Epidemiological analysis of worldwide bovine, canine and human clinical Prototheca isolates by PCR genotyping and MALDI-TOF mass spectrometry proteomic phenotyping.

This study presents information on the phenotypic and genotypic characterization of clinical Prototheca spp. isolates obtained from different geographic regions. Of 350 isolates studied, 342 came from cattle, six from canines and two from humans. Phenotypic characterization was carried out by a matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) proteomic analysis. The peptide extraction that was used for this analysis included the additional steps of washing and sonication to increase the yield of peptide. Genotypic analysis was conducted using species- and genotype-specific primers. The study revealed that among the cattle isolates, 310 (90.6%) belonged to Prototheca zopfii genotype 2, 30 (8.8%) to P. blaschkeae, and two (0.6%) to P. zopfii genotype 1. P. zopfii genotype 2 is the principal etiological agent of protothecal mastitis in cattle regardless of the geographic region. Similarly, all canine and human isolates also belonged to the P. zopfii genotype 2, suggesting that this is probably the most virulent species of the genus. The role of P. blaschkeae needs further epidemiologic studies to ascertain its etiologic role in bovine mastitis. To the best of our knowledge, this is the first comprehensive study on phenotypic and genotypic characterization of P. zopfii and P. blaschkeae isolates originating from diverse clinical specimens from different countries.

[Genotypical differentiation of Prototheca isolates of milk samples from mastitis affected cattle in Germany]. / Genotypische Differenzierung von Prototheca-Isolaten aus Milchproben mastitiskranker Rinder in Deutschland.

To elucidate the spectra of Prototheca species and P. zopfii genotypes in milk samples from mastitis affected cattle, 200 Prototheca-isolates of 57 German dairy cattle barns were analyzed by species- and genotype- specific PCR and RFLP analysis. The investigations showed that 177 (88.5%) of all isolates represented P. zopfii genotype 2, 21 (10.5%) isolates represented P. blaschkeae and two (1.0%) P. zopfii genotype 1. Therefore mainly P. zopfii genotype 2 but also P. blaschkeae act in the epidemiology of mastitis in cattle, whereas the role of P. blaschkeae in development of bovine protothecal mastitis should be further explored in epidemiological studies. In this study P. blaschkeae was demonstrated for the first time in association with bovine mastitis in Germany.

Genotyping of bovine Prototheca mastitis isolates from Poland.

Bovine mastitis due to unicellular, achlorophyllous algae of the genus Prototheca is a serious and complex ailment that accounts for high economic losses in the dairy industry. Bovine protothecal mastitis has been almost exclusively associated with only one species Prototheca zopfii. Recently, based on the 18S rDNA sequence analysis, P. zopfii has been divided into three genotypes (1-3), of which P. zopfii genotype 3 has been given the status of a new species Prototheca blaschkeae sp. nov. The aim of this study was to investigate the genotypic composition of the population of P. zopfii bovine mastitis isolates from Poland, according to that threefold classification. Of the 44 isolates tested, 43 (98%) were identified as P. zopfii genotype 2. Only one isolate was assigned to the newly established P. blaschkeae species. These findings clearly show the predominance of the P. zopfii genotype 2 in the etiology of bovine mammary protothecosis in Poland, but also underline the involvement of P. blaschkeae in the disease. The study is the first to provide molecular characterization of Polish P. zopfii mastitis isolates, collected from different regions of Poland.