scholarly journals Bacteria isolated from abscesses of small ruminants inspected in the semiarid region of Brazil

2016 ◽  
Vol 37 (3) ◽  
pp. 1337
Author(s):  
Wellington Erasmo Lima e Silva ◽  
Gisele Veneroni Gouveia ◽  
Maria da Conceição Aquino de Sá ◽  
João José De Simoni Gouveia ◽  
Rodolfo De Moraes Peixoto ◽  
...  
Keyword(s):  
Small Ruminants ◽  
Northeastern Brazil ◽  
Semiarid Region ◽  
Microbiological Analysis ◽  
Bacterial Isolation ◽  
Tuberculosis Complex ◽  
Chain Reaction ◽  
Lowenstein Jensen ◽  
Polymerase Chain ◽  
Mycobacterium Spp

Loss in the supply chain of small ruminants owing to condemnations of carcasses in the abattoirs and slaughterhouses is common in northeastern Brazil. This study aims to identify bacterial agents, including Mycobacterium spp., in the abscesses found in the postmortem analysis of the carcasses of sheep and goats bred in northeastern Brazil. Our analysis involved 679 goats and 1,838 sheep carcasses. Abscess samples were extracted and inoculated on blood agar and Lowenstein Jensen with pyruvate or glycerol for bacterial isolation. We then performed polymerase chain reaction of the hps 65 gene; samples positive for Mycobacterium spp. were subjected to DNA sequencing. Relative frequencies of abscesses in goats and sheep were 5.44 and 3.26%, respectively. Microbiological analysis revealed 87.7% bacterial growth in the inoculated samples. Among these, Corynebacterium pseudotuberculosis represented 67.7% of the isolates. We observed 1.9% mycobacteria growth in the abscess samples inoculated on Lowenstein-Jensen medium. PCR of DNA extracted from abscesses samples showed amplification of 0.9% of samples. After sequencing, Mycobacterium spp. isolate was identified as M. novocastrense. C. pseudotuberculosis was the main agent responsible for the formation of abscesses in the examined animals, and we did not identify any species of the M. tuberculosis complex in the examined small ruminants.

scholarly journals UTILITY OF ANTIGEN DETECTION TEST AND POLYMERASE CHAIN REACTION IN THE DIFFERENTIATION OF TUBERCULOUS AND NON-TUBERCULOUS MYCOBACTERIA

2017 ◽  
Vol 10 (6) ◽  
pp. 289
Author(s):  
Yogita Singh ◽  
Raji Vasanth ◽  
Shrikala Baliga ◽  
Dhanashree B
Keyword(s):  
Polymerase Chain Reaction ◽  
False Negative ◽  
Diagnostic Methods ◽  
Clinical Samples ◽  
Chain Reaction ◽  
College Hospital ◽  
Medical College ◽  
Negative Results ◽  
Polymerase Chain ◽  
Mycobacterium Spp

Objectives: Cultivation and identification of mycobacteria to species level remains difficult and time-consuming. Hence, easy and rapid diagnostic methods are necessary for the differentiation of Mycobacterium tuberculosis (MTB) from non-tuberculous mycobacteria (NTM). The present study aims to detect and differentiate MTB from NTM isolated from clinical samples by immunochromatographic test (ICT) and polymerase chain reaction (PCR). Methods: Over a period of 1 year, clinical samples (n=496) received from suspected cases of TB, at the Department of Microbiology, Kasturba Medical College Hospital, Mangalore were cultured to isolate Mycobacterium spp. Identification of all the isolates was done by conventional biochemical technique, ICT, and PCR. Results: Among the 496 samples processed, 49 (9.87%) were acid-fast bacilli smear positive and 59 (11.89%) samples showed the growth of Mycobacterium spp. Among these, 10 were rapid growers, 49 were slow-growing mycobacteria, out of which 30 were MTB as identified by conventional biochemical reaction. Out of 59 Mycobacterial isolates subjected to ICT for the detection of MPT 64 antigen, only 28 were identified as MTB. However, all the 30 isolates were correctly identified as MTB by PCR. Conclusion: Hence, PCR is essential for rapid differentiation of non-tuberculous Mycobacterium from MTB. False negative results seen with immunochromatographic MPT 64 antigen assay could be due to mutations within the mpt64 gene. Further studies are necessary to characterize these PCR-positive and immunochromatographic assay negative MTB isolates.

scholarly journals Isolation ofMycobacterium aviumsubspeciesparatuberculosisfrom non-ruminant wildlife living in the sheds and on the pastures of Greek sheep and goats

2007 ◽  
Vol 136 (5) ◽  
pp. 644-652 ◽  
Author(s):  
M. FLOROU ◽  
L. LEONTIDES ◽  
P. KOSTOULAS ◽  
C. BILLINIS ◽  
M. SOFIA ◽  
...  
Keyword(s):  
Type Strain ◽  
Insertion Sequence ◽  
Small Ruminants ◽  
Dairy Sheep ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Sheep And Goats ◽  
Faecal Samples ◽  
Polymerase Chain ◽  
Type Strains

SUMMARYThis study aimed to: (1) investigate whether non-ruminant wildlife interfacing with dairy sheep and goats of four Greek flocks endemically infected withMycobacterium aviumsubspeciesparatuberculosis(MAP) harboured MAP and (2) genetically compare the strains isolated from the wildlife to those isolated from the small ruminants of these flocks. We cultured and screened, by polymerase chain reaction (PCR), pooled-tissue samples from 327 wild animals of 11 species for the MAP-specific IS900insertion sequence. We also cultured faecal samples from 100 sheep or goats from each of the four flocks. MAP was detected in samples from 11 sheep, 12 goats, two mice, two rats, a hare and a fox. Only one rat had histopathological findings. Genetic typing categorized 21 isolates as cattle-type strains and two, from a house mouse and a goat respectively, as sheep-type strains; this is the first report of a rodent harbouring a sheep-type strain. The MAP types that were most frequently isolated amongst the sheep and goats of each flock were also the ones isolated from sympatric rodents; those isolated from the fox and hare also belonged to the predominant ruminant strains.

scholarly journals TRC4 Gene Based PCR Assay in Diagnosis of Tuberculous Meningitis

2017 ◽  
Vol 8 (2) ◽  
pp. 19-22
Author(s):  
Abu Naser Ibne Sattar ◽  
Sanjida Khondakar Setu ◽  
Ahmed Abu Saleh ◽  
Sharmeen Ahmed
Keyword(s):  
Polymerase Chain Reaction ◽  
Clinical Manifestation ◽  
Tuberculous Meningitis ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Early Stages ◽  
Lowenstein Jensen ◽  
Polymerase Chain ◽  
Tb Pcr ◽  
Is6110 Element

The prevalence of Tuberculous meningitis remains largely underestimated due to nonspecific clinical manifestation at early stages. A total of 20 CSF samples were studied from clinically suspected cases of tuberculous meningitis. All the samples were processed for ZN staining, TB culture on LJ media and TB-PCR using IS6110 and TRC4 primers by standard protocols. Of the total 20 CSF samples, 12 cases were diagnosed as TB meningitis. Most of the tuberculous meningitis cases were found positive by PCR using TRC4 primers (83.33%) followed by IS6110 primer (66.67%) and culture on L-J media(8.33%). None were found positive by ZN staining. TB-PCR usingTRC4 primer showed higher positivity than using IS6110 in detecting tuberculous meningitis, since some strains of MTB may lack the IS6110 element in their genome. PCR assay using TRC4 primer is superior in diagnosing tuberculous meningitis. This study was aimed to evaluate the diagnostic potentials of CSF polymerase chain reaction (PCR) by using TRC4 and IS6110 primers. Further the results were also compared with culture on Lowenstein-Jensen (LJ) media and AFB smear by Zeihl-Nelson (ZN) staining.Bangladesh J Med Microbiol 2014; 08 (02): 19-22

Biotechnical use of polymerase chain reaction for microbiological analysis of biological samples

2000 ◽  
pp. 87-130 ◽  
Author(s):  
Pär-G Lantz ◽  
Waleed Abu Al-Soud ◽  
Rickard Knutsson ◽  
Bärbel Hahn-Hägerdal ◽  
Peter Rådström
Keyword(s):  
Polymerase Chain Reaction ◽  
Biological Samples ◽  
Microbiological Analysis ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Bovine tuberculosis: Diagnosis by PCR technique in bovine milk samples

2013 ◽  
Vol 37 (2) ◽  
pp. 156-160
Author(s):  
Mohammed J. Alwan
Keyword(s):  
Bovine Milk ◽  
Culture Method ◽  
Bacterial Isolation ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Culture Methods ◽  
Milk Samples ◽  
Tuberculosis Diagnosis ◽  
Pcr Method ◽  
Polymerase Chain

     The aim of this study was to determine the percentage of borne tuberculin infection in milk sample by using PCR.  (102) milk samples were collected from  cows , AL-Dejella (39) samples,  AL-Suara (20) samples cow station, AL-Fthalia (20) samples,  AL-Azezia (11) samples and AL-Twarege (12) samples (Iraq) during the period  July 10th   2010 to  Nov.30th   2010. The samples were examined by direct smear stained by Ziehle-Neelson stain, culture methods and confirmed the isolates by Polymerase Chain Reaction (PCR) assay. The results showed that  5, 102 (4.9%) milk samples were M. bovis positive that were detected by direct milk smear method and 10 out 102(9.8%) M.bovis +ve were detected by culture method and PCR assay. The results also showed that high percentage of bacterial isolates from milk samples AL-Dejella city show (12.8%) by culturing and PCR method followed by AL-Suara (10%), AL-Fthelia (10%), Al-Twarege (8.3%) but no bacterial isolation was recorded in AL-Azezia milk samples. This study concluded that M.bovis infection was spreading in dairy cow within the mentioned areas and PCR was more sensitive, rapid, and accurate technique for M.bovis infection diagnosis.

scholarly journals Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis

2010 ◽  
Vol 30 (12) ◽  
pp. 1031-1035 ◽  
Author(s):  
Ana C.M. Groff ◽  
Jackeline K. Kirinus ◽  
Mariana Sá e Silva ◽  
Gustavo Machado ◽  
Mateus M. Costa ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Clinical Samples ◽  
Hexadecyltrimethylammonium Bromide ◽  
Bacterial Isolation ◽  
Chain Reaction ◽  
Guanidine Isothiocyanate ◽  
Campylobacter Fetus ◽  
Polymerase Chain ◽  
Bovine Genital Campylobacteriosis

Bovine genital campylobacteriosis is a common venereal disease of cattle; the prevalence of this disease can be underestimated mostly because of the nature of the etiological agent, the microaerobic Campylobacter fetus subspecies venerealis. The purpose of the current study was to evaluate the utilization of polymerase chain reaction (PCR) in the diagnosis of genital campylobacteriosis in samples obtained from bull prepuce aspirate, cow cervical mucus, and abomasum contents of aborted fetuses, collected into enrichment medium. Five different DNA extraction protocols were tested: thermal extraction, lysis with proteinase K, lysis with guanidine isothiocyanate, lysis with DNAzol, and lysis with hexadecyltrimethylammonium bromide (CTAB). The specificity, sensitivity, and technical application of the PCR assay were also evaluated with clinical samples and compared to bacterial isolation by standard culture. DNA extraction by the CTAB protocol provided better results in PCR, and it was able to detect 63 colony-forming units per ml of C. fetus. Out of 277 clinical samples tested, 68 (24%) were positive for Campylobacter fetus using PCR, while only 8 (2.8%) of the samples were positive by bacterial isolation in solid medium, proving the superiority of the PCR technique when compared to the standard isolation method, and providing evidence for its usefulness as a better screening test in cattle for the diagnosis of bovine genital campylobacteriosis.

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