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Item Development of a selective culture medium for bifidobacteria, Raffinose-Propionate Lithium Mupirocin (RP-MUP) and assessment of its usage with Petrifilm™ Aerobic Count plates(Food Microbiology, 2014-05) Miranda, Rodrigo Otávio; Carvalho, Antonio Fernandes de; Nero, Luís AugustoThis study aimed to develop a selective culture media to enumerate bifidobacteria in fermented milk and to assess this medium when used with Petrifilm™ AC plates. For this purpose, Bifidobacterium spp., Lactobacillus spp. and Streptococcus thermophilus strains were tested to verify their fermentation patterns for different carbohydrates. All bifidobacteria strains were able to use raffinose. Based on these characteristic, a selective culture medium was proposed (Raffinose-Propionate Lithium Mupirocin, RP-MUP), used with Petrifilm™ AC plates, and was used to enumerate bifidobacteria in fermented milk. RP-MUP performance was assessed by comparing the results with this medium to reference protocols and culture media for bifidobacteria enumeration. RP-MUP, whether used or not with Petrifilm™ AC, presented similar performance to TOS-MUP (ISO 29981), with no significant differences between the mean bifidobacteria counts (p < 0.05) and with high correlation indices (r = 0.99, p < 0.05). As an advantage, reliable results were obtained after just 48 h of incubation when RP-MUP was used with Petrifilm™ AC, instead of the 72 h described in the ISO 29981 protocol.Item Enumeration of bifidobacteria using Petrifilm™ AC in pure cultures and in a fermented milk manufactured with a commercial culture of Streptococcus thermophilus(Food Microbiology, 2011-07-21) Miranda, Rodrigo Otávio; Gama Neto, Gabriel; Freitas, Rosangela de; Carvalho, Antônio Fernandes de; Nero, Luís AugustoBifidobacteria are probiotic microorganisms that are widely used in the food industry. With the aim of using of Petrifilm™ Aerobic Count (AC) plates associated with selective culture media, aliquots of sterile skim milk were inoculated separately with four commercial cultures of bifidobacteria. These cultures were plated by both the conventional method and Petrifilm™AC, using the culture media NNLP and ABC. The cultures were incubated under anaerobiosis at 37 °C for 24, 48 and 72 h. No significant differences (p > 0.05) were observed between the obtained counts at 48 and 72 h. Bifidobacteria counts in ABC were usually higher than in NNLP, independent of the plating method. Subsequently, fermented milk was prepared with a Streptococcus thermophilus strain, and aliquots were inoculated with the same bifidobacteria. Then, the fermented milks were submitted to microbiological analysis for bifidobacteria enumeration using the same culture media and methodologies previously described, incubated under anaerobiosis at 37 °C for 48 h. Again, bifidobacteria counts in ABC were higher than in NNLP, with significant differences for some cultures (p < 0.05). The counts obtained by both methodologies presented significant correlations (p < 0.05). The results indicate the viability of Petrifilm™AC as an alternative method for bifidobacteria enumeration when associated to specific culture media, specially the ABC.Item Enumeration of bifidobacteria using PetrifilmTM AC in pure cultures and in a fermented milk manufactured with a commercial culture of Streptococcus thermophilus(Food Microbiology, 2011-07-21) Miranda, Rodrigo Otávio; Freitas, Rosangela de; Carvalho, Antônio Fernandes de; Nero, Luís Augusto; Gama Neto, GabrielBifidobacteria are probiotic microorganisms that are widely used in the food industry. With the aim of using of PetrifilmÔ Aerobic Count (AC) plates associated with selective culture media, aliquots of sterile skim milk were inoculated separately with four commercial cultures of bifidobacteria. These cultures were plated by both the conventional method and PetrifilmÔAC, using the culture media NNLP and ABC. The cultures were incubated under anaerobiosis at 37 C for 24, 48 and 72 h. No significant differences (p > 0.05) were observed between the obtained counts at 48 and 72 h. Bifidobacteria counts in ABC were usually higher than in NNLP, independent of the plating method. Subsequently, fermented milk was prepared with a Streptococcus thermophilus strain, and aliquots were inoculated with the same bifidobacteria. Then, the fermented milks were submitted to microbiological analysis for bifidobacteria enumeration using the same culture media and methodologies previously described, incubated under anaerobiosis at 37 C for 48 h. Again, bifidobacteria counts in ABC were higher than in NNLP, with significant differences for some cultures (p < 0.05). The counts obtained by both methodologies presented significant correlations (p < 0.05). The results indicate the viability of PetrifilmÔAC as an alternative method for bifidobacteria enumeration when associated to specific culture media, specially the ABC.Item Development of a selective culture medium for bifidobacteria, Raffinose-Propionate Lithium Mupirocin (RP-MUP) and assessment of its usage with PetrifilmÔ Aerobic Count plates(Food Microbiology, 2013-11-26) Miranda, Rodrigo Otávio; Carvalho, Antonio Fernandes de; Nero, Luís AugustoThis study aimed to develop a selective culture media to enumerate bifidobacteria in fermented milk and to assess this medium when used with PetrifilmÔ AC plates. For this purpose, Bifidobacterium spp., Lactobacillus spp. and Streptococcus thermophilus strains were tested to verify their fermentation patterns for different carbohydrates. All bifidobacteria strains were able to use raffinose. Based on these characteristic, a selective culture medium was proposed (Raffinose-Propionate Lithium Mupirocin, RP-MUP), used with PetrifilmÔ AC plates, and was used to enumerate bifidobacteria in fermented milk. RP-MUP performance was assessed by comparing the results with this medium to reference protocols and culture media for bifidobacteria enumeration. RP-MUP, whether used or not with PetrifilmÔ AC, presented similar performance to TOS-MUP (ISO 29981), with no significant differences between the mean bifi dobacteria counts (p < 0.05) and with high correlation indices (r 1⁄4 0.99, p < 0.05). As an advantage, reliable results were obtained after just 48 h of incubation when RP-MUP was used with PetrifilmÔ AC, instead of the 72 h described in the ISO 29981 protocol.