Abattoir survey of bovine tuberculosis in tanta, centre of the Nile delta, with in silico analysis of gene mutations and protein–protein interactions of the involved mycobacteria

Abstract

Bovine tuberculosis is a transboundary disease of high economic and public health burden worldwide. In this study, post‐mortem examination of 750 cattle and buffalo in Tanta abattoir, Centre of the Nile Delta, revealed visible TB in 4% of animals and a true prevalence of 6.85% (95% CI: 5.3%–8.9%). Mycobacterial culture, histopathology and RT‐PCR targeting all members of M. tuberculosis complex were performed, upon which 85%, 80% and 100% of each tested lesions were confirmed as TB, respectively. Mpb70‐targeting PCR was conducted on ten RT‐PCR positive samples for sequencing and identified nine Mycobacterium (M.) bovis strains and, interestingly, one M. tuberculosis (Mtb) strain from a buffalo. Bioinformatics tools were used for prediction of mutations, nucleotide polymorphisms, lineages, drug resistance and protein–protein interactions (PPI) of the sequenced strains. The Mtb strain was resistant to rifampicin, isoniazid and streptomycin, and to the best of our knowledge, this is the first report of multidrug resistant (MDR)‐Mtb originating from buffaloes. Seven M. bovis strains were resistant to ethambutol and ethionamide. Such resistances were associated with KatG, rpoB, rpsL, embB and ethA genes mutations. Other mutations and nucleotide polymorphisms were also predicted, some are reported for the first time and require experimental work for validation. PPI revealed more interactions than what would be expected for a random set of proteins of similar size and had dense interactions between nodes that are biologically connected, as a group. Two M. bovis strains belonged to BOV AFRI lineage (Spoligotypes BOV 1; BOV 2) and eight strains belonged to East‐Asian (Beijing) lineage. In conclusion, visible TB was prevalent in the study area, RT‐PCR is the best to confirm the disease, MDR‐Mtb is associated with buffalo TB, and mycobacteria of different lineages carry many resistance genes to chemotherapeutic agents used in treatment of human TB constituting a major public health risk.

Bacteriological and Molecular studies on Mycobacterium Bovis in Cattle, with Special Reference to its Antimicrobial Resistance

“…From Table 1, it appears that results of direct smear were less than those reported by Sohair and Riad (2002) who detected 69% positive cases but higher than 46.88% and 48.04% reported by Silva et al (2018) and Abdelsadek et al (2020). Regarding culture, the obtained results in Table 1 are consistent with those reported by Silva et al (2018) and Hamed et al (2021) who detected M. bovis in 68.75% and 64.9% of tested samples, respectively, but were less than 80%, 71.4% and 85% reported by Nasr et al (2016), Mohammed (2017) and Borham et al (2021), respectively. In contrast, the isolation rate was higher than 51.59% reported by Abdelsadek et al (2020).…”

Bayesian Estimation of Diagnostic Accuracy of Three Diagnostic Tests for Bovine Tuberculosis in Egyptian Dairy Cattle Using Latent Class Models

“…Cattle importation, which is responsible for the emergence of new strains of M. bovis in Egypt, may have a role in the high frequency of bTB found in this study [50]. The animals used in this investigation were females, and the incidence of bTB infection among females was much higher than that of males, owing to the fact that females are typically bred for longer periods of time than males and therefore subjected to repeated infection [52]. Furthermore, the Egyptian bTB eradication program uses the SCT, which is affected by the presence of other infectious diseases such as Johnes disease [53] and the high number of cows in the late phase of the disease [54].…”

Review on Bovine Tuberculosis: An Emerging Disease Associated with Multidrug-Resistant Mycobacterium Species

“…However, such type of analysis on bTB is still limited worldwide, and is nearly absent in Egypt, except for one study by Abdelaal et al [ 120 ]. Recently, Borham et al in Egypt used bioinformatics tools for prediction of mutations, nucleotide polymorphisms, lineages, drug resistance, and protein–protein interactions (PPI) of ten Mycobacterium sequenced strains [ 139 ]. Future in silico analysis studies are strongly required for bTB and its related isolates in Egypt as well as worldwide to understand many mysterious aspects of bTB.…”

“…In addition, MBTC isolates were highly resistant to the second-line drugs; KAN and AMK in a percentage of 82.3% and 80.7%, respectively. Borham et al reported one Mtb strain that was MDR to RIF, INH, and streptomycin, this was the first report of multidrug resistant (MDR)- Mtb originating from buffaloes [ 139 ]. In addition, the authors reported seven M. bovis strains that were resistant to ethambutol and ethionamide.…”