regency, and Surabaya region, Indonesia.

Bovine tuberculosis (TB) is a zoonotic disease of great public health importance, particularly in Indonesia, where control measures are limited or are not implemented. This study aimed to detect the presence of Mycobacterium pathogens in milk samples from dairy cattle in Pasuruan regency and Surabaya City, East Java, using Ziehl-Neelsen acid-fast staining and polymerase chain reaction (PCR). Milk samples were aseptically collected from 50 cattle in the Lekok Subdistrict, Pasuruan Regency, and 44 from dairy farms in the Lakarsantri Subdistrict, Wonocolo Subdistrict, Mulyorejo Subdistrict, and Kenjeran Subdistrict, Surabaya, East Java. To detect Mycobacteria at the species level, each sample was assessed by Ziehl-Neelsen staining and PCR using the RD1 and RD4 genes. The results of PCR assay from 50 samples in Lekok Subdistrict, Pasuruan Regency showed that 30 samples (60%) were positive for Mycobacterium tuberculosis and two samples (4%) were positive for Mycobacterium bovis, although Ziehl-Neelsen staining did not show the presence of Mycobacterium spp. In the Surabaya region, 31 samples (70.45%) were positive for M. tuberculosis and three samples (6.8%) were positive for M. bovis. Six samples (13.63%) from all PCR-positive samples could be detected microscopically with Ziehl-Neelsen. The presence of bovine TB in this study supports the importance of using a molecular tool alongside routine surveillance for a better understanding of the epidemiology of bovine TB in East Java.

Summary of the clinical characteristics of tuberculous otitis media

Objective:The purpose of this study was to analyze and summarize the clinical characteristics and diagnostic methods of tuberculous otitis media（TOM）, to enrich clinical experience in diagnosis and treatment of tuberculous otitis media, so as to reduce missed diagnosis and misdiagnosis, and facilitate timely and effective therapy for better prognosis. Methods:This study retrospectively analyzed the clinical data of patients with tuberculous otitis media who were hospitalized in the Ear ward of our hospital and received surgical treatment from 2008 to 2022. The data of patients' clinical characteristics, radiological examination, intraoperative findings and therapeutic strategies were recorded and summarized. Results:A total of 23 cases （26 ears） of tuberculous otitis media were included in this retrospective study. The most common clinical symptoms were otorrhea（thin odorless fluid）（100%） and conductive hearing loss（100%）, with a high incidence of facial paralysis（23.1%）. It was not sensitive to traditional antibiotic treatment, eg. Levofloxacin （50% effective rate only）, and relapsed soon after drug withdrawal. It was revealed that all the surgical views had gray and white tough granulation tissue hyperplasia（100%）, and 23.1% with caseous necrosis. The purpose of surgery was to clear the lesion, reduce the recurrence rate of suppurative infection, and repair the function （hearing reconstruction or facial nerve decompression） as appropriate. The paraffin pathology of granulation tissue were reported as typical granulomatous inflammation and caseous necrosis with positive acid-fast staining, which was consistent with tuberculosis. Conclusion:It was easily confused by the clinical manifestations of tuberculous otitis media and common chronic suppurative otitis media. When met with the following conditions, we should pay highly attention to suspect tuberculous otitis media: The severity of local manifestations did not match with the length of the disease; with poor tympanic membrane at the early stage with no obvious cholesteatomas, with facial paralysis or hearing loss early onset; insensitive to traditional antibiotic treatment; with extensive granulation appeared in the tympanum and or mastoid cavity, with or without caseous necrosis or dead bone in the early days. The diagnosis should be confirmed based on the acid-fast staining of the histopathological section to detect positive acid-fast bacilli. Meanwhile, multiple laboratory examination methods（such as T-spot and PCR） should be integrated synchronously to help support the diagnosis.

Management of newborns and healthcare workers exposed to isoniazid-resistant congenital tuberculosis in the neonatal intensive care unit

Management of newborns and healthcare workers (HCWs) exposed to congenital tuberculosis (TB) in the neonatal intensive care unit (NICU) has been reported rarely. To outline a contact investigation process for individuals exposed to congenital TB in the NICU and investigate nosocomial transmission. Additionally, to assess the efficacy and safety of window prophylaxis in exposed newborns. A baby, born at a gestational age of 28+ 1 weeks, was diagnosed with isoniazid-resistant congenital TB on the 39th day of admission to the level IV NICU. Newborns and HCWs exposed cumulatively for ≥8h underwent contact investigation and follow-up for a year. Eighty-two newborns underwent contact investigation. All newborns displayed normal chest X-rays, and 42 hospitalized newborns tested negative for acid-fast bacilli stain and Xpert® MTB/RIF assay in their endotracheal sputum or gastric juices. Eighty received window prophylaxis: six of 75 on rifampin experienced mild adverse events, and none of the five on levofloxacin. After 12 weeks, five (6.1%) had a positive tuberculin skin test, all of whom had already received the Bacillus Calmette-Guérin vaccine and tested negative on TB interferon-gamma releasing assay. Of 119 exposed HCWs, three (2.5%) were diagnosed with latent TB infection and completed a four-month rifampin therapy. There was no active TB disease among exposed newborns and HCWs during a one-year follow-up. Timely diagnosis of congenital TB is crucial for minimizing transmission among exposed neonates and HCWs in the NICU setting. In cases of isoniazid-resistant index patients, even premature newborns may consider the use of rifampin or levofloxacin for window prophylaxis.

Pyroptosis Tuning in Intestinal Cryptosporidiosis via the Natural Histone Deacetylase Inhibitor Romidepsin.

Cryptosporidium is an opportunistic protozoan, with many species of cross-human infectivity. It causes life-threatening diarrhoea in children and CD4-defective patients. Despite its limited efficacy, nitazoxanide remains the primary anti-cryptosporidial drug. Cryptosporidium infects the intestinal brush border (intracellular-extracytoplasmic) and down-regulates pyroptosis to prevent expulsion. Romidepsin is a natural histone deacetylase inhibitor that triggers pyroptosis. Romidepsin's effect on cryptosporidiosis was assessed in immunocompromised mice via gasdermin-D (GSDM-D) immunohistochemical expression, IFN-γ, IL-1β and IL-18 blood levels by ELISA, and via parasite scanning by modified Ziehl-Neelsen staining and scanning electron microscopy (SEM). Oocyst deformity and local cytokines were also assessed in ex vivo ileal explants. Following intraperitoneal injection of romidepsin, oocyst shedding significantly reduced at the 9th, 12th and 15th d.p.i. compared with infected-control and drug-control (nitazoxanide-treated) mice. H&E staining of intestinal sections from romidepsin-treated mice showed significantly low intestinal scoring with marked reduction in epithelial hyperplasia, villous blunting and cellular infiltrate. SEM revealed marked oocyst blebbing and paucity (in vivo and ex vivo) after romidepsin compared with nitazoxanide. Regarding pyroptosis, romidepsin triggered significantly higher intestinal GSDM-D expression in vivo, and higher serum/culture IFN-γ, IL-1β and IL-18 levels in romidepsin-treated mice than in the control groups. Collectively, in cryptosporidiosis, romidepsin succeeded in enhancing pyroptosis in the oocysts and infected epithelium, reducing infection and shifting the brush border towards normalisation.

Predictors for Concurrent Diabetes in Tuberculosis Patients. Perspectives from Two Mining Districts of Eastern Tribal State Jharkhand, in India.

Tuberculosis and diabetes both diseases are present in large numbers in the country and we are major contributors to both globally. With the objective to understand the various traits of patients having both tuberculosis and diabetes and to ascertain various possible predictors for such occurrence based on the public health database we carried out this study. We seek answers to questions like they have any effects? Are they having any additive role to play? One-year data from the NIKSHAY portal of both districts were analyzed to look for possible associations and other variable traits. Data were analyzed using standard methods to express data in frequency and percentage. Chi-square test was used to establish association, while step-wise approach was used to calculate univariate and multivariate logistic regression analysis for knowing various predictors. P-value of <0.05 was considered statistically significant. Concurrent diabetes in tuberculosis patients was close to 294 (6%) in the 4933 individuals. In total, 65.2% of the study population were male. Diagnosis of tuberculosis was made most of the time by chest X-ray (49.4%) followed by Microscopy ZN staining and cartridge-based nucleic acid amplification test (CBNAAT). Death was more among diabetics (4.4%) as compared to nondiabetics (3.5%). Diabetes is increasing in tuberculosis patients; improvement in data quality is needed. More research is required to reveal various other reasons that make tuberculosis patients more prone to develop diabetes.

Diagnostic advancements: Isolating Mycobacterium avium ssp. paratuberculosis and unveiling its molecular identity with nested-PCR.

Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of paratuberculosis, which is currently prevalent in many parts of Iran and produces severe economic loss. It is hence necessary to identify and isolate the animals infected with this bacterium, so this research aimed to isolate MAP from milk and fecal samples of ELISA-positive animals and determine the molecular identity of isolates. After performing ELISA on 3,700 bovine blood samples, 115 samples of milk and feces were taken from ELISA-positive cattle and were cultured on Herald's egg yolk medium with and without mycobactin-J and then the acid-fastness of positive samples was determined using Ziehl-Neelsen staining. The 16S rRNA-PCR test was performed after DNA extraction to determine the molecular identity of isolates. Primers IS6110 and IS901 were employed to ensure that the isolates were not related to members of M. tuberculosis complex and M. avium, respectively. Primer IS900 was also used to determine the molecular identity of MAP isolates. Also, expression levels of MAP-related genes (IS900, ISMAP02, F57, MAP2191, MAP4027) were evaluated via qPCR. Finally, positive samples were confirmed based on the Nested-PCR. Results showed that a total of 9 isolates were obtained from the culture of 90 ELISA-positive samples. The results revealed that all grown samples were positive for acid-fastness. The 16S rRNA-PCR test revealed the 543 bp band, which confirms the presence of Mycobacterium in the samples. The PCR test with Primer IS900 generated the 398 bp fragment in the first step and the 298 bp fragment in the second step, indicating the presence of MAP in samples. Also, relative expression analysis revealed that MAP-related genes were significantly higher in ELIZA-positive samples than in negative ones. Based on the study findings, it can be concluded that MAP-infected animals can be identified by ELISA. In addition, mycobacterium can be isolated by culturing the samples on appropriate media and then its molecular identity can be determined by using nested-PCR.

Fullertubes inhibit mycobacterial viability and prevent biofilm formation by disrupting the cell wall.

Mycobacterium tuberculosis and nontuberculous mycobacteria such as Mycobacterium abscessus cause diseases that are becoming increasingly difficult to treat due to emerging antibiotic resistance. The development of new antimicrobial molecules is vital for combating these pathogens. Carbon nanomaterials (CNMs) are a class of carbon-containing nanoparticles with promising antimicrobial effects. Fullertubes (C90 ) are novel carbon allotropes with a structure unique among CNMs. The effects of fullertubes on any living cell have not been studied. In this study, we demonstrate that pristine fullertube dispersions show antimicrobial effects on Mycobacterium smegmatis and M. abscessus. Using scanning electron microscopy, light microscopy, and molecular probes, we investigated the effects of these CNMs on mycobacterial cell viability, cellular integrity, and biofilm formation. C90 fullertubes at 1 µM inhibited mycobacterial viability by 97%. Scanning electron microscopy revealed that the cell wall structure of M. smegmatis and M. abscessus was severely damaged within 24 h of exposure to fullertubes. Additionally, exposure to fullertubes nearly abrogated the acid-fast staining property of M. smegmatis. Using SYTO-9 and propidium iodide, we show that exposure to the novel fullertubes compromises the integrity of the mycobacterial cell. We also show that the permeability of the mycobacterial cell wall was increased after exposure to fullertubes from our assays utilizing the molecular probe dichlorofluorescein and ethidium bromide transport. C90 fullertubes at 0.37 µM and C60 fullerenes at 0.56 µM inhibited pellicle biofilm formation by 70% and 90%, respectively. This is the first report on the antimycobacterial activities of fullertubes and fullerenes.

Digital Pathology in the Detection of Infectious Microorganisms: An Evaluation of Its Strengths and Weaknesses Across a Panel of Immunohistochemical and Histochemical Stains Routinely Used in Diagnostic Surgical Pathology.

The diagnosis of some infectious diseases requires their identification in tissue specimens. As institutions adopt digital pathology for primary diagnosis, the limits of microorganism detection from digital images must be delineated. To assess the reliability of microorganism detection from digitized images of histochemical and immunohistochemical stains commonly used in pathology. Original glass slides from 620 surgical pathology cases evaluated for the presence of infectious microorganisms were digitized. Immunohistochemical stains included those for herpes simplex virus (n = 100), cytomegalovirus (n = 100), Helicobacter pylori (n = 100), and spirochetes (n = 80). Histochemical stains included mucicarmine for Cryptococcus spp (n = 20), Grocott methenamine silver for fungi (n = 100), Giemsa for H pylori (n = 100), and Ziehl-Neelsen for acid-fast bacilli (n = 20). The original diagnosis based on the glass slides was regarded as the reference standard. Six pathologists reviewed the digital images. Digital review was generally associated with high (ie, ≥90%) specificity and positive predictive value owing to a low percentage of false positive reads, whereas a high percentage of false negatives contributed to low sensitivity and negative predictive value for many stains. Fleiss κ showed substantial interobserver agreement in the interpretation of Grocott methenamine silver and immunostains for herpes simplex virus, H pylori, and cytomegalovirus; moderate agreement for spirochete, Ziehl-Neelsen, and mucicarmine; and poor agreement for Giemsa. Digital immunohistochemistry generally outperforms histochemical stains for microorganism detection. Digital interpretation of Ziehl-Neelsen and mucicarmine stains is associated with low scores for interrater reliability, accuracy, sensitivity, and negative predictive value such that it should not substitute for conventional review of glass slides.