Histopathological and Molecular Investigation of Natural Cases of Bovine Tuberculosis Infection in Cattle

Bovine tuberculosis (bTB), a chronic infection in cattle caused by Mycobacterium tuberculosis/bovis, that impacts productivity and represents a major public health threat. Although the considerable economic costs and zoonotic risk consequences associated with the disease, accurate estimates of bTB prevalence are lacking in many countries, including India. Therefore, in the current study for collection of tubercular lesions the postmortem examination of 100 cattle was conducted. All major viscera and regional lymph nodes were examined and incised. Histopathology was performed in the cases where gross lesions were suggestive of tuberculosis. PCR was performed on the tissue and faecal samples by using IS6110 insertion sequence, Mycobacterium tuberculosis/bovis complex PCR kit. In 12 animals, nodular lesions with casseating mass suggestive of tuberculosis were observed in the lung tissue. All the 12 lung impression smear and only five faecal smear showed acid fast bacilli stained by Ziehl-Neelsen staining. Histologic features comprised a classic granuloma as a characteristic lesion of tuberculosis composed of a central caseous necrosis with mantle of macrophages, lymphocytes, plasma cells, epithelioid macrophages and Langhan’s giant cells and were observed in all 12 cases. All the tissue samples and 11 faecal samples were positive for the Mycobacterium tuberculosis complex using IS6110 sequence. 8 tissue samples and 4 faecal samples were positive by using Mycobacterium tuberculosis/bovis complex PCR kit. It can be concluded that there was good agreement between histopathology, acid fast staining and PCR. It can also be concluded that faecal samples which are easier to collect should be preferred for diagnosis of TB by PCR in cattle

Evaluation of a nested-pcr for Mycobacterium tuberculosis detection in blood and urine samples

The polymerase chain reaction (PCR) and its variations, such as the nested-PCR, have been described as promising techniques for rapid diagnosis of tuberculosis (TB). With the aim of evaluating the usefulness of a nested-PCR method on samples of blood and urine of patients suspected of tuberculosis we analyzed 192 clinical samples, using as a molecular target the insertion element IS6110 specific of M. tuberculosis genome. Nested-PCR method showed higher sensitivity in patients with extrapulmonary tuberculosis (47.8 percent and 52 percent in blood and urine) when compared to patients with the pulmonary form of the disease (sensitivity of 29 percent and 26.9 percent in blood and urine), regardless of the type of biological sample used. The nested-PCR is a rapid technique that, even if not showing a good sensitivity, should be considered as a helpful tool especially in the extrapulmonary cases or in cases where confirmatory diagnosis is quite difficult to be achieved by routine methods. The performance of PCR-based techniques should be considered and tested in future works on other types of biological specimens besides sputum, like blood and urine, readily obtainable in most cases. The improving of M. tuberculosis nested-PCR detection in TB affected patients will give the possibility of an earlier detection of bacilli thus interrupting the transmission chain of the disease.