Márcia Maria dos Anjos

Evaluation of the antibacterial activity of Piperaceae extracts and nisin on Alicyclobacillus acidoterrestris.

Alicyclobacillus acidoterrestris is a gram-positive aerobic bacterium. This bacterium resists pasteurization temperatures and low pH and is usually involved in the spoilage of juices and acidic drinks. The objective of this study was to evaluate the antibacterial activities of nisin and the species Piper (Piperaceae) on A. acidoterrestris. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined by the broth microdilution method. The species Piper aduncum had the lowest MIC and an MBC of 15.6 μg/mL and was selected for fractionation. Six fractions were obtained, and the dichloromethane fraction (F.3) had the lowest MIC/MBC (7.81 μg/mL). The dichloromethane fraction was again fractionized, and a spectral analysis revealed that the compound was prenylated chromene (F.3.7). The checkerboard method demonstrated that the crude extract (CE) of P. aduncum plus nisin had a synergistic interaction (fractional inhibitory concentration [FIC] = 0.24). The bactericidal activity of (F.3.7) was confirmed by the time-kill curve. P. aduncum, nisin, and prenylated chromene exhibited strong antibacterial activity against the spores and vegetative cells of A. acidoterrestris. The results of this study suggest that extracts of the genus Piper may provide an alternative to the use of thermal processing for controlling A. spoilage.

Antibacterial activity of papain and bromelain on Alicyclobacillus spp.

Alicyclobacillus spp. are spore forming bacteria that are often related to the deterioration of acidic products such as beverages and citrus juices. After the process of industrial pasteurization, the spore produced by the bacteria can germinate and the microorganism can grow, causing sensory abnormalities in the product. Alternative biopreservatives, such as the antimicrobial compounds, are of considerable importance to the food industry. Papain and bromelain are proteolytic enzymes derived frompapaya and pineapple, respectively. These enzymes are widely used in medicine and in the pharmaceutical and food industries, but while some studies have described their antibacterial action, no studies of the Alicyclobacillus spp. exist. The aimof this studywas to analyze the antibacterial effect of papain and bromelain on Alicyclobacillus spp. through 1) determining minimum inhibitory and bactericidal concentration (MIC and MBC); 2) determining the death time curve of the micro-organism in the presence and absence of enzymes; and 3) investigating the enzymatic mechanism on the microorganism. The antibacterial activity of enzymes in combination with nisin was also evaluated. The results showed that for the Alicyclobacillus acidoterrestris strain, the MIC of papain was 0.98 μg/mL and the MBC was 3.91 μg/mL, while theMIC of bromelain was 62.5 μg/mL and the MBCwas 250 μg/mL. The concentration of 4 ×MIC for both the enzymes was sufficient to eliminate 4 logs of the micro-organism after 24 h of incubation. Through the use of enzyme inhibitors specific for cysteine proteases, it was found that the antibacterial activity of papain and bromelain is not related to its proteolytic activity, butmay be related to other activities, such as amidse and esterase. The synergistic activity of the enzymes revealed a fractional inhibitory concentration (FIC) level of 0.16. Combination with nisin revealed an FIC of 0.25 for papain and 0.19 for bromelain, indicating synergism between both compounds. The application of enzymes in reconstituted orange juice contaminated with A. acidoterrestris was found to be effective, as after 48 h of incubation, at three different temperatures, the initial microbial population was eliminated. This study showed that the enzymes papain and bromelain have an antibacterial effect on A. acidoterrestris.

Resistance of Alicyclobacillus acidoterrestris spores and biofilm to industrial sanitizers.

This study evaluated the adhesion and biofilm formation of Alicyclobacillus acidoterrestris on industrial orange juice processing equipment and the bactericidal efficacy of peracetic acid, sodium hypochlorite, and quaternary ammonia after biofilm formation. The efficacy of these sanitizers against the spores of this microorganism was also evaluated. Stainless steel and nylon surfaces exhibited higher cell adhesion levels than did polyvinyl chloride surfaces. Peracetic acid was the most effective in removing biofilms from all surfaces (P < 0.05) and also reduced bacterial counts by 3 log CFU/cm² on the surface of polyvinyl chloride, but the other sanitizers also reduced the bacterial counts by 2 log CFU/cm². Quaternary ammonia exhibited the optimal minimum sporicidal concentration, preventing spore germination after only 15 s of contact at a concentration of 82 ppm. The flow cytometry results indicated that the spores and cells had low incidences of plasma membrane lysis after treatment with sanitizer, suggesting that lysis is not the principal mode of action for these sanitizers on A. acidoterrestris.

Avaliação da Atividade Antibacteriana de Enzima Natural em Alicyclobcillus Acidoterrestris

Marcia Maria dos Anjos, Angela Aparecida da Silva, Isabella Carolini Pascoli, Jane Graton-Mikcha, Jane GratonMikcha, Miguel Machinski-Junior, Celso Vataru Nakamura, Benicio Alves de Abreu-Filho. Avaliacao da Atividade Antibacteriana de Enzima Natural em Alicyclobcillus Acidoterrestris. In: Anais do 12o Congresso Latinoamericano de Microbiologia e Higiene de Alimentos MICROAL 2014 [= Blucher Food Science Proceedings, num.1, vol.1]. Sao Paulo: Editora Blucher, 2014. DOI 10.5151/foodsci-microal-285 Avaliacao da Atividade Antibacteriana de Enzima Natural em Alicyclobcillus Acidoterrestris

Avaliação Quantitativa da Qualidade Microbiológica do Ar de Ambientes Climatizados de Manipulação de Alimentos da Universidade Estadual de Maringá.

Jessica Marchiori Bicalho, Marcia Maria dos Anjos, Angela Aparecida da Silva, Isabella Carolini Pascoli, Celso Vataru Nakamura, Benicio Alves de Abreu-Filho. Avaliacao Quantitativa da Qualidade Microbiologica do Ar de Ambientes Climatizados de Manipulacao de Alimentos da Universidade Estadual de Maringa.. In: Anais do 12o Congresso Latinoamericano de Microbiologia e Higiene de Alimentos MICROAL 2014 [= Blucher Food Science Proceedings, num.1, vol.1]. Sao Paulo: Editora Blucher, 2014. DOI 10.5151/foodsci-microal-150 Avaliacao Quantitativa da Qualidade Microbiologica do Ar de Ambientes Climatizados de Manipulacao de Alimentos da Universidade Estadual de Maringa.

Avaliação da Atividade Antibacteriana do Óleo Essencial de Copaifera Multijuga Frente Às Células Vegetativas de Alicyclobacillus Acidoterrestris.

Angela Aparecida Silva, Idinea Fernandes Santos, Marcia Maria Anjos, Jane Martha Graton Mikcha, Miguel Machinski Junior, Celso Vataru Nakamura, Benicio Alves Abreu Filho. Avaliacao da Atividade Antibacteriana do Oleo Essencial de Copaifera Multijuga Frente As Celulas Vegetativas de Alicyclobacillus acidoterrestris. In: Anais do 12o Congresso Latinoamericano de Microbiologia e Higiene de Alimentos MICROAL 2014 [= Blucher Food Science Proceedings, num.1, vol.1]. Sao Paulo: Editora Blucher, 2014. DOI 10.5151/foodsci-microal-276 Avaliacao da Atividade Antibacteriana do Oleo Essencial de Copaifera Multijuga Frente as Celulas Vegetativas de Alicyclobacillus acidoterrestris.

Atividade Antimicrobiana do Óleo Essencial de Copaifera Guianensis Frente À Alicyclobacillus Acidoterrestris.

Angela Aparecida Silva, Idinea Fernandes Santos, Marcia Maria Anjos, Isabela Carolini Pascoli, Daniela Biral Prado, Celso Vataru Nakamura, Jane Martha Graton Mikcha, Miguel Machinski Junior, Benicio Alves Abreu Filho. Atividade Antimicrobiana do Oleo Essencial de Copaifera guianensis Frente A Alicyclobacillus acidoterrestris.. In: Anais do 12o Congresso Latinoamericano de Microbiologia e Higiene de Alimentos MICROAL 2014 [= Blucher Food Science Proceedings, num.1, vol.1]. Sao Paulo: Editora Blucher, 2014. DOI 10.5151/foodsci-microal-298 Atividade Antimicrobiana do Oleo Essencial de Copaifera guianensis Frente a Alicyclobacillus acidoterrestris.

Evaluation of different culture media and enrichment in orange juice upon the growth of Alicyclobacillusspp.

Bacteria of the genus Alicyiclobacillus spp. form spores and develop in acid media, leading to the spoilage of citrus juices. Brazil is the largest exporter of orange juice concentrate, and yet, it has been extensively studied due to changes in taste and smell. Several investigations have reported different culture media used to detect and enumerate Alicyiclobacillus spp. The objective of the present study was to evaluate the recovery of Alicyiclobacillus spp. spores grown in ALI, BAT, K agar and YSG media using the methodology suggested by ABECitrus. Five inocula were used, two from reference strains and three from pasteurized concentrated orange juice. Cell recovery after the enrichment in reconstituted orange juice was also analyzed. An initial population of 6 log CFU/mL was inoculated. ALI, BAT and YSG media were able to recover the initial population of all different inocula, with no significant differences between the results. When compared to BAT, however, the preparation of ALI and YSG media was simpler and had more advantages. The recovery with K agar medium was lower than the other media for all the tested inocula, with significant differences found for Alicyclobacillus acidocaldarius 0298T (3.66 log CFU/mL) and Alicyclobacillus pomorum-like CBMAI 0278 (4.11 log CFU/mL).