Using Ultrasound Guided Needle Biopsy in Combination With GeneXpert MTB/RIF to Diagnose Epididymal Tuberculosis: a Study of 12 Cases

Objective: This study aims to investigate the diagnostic utility of percutaneous ultrasound-guided needle biopsy in combination with GeneXpert MTB/RIF for epididymal tuberculosis.Methods: Between March 2019 to March 2021, specimens obtained from 12 patients with confirmed epididymal tuberculosis by ultrasound guided needle biopsy were sent for pathology and laboratory examination at the Shandong Public Health Clinical Center. The laboratory examination included acid-fast staining, Mycobacterium tuberculosis culture by BACTEC MGIT 960, and the GeneXpert MTB/RIF test. The diagnosis and complications were comprehensively analyzed.Results: Among the 12 cases, seven cases had granulomatous inflammation and necrotic tissue, four cases had chronic inflammatory cells and necrotic tissue, and one case had chronic inflammatory cell infiltration. Furthermore, among the 12 patients, five patients were positive for Mycobacterium tuberculosis culture, two patients were positive for the acid-fast staining, and twelve patients were positive for the GeneXpert MTB/RIF assay. The sensitivity and specificity of the acid-fast staining, Mycobacterium tuberculosis culture and GeneXpert MTB/RIF in the diagnosis of epididymis tuberculosis were 16.67% and 100.00%, 41.67% and 100.00%, and 100.00% and 50.00%, respectively. The diagnostic value analysis of the three detection techniques indicated that the GeneXpert MTB/RIF technique (Kappa=0.63, AUC=0.75) is superior to the Mycobacterium tuberculosis culture (Kappa=0.17, AUC =0.71) and acid-fast staining (Kappa=0.05, AUC=0.58). Conclusion: Ultrasound guided percutaneous biopsy combined with GeneXpert MTB/RIF assay is extremely useful for diagnosing epididymitis tuberculosis and determining rifampin resistance, with superior sensitivity, specificity and AUC value.

Detection of Cryptosporidiosis in Dogs of Veterinary Clinics in Surabaya City Using Acid-Fast Staining and PCR

The need for maintaining pets, such as dogs, is increasing along with the human population. When individuals keep dogs as their pets, they must be aware of disease transmission from dogs. One of the disease agents transmitted from pets to their owners is Cryptosporidium spp. causing cryptosporidiosis. The aim of the present study was to detect Cryptosporidium spp. infection in dogs through a fecal examination using the acid-fast staining method (Ziehl Neelsen) confirmed with the molecular examination of Polymerase Chain Reaction (PCR). Detection of Cryptosporidium sp. in feces of dogs was set up by using an acid-fast staining method. Positive results of the acid-fast staining were further confirmed using PCR. Polymerase Chain Reaction used primary AB210854 specific to the Cryptosporidium canis and S139-S141 genes which were specific primary for the Cryptosporidium parvum gene. Results of the acid-fast staining showed that 80% of the samples (40 samples from total samples) were infected with Cryptosporidium spp. Further detection using PCR showed that four samples were positive for Cryptosporidium canis infection, and two samples showed positive results of Cryptosporidium parvum infection. Dog samples were mostly infected with Cryptosporidium spp. including Cryptosporidium canis and Cryptosporidium parvum through a fecal examination using acid-fast staining and PCR. Keywords: Acid-fast staining, Cryptosporidium spp., Dogs, PCR

Gross and histopathological lesions associated with tuberculosis in two sloth bears (Melursus ursinus) in India

Post-mortem examination of two sloth bears which died in Bannerghatta Bear Rescue Centre, Bengaluru, Karnataka, were performed. Both the animals were anorectic and had considerable weight loss before death. Representative lung tissue samples were subjected to histopathology and staining. The lung tissues of the animals had diffuse congestion and subpleural petechial hemorrhages. In addition, small nodules of varied diameters were seen scattered on the lung lobes of both animals. On histopathological examination, the lung tissue of one of the animals showed extensive proliferation of blood vessels. Congestion and subpleural hemorrhages were seen in both cases. Few macrophages and epithelioid cells were seen scattered adjacent to a bronchiole. Kinyoun’s acid fast staining of the histological sections revealed numerous acid fast bacilli indicative of tuberculosis.

Prevalence and genetic characterization of Cryptosporidium in pre-weaned cattle in Urmia (Northwestern Iran)

Introduction: Cryptosporidiosis is a zoonotic disease causing digestive problems in pre-weaned calves. Considering the zoonosis of the parasite and its importance in veterinary medicine, we evaluated the prevalence and genotyping of Cryptosporidium spp. in diarrheic pre-weaned calves in the northwest of Iran. Methodology: A total of 100 stool samples of the infant calves with diarrhea were collected from industrial and conventional livestock farms in Urmia City. All the samples were tested with acid-fast staining, ELISA, and PCR. Positive samples of the PCR method were sequenced to determine the Cryptosporidium species. The obtained results were compared for the mentioned methods based on statistical factors, sensitivity, specificity, positive and negative predictive values, as well as duration of the experiment and the costs of testing. Results: The results of this study showed that the prevalence of Cryptosporidium spp. in diarrheic infant calves in Urmia city was 5%, and C. parvum species of Cryptosporidium was detected in all the sequenced samples. According to the findings of the current study, the most appropriate method for the detection of the parasite is the ELISA that has a higher sensitivity and predictive value than acid-fast staining method and should be used in veterinary laboratories. Conclusions: In the current investigation, C. parvum was identified as the only infectious agent in the region and could be the main cause of human infection. More studies are needed to find the source of infection for establishing the control measures.

The value of GeneXpert MTB/RIF in bronchoalveolar lavage fluid in the diagnosis of smear-negative pulmonary tuberculosis

This study aims to evaluate the diagnostic value of Xpert MTB/ RIF assay in bronchoalveolar lavage fluid (BALF) in subjects with smear-negative pulmonary tuberculosis. From January 2019 to December 2019, 197 patients with suspected pulmonary tuberculosis were recruited, and bronchoalveolar lavage fluid was collected for acid-fast staining smear, liquid culture of Mycobacterium combined drug sensitivity and Xpert MTB/RIF detection. The sensitivity, specificity, positive predictive value and negative predictive value of Xpert MTB/RIF in bronchoalveolar lavage fluid (BALF) were calculated with smear-negative pulmonary tuberculosis as the reference standard. The consistency of xpert MTB/RIF in the diagnosis of rifampicin resistance was evaluated, with the results of Mycobacterium liquid culture drug sensitivity test and drug sensitivity test as the gold standards. The results showed that among 197 suspected tuberculosis patients, 55 patients were not diagnosed with tuberculosis and 142 patients were diagnosed with smear-negative pulmonary tuberculosis. One hundred and twenty three cases (86.62%) were positive for Xpert MTB/ RIF in bronchoalveolar lavage fluid, 15 cases (10.56%) were positive by acid-fast staining smear method, and 88 cases (61.97%) were positive by the liquid culture method. The positive rate of Xpert MTB / RIF was 93.18% (82 / 88), which was higher than that of 75.93% (41 / 54) of the negative BALF mycobacterium culture (χ 2 = 8.598, P<0.01). The sensitivity and specificity of Xpert MTB/RIF for rifampicin resistance were 100.00% and 97.30%, respectively. Therefore, the diagnostic value of Xpert MTB/RIF in bronchoalveolar lavage fluid for bacterialnegative pulmonary tuberculosis is superior to the acid-fast staining smear of lavage fluid and the mycobacterium culture method.

A Case Study of Multiple Parasitisms in a Calf Buffalo (Bubalus bubalis)

Background: Gastrointestinal (GI) parasitism by protozoan and helminth parasites exists as one of the major limiting factors in the buffalo industry, especially in the wellbeing of the calves around the developing countries like Nepal. During a field survey on buffaloes, we encountered a two- and half month ill male calf suffering from various illnesses for 14 days. Methods: We collected its stool sample for three days and processed through a direct wet mount, sedimentation, floatation and acid-fast staining techniques and observed via a compound microscope. Result: We detected the multiple patterns of infections of GI parasites and even counted the oocysts of coccidia and eggs of nematodes released per gram of the feces and discussed that co-infection was associated with the pathologic consequences. Coprological surveys with appropriate egg/oocyst counting techniques are useful in the treatment and preventive options of GI infections in calves.