Identification of Corynebacterium pseudotuberculosis isolates from sheep and goats by PCR (original) (raw)
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Arquivos do Instituto Biológico, 2015
Caseous lymphadenitis is a chronic infectious disease caused by Corynebacterium pseudotuberculosis , which is a bacterium responsible for a great number of economic losses on goat and sheep production. It is characterized by the formation of abscesses in superficial lymph nodes and in internal organs and lymph nodes. This study aimed at determining the agreement between microbiological culture and PCR in the identification of C. pseudotuberculosis , in samples collected from animals in slaughterhouses and in animals that presented lymph node enlargement in field conditions. From the 202 samples analyzed through microbiological culture, 113 (56%) were positive for Corynebacterium sp.; from these positive samples, 38 (34%) were identified as C. pseudotuberculosis by microbiological culture. From the amount of samples, 110 (54%) were positive and 92 (46%) were negative in the PCR. Kappa index (0.193) presented a weak agreement between PCR and microbiological culture. We concluded that ...
2021
Article history: Received March 7, 2020 Accepted April 28, 2020 Available online February 15, 2021 Caseous lymphadenitis is a chronic infection caused by Corynebacterium pseudotuberculosis that affects the lymphatic system, resulting in abscesses in the lymph nodes and internal organs. This study was conducted to determine the prevalence rate of caseous lymphadenitis among slaughtered sheep in Duhok abattoirs, Duhok, Iraq and to investigate to what extend rabbits can develop caseous lymphadenitis just like that in sheep. A total of 1052 carcasses of sheep were randomly selected (939 males and 113 females) and carefully inspected for detection any suspected caseous lymph adenitis. The isolated Corynebacterium pseudotuberculosis from lymph nodes of sheep carcasses were identified by PCR before used in in vivo infection in rabbits. Data revealed that the prevalence rate of caseous lymphadenitis was at 1.9% among the slaughtered sheep and the prevalence rates were higher in females and ...
The Veterinary Journal, 2013
The biochemical pattern of Cowan and Steel (BPCS) was compared with a commercial biochemical strip for the identification of Corynebacterium pseudotuberculosis isolated from small ruminants. On 16S rRNA gene sequencing, 40/78 coryneform isolates from the lymph nodes of sheep and goats with lesions resembling caseous lymphadenitis were identified as C. pseudotuberculosis. The sensitivities of the BPCS and the commercial biochemical strip relative to 16S rRNA sequencing were 80% and 85%, and their specificities were 92.1% and 94.7%, respectively; the level of agreement between the BPCS and the commercial biochemical strip was high (j = 0.82). Likelihood ratios for positive and negative results were 10.0 and 0.22 for the BPCS, and 16.0 and 0.16 for the commercial biochemical strip, respectively. These results indicate that the BPCS and the commercial biochemical strip are both useful for identifying C. pseudotuberculosis in veterinary microbiology laboratories.
Animals
Caseous lymphadenitis (CLA) is a bacterial infection caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis) that affects sheep and goats, leading to abscess formation in their lymph nodes. The present study aimed to isolate and identify C. pseudotuberculosis from CLA in smallholder sheep and goats, and determine the resistance patterns, virulence, and resistance genes of the isolates. Additionally, genotypic and phylogenetic analysis of the isolates was conducted using ERIC-PCR and DNA sequencing techniques. A cross-sectional study examined 220 animals (130 sheep and 90 goats) from 39 smallholder flocks for clinical signs of CLA. Fifty-four (24.54%) animals showed CLA-compatible lesions, confirmed by C. pseudotuberculosis isolation and PCR identification. Sheep had a lower infection rate of CLA (18.46%) compared with goats (33.3%). Antimicrobial susceptibility testing of 54 C. pseudotuberculosis isolates to 24 antimicrobial drugs revealed that they were 100% resistant ...
Corynebacterium pseudotuberculosis and its Role in Ovine Caseous Lymphadenitis
Caseous lymphadenitis (CLA) of sheep, caused by Corynebacterium pseudotuberculosis, has been a signi¢cant disease in themajority of sheep-rearing regions for over a century. Because of the chronic and often sub-clinical nature of the infection, it has proved di⁄cult to control and prevalence is high in many parts of the world, which in turn leads to signi¢cant economic losses for farmers.This review describes the important characteristics of C. pseudotuberculosis and examines the pathogenesis and epidemiology of the infection in sheep.The review also discusses the immune response to infection and describes themethods that have been developed to control CLA, with particular emphasis on the use of vaccination and serological testing.
Brazilian Journal of Microbiology, 2020
Caseous lymphadenitis (CL) in sheep is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, commonly characterized by abscess formation in peripheral lymph nodes and disseminated infections. Nonetheless, other microorganisms, including with zoonotic relevance, can be isolated from CL-resembling lymph nodes. Currently, mycobacteria have been reported in visceral granulomatous lesions in small ruminants, a fact that poses a public health issue, particularly in slaughtered sheep intended for human consumption. Cytology using fine needle aspiration and microbiological culturing are suitable tests for routine diagnostic, whereas present drawbacks and molecular methods have been confirmatory. Data about the occurrence of mycobacteria in both lymph nodes with aspect of CL and apparently healthy visceral nodes of sheep slaughtered for human consumption are scarce. In this study, 197 visceral lymph nodes of sheep showed lymphadenitis and 202 healthy visceral lymph nodes of slaughtered sheep intended for human consumption were submitted to conventional bacteriological diagnosis, mycobacteria culturing, and cytological evaluation. Compatible Corynebacterium isolates were subjected to multiplex PCR targeting 16S rRNA, rpoB, and pld genes to detect C. pseudotuberculosis. Based on microbiological identification, C. pseudotuberculosis (86/197; 43.7%), streptococci γ-hemolytic (17/197; 8.6%), and Trueperella pyogenes (12/197; 6.1%) were prevalent in lymph nodes with abscesses, as opposed to staphylococci (53/202; 26.2%) in apparently healthy lymph nodes. No mycobacteria were isolated. Cytology identified 49.2% (97/197) Gram-positive pleomorphic organisms (coryneform aspect). Multiplex PCR confirmed genetic material of C. pseudotuberculosis in 74.4% (64/86) of the samples with C. pseudotuberculosis isolation and 66% (64/97) samples with cytological coryneform aspect (κ = 86.78%; 95% CI = 79.87-93.68%). These findings emphasize the prevalence of C. pseudotuberculosis in abscess formation among peripheral lymph nodes of sheep. Other bacteria were also identified in lymph nodes sampled that resembling C. pseudotuberculosis-induced infections that may difficult the diagnosis. Multiplex PCR revealed a valuable assay to detect C. pseudotuberculosis, in addition to routine methods applied to CL-diagnosis. No mycobacteria were identified in lymph nodes sampled, with and without apparent lesions. Nonetheless, due to
BMC veterinary research, 2014
Background Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis, a common disease in small ruminant populations throughout the world and responsible on a significant economic impact for producers.Case presentationTo our knowledge, this is the first characterization of C. pseudotuberculosis from caseous lymphadenitis lesions in Black Alentejano pig (Sus scrofa domesticus). In this study, phenotypic and genotypic identification methods allocated the swine isolates in C. pseudotuberculosis biovar ovis. The vast majority of the isolates were able to produce phospholipase D and were susceptible to most of the antimicrobial compounds tested. Macrorestriction patterns obtained by Pulsed Field Gel Electrophoresis (PFGE) grouped the C. pseudotuberculosis in two clusters with a high similarity index, which reveals their clonal relatedness. Furthermore, swine isolates were compared with C. pseudotuberculosis from caprines and PFGE patterns also showed high similar...
Zagazig Veterinary Journal Volume 44, Number 1, p. 1-8, 2016
Caseous lymphadenitis is a highly prevalent worldwide disease of sheep and goats caused by C. pseudotuberculosis, resulting in significant economic losses. In order to investigate the prevalence as well as the molecular characterization of C. pseudotuberculosis and monitoring the antibiotic susceptibility of the isolated strains, a total of 126 pus samples were collected aseptically from 80 sheep and 46 goats suffering from abscessation. The collected samples were subjected to bacteriological examination where the prevalence of C. pseudotuberculosis was 43.8% and 36.9% in diseased sheep and goats, respectively. The antibiotic sensitivity test was carried out using disc diffusion method, most of the isolates were resistant to penicillin (96.2%) and erythromycine (92.3%), highly sensitive to ciprofloxacin (96.2%), amikacin (90.4%) and neomycin (88.5%). Polymerase chain reaction was applied on 30 C. pseudotuberculosis isolates for the amplification and detection of both 16S rRNA and the RNA polymerase β-subunit gene (rpoB) genes to confirm the diagnosis of the isolates. The results revealed that all the tested isolates were positive for both genes.
Bacteriological characterization of Corynebacterium pseudotuberculosis in Venezuelan goat flocks
Abacteriological characterization of strains of Corynebacterium pseudotuberculosis, etiological agent of caseous lymphadenitis, was made in goat flocks from Venezuela Northwestern region where goat breeding is the main way of living. From 18 goat farms, 65 animals presented abscesses. Of the total number of goats affected, C. pseudotuberculosis was isolated in 67.69% of them; other microorganisms in 27.71%. C. pseudotuberculosis grew forming cream white, dry, waxy colonies with a narrow zone of -haemolysis; pellicle and sediment were seen in BHI broth but did not become turbid. All the isolates were similar in morphological, culture and biochemical characteristics to the C. pseudotuberculosis reference strain ATCC 19410, but some isolates showed variations in the minimum inhibitory concentration values with antibiotics. The frequency of isolation of the microorganism was higher in animals older than a year. According to these results, it is established that C. pseudotuberculosis is actually widely spread through goat flocks in the Northwestern region of Venezuela.