Hostname: page-component-8448b6f56d-c47g7 Total loading time: 0 Render date: 2024-04-24T15:42:17.063Z Has data issue: false hasContentIssue false
  • English
  • Français

Pseudomonas fluorescens and Pseudomonas putida from refrigerated raw milk: genetic diversity and lipoproteolytic activity

Published online by Cambridge University Press:  11 February 2022

Rosana De Longhi ,
Selma de Souza Correia ,
Samera Rafaela Bruzaroski ,
Regina Celia Poli-Frederico ,
Rafael Fagnani  and
Elsa Helena Walter de Santana
Rosana De Longhi
Affiliation:
Universidade Pitágoras Unopar (UNOPAR), Mestrado em Ciência e Tecnologia de Leite e Derivados, Londrina, Paraná, Brazil
Selma de Souza Correia
Affiliation:
UNOPAR, Mestrado em Saúde e Produção Animal, Arapongas, Paraná, Brazil
Samera Rafaela Bruzaroski
Affiliation:
Universidade Pitágoras Unopar (UNOPAR), Mestrado em Ciência e Tecnologia de Leite e Derivados, Londrina, Paraná, Brazil
Regina Celia Poli-Frederico
Affiliation:
UNOPAR, Mestrado e Doutorado em Ciência da Reabilitação, Londrina, Paraná, Brazil
Rafael Fagnani
Affiliation:
Universidade Estadual de Londrina, Londrina, Paraná, Brazil
Elsa Helena Walter de Santana*
Affiliation:
Universidade Pitágoras Unopar (UNOPAR), Mestrado em Ciência e Tecnologia de Leite e Derivados, Londrina, Paraná, Brazil UNOPAR, Mestrado em Saúde e Produção Animal, Arapongas, Paraná, Brazil
*
Author for correspondence: Elsa Helena Walter de Santana, Email: elsahws@hotmail.com
Get access Rights & Permissions [Opens in a new window]

Abstract

In this research communication the genetic diversity of Pseudomonas fluorescens (n = 67) and Pseudomonas putida (n = 44) isolated from refrigerated raw milk from bulk tank trucks were verified. The relationship between the genetic profile of the isolates and their lipoproteolytic potential was evaluated using skim milk agar and tributyrin agar (21°C/72 h). The lipoproteolytic potential (low or high), evaluated by the diameter of the halos (cm), was correlated with the number of milk producing properties that contributed to each sample (one sample = one bulk tank truck; 8−80 producers/sample) and the distance between the dairy properties and the processing plant (21−370 km). P. fluorescens was confirmed in all samples, while P. putida in 60% samples. For both species, two clusters (I and II) were observed, and the first one showed lower genotypic diversity and the presence of isolates with 100% similarity. P. fluorescens isolates presenting at least 70% similarity were 83.9% in Cluster I (n = 31) and 44.4% in Cluster II. In both clusters (I and II) observed in the P. fluorescens dendrogram, the occurrence of high proteolytic and lipolytic potential were equivalent. The higher the number of farms per milk sample, the greater the lipoproteolytic intensity of P. fluorescens isolates. In relation to P. putida isolates, 74% presented at least 50% similarity in Cluster I (n = 27) and only 35% in Cluster II (n = 17). The occurrence of high proteolysis linked to P. putida was proportional between both Clusters, but the occurrence of high lipolysis was greater in Cluster II. No significant association was detected between P. putida isolates and the variables studied. The results indicate the circulation of P. putida and P. fluorescens with 100% similarity in different milk producing regions. The level of genetic diversity was related only to the lipolytic capacity of P. putida.

Keywords

EnzymespsychrotrophicqualityRep PCR
Type
Research Article
Information
Journal of Dairy Research , Volume 89 , Issue 1 , February 2022 , pp. 86 - 89
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aguiar, BM, Longhi, R, Poli-Frederico, RC, Fagnani, R and Santana, EHW (2019) Lipoproteolytic capacity and potential of Pseudomonas spp. Isolated from cold raw milk. Journal of Dairy Research 86, 467–469.CrossRefGoogle Scholar
Almeida, KM, Bruzaroski, SR, Zanol, D, Melo, M, Santos, JS, Aragon- Alegro, LC, Botaro, BG and Santana, EHW (2017) Pseudomonas spp. and P. fluorescens: population in refrigerated raw milk.. Ciência Rural 47, 2–6.CrossRefGoogle Scholar
Chen, L, Daniel, RM and Coolbear, T (2003) Detection and impact of protease and lipase activities in milk and milk powders. International Dairy Journal 13, 255–275.CrossRefGoogle Scholar
Decimo, M, Morandi, S, Silvetti, T and Brasca, M (2014) Characterization of Gram negative psychrotrophic bacteria isolated from Italian bulk tank milk. Journal of Food Science 79, 2081–2090.CrossRefGoogle ScholarPubMed
Ercolini, D, Russo, F, Ferrocino, F and Villani, F (2009) Molecular identification of mesophilic and psychrotrophic bacteria from raw cow's milk. Food Microbiology 26, 228–231.CrossRefGoogle ScholarPubMed
Frank, JF and Yousef, AE (2004) Tests for groups of microorganisms. In Weir MH (ed.), Standard Methods for the Examination of Dairy Products. Nova York: American Public Health Association, pp. 227–247.Google Scholar
Kumaresan, G, Annalvilli, R and Sivakumar, K (2007) Psychrotrophic spoilage of raw milk at different temperatures of storage. Journal of Applied Sciences Research 3, 1383–1387.Google Scholar
Louws, FJ, Fulbright, DW, Stephens, CT and Debruijn, FJ (1994) Specific -genomic fingerprints of phytopathogenic Xanthomonas and Pseudomonas pathovars and strains generated with repetitive sequences and PCR. Applied and Environmental Microbiology 60, 2286–2295.CrossRefGoogle ScholarPubMed
Mahieu, H (1991) Modificaciones de la leche después de su recogida. In Luquet, FM (ed.), Leche y productos lácteos. la leche de la mama a la lechería. Zaragoza: Acribia, pp. 22–181.Google Scholar
Scarpellini, M, Franzetti, L and Galli, A (2004) Development of PCR assay to identify Pseudomonas fluorescens and its biotype. FEMS Microbiology Letters 236, 257–260.CrossRefGoogle ScholarPubMed
Spilker, T, Coenye, T, Vandame, P and Lipuma, JJ (2004) PCR-Based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. Journal of Clinical Microbiology 42, 2074–2079.CrossRefGoogle ScholarPubMed
Vidal, AMC, Netto, AS, Vaz, ACN, Capodifóglio, E, Gonçalves, ACS, Rossi, GAM, Figueiredo, AS and Ruiz, VLA (2017) Pseudomonas spp.: contamination sources in bulk tanks of dairy farms. Pesquisa Veterinaria Brasileira 37, 941–948.CrossRefGoogle Scholar
Yamamoto, S and Harayama, S (1995) Amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Applied and Environmental Microbiology 61, 1104–1109.CrossRefGoogle Scholar