title: Lipolytic and proteolytic activity of Pseudomonas spp. isolated during milking and storage of refrigerated raw milk.
abstract: The aim of this study was to verify the presence of lipolytic and proteolytic Pseudomonas spp. during milking and storage of refrigerated raw milk. We also intended to compare samples collected during rainy and dry seasons, from farms with manual and mechanical milking systems. For this, samples of milkers' hands, cows' teats, water, expansion tanks, equipment, and utensils used during milking were analyzed regarding Pseudomonas spp.
  
  
  METHODS
  Positive samples were tested for the production of lipolytic and proteolytic enzymes. Microorganisms of the genus Pseudomonas were isolated from all sampling points. A higher isolation rate of the bacterium was found in the rainy season except for 6 sampling points, with all of these associated with mechanical milking systems. Pseudomonas spp. exhibiting lipolytic activity were found to be predominant during the dry season, since no activity was detected during the rainy season in 26 of the 29 sampling sites. The highest number of lipolytic Pseudomonas isolates was obtained from water. Presence of lipase-producing Pseudomonas spp. was verified in 7 and 36% of the samples collected from farms with manual and mechanical milking, respectively. When analyzing raw milk collected from expansion tanks immediately (0 h) and 24h after milking, we observed that for dairy properties with manual milking process, 10% of the Pseudomonas isolates were positive for lipolytic activity. The percentage increased to 12% 48h after milking. Mean averages were 32, 33, and 39% immediately after, 24 and 48h after milking, respectively, for farms with mechanical milking. All sampling points showed the presence of proteolytic strains of Pseudomonas. The highest proteolytic activity was found during the rainy season, except for the samples collected from milkers' hands before milking, buckets, and teat cup inner surfaces after milking and from the water in dairy farms with mechanical milking system. Of these samples, 72, 56, and 50%, respectively, were positive for proteolysis during the dry season. For the water samples, a statistical difference was observed between mechanical (50%) and manual (7%) milking systems in the percentage of proteolytic activity. No production of proteolytic enzyme was detected in the samples from milkers' hands taken after milking and no statistically significant difference was found among manual (19.91%) and mechanical (47.85%) milking. During the rainy months, no proteolysis was detected in the samples taken from cows' teats after the predipping. It is evident, therefore, that preventive measures capable of minimizing the contamination with Pseudomonas spp. during milking and storage of refrigerated raw milk are needed, regardless of season.
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Capodifoglio
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Ana Maria Centola
authors.initials: AMC
authors.lastname: Vidal
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Joyce Aparecida Santos
authors.initials: JAS
authors.lastname: Lima
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Fernanda
authors.initials: F
authors.lastname: Bortoletto
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Léa Furlan
authors.initials: LF
authors.lastname: D'Abreu
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Ana Carolina Siqueira
authors.initials: ACS
authors.lastname: Gonçalves
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Andreia Cristina Nakashima
authors.initials: ACN
authors.lastname: Vaz
authors.affiliation: School of Veterinary Medicine and Animal Science, University of Sao Paulo, Pirassununga 13635-900, Brazil.
authors.forename: Julio Cesar de Carvalho
authors.initials: JCC
authors.lastname: Balieiro
authors.affiliation: School of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga 13635-900, Brazil. Electronic address: saranetto@usp.br.
authors.forename: Arlindo Saran
authors.initials: AS
authors.lastname: Netto
mesh_terms: D000818:Animals; D003612:Dairying / Q000379:methods*; D061353:Food Storage / Q000379:methods*; D008066:Lipolysis; D008892:Milk / Q000737:chemistry* / Q000382:microbiology; D059748:Proteolysis; D011549:Pseudomonas / Q000378:metabolism*; D012034:Refrigeration; D012621:Seasons
keywords: milk; milking system; psychrotrophic; storage