scholarly journals Detection of Mycobacterium bovis DNA in oronasal swabs from infected African buffaloes (Syncerus caffer).

2021 ◽  
Author(s):  
Charlene Clarke ◽  
David V Cooper ◽  
Michele Ann Miller ◽  
Wynand Johan Goosen
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Bovis ◽  
Causative Agent ◽  
Molecular Transport ◽  
High Specificity ◽  
Qpcr Assay ◽  
Post Mortem ◽  
Syncerus Caffer ◽  
Tissue Samples ◽  
Sensitive Method

Abstract Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex (MTBC), is the causative agent of bovine TB (bTB) in animals. Spread occurs through inhalation or ingestion of bacilli transmitted from infected individuals. Early and accurate detection of infected African buffaloes shedding M. bovis is essential for interrupting transmission. In this study, we determined if M. bovis shedding could be detected in buffalo oronasal secretions using a molecular transport media (PrimeStore MTM) with oronasal swabs and a rapid qPCR assay (Xpert MTB/RIF Ultra). Bovine TB test-positive buffaloes were culled, then tissue samples and oronasal swabs collected post-mortem for mycobacterial culture and Ultra testing, respectively. The Ultra detected MTBC DNA in 5/12 swabs from M. bovis culture-confirmed buffaloes. Oronasal swabs from M. bovis negative buffaloes (n = 20) were negative on Ultra, indicating the high specificity of this test. This study showed that MTM can successfully preserve MTBC DNA in oronasal swabs. The proportion of MTBC positive oronasal swabs was higher than expected and suggests that the Ultra may be a sensitive method for identifying infected buffaloes. Further studies are needed to confirm the utility of the Ultra assay with oronasal swabs as a promising assay to evaluate MTBC shedding in buffaloes.

scholarly journals Novel molecular transport medium used in combination with Xpert MTB/RIF ultra provides rapid detection of Mycobacterium bovis in African buffaloes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Charlene Clarke ◽  
Katrin Smith ◽  
Samantha J. Goldswain ◽  
Christopher Helm ◽  
David V. Cooper ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Human Health Risk ◽  
Molecular Transport ◽  
Sampling Technique ◽  
High Specificity ◽  
Negative Control ◽  
Post Mortem ◽  
Tissue Samples ◽  
Transport Medium ◽  
Tissue Homogenates

AbstractMycobacterium bovis is the causative agent of bovine tuberculosis (bTB) in wildlife. Confirmation of M. bovis infection relies on mycobacterial culture, which is time-consuming. Collection and transportation of infectious material also pose a human health risk. PrimeStore Molecular Transport Medium (MTM) has been shown to effectively inactivate infectious organisms, making it a safe method for handling infectious samples. This study investigated an in-field sampling technique for rapid, safe detection of M. bovis in buffalo tissues. Potentially infected tissues from bTB test-positive buffaloes were swabbed at post-mortem examination and stored in PrimeStore MTM at ambient temperature until Xpert MTB/RIF Ultra testing was performed. Additionally, tissue samples were frozen and transported before homogenisation for culture and Ultra testing. Oral swabs were collected from M. bovis-unexposed buffaloes as a negative control cohort. Mycobacterium tuberculosis complex (MTBC) DNA was detected by Ultra in 13/16 tissue swabs and 9/16 matched tissue homogenates from culture-confirmed M. bovis-positive buffalo tissues. MTBC DNA was not detected in swabs from M. bovis-unexposed animals, showing the potentially high specificity of Ultra with PrimeStore swabs. PrimeStore MTM sample processing, in combination with the Ultra assay, has the potential to provide a safe, rapid post-mortem screening test for M. bovis in buffaloes.

scholarly journals Use of PrimeStore® Molecular Transport Medium and Xpert MTB/RIF Ultra for Rapid Detection of Mycobacterium Bovis in African buffaloes (Syncerus Caffer).

2021 ◽  
Author(s):  
Charlene Clarke ◽  
Katrin Smith ◽  
Samantha J Goldswain ◽  
Christopher Helm ◽  
David V Cooper ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Human Health Risk ◽  
Molecular Transport ◽  
Sampling Technique ◽  
High Specificity ◽  
Negative Control ◽  
Post Mortem ◽  
Tissue Samples ◽  
Transport Medium ◽  
Tissue Homogenates

Abstract Mycobacterium bovis is the causative agent of bovine tuberculosis (bTB) in wildlife. Confirmation of M. bovis infection relies on mycobacterial culture, which is time-consuming. Collection and transportation of infectious material also pose a human health risk. PrimeStore® Molecular Transport Medium (MTM) has been shown to effectively inactivate infectious organisms, making it a safe method for handling infectious samples. This study investigated an in-field sampling technique for rapid, safe detection of M. bovis in buffalo tissues. Potentially infected tissues from bTB test-positive buffaloes were swabbed at post-mortem examination and stored in PrimeStore® MTM at ambient temperature until Xpert® MTB/RIF Ultra testing was performed. Additionally, tissue samples were frozen and transported before homogenisation for culture and Ultra testing. Oral swabs were collected from M. bovis-unexposed buffaloes as a negative control cohort. Mycobacterium tuberculosis complex (MTBC) DNA was detected by Ultra in 13/16 tissue swabs and 9/16 matched tissue homogenates from culture-confirmed M. bovis-positive buffalo tissues. MTBC DNA was not detected in swabs from M. bovis-unexposed animals, showing the potentially high specificity of Ultra with PrimeStore® swabs. PrimeStore® MTM sample processing, in combination with the Ultra assay, has the potential to provide a safe, rapid post-mortem screening test for M. bovis in buffaloes.

scholarly journals Characterization of Exochelins of theMycobacterium bovis Type Strain and BCG Substrains

1999 ◽  
Vol 67 (4) ◽  
pp. 2035-2039 ◽  
Author(s):  
Jovana Gobin ◽  
Diane K. Wong ◽  
Bradford W. Gibson ◽  
Marcus A. Horwitz
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Bovis ◽  
Causative Agent ◽  
Type Strain ◽  
Binding Proteins ◽  
Iron Binding ◽  
Iron Binding Proteins ◽  
Do So

ABSTRACT Pathogenic mycobacteria must acquire iron in the host in order to multiply and cause disease. To do so, they release abundant quantities of siderophores called exochelins, which have the capacity to scavenge iron from host iron-binding proteins and deliver it to the mycobacteria. In this study, we have characterized the exochelins of Mycobacterium bovis, the causative agent of bovine and occasionally of human tuberculosis, and the highly attenuated descendant of M. bovis, bacillus Calmette-Guérin (BCG), widely used as a vaccine against human tuberculosis. The M. bovis type strain, five substrains ofM. bovis BCG (Copenhagen, Glaxo, Japanese, Pasteur, and Tice), and two strains of virulent Mycobacterium tuberculosis all produce the same set of exochelins, although the relative amounts of individual exochelins may differ. Among these mycobacteria, the total amount of exochelins produced is greatest in M. tuberculosis, intermediate in M. bovis, and smallest in M. bovis BCG.

Postmortem small babesia-like morphology of Babesia canis — Short communication

2011 ◽  
Vol 59 (4) ◽  
pp. 427-432 ◽  
Author(s):  
Zoltán Demeter ◽  
Elena Palade ◽  
Éva Balogh ◽  
Csaba Jakab ◽  
Róbert Farkas ◽  
...  
Keyword(s):  
Causative Agent ◽  
18S Rrna Gene ◽  
Severe Anaemia ◽  
Rrna Gene ◽  
Babesia Canis ◽  
Post Mortem ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Sequence Identity ◽  
Babesia Canis Canis

Here we report a case of canine babesiosis with unusual morphology of the causative agent. A male, seven-week-old Labrador retriever puppy, exhibiting severe anaemia and haemoglobinuria, was presented at the Clinic of Internal Medicine in February 2011. The puppy was euthanised. The most relevant pathological changes were icterus, severe splenomegaly, generalised lymphadenopathy and haemoglobin nephrosis. Samples were collected from various organs for histology within one hour post mortem. Impression smears were also prepared from the spleen after overnight storage at 4 °C. Tissue sections and smears showed the presence of multiple, coccoid intraerythrocytic bodies that measured 1–2 μm and resembled small babesiae. No large piroplasms were seen. DNA was extracted from the spleen, and a conventional PCR was performed for the amplification of a 450-bp region of the 18S rRNA gene of piroplasms. The causative agent was identified as Babesia canis canis, with 99% sequence identity to other European isolates. Sequence identity to B. gibsoni was only 91%. This is the first account to verify that the morphology of the large canine piroplasm, B. canis, can be uniformly small babesia-like post mortem or following the storage of tissue samples.

Isolation and identification of Mycobacterium bovis and Mycobacterium tuberculosis from animal tissues by conventional and molecular method

2015 ◽  
Author(s):  
Abdul Basit ◽  
Mubbashir Hussain ◽  
Sultan Ayaz ◽  
Muhammad Shahid ◽  
Kashif Rahim ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sensitivity And Specificity ◽  
Mycobacterium Bovis ◽  
Smear Microscopy ◽  
Reference Laboratory ◽  
Public Health Importance ◽  
Isolation And Identification ◽  
Awareness Campaign ◽  
Tissue Samples ◽  
Worldwide Distribution

Tuberculosis is a worldwide contagious and chronic disease of human as well as domestic animals with zoonotic potential. The Mycobacterium bovis and Mycobacterium tuberculosis are the main cause of tuberculosis. It has worldwide distribution with significant effect on animals and has public health importance. Therefore the present study was conducted to determine the prevalence of Mycobacterium bovis and Mycobacterium tuberculosis among the ruminant of district Kohat, Khyber PakhtunKhwa and also to evaluate the sensitivity and specificity of Microscopy and PCR. A total 200 tissue samples of lungs, lymph nodes and liver from cattle, buffaloes, sheep and goats were collected from Abattoir Kohat. All the tissue were first examined by direct smear microscopy by Ziehl Neelsen staining and then subjected to the PCR for the detection of M. bovis and M. tuberculosis. The overall prevalence of tuberculosis was 6.5% by PCR.Prevalence of tuberculosis was recorded in7.87% of lungs samples followed by 5.26% lymph node. Moreover the prevalence was found 5.2%, 4%, 10.6% and 6.5% in cattle, buffalos, Goats and sheep respectively. Furthermorethe sensitivity and specificity of PCR and microscopy in term of detection of TB was determined that PCR was found less sensitive then microscopy because of other species which was not amplified due to non availability of specific primer and were found positive in microscopy.In conclusion PCR is more reliable diagnostic tool for diagnosis of bovine tuberculosis. It is recommended that PCR based diagnostic reference laboratory may be established at district level and Tuberculosis awareness campaign must be arranged.

scholarly journals Protection Elicited by Two Glutamine Auxotrophs of Mycobacterium tuberculosis and In Vivo Growth Phenotypes of the Four Unique Glutamine Synthetase Mutants in a Murine Model

2006 ◽  
Vol 74 (11) ◽  
pp. 6491-6495 ◽  
Author(s):  
Sunhee Lee ◽  
Bo-Young Jeon ◽  
Svetoslav Bardarov ◽  
Mei Chen ◽  
Sheldon L. Morris ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Glutamine Synthetase ◽  
Mycobacterium Bovis ◽  
Murine Model ◽  
Triple Mutant ◽  
Auxotrophic Mutants ◽  
Bcg Vaccination ◽  
In Vivo Growth ◽  
Subcutaneous Immunization

ABSTRACT We generated four individual glutamine synthetase (GS) mutants (ΔglnA1, ΔglnA2, ΔglnA3, and ΔglnA4) and one triple mutant (ΔglnA1EA2) of Mycobacterium tuberculosis to investigate the roles of GS enzymes. Subcutaneous immunization with the ΔglnA1EA2 and ΔglnA1 glutamine auxotrophic mutants conferred protection on C57BL/6 mice against an aerosol challenge with virulent M. tuberculosis, which was comparable to that provided by Mycobacterium bovis BCG vaccination.

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