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      Survey of molds, yeast and Alicyclobacillus spp. from a concentrated apple juice productive process

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          Abstract

          Bacteria and molds may spoil and/or contaminate apple juice either by direct microbial action or indirectly by the uptake of metabolites as off-flavours and toxins. Some of these microorganisms and/or metabolites may remain in the food even after extensive procedures. This study aim to identify the presence of molds (including heat resistant species) and Alicyclobacillus spp., during concentrated apple juice processing. Molds were isolated at different steps and then identified by their macroscopic and microscopic characteristics after cultivation on standard media at 5, 25 and 37ºC, during 7 days. Among the 19 isolated found, 63% were identified as Penicillium with 50% belonging to the P. expansum specie. With regards to heat resistant molds, the species Neosartorya fischeri, Byssochlamys fulva and also the genus Eupenicillium sp., Talaromyces sp. and Eurotium sp. were isolated. The thermoacidophilic spore-forming bacteria were identified as A. acidoterrestris by a further investigation based on 16S rRNA sequence similarity. The large contamination found indicates the need for methods to eliminate or prevent the presence of these microorganisms in the processing plants in order to avoid both spoilage of apple juice and toxin production.

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          Most cited references56

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          Heat-resistant fungi of importance to the food and beverage industry.

          V Tournas (1993)
          Spoilage of pasteurized and canned fruit and fruit products caused by heat-resistant molds have been reported repeatedly in recent years. Species most commonly implicated in fruit and fruit product disintegration are Byssochlamys fulva, Byssochlamys nivea, Neosartorya fischeri, Talaromyces flavus, and Eupenicillium brefeldianum. These organisms are saprophytic rather than parasitic and usually contaminate fruits on or near the ground. They can survive heat treatments used for fruit processing and can grow and spoil the products during storage at room temperature, which results in great economic losses. Mold heat resistance is attributed to the formation of sexual spores, ascospores. Ascospores have a wide range of heat resistance, depending on species, strain, age of organism, heating medium, pH, presence of sugars, fats, and acids in heating medium, growth conditions, etc. The mechanism(s) of thermoresistance are not clear; probably some very stable compound(s) critical to germination and outgrowth are present in the heat-resistant ascospores. Besides spoilage, the heat-resistant molds produce a number of toxic secondary metabolites, such as byssotoxin A; byssochlamic acid; the carcinogen, patulin, the tremorgenic substances, fumitremorgin A and C, and verruculogen; fischerin, which caused fatal peritonitis in mice; and eupenifeldin, a compound possessing cytotoxicity as well as in vivo antitumor activity. Growth of heat-resistant fungi can be controlled by lowering the water activity, adding sulfur dioxide, sorbate, or benzoate; washing of fruits in hypochlorite solution before heat treatment reduces the number of ascospores and makes the heat destruction more successful. More research is needed to elucidate the mechanism(s) of thermoresistance and develop new methods for the complete inactivation of resistant ascospores.
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            Influence of package, type of apple juice and temperature on the production of patulin by Byssochlamys nivea and Byssochlamys fulva.

            Although the production of patulin in apple fruits is mainly by Penicillium expansum, there is no information on the ability of heat resistant moulds that may survive pasteurization to produce this mycotoxin in juice packages during storage and distribution. In this study, the production of patulin by Byssochlamys spp (Byssochlamys nivea FRR 4421, B. nivea ATCC 24008 and Byssochlamys fulva IOC 4518) in cloudy and clarified apple juices packaged in laminated paperboard packages or in polyethylene terephthalate bottles (PET) and stored at both 21 degrees C and 30 degrees C, was investigated. The three Byssochlamys strains were able to produce patulin in both cloudy and clarified apple juices. Overall, the lower the storage temperature, the lower the patulin levels and mycelium dry weight in the apple juices (p<0.05). The greatest variations in pH and degrees Brix were observed in the juices from which the greatest mycelium dry weights were recovered. The maximum levels of patulin recovered from the juices were ca. 150 microg/kg at 21 degrees C and 220 microg/kg at 30 degrees C. HPLC-UV, HPCL-DAD and mass spectrometry analyses confirmed the ability of B. fulva IOC 4518 to produce patulin. Due to the heat resistance of B. nivea and B. fulva and their ability to produce patulin either in PET bottles or in laminated paperboard packages, the control of contamination and the incidence of these fungi should be a matter of concern for food safety. Control measures taken by juice industries must also focus on controlling the ascospores of heat resistant moulds.
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              Alicyclobacillus acidoterrestris in pasteurized exotic Brazilian fruit juices: isolation, genotypic characterization and heat resistance.

              In this study, the population of Alicyclobacillus spp. was estimated in pasteurized exotic Brazilian fruit juices using the most probable number (MPN) technique followed by biochemical tests. Pasteurized passion fruit (n = 57) and pineapple (n = 50) juices were taken directly from Brazilian manufacturers. While Alicyclobacillus spp. was isolated from passion fruit juice, the microorganism was not found in any pineapple juice samples. A higher incidence of Alicyclobacillus was observed in samples taken in June and July (dry months in Brazil) in comparison to the other months (March, April, May and August), and the highest Alicyclobacillus counts were recovered from these samples(>23 MNP/100 mL). Sixteen (n = 16) Alicyclobacillus strains were typed using the randomly amplified polymorphic DNA method (RAPD-PCR). RAPD-PCR revealed great genetic similarity between the passion fruit juice strains and Alicyclobacillus acidoterrestris DSM 2498. The heat resistance of three isolates was determined, and the mean D(95°) (1.7 min) and z (7.6 °C) values in the passion fruit juice were not significantly different (p > 0.05) from those obtained for the DSM 2498 strain (D(95°) = 1.5 min and z = 7.1 °C). This is the first report on the isolation of A. acidoterrestris from exotic fruit juices such as passion fruit juice. It is worth pointing out the importance of applying good agricultural practices in the field and applying controls for the fruit selection and washing steps, as well as controlling the time/temperature conditions for pasteurization so as to reduce the incidence and chances of A. acidoterrestris spoilage in these juices.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                bjm
                Brazilian Journal of Microbiology
                Braz. J. Microbiol.
                Sociedade Brasileira de Microbiologia (São Paulo )
                1678-4405
                2014
                : 45
                : 1
                : 49-58
                Affiliations
                [1 ] Universidade Federal do Rio Grande do Norte Brazil
                [2 ] Universidade do Oeste de Santa Catarina Brazil
                [3 ] Universidade Federal de Santa Catarina Brazil
                Article
                S1517-83822014000100008
                10.1590/S1517-83822014000100008
                f1f396e1-3f57-4947-a63a-0971819ed8ab

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1517-8382&lng=en
                Categories
                MICROBIOLOGY

                Microbiology & Virology
                concentrated apple juice,Alicyclobacillus acidoterrestris,heat resistant mold,Penicillium expansum

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