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Teider Junior P.I., Ribeiro Júnior J.C., Ossugui E.H., Tamanini R., Ribeiro J., Santos G.A., Alfieri A.A. & Beloti V. 2019. Pseudomonas spp. and other psychrotrophic microorganisms in inspected and non-inspected Brazilian Minas Frescal cheese: proteolytic, lipolytic and AprX production potential. Pesquisa Veterinária Brasileira 39(10):807-815. Instituto Nacional de Ciência e Tecnologia para a Cadeia Produtiva do Leite, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid PR-445 Km 380, Cx. Postal 10.011, Campus Universitário, Londrina, PR 86057-970, Brazil. E-mail: ribeirojuniorjc@gmail.com The most consumed cheese in Brazil, Minas Frescal cheese (MFC) is highly susceptible to microbial contamination and clandestine production and commercialization can pose a risk to consumer health. The storage of this fresh product under refrigeration, although more appropriate, may favor the growth of spoilage psychrotrophic bacteria. The objective of this study was to quantify and compare Pseudomonas spp. and other psychrotrophic bacteria in inspected and non-inspected MFC samples, evaluate their lipolytic and proteolytic activities and their metalloprotease production potentials. Twenty MFC samples were evaluated: 10 inspected and 10 non-inspected. Counts of psychrotrophic bacteria and Pseudomonas spp., evaluation of the proteolytic and lipolytic potential of the isolates, and identification of potential producers of alkaline metalloprotease (AprX) were assessed. The mean total psychrotrophic counts were 1.07 (±2.18) × 109CFU/g in the inspected samples and 4.5 (±5.86) × 108CFU/g in the non-inspected, with no significant difference (p=0.37). The average score of Pseudomonas spp. was 6.86 (±18.6) × 105 and 2.08 (±3.65) × 106 CFU/g for the inspected and non-inspected MFC samples, respectively, with no significant difference (p=0.1). Pseudomonas spp. represented 0.06% and 0.004% of psychrotrophic bacteria found in inspected and non-inspected MFC samples, respectively. Collectively, 694 psychrotrophic strains and 47 Pseudomonas spp. were isolated, of which 59.9% and 68.1% were simultaneously proteolytic and lipolytic, respectively. Of the 470 psychrotrophs isolated from inspected and 224 from non-inspected cheese samples, 5.74% and 2.23% contained aprX, respectively, while 100 and 86.96% of the Pseudomonas spp. isolates in inspected and non-inspected cheese samples contained the gene. The production potential of AprX did not, however, determine the proteolytic activity on plates of these isolates under the conditions evaluated in this study. Of total, 65.63% of the psychrotrophs that contained aprX gene were confirmed as Pseudomonas spp., using genus-specific PCR. Phylogenetic analysis of the 16S rRNA gene of the other psychrotrophs that were potential producers of AprX identified them as Serratia spp. (n=7), Raoultella ornithinolytica (n=1), and Acinetobacter schindleri (n=1) in the inspected samples and Psychrobacter sanguinis (n=1) and Leuconostoc mesenteroides (n=1) in the non-inspected PVB-6037 LD Pseudomonas spp. and other psychrotrophic microorganisms in inspected and non-inspected Brazilian Minas Frescal cheese: proteolytic, lipolytic and AprX production potential1 Pedro I. Teider Junior2, José C. Ribeiro Júnior3* , Eric H. Ossugui2, Ronaldo Tamanini2, Juliane Ribeiro2, Gislaine A. Santos2, Amauri A. Alfieri2  and Vanerli Beloti2 1 Received on February 18, 2019. Accepted for publication on April 1, 2019. 2 Instituto Nacional de Ciência e Tecnologia para a Cadeia Produtiva do Leite (INCT-Cadeia do Leite), Universidade Estadual de Londrina (UEL), Rodovia Celso Garcia Cid PR-445 Km 380, Cx. Postal 10.011, Campus Universitário, Londrina, PR 86057-970, Brazil. 3 Escola de Medicina Veterinária e Zootecnia, Universidade Federal do Tocantins (UFT), BR-153 Km 112, Zona Rural, Araguaína, TO 77804-970, Brazil. *Corresponding author: ribeirojuniorjc@gmail.com Pseudomonas spp. and other psychrotrophic microorganisms in inspected and noninspected Brazilian Minas Frescal cheese: proteolytic, lipolytic and AprX production

pesquisa veterinária brasileira

Pseudomonas spp. and other psychrotrophic microorganisms in inspected and non-inspected Brazilian Minas Frescal cheese: proteolytic, lipolytic and AprX production potential