Identification and epidemiological characterization of Streptococcus uberis isolated from bovine mastitis using conventional and molecular methods

Khan, Izhar U. H.; Hassan, Abdulwahed Ahmed; Abdulmawjood, Amir; Lämmler, Christoph; Wolter, W.; Zschöck, M.

doi:10.4142/jvs.2003.4.3.213

Cited by 64 publications

(69 citation statements)

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“…Corresponding to previous studies, where Hassan et al. (2001) and Khan et al. (2003) differentiated both species with oligonucleotide primers derived from species specific parts of the 16S rDNA, the 23S rDNA and the 16S–23S rDNA intergenic spacer region, the results of the present study confirmed that the occurrence of S. parauberis as mastitis causing pathogen appears to be rare.…”

Section: Discussionsupporting

confidence: 89%

“…The S. uberis and S. parauberis were characterized biochemically, serologically and by molecular methods (Lämmler, 1991; Lämmler et al., 1998; Hassan et al., 2001; Khan et al., 2003). The remaining species were characterized by conventional, partly by molecular methods (Abdulmawjood et al., 1998; Abdulmawjood and Lämmler, 1999, 2000; Hassan et al., 2000, 2003a,b,c; Vossen et al., 2004).…”

Section: Bacterial Isolatesmentioning

confidence: 99%

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Polymerase Chain Reaction Mediated Identification of Streptococcus uberis and Streptococcus parauberis Using Species‐specific Sequences of the Genes Encoding Superoxide Dismutase A and Chaperonin 60*

Alber

El-Sayed

Lämmler

et al. 2004

Journal of Veterinary Medicine, Series B

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Streptococcus uberis, a well-known bacterial pathogen associated with bovine mastitis, appears to be biochemically and serologically almost indistinguishable from the closely related species Streptococcus parauberis. In the present study, species-specific oligonucleotide primers were designed using internal parts of the genes sodA, encoding superoxide dismutase A, and cpn60 encoding chaperonin 60 of S. uberis and S. parauberis, respectively. The two oligonucleotide primer pairs allowed a rapid and reliable PCR-mediated identification and differentiation of both species. These studies, performed with S. uberis and S. parauberis reference cultures and clinical isolates from routine diagnostics, revealed that the occurrence of S. parauberis as causative agent of bovine mastitis appears to be rare. In addition the sodA and cpn60 sequence data confirmed that both species could taxonomically be classified to the pyogenic group of genus Streptococcus.

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Section: Discussionsupporting

confidence: 89%

Section: Bacterial Isolatesmentioning

confidence: 99%

Polymerase Chain Reaction Mediated Identification of Streptococcus uberis and Streptococcus parauberis Using Species‐specific Sequences of the Genes Encoding Superoxide Dismutase A and Chaperonin 60*

Alber

El-Sayed

Lämmler

et al. 2004

Journal of Veterinary Medicine, Series B

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“…In the present study, 94.9 % of the strains studied carried the pauA gene. The prevalence of the pauA gene was previously reported as ranging from 61.5 % in Argentina to 100 % in India 17,33,36 . To get further insight, we studied the nucleotide and amino acid sequences from 20 selected S. uberis mastitis isolates.…”

Section: Discussionmentioning

confidence: 94%

Genotyping and study of the pauA and sua genes of Streptococcus uberis isolates from bovine mastitis

Perrig

Ambroggio

Buzzola

et al. 2015

Revista Argentina de Microbiología

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This study aimed to determine the clonal relationship among 137 Streptococcus uberis isolates from bovine milk with subclinical or clinical mastitis in Argentina and to assess the prevalence and conservation of pauA and sua genes. This information is critical for the rational design of a vaccine for the prevention of bovine mastitis caused by S. uberis. The isolates were typed by random amplified polymorphic DNA (RAPD) analysis and by pulsed-field gel electrophoresis (PFGE). The 137 isolates exhibited 61 different PFGE types and 25 distinct RAPD profiles. Simpson's diversity index was calculated both for PFGE (0.983) and for RAPD (0.941), showing a high discriminatory power in both techniques. The analysis of the relationship between pairs of isolates showed 92.6% concordance between both techniques indicating that any given pair of isolates distinguished by one method tended to be distinguished by the other. The prevalence of the sua and pauA genes was 97.8% (134/137) and 94.9% (130/137), respectively. Nucleotide and amino acid sequences of the sua and pauA genes from 20 S. uberis selected isolates, based on their PFGE and RAPD types and geographical origin, showed an identity between 95% and 100% with respect to all reference sequences registered in GenBank. These results demonstrate that, in spite of S. uberis clonal diversity, the sua and pauA genes are prevalent and highly conserved, showing their importance to be included in future vaccine studies to prevent S. uberis bovine mastitis.

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“…These findings are in good agreement with the results of Odierno et al [31] who achieved correct identification in 94% of the cases by use of 11 biochemical tests. Khan et al [32] described the cultural, biochemical, serological and molecular properties of 132 strains of Strep. uberis .…”

Section: Discussionmentioning

confidence: 99%

Phenotypic and genotypic identification of streptococci and related bacteria isolated from bovine intramammary infections

et al. 2013

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BackgroundStreptococcus spp. and other Gram-positive, catalase-negative cocci (PNC) form a large group of microorganisms which can be found in the milk of cows with intramammary infection. The most frequently observed PNC mastitis pathogens (major pathogens) are Streptococcus uberis, Strep. dysgalactiae, and Strep. agalactiae. The remaining PNC include a few minor pathogens and a large nonpathogenic group. Improved methods are needed for the accurate identification and differentiation of PNC.A total of 151 PNC were collected from cows with intramammary infection and conclusively identified by 16S rRNA sequencing as reference method. Nine phenotypic microbiological tests (alpha-hemolysis, CAMP reaction, esculin hydrolysis, growth on kanamycin esculin azide agar and on sodium chloride agar, inulin fermentation, hippurate hydrolysis, leucine aminopeptidase and pyrrolidonyl peptidase activity), multiplex PCR for the three major pathogens (target genes for Strep. uberis, Strep. dysgalactiae and Strep. agalactiae: pauA, 16S rRNA, and sklA3, respectively), and mass spectroscopy using the matrix-assisted laser desorption ionization-time of flight (MALDI-TOF MS) were evaluated for the diagnosis and discrimination of the three clinically most relevant PNC.ResultsThe probability that a strain of Strep. uberis, Strep. dysgalactiae and Strep. agalactiae was correctly identified by combining the results of the 9 phenotypic tests was 92%, 90%, and 100%, respectively. Applying the multiplex PCR, all strains of the three major pathogens were correctly identified and no false positive results occurred. Correct identification was observed for all strains of Strep. uberis and Strep. agalactiae using MALDI-TOF MS. In the case of Strep. dysgalactiae, some variability was observed at the subspecies level, but all strains were allocated to one single cluster.ConclusionsThe results of the present study show that reliable identification of the clinically most relevant PNC (Strep. uberis, Strep. agalactiae and Strep. dysgalactiae) can be obtained by use of a combination of colony morphology, hemolysis type and catalase reaction, and a multiplex PCR with specific primers restricted to these 3 pathogens. The MALDI-TOF MS is a fast method that shows promising results, although identification of Strep. dysgalactiae at the subspecies level is not yet satisfactory.

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Identification and epidemiological characterization of Streptococcus uberis isolated from bovine mastitis using conventional and molecular methods

Cited by 64 publications

References 0 publications

Polymerase Chain Reaction Mediated Identification of Streptococcus uberis and Streptococcus parauberis Using Species‐specific Sequences of the Genes Encoding Superoxide Dismutase A and Chaperonin 60*

Polymerase Chain Reaction Mediated Identification of Streptococcus uberis and Streptococcus parauberis Using Species‐specific Sequences of the Genes Encoding Superoxide Dismutase A and Chaperonin 60*

Genotyping and study of the pauA and sua genes of Streptococcus uberis isolates from bovine mastitis

Phenotypic and genotypic identification of streptococci and related bacteria isolated from bovine intramammary infections

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