Orientador: Pilar Rodriguez de Massaguer / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-04T02:27:14Z (GMT). No. of bitstreams: 1 TamegaJunior_WilsonPedro_M.pdf: 3655390 bytes, checksum: 7393715ee9c1c652503290358f0b2729 (MD5) Previous issue date: 2005 / Mestrado / Ciência de Alimentos / Mestre em Ciência de Alimentos

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Orientador: Fumio Yokoya / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-22T20:33:42Z (GMT). No. of bitstreams: 1 Conti_MariaJosiane_M.pdf: 10300128 bytes, checksum: c571b2db431105550b581c8203ff0630 (MD5) Previous issue date: 2013 / Resumo: O Alicyclobacillus acidoterrestris, é uma bactéria acidotermofílica, não patogênica, de origem ambiental. Este organismo é um potencial agente deteriorante em polpas e sucos de frutas ácidos. Devido à presença de endosporos de elevada resistência térmica, torna-se difícil sua inativação pelos tratamentos térmicos convencionais de pasteutização. Esta pesquisa visou extrair e processar termicamente o suco de seis tipos de frutas tropicais, de caráter ácido (abacaxi, acerola, caju, goiaba, manga e maracujá), e avaliar se estes apresentam condições para que o A. acidoterrestris se desenvolva em sua temperatura ótima (45°C) e também em temperatura de abuso de processamento (35°C), temperatura que favorece o crescimento de microrganismos mesófilos. Além disso, foi analisada a concentração mínima inicial, do microrganismo em estudo, capaz de sobreviver e se desenvolver no suco de manga, aquele que apresentou melhores condições de desenvolvimento na primeira fase do estudo. Neste estudo foi também incluída a análise qualitativa de guaiacol como indicador de deterioração. Foi preparada uma suspensão com a mistura de esporos de quatro cepas de A. acidoterrestris em mesma concentração, padronizada e inoculada nos sucos, previamente extraídos e tratados termicamente, apresentando carga inicial no suco de 102 UFC/mL. O desenvolvimento do microrganismo foi acompanhado através de plaqueamentos em meio de cultura próprio, por até 30 dias, ou até que atingissem uma contagem constante. Ao final, foram avaliados o desenvolvimento máximo da bactéria, em cada suco, na respectiva condição de cultivo, e o período que levou para atingir estes determinados valores. Apenas no suco de caju o pool de bactérias não apresentou desenvolvimento, no prazo máximo estipulado do ensaio (30 dias). O microrganismo apresentou desenvolvimento em todos os demais sucos, alcançando índices de produção de guaiacol e outros subprodutos / Abstract: Alicyclobacillus acidoterrestris is an acidophilic bacterium, nonpathogenic, of environmental origin. This organism is a potential spoiler acid frutits, pulps and juices. Due to the presence of endospores of high heat resistance, it becomes difficult to inactivate it by pasteurization conventional thermal treatment. This research aimed to extract and process thermally the juice of six kinds of tropical fruit, acid character (pineapple, acerola, cashew, guava, mango and passion fruit) and to evaluate if these have conditions for Alicyclobacillus acidoterrestris to develop in optimum temperature (45°C) and also in processing abuse temperature (35°C). Besides, it was analyzed the minimum initial concentration of the microorganism under study, which is able to survive and develop in the mango juice, which presented better conditions of development, according to the first phase of the study. A suspension was prepared with the spores mixture of four strains of A. acidoterrestris in the same concentration, because in preliminary studies it was possible to verify that some strains did not develop in all types of juice. The suspension was standardized and inoculated into the previously extracted and heart treated juices, showing initial concentration of 102 CFU/mL in the juice. The development of the microorganism was accompanied through plating in specific culture medium, for around 30 days or until it reached a constant counting. In the end, bacterium maximum growth was evaluated in each juice, in its respective growing condition and the time it took to achieve these values. The pool of bacteria didn't grow only in the cashew juice. All other juices showed development of the microorganism, reaching production rates of guaiacol and other byproducts, in accordaner with to previous studies / Mestrado / Ciência de Alimentos / Mestra em Ciência de Alimentos

Made available in DSpace on 2016-12-23T13:50:03Z (GMT). No. of bitstreams: 1 Glaucia Aparecida Mataveli Ferreira.pdf: 3542817 bytes, checksum: 07aaaa82697b511556deff5cd7ccaf87 (MD5) Previous issue date: 2013-08-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Byssochlamys nívea e Alicyclobacillus acidoterrestris são micro-organismos deterioradores de grande importância para a indústria de suco de frutas, por serem capazes de produzir esporos que por sua vez suportam os tratamentos térmicos, normalmente utilizados na pasteurização de sucos. Esses esporos podem ser ativados pelo calor antecipando a deterioração desses alimentos e causando grandes prejuízos à indústria de suco de frutas. Assim, a busca por novas estratégias para minimizar esses problemas tem sido objeto de pesquisa. O estudo da associação entre tecnologias térmicas e não térmicas vem se destacando, na tentativa de controlar ou inibir a presença desses micro-organismos. Uma das estratégias é a inativação dos micro-organismos em duas etapas. Na primeira, um pré-tratamento visa sensibilizar os microorganismos por meio de ativação da germinação. Na segunda etapa, o tratamento teria como objetivo inativar os micro-organismos sensibilizados. Neste estudo, utilizou-se a combinação de temperatura seguido de alta pressão, a fim de testar o comportamento de esporos de B. nivea e A. acidoterrestris. As temperaturas variaram de 46,6-63,4 °C durante vinte minutos, seguidos de alta pressão que variaram entre 300 e 500 MPa, por tempos de 5 a 15 minutos, constituindo o delineamento composto central rotacional (DCCR). Para a germinação de B. nivea, o fator mais significativo (p<0,05) foi a alta pressão. Quanto mais alta a pressão, maior a germinação, atingindo um nível máximo a 500 MPa. Para a germinação de esporos de A. acidoterrestris, a temperatura foi o fator mais significativo (p<0,05), seguindo a tendência de quanto mais elevada a temperatura, maior a germinação dos esporos / Byssochlamys nivea and Alicyclobacillus acidoterrestris are spoilage microorganisms of great importance to the juice industry, they are able to produce spores. In addition to supporting the heat treatment commonly used in pasteurization of juices, these spores can also be activated by heat anticipating the deterioration of these foods and causing major damage to the fruit juice industry. New technologies have emerged for food processing. One is the use of high pressure. However, even with this technology, these microorganisms are difficult to control. Therefore, new strategies have been search to minimize these problems. One of such strategy is the inactivation of these microorganismos in two steps. First, a pre-treatment aims to sensitize the microorganisms through activation of germination. . In the second stage, the treatment would aim to inactivate microorganisms sensitized. However, data are needed to describe the levels of temperature and pressure necessary to sensitize these microorganisms. In this study, we used a combination of temperature followed by high pressure in order to test the behavior of B. nivea and A.acidoterrestris spores. Temperatures ranging from 46.6 to 63.4 °C for twenty minutes, followed by high-pressure ranging between 300 and 500 MPa for 5 to 15 minutes constituted the central composite rotational design (CCRD). For germination of B. nivea, the most significant factor (p<0.05) was high pressure, higher the pressure, greater the germination, reaching a maximum at 500 MPa. For germination of Alicyclobacillus acidoterrestris spores, temperature was the most significant factor (p<0.05), following the trend of higher the temperature, higher the germination of spores

Made available in DSpace on 2015-03-26T13:51:43Z (GMT). No. of bitstreams: 1 01 - capa_abstract.pdf: 73887 bytes, checksum: 3f348de8ab7b9bec00479392ed9cc128 (MD5) Previous issue date: 2008-06-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Aguardando liberação do SIGILO / Aguardando liberação do SIGILO

Thesis (MSc Food Sc (Food Science))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: Alicyclobacillus species are thermo-acidophilic bacteria that produce highly resistant endospores able to survive the processing temperatures of fruit concentrate manufacturing, including evaporation and conventional pasteurisation (86 ° - 96 °C for ± 2 min). Alicyclobacillus endospores retain their viability in juice concentrates which, under favourable conditions, could germinate and multiply to numbers high enough to cause spoilage and product deterioration through the production of chemical taint compounds. This thesis reports on the distribution of Alicyclobacillus in the fruit concentrate processing environment and the effect of current manufacturing practices on the accumulation of Alicyclobacillus in fruit concentrates. These practices include the recirculation (recycling) of flume water as a means of water conservation, as well as continuous process running times when facilities operate at full capacity. This thesis also reports on the effect of fruit variety and skin type on the occurrence of Alicyclobacillus in fruit concentrates. Alicyclobacillus was monitored at nine processing stages of fruit concentrate manufacturing during the functioning of either a recirculating or a one-pass (not recirculated) flume water system. Significantly higher Alicyclobacillus levels were recovered in fruit mash, single strength juice, concentrate and the final pasteurised product (± 30 °Brix) during the functioning of a re circulating flume system, compared to when a one-pass flume system was functional (P < 0.05). Irrespective of the flume system, high Alicyclobacillus levels were recovered from the concentrate and condensate water (a by-product of juice concentration) from the evaporator, which makes this a point of concern during concentrate manufacturing. Manufacturing practices such as the recirculation of flume water and the recovery of condensate water for fruit washing purposes pose a potential risk of Alicyclobacillus contamination and accumulation in fruit concentrates and the processing environment. The effect of continuous process running times on Alicyclobacillus was monitored in a facility that was operating at full capacity. Sampling occurred every 12 h at four processing stages, during a processing tempo of 1.8 - 2.0 t h-1 for 108 h. Vegetative cells increased significantly (P < 0.05) in single strength juice and condensate water after 84 h of processing, with 3.15 and 3.85 log10 cfu mL-1 recovered, respectively. Similar accumulation patterns of vegetative cells were observed in concentrate and the final pasteurised product. Endospores in single strength juice, concentrate and the final product were also the highest after 84 h of processing with 1.32, 1.59 and 1.64 log10 cfu mL-1, respectively. When fruit concentrate manufacturing facilities process at full capacity, a restriction in the continuous process running time to under 84 h in between CIP procedures, along with good manufacturing practices, can minimise Alicyclobacillus accumulation in fruit concentrates. The effect of fruit skin type, specifically hairy-skinned stone fruits (peach and apricot) and smooth-skinned pome fruits (apple and pear) on the occurrence of Alicyclobacillus in concentrates were examined. Apple concentrate samples had the highest occurrence (average %) of vegetative Alicyclobacillus cells (50%), followed by apricot (40%), peach (15%) and pear (10%) concentrates. The occurrence of Alicyclobacillus endospores in fruit concentrate samples were also the highest in apple (50%), followed by pear (25%), apricot (20%), and peach (10%) concentrates. The occurrence of Alicyclobacillus vegetative cells and endospores did not differ significantly (P > 0.05) between concentrates from hairy-skin and smooth-skin fruit varieties. Thus it was concluded that fruit washing steps prior to processing was more critical for the control of Alicyclobacillus than the type of fruit skin being processed. / AFRIKAANSE OPSOMMING: Alicyclobacillus spesies is termo-asidofiliese bakterieë wat hoogs bestande endospore produseer met die vermoë om prosesseringstemperature, insluitend verdamping en konvensionele pasteurisasie temperature (86 ° - 96 °C vir ± 2 min), tydens die vervaardiging van vrugtekonsentraat te oorleef. Alicyclobacillus endospore behou hul lewensvatbaarheid in vrugtekonsentrate en kan in gunstige toestande ontkiem en vermeerder tot getalle wat wansmake in produkte kan veroorsaak weens die produksie van chemiese verbindings. Hierdie tesis doen verslag oor die verspreiding van Alicyclobacillus in die vrugtekosentraat prosesseringsomgewing en oor die effek van huidige produksie praktyke op die akkumulasie van Alicyclobacillus in vrugtekonsentrate. Die praktyke sluit in, die hersirkulering van leigeut (transport) water as ‘n wyse van waterbesparing, asook aaneenlopende prosesseringstye wanneer vrugtekosentraat fabrieke teen ‘n volle kapasiteit prosesseer. Daar word ook verslag gedoen oor die effek van verskillende vrug variëteite en vel tipes op die voorkoms van Alicyclobacillus in vrugtekonsentrate. Alicyclobacillus was gemonitor by nege verskillende stadiums van ‘n vrugtekosentraat prosesseringsfabriek tydens die funksionering van óf 'n hersirkulerende óf ‘n deurlopende (nie-hersirkulerende) leigeut waterstelsel. Alicyclobacillus vlakke was beduidend hoër in gemaalde vrugte, enkelsterkte sap, konsentraat en die finale gepasteuriseerde produk (± 30 °Brix), gedurende die funksionering van 'n hersirkulerende leigeutstelsel, in vergelyking met die funksionering van ‘n deurlopende leigeutstelsel (P < 0.05 ). Ongeag van die leigeutstelsel, is hoë vlakke Alicyclobacillus gevind in konsentraat en kondensaat water ('n by-produk van die sap konsentrasie porses) vanuit die verdamper, en maak dit dus ‘n punt van belang tydens die vervaardiging van vrugtekonsentraat. Daar is gevind dat vervaardigingspraktyke soos die hersirkulasie van leigeut water en die herwinnig van kondensaat water moontlike risiko’s inhou vir die besoedeling en akkumulasie van Alicyclobacillus in vrugtekosentrate en die prosesseringsomgewing. Die effek van aaneenlopende prosesseringstye op Alicyclobacillus was gemonitor in 'n vrugtekosentraat prosesseringsfabriek wat teen volle kapasiteit prosesseer. Steekproefneming het elke 12 h by vier prosesseringsstadiums geskied, tydens 'n prosesseringstempo van 1.8 - 2.0 t h-1 vir 108 h. Vegetatiewe selle het beduidend toegeneem (P < 0.05) in die enkelsterkte sap en kondensaat water na 84 uur van prosessering, met 3.15 en 3.85 log10 kve mL-1, onderskeidelik verhaal. Soortgelyke akkumulasiepatrone vir vegetatiewe selle was waargeneem in konsentraat en die finale gepasteuriseerde produk. Endospore in enkelsterkte sap, konsentaat en die finale produk was ook die hoogste na 84 uur van prosessering, met 1.32, 1.59 en 1.64 log10 kve mL-1, onderskeidelik. Wanneer vrugtekonsentraat fabrieke teen volle kapasiteit prosesseseer, kan 'n beperking in aaneenlopende prosesseringstye tot onder 84 h tussen CIP prosedures, gepaard met goeie vervaardigingspraktyke, die akkumulasie van Alicyclobacillus in vrugte konsentate verminder. Die effek van verskillende vrug variëteite se vel tipes, spesifiek harige-vel steenvrugte (perske en appelkoos) en gladde-vel kernvrugte (appel en peer) op die voorkoms van Alicyclobacillus in vrugtekonsentrate was ondersoek. Appel konsentaat monsters het die hoogste voorkoms van vegetatiewe Alicyclobacillus selle gehad (gemiddelde %), met (50%), gevolg deur appelkoos (40%), perske (15%) en peer (10%) konsentraat. Die voorkoms van Alicyclobacillus endospore in vrugte konsentraat monsters was weer die hoogste in appel (50%), gevolg deur peer (25%), appelkoos (20%), en perske (10%) konsentraat. Die voorkoms van Alicyclobacillus vegetatiewe selle en endospore het nie betekenisvol tussen konsentrate van harige-vel en gladdevel vrug variëteite verskil nie (P > 0.05). Die gevolgtrekking was dat vrugte wasstappe, voor die prosessering van vrugtekonsentraat, van meer belang is vir die beheer van Alicyclobacillus as die vel tipe van die vrug variëteit wat geprosesseer word.

Orientador: Pilar Rodriguez de Massaguer / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-06T19:18:35Z (GMT). No. of bitstreams: 1 Spinelli_AnaClaudiaNevesFranco_M.pdf: 3963667 bytes, checksum: 1c82e340e7fafa80d72db43568baa2bb (MD5) Previous issue date: 2006 / Resumo: Os tratamentos térmicos de enchimento a quente não são suficientes para eliminar Alicyclobacillus acidoterrestris no suco de laranja, devido a sua alta resistência térmica neste produto. O controle deste microrganismo apenas pode acontecer sob adequada estocagem do produto. Com o intuito de analisar as condições térmicas de estocagem na germinação do Alicyclobacillus acidoterrestris em suco de laranja enchido a quente foi desenvolvida uma metodologia abrangendo desde o processamento em unidade Microthermics UHT (Ultra High Temperature), passando pelo monitoramento de vida de prateleira, até o alcance da fase estacionária do microrganismo, através de contagem das formas vegetativas utilizando o meio YSG (Yeast Extract Soluble Starch Glucose). A vazão aplicada foi de 1,7 L/min., ao passo que diversos termosensores foram estrategicamente posicionados para acompanhamento termométrico de todo o procedimento. Os parâmetros de crescimento foram analisados por meio da utilização do software DMFit, que ajusta os modelos primários de Baranyi & Roberts e Gompertz modificado. O objetivo central desta pesquisa foi observar possíveis inibições do crescimento em função das condições impostas, permitindo esclarecer qual é a melhor condição de estocagem do suco de laranja enchido a quente para evitar a germinação, crescimento e produção de guaiacol. Suco de laranja (11º Brix, pH 3,5) foi intencionalmente inoculado com esporos de Alicyclobacillus acidoterrestris CRA 7152 reconhecido como produtor de guaiacol (responsável pela deterioração). O suco envasado em garrafa PET (500 mL) foi mantido a 85ºC por 150 segundos em banho termostático. Para verificar o número de reduções decimais foram realizados seis experimentos, simulando a condição da indústria (Hot Fill), ou seja, processamento a 92ºC por 10 segundos, seguido de envase a 85ºC, com manutenção a esta temperatura por 150 segundos e resfriamento por aspersão até 35ºC em 30 minutos. Para avaliação do número de pontos significativos na descrição da curva microbiana de crescimento de Alicyclobacillus acidoterrestris foram realizados dois experimentos de enchimento a quente com resfriamento a 25ºC por 48 horas com inóculos (102 e 103 esporos/mL de suco). A evolução da população de Alicyclobacillus acidoterrestris em suco de laranja foi monitorada sob cinco diferentes temperaturas de abuso de resfriamento, após pasteurização (92ºC/10s), manutenção a 85ºC por 150 segundos e resfriamento por aspersão de água (85ºC a 35ºC em 30 minutos). Os tratamentos, com três níveis diferentes de inóculo cada (100, 101 esporos/mL de suco e sem inóculo), foram: i. 30ºC no ponto frio da garrafa, ii. 30ºC por 48 horas, iii. 25ºC no ponto frio da garrafa, iv. 25ºC por 48 horas, todos seguidos de incubação a 35ºC, e v. manutenção constante a 20ºC (controle). Foi mostrado que, independente do nível de inóculo, o processo de pasteurização não inativa os esporos de Alicyclobacillus acidoterrestris, causando redução decimal (g) inexpressiva, inferiores a 0,5 reduções logarítmicas. Cerca de quinze pontos de contagem foram estabelecidos para descrever com precisão cada curva de crescimento, as quais foram acompanhadas por cerca de 260 horas. Em relação aos tratamentos de resfriamento, a condição v (manutenção a 20ºC) foi a mais eficiente, pois inibe completamente a germinação do microrganismo. Enquanto o tratamento iv (25ºC por 48 horas) para inóculo 100 esporos/mL, mostrou maior tempo de adaptação (100,4 horas), e conseqüente maior tempo para atingir 104UFC/mL (132 horas), condição crítica para iniciar a produção de guaiacol, o tratamento iii (25º no ponto frio) para inóculo 101 esporos/mL, resultou em menor população máxima (logN/N0 = 2,69). Nesse último caso, as garrafas resfriadas até 25ºC apresentaram uma menor população máxima em relação àquelas resfriadas até 30ºC com o mesmo inóculo (logN/N0 = 3,26). O tempo para início da produção detectável de guaiacol foi determinado por meio da utilização do kit Kirin, que é baseado em julgamento visual. Enquanto as predições feitas a partir da curvas de crescimento propiciaram para o tratamento ii a estimativa do defeito entre 100-108 horas, o primeiro resultado positivo obtido através do kit foi às 144 horas, o que pode ser explicado pela baixa produção inicial de guaiacol que é inferior ao tempo de aparecimento de defeito do kit a partir de 25ppm / Abstract: Hot-Fill thermal treatments of orange juice are not enough to eliminate Alicyclobacillus acidoterrestris due to its high thermal resistance. Therefore the microbial control of this microorganism can only occur by adequate product storage conditions. In order to evaluate the effects of the thermal conditions of storage on germination of Alicyclobacillus acidoterrestris in hot-filled orange juice, experiments were carried out involving processing in a Microthermics UHT (Ultra High Temperature) unit and shelf life monitoring until the microorganism reached the stationary phase, by counting in YSG medium (Yeast extract, Soluble Starch, Glucose). The UHT unit was supplied with thermosensors and data loggers for thermal data acquisition of the whole processing. The flow rate applied was 1.7 L/min. Growth data were analyzed by the DMFit program, which fits both the Baranyi & Roberts and the modified Gompertz primary models. The main purpose of this research was to observe possible growth inhibitions as a function of the conditions imposed, permitting to clarify which is the best condition of storage for hot-filled orange juice amongst those tested. Such a condition should avoid or minimize germination, growth and guaiacol production. Orange juice (11º Brix, pH 3.5) was intentionally inoculated with Alicyclobacillus acidoterrestris CRA 7152 spores, recognized as guaiacol producers. In order to simulate surge tank maintenance before filling, juice (filled in 500 mL PET bottles) was kept at 85°C for 150s. To verify the number of decimal reductions (g), six experiments were carried out. Industrial conditions were simulated by processing at 92°C for 10 seconds, followed by filling at 85°C, with maintenance at 85°C for 150 seconds and cooling to 35°C in about 30 minutes by spraying with water. Two Hot-Filling experiments with cooling maintained at 25°C for 48 hours were performed, to determine the number of significant points needed to describe the growth curve behavior of Alicyclobacillus acidoterrestris. The inoculum levels were 102 and 103 spores/mL. The evolution of the Alicyclobacillus acidoterrestris population was also monitored under 5 different cooling abuse conditions, after pasteurization (92ºC/10s), maintenance at 85°C for 150 seconds, and cooling with water spray to 35°C in about 30 minutes. The treatments were: i. 30°C for the bottle cold point and storage at 35°C; ii. 30°C for 48h and storage at 35°C; iii. 25°C for the bottle cold point and storage at 35°C; iv. 25°C for 48h and storage at 35°C; v. storage at 20°C (control). Three different levels of inoculum were applied: 100, 10¹ spores/mL of orange juice and a total absence of inoculum. It was shown that, no matter what the inoculum level, the process did not inactivate the spores of Alicyclobacillus acidoterrestris and caused no expressive reduction in the microorganism population (g < 0.5). About fifteen points were established for every condition studied to accurately describe the growth curves. Each curve was monitored for about 260 hours. Concerning the cooling treatments, it was concluded that treatment v (storage at 20°C) was more efficient than any of the others, since in this case the population remained inhibited. Whilst treatment iv (25°C for 48 hours) with 100 spores/mL, showed a longer lag time (100.4 hours), and consequently longer time to reach 104 CFU/mL, the critical count for guaiacol production (132 hours), treatment iii (25°C for the bottle cold point) for 101 spores/mL, resulted in a lower maximum population ratio (logN/N0 = 2.69). In this latter case, the bottles that were cooled to 25ºC showed a lower maximum population ratio than those cooled to 30ºC, as can be seen in treatment i with the same inoculum 101spores/mL, where the maximum population ratio was 3.26. In addition, the time taken to initiate the detection of guaiacol was determined using the Kirin kit, which is based on a visual examination. Although the estimate for guaiacol production obtained from the growth curves for treatment ii. (30ºC for 48 hours) was between 100-108 hours, the first positive result using the kit was 144 hours, explained by the fact that a visual judgment is only possible at over 25 ppm of guaiacol / Mestrado / Mestre em Ciência de Alimentos

Thesis (Msc Food Sc (Food Science))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Bacteria belonging to the genus Alicyclobacillus are thermo-acidophilic spore-formers that are able to spoil acidic food and beverage products through the production of guaiacol and other taint compounds, which causes a medicinal off-flavour and/or odour in the products. This thesis reports on the comparison of methods used for the isolation of species of Alicyclobacillus, as well as the growth behaviour and guaiacol production of different strains isolated from the South African fruit processing environment. Two methods for guaiacol detection were also evaluated and compared. Three isolation methods frequently used by South African fruit processors were compared with regards to their ability to isolate a strain of A. acidoterrestris from diluted peach juice concentrate. Method 1, the International Federation of Fruit Juice Producers (IFU) Method No. 12, makes use of spread plating onto Bacillus acidoterrestris (BAT) agar plates; Method 2 involves pour plating using acidified potato dextrose agar (PDA); and Method 3 makes use of membrane filtration and incubation of the membrane on K agar. The IFU Method No. 12 was the most effective method for the isolation of A. acidoterrestris, with a recovery of 75.97%. These results support the use of the IFU Method No. 12 as a standard international method for the isolation and detection of species of Alicyclobacillus. Seven strains of Alicyclobacillus, including the type strains A. acidoterrestris DSM 3922T and A. acidocaldarius DSM 446T and five strains isolated from a South African fruit processing plant, A. acidoterrestris FB2, FB14, FB32, FB38 and A. acidocaldarius FB19, were analysed based on their growth characteristics and guaiacol production under optimum conditions. Strains were inoculated into BAT medium at pH 4.00, supplemented with 100 mg.L-1 vanillin, and incubated at 45°C for 7 d. All the strains had similar growth patterns, with cell concentrations increasing rapidly from 0-24 h, followed by a stabilisation around maximum cell concentrations of 105-107 cfu.mL-1. Cell concentrations after heat shock, measured as an indication of spore formation, increased to maximum values of 105-107 cfu.mL-1, indicating an increase in spores as the cell density and competition for resources increased. All the strains were able to produce guaiacol in detectable concentrations [as measured by the peroxidase enzyme colourimetric assay (PECA)], and, therefore, possess the potential to cause product spoilage. Bacteria belonging to the genus Alicyclobacillus are thermo-acidophilic spore-formers that are able to spoil acidic food and beverage products through the production of guaiacol and other taint compounds, which causes a medicinal off-flavour and/or odour in the products. This thesis reports on the comparison of methods used for the isolation of species of Alicyclobacillus, as well as the growth behaviour and guaiacol production of different strains isolated from the South African fruit processing environment. Two methods for guaiacol detection were also evaluated and compared. Three isolation methods frequently used by South African fruit processors were compared with regards to their ability to isolate a strain of A. acidoterrestris from diluted peach juice concentrate. Method 1, the International Federation of Fruit Juice Producers (IFU) Method No. 12, makes use of spread plating onto Bacillus acidoterrestris (BAT) agar plates; Method 2 involves pour plating using acidified potato dextrose agar (PDA); and Method 3 makes use of membrane filtration and incubation of the membrane on K agar. The IFU Method No. 12 was the most effective method for the isolation of A. acidoterrestris, with a recovery of 75.97%. These results support the use of the IFU Method No. 12 as a standard international method for the isolation and detection of species of Alicyclobacillus. Seven strains of Alicyclobacillus, including the type strains A. acidoterrestris DSM 3922T and A. acidocaldarius DSM 446T and five strains isolated from a South African fruit processing plant, A. acidoterrestris FB2, FB14, FB32, FB38 and A. acidocaldarius FB19, were analysed based on their growth characteristics and guaiacol production under optimum conditions. Strains were inoculated into BAT medium at pH 4.00, supplemented with 100 mg.L-1 vanillin, and incubated at 45°C for 7 d. All the strains had similar growth patterns, with cell concentrations increasing rapidly from 0-24 h, followed by a stabilisation around maximum cell concentrations of 105-107 cfu.mL-1. Cell concentrations after heat shock, measured as an indication of spore formation, increased to maximum values of 105-107 cfu.mL-1, indicating an increase in spores as the cell density and competition for resources increased. All the strains were able to produce guaiacol in detectable concentrations [as measured by the peroxidase enzyme colourimetric assay (PECA)], and, therefore, possess the potential to cause product spoilage. iv The influence of temperature on the growth and guaiacol production of the Alicyclobacillus strains was also investigated and two guaiacol detection methods, the PECA and headspace gas-chromatography mass-spectrometry (HS GC-MS), were compared with regards to their ability to detect guaiacol. The strains were incubated at 25°C and 45°C for 6 d and samples analysed every 24 h. Growth of the A. acidoterrestris strains was slower at 25°C, and maximum cell concentrations were lower than at 45°C. A decrease in cell concentrations was observed in the A. acidocaldarius strains at 25°C, as this temperature is below their growth temperature range. All the strains were able to produce guaiacol at 45°C, with guaiacol only being detected once a cell concentration of 104-105 cfu.mL-1 had been reached. The maximum guaiacol concentrations detected at 45°C in the samples containing A. acidoterrestris were significantly higher than those detected in the A. acidocaldarius samples. At 25°C there was a longer lag phase before guaiacol was detected in the A. acidoterrestris samples, while no guaiacol was detected in the samples containing A. acidocaldarius. Because guaiacol is produced at ambient temperatures, cooling of products is recommended to control spoilage by A. acidoterrestris. The sensitivity of the two guaiacol detection methods also differed significantly and, therefore, the PECA is recommended for presence/absence detection of guaiacol, while HS GCMS is recommended where accurate quantification of guaiacol is required. Alicyclobacillus acidoterrestris FB2 was investigated for its ability to grow and produce guaiacol in white grape juice supplemented with vanillin at different concentrations. Alicyclobacillus acidoterrestris FB2 was inoculated into white grape juice concentrate diluted 1:10 with distilled water containing 0-500 mg.L-1 vanillin and incubated at 45°C for 6 d. Similar growth patterns were observed in all the samples, except in the sample containing 500 mg.L-1 vanillin, which had a longer lag phase of growth. Guaiacol concentrations, detected using the PECA, increased as the vanillin concentration increased, with the exception of the sample containing 500 mg.L-1 vanillin, where less guaiacol was detected than in the sample containing 250 mg.L-1 vanillin, due to growth inhibition caused by the higher vanillin concentration. A number of conditions need to be favourable for detectable guaiacol production to occur and it could, therefore, be possible to minimise or prevent guaiacol production by controlling or eliminating some of these factors. Good manufacturing practices should be employed in order to minimise contamination and, therefore, spoilage, by Alicyclobacillus species. / AFRIKAANSE OPSOMMING: Bakterieë wat aan die genus Alicyclobacillus behoort, is termo-asidofiliese spoorvormers wat suur voedsel en drank produkte kan bederf deur die produksie van guaiakol en ander bederf verbindings, wat ‘n medisinale geur en/of reuk in die produkte veroorsaak. Hierdie tesis doen verslag oor die vergelyking van metodes wat vir die isolasie van spesies van Alicyclobacillus gebruik word, sowel as die groei kenmerke en guaiakol produksie van verskillende stamme wat uit die Suid- Afrikaanse vrugte prosesseringsomgewing geïsoleer is. Twee metodes vir die deteksie van guaiakol is ook geëvalueer en vergelyk. Drie isolasie metodes wat algemeen deur Suid-Afrikaanse vrugteprosesseerders gebruik word, is vergelyk ten opsigte van hul vermoë om H A. acidoterrestris stam uit verdunde perskesap konsentraat te isoleer. Metode 1, die Internasionale Federasie van Vrugtesap Produseerders (IFU) Metode No. 12, maak gebruik van spreiplating op Bacillus acidoterrestris (BAT) agar plate; Metode 2 behels gietplating met aartappel dekstrose agar (PDA) and Metode 3 maak gebruik van membraan filtrasie en inkubasie van die membraan op K agar. Die IFU Metode No. 12 was die mees effektiewe metode vir die isolasie van A. acidoterrestris, met H sel herwinning van 75.97%. Hierdie resultate ondersteun die gebruik van die IFU Metode No. 12 as H standaard internasionale metode vir die isolasie en deteksie van spesies van Alicyclobacillus. Sewe Alicyclobacillus stamme, insluitende die tipe stamme A. acidoterrestris DSM 3922T en A. acidocaldarius DSM 446T en vyf stamme geïsoleer uit ‘n Suid- Afrikaanse vrugte prosesseringsaanleg, A. acidoterrestris FB2, FB14, FB32, FB38 en A. acidocaldarius FB19, is geanaliseer met betrekking tot hul groei kenmerke en guaiakol produksie onder optimum toestande. Stamme is in BAT medium by pH 4.00, aangevul met 100 mg.L-1 vanillin, geïnokuleer en geïnkubeer teen 45°C vir 7 d. Al die stamme het soortgelyke groeipatrone getoon, met selgetalle wat vinnig toegeneem het van 0-24 h, gevolg deur ‘n stabilisering rondom maksimum selgetalle van 105-107 kve.mL-1. Selgetalle na hitte behandeling, gemeet as H aanduiding van spoorvorming, het toegeneem tot maksimum waardes van 105-107 kve.mL-1, wat aandui dat spore toegeneem het soos die seldigtheid en kompetisie vir voedingsbronne toegeneem het. Al die stamme kon guaiakol in bespeurbare konsentrasies produseer [soos gemeet deur die peroksidase ensiem kolorimetriese bepaling (PEKB)] en besit dus die potensiaal om produkte te bederf. Die invloed van temperatuur op groei en guaiakol produksie van die Alicyclobacillus stamme is ook ondersoek en twee guaiakol deteksie metodes, die PEKB en topspasie gas-kromatografie massa-spektrometrie (TS GK-MS) is vergelyk ten opsigte van hul vermoë om guaiakol op te spoor. Die stamme is geïnkubeer teen 25°C en 45°C vir 6 d en monsters is elke 24 h geanaliseer. Groei van die A. acidoterrestris stamme was stadiger by 25°C en maksimum selgetalle was laer as by 45°C. H Vermindering in selgetalle is waargeneem in die A. acidocaldarius stamme by 25°C, aangesien hierdie temperatuur buite hul groei temperatuur grense val. Al die stamme kon guaiakol produseer by 45°C, met guaiakol deteksie wat eers H aanvang geneem het nadat H sel konsentrasie van 104-105 kve.mL-1 bereik is. Die maksimum guaiakol konsentrasies wat by 45°C in die monsters met A. acidoterrestris opgespoor is, was beduidend hoër as die konsentrasies wat in die A. acidocaldarius monsters opgespoor is. By 25°C was daar H langer sloerfase voor guaiakol opgespoor is in die A. acidoterrestris monsters, terwyl geen guaiakol opgespoor is in die monsters wat A. acidocaldarius bevat het nie. Aangesien guaiakol by kamertemperatuur geproduseer word, word verkoeling van produkte aanbeveel ten einde bederf deur A. acidoterrestris te beheer. Die sensitiwiteit van die twee guaiakol deteksie metodes het ook beduidend verskil en dus word die gebruik van die PEKB aanbeveel vir teenwoordigheid/afwesigheid deteksie van guaiakol, terwyl TS GK-MS aanbeveel word waar akkurate kwantifisering van guaiakol vereis word. Ondersoek is ingestel na die vermoë van A. acidoterrestris FB2 om te groei en guaiakol te produseer in witdruiwesap aangevul met verskillende vanillin konsentrasies. Alicyclobacillus acidoterrestris FB2 is geïnokuleer in witdruiwesap konsentraat 1:10 verdun met gedistilleerde water wat 0-500 mg.L-1 vanillin bevat het en is geïnkubeer teen 45°C vir 6 d. Soortgelyke groeipatrone is waargeneem in al die monsters, behalwe die monster wat 500 mg.L-1 vanillin bevat het, wat H langer sloerfase van groei gehad het. Guaiakol konsentrasies, soos gemeet deur die PEKB, het toegeneem soos die vanillin konsentrasie toegeneem het, met die uitsondering van die monster wat 500 mg.L-1 vanillin bevat het, waar minder guaiakol opgespoor is as in die monster wat 250 mg.L-1 bevat het as gevolg van groei inhibisie veroorsaak deur die hoër vanillin konsentrasie. H Aantal toestande moet gunstig wees vir guaiakol produksie om plaas te vind en dit kan dus moontlik wees om guaiakol produksie te minimaliseer of te voorkom deur die beheer of uitskakeling van sommige van hierdie faktore. Goeie vervaardigingspraktyke moet in plek gestel word ten einde kontaminasie en bederf deur Alicyclobacillus spesies tot H minimum te beperk.

Orientador: Pilar Rodriguez de Massaguer / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-04T11:48:31Z (GMT). No. of bitstreams: 1 LueraPena_WilmerEdgard_D.pdf: 1783533 bytes, checksum: 59fcd5288937a1e607c7124e74e4aef5 (MD5) Previous issue date: 2005 / Doutorado / Ciência de Alimentos / Doutor em Ciência de Alimentos

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