RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections

Stefańska, Ilona; Kwiecień, Ewelina; Górzyńska, Małgorzata; Sałamaszyńska-Guz, Agnieszka; Rzewuska, Magdalena

doi:10.3390/pathogens11050562

Cited by 11 publications

(12 citation statements)

References 26 publications

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“…In this study, the RAPD-PCR method with the M13 primer was successfully used for the typing of T. pyogenes strains obtained from European bison from different locations in Poland. It should be highlighted that the method applied in this study was previously designed and optimized for the investigation of the genetic diversity of T. pyogenes strains of different origins, including isolates from European bison (however, those isolates were other than tested in this research) [ 25 ]. This typing method proved to be suitable for assessment of genetic diversity among T. pyogenes strains collected from European bison who inhabited various regions.…”

Section: Discussionmentioning

confidence: 99%

“…The RAPD-PCR typing was performed with the M13 primer (5′-GAGGGTGGCGGTTCT-3′) (Eurofins Genomics, Ebersberg, Germany) according to our previous optimization study [ 25 ]. Each reaction mixture contained 12.5 µL of DreamTaq Green PCR Master Mix (Thermo Fisher Scientific, Waltham, MA, USA), 3.5 mM MgCl 2 (Thermo Fisher Scientific, Waltham, MA, USA), 0.8 mM of each dNTP (Thermo Fisher Scientific, Waltham, MA, USA), 20 pmol of the M13 primer, 20 ng of the genomic DNA and nuclease-free water (Thermo Fisher Scientific, Waltham, MA, USA) up to a final volume of 25 µL.…”

Section: Methodsmentioning

confidence: 99%

“…The evaluation of genetic diversity and relationships among bacterial strains is a very important aspect of epidemiological studies. So far, few molecular methods were applied for typing T. pyogenes strains isolated from various hosts [ 17 , 20 , 21 , 22 , 23 , 24 , 25 ]. However, the knowledge about the genetic diversity among T. pyogenes strains found in European bison is insufficient.…”

Section: Introductionmentioning

confidence: 99%

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Prevalence and Genetic Diversity of Trueperella pyogenes Isolated from Infections in European Bison (Bison bonasus)

Kwiecień

Stefańska

Kizerwetter-Świda

et al. 2022

Animals

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Trueperella pyogenes is a Gram-positive bacterium causing purulent infections in many animal species, including the European bison. However, the data about the virulence and genetic relationships of T. pyogenes strains isolated from these wild ruminants are strongly limited. The aim of this study was to investigate the prevalence of T. pyogenes infections in the European bison, and to evaluate the genetic diversity of isolates from these animals. In the time span of10 years, 328 European bison from 16 different locations were examined. The standard bacteriological methods were used for T. pyogenes isolation and identification from clinical specimens obtained from urogenital tract infections and abscesses of different locations. The presence of genes encoding known virulence factors was investigated by PCR, and the genetic diversity of T. pyogenes strains was examined with the RAPD-PCR method. The prevalence of T. pyogenes infections was 14.6%, and the pathogen was isolated from both female (47.9% of isolates) and male (52.1% of isolates) European bison. It should be highlighted that a considerable number of strains were isolated from the prepuce and penis infections. Therefore, the role of T. pyogenes in the pathogenesis of balanoposthitis should be seriously perceived. A total of 39 T. pyogenes strains were subjected to genetic characterization. All studied strains carried the plo gene, while the nanH (25.6%), nanP (23.1%), cbpA (7.7%), fimA (97.4%), fimC (69.2%), fimE (92.3%) and fimG (15.4%) genes were present with a variable frequency among the tested strains. The virulence genotype plo/fimA/fimC/fimE was dominant. RAPD-PCR typing showed a high level of genetic diversity among European bison T. pyogenes strains, and a total of 31 different RAPD profiles were distinguished. In a few cases, the same RAPD profile was found in strains obtained from animals living in the same area. This study provided the first data about the prevalence and genetic relationships of T. pyogenes in the Polish population of European bison. However, further epidemiological investigations are needed to understand the routes of transmission and dissemination of the pathogen in these wild animals.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Prevalence and Genetic Diversity of Trueperella pyogenes Isolated from Infections in European Bison (Bison bonasus)

Kwiecień

Stefańska

Kizerwetter-Świda

et al. 2022

Animals

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“…The most-often applied methods from the MAAP group are Arbitrary Primed Polymerase Chain Reaction (AP-PCR, which uses accidental sequences starters) and Random Amplified Polymorphic DNA (RAPD; the accidental amplification of polymorphic DNA fragments) [ 8 , 9 , 10 ], which are described as “fingerprinting” methods [ 2 , 7 , 11 , 12 , 13 ]. The differences between these methods concern the length of the primers and the way the resulting products are separated.…”

Section: Introductionmentioning

confidence: 99%

Improved RAPD Method for Candida parapsilosis Fingerprinting

Wojciechowska‐Koszko

Mnichowska-Polanowska

Roszkowska

et al. 2023

Genes

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Recently, methods based on the analysis of arbitrarily amplified target sites of genome microorganisms have been extensively applied in microbiological studies, and especially in epidemiological studies. The range of their application is limited by problems with discrimination and reproducibility resulting from a lack of standardized and reliable methods of optimization. The aim of this study was to obtain optimal parameters of the Random Amplified Polymorphic DNA (RAPD) reaction by using an orthogonal array as per the Taguchi and Wu protocol, modified by Cobb and Clark for Candida parapsilosis isolates. High Simpson’s index values and low Dice coefficients obtained in this study indicated a high level of interspecies DNA polymorphism between C. parapsilosis strains, and the optimized RAPD method proved useful in the microbiological and epidemiological study.

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“…Though, RAPDs have low reproducibility and result can be difficult to interpret, among several PCR-based tools, Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) is a simple, robust, rapid and cost-effective tool (ReshmaRaj and Das, 2021). Therefore, highly standardized laboratory protocols are required because of their sensitivity to reaction conditions (Ador et al, 2022;Stefanska et al, 2022).The quality and concentration of template DNA, concentrations of PCR components, and the PCR cycling conditions greatly influence the outcome of RAPD. Also, to ensure reproducibility, purified and high molecular weight DNA is required for RAPD analyses (ReshmaRaj and Das, 2021).…”

Section: Introductionmentioning

confidence: 99%

Genotoxic potentials of some selected heavy metals exposure on Clarias gariepinus (Burchell, 1822) and Oreochromis niloticus (Linnaeus, 1758) using RAPD-PCR technique

Ejikeme¹,

Ayoola

Amusa³

2022

Aceh J. Anim. Sci.

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Fish health and the aquatic ecosystem are strongly interwoven and interrelated. The aquatic ecosystem receives a range of anthropogenic chemicals which have toxicological or lethal health effects on the aquatic animals. The aim of this study was to determine the level of induced mutation and genomic stability of different sub-lethal doses (25%, 50% and 75% LC50) of heavy metal [Nickel (Ni), Mercury (Hg), Lead (Pb)and Zinc (Zn)] in Clarias gariepinus (Burchell, 1822) andOreochromis niloticus (Linnaeus, 1758) using RAPD-PCR technology over a period of 21 days. Four highly polymorphic RAPD markers were evaluated on isolated DNA from both heavy metal exposed fishes and control fishes after exposure. Observation of RAPD profiles in C. gariepinus revealed more damaging effect as sub lethal doses increased with zinc (a + b = 44 bands) mercury (a + b = 41 bands) nickel (a + b = 37 bands) lead (a + b = 35 bands) than with nickel (a + b = 37 bands) zinc (a + b = 34 bands) lead (a + b = 32 bands) mercury (a + b = 31 bands) for O. niloticus when compare to the control groups. Although, the genomic stability template decreased sub lethal heavy metal doses, higher stability was observed in O. niloticus (GTSPb = 26.32% GTSHg = 18.42% GTSZn = 10.53% GTSNi = 2.63%) than in C. gariepinus (GTSPb = 5.41% GTSNi = 0.0% GTSHg = -10.81% GTSZn = -18.92%). The results obtained showed differential variation in heavy metal induced genetic mutation and genomic stabilty in C. gariepinus and O. niloticus. These observable differences might be due to the physiological structure of the fish species evaluated. This study also confirms that RAPD–PCR technology is a useful tools in detecting the genotoxic effect of heavy metals in aquatic organisms but due to low reproducibility of RAPD results, it is recommended that this technology should be used along with other molecular techniques in fish genotoxicity studies.Keywords: Heavy metals, Induced, Genotoxicity, Fish, RAPD-PCR Technique

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RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections

Cited by 11 publications

References 26 publications

Prevalence and Genetic Diversity of Trueperella pyogenes Isolated from Infections in European Bison (Bison bonasus)

Prevalence and Genetic Diversity of Trueperella pyogenes Isolated from Infections in European Bison (Bison bonasus)

Improved RAPD Method for Candida parapsilosis Fingerprinting

Genotoxic potentials of some selected heavy metals exposure on Clarias gariepinus (Burchell, 1822) and Oreochromis niloticus (Linnaeus, 1758) using RAPD-PCR technique

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