Microbiological characterization of kombucha and biocellulose film produced with black tea and cocoa bean shell infusion.

The food industry is increasingly striving to produce probiotics-based food and beverages using sustainable processes. Therefore, the use of by-products in product development has been investigated by several authors. The aim of this work was to investigate the effects of cocoa bean shell infusion in the production of kombucha through microbiological and genetic characterization. Three beverage formulations were prepared, one based on black tea (KBT), one based on cocoa bean shell infusion (KCS) and one containing 50 % black tea and 50 % cocoa shell infusion (KBL). The infusions were prepared with water, filtered, and sucrose was added. They were then homogenized and a portion of finished kombucha and SCOBY (symbiotic culture of bacteria and yeast) were added. Fermentation took place for 13 days and aliquots were collected every three days for physicochemical and microbial count analyses. Samples from the last day of fermentation were sent for DNA sequencing, extraction and quantification. The results were subjected to analysis of variance and compared by using Tukey's test (p < 0.05). The results show that there was a significant decrease in pH over time in all samples, while the titratable acidity increased, indicating an acidification of the beverage due to the production of organic acids. There was an increase in lactic acid bacterial colonies in all the formulations, which have a probiotic nature and are not always found in this type of beverage. Regarding the taxonomic classification of the samples, microorganisms of the kingdoms Fungi and Bacteria, of the families Saccharomycetaceae and Acetobacteraceae, were found in KBT, KCS and KBL, but with different microbiological compositions, with different amounts of yeasts and bacteria. Therefore, the use of by-products such as cocoa bean shell in the production of kombucha can contribute to the reduction of waste in the food industry and, at the same time, accelerate fermentation increasing the presence of lactic acid bacteria when compared to black tea.

Kombuchas from Green and Black Tea Modulate the Gut Microbiota and Improve the Intestinal Health of Wistar Rats Fed a High-Fat High-Fructose Diet.

The Western diet can negatively affect the gut microbiota and is associated with metabolic disorders. Kombucha, a tea fermented by a symbiotic culture of bacteria and yeast (SCOBY), is known for its bioactive properties and has become popular in the last years. In this study, we evaluated the effects of regular kombucha consumption on the gut microbiota and on outcomes related to the intestinal health of Wistar rats fed a high-fat high-fructose diet. After eight weeks receiving a standard diet (AIN-93M) (n = 10) or a high-fat and high-fructose diet (HFHF) (n = 30) to induce metabolic disorders, the animals were subdivided into four groups: AIN-93M (n = 10); HFHF (n = 10); GTK (HFHF + green tea kombucha (n = 10); and BTK (HFHF + black tea kombucha; n = 10) for 10 weeks. Although body composition did not differ among the groups, the HFHF diet was associated with metabolic alterations, and stimulated the growth of gram-negative bacteria such as Proteobacteria and Bacteroides. Kombucha ingestion could somewhat modulate the gut microbiota, attenuating the effects of a Western diet by increasing propionate production and favoring the growth of beneficial bacteria, such as Adlercreutzia in the GTK group. Our results suggest that regular kombucha consumption may be beneficial to intestinal health, which can be mostly attributed to its high content and diversity of phenolic compounds.

Enzymatic Electroanalytical Biosensor Based on Maramiellus colocasiae Fungus for Detection of Phytomarkers in Infusions and Green Tea Kombucha.

In this work, we developed an enzymatic voltammetric biosensor for the determination of catechin and gallic acid in green tea and kombucha samples. The differential pulse voltammetry (DPV) methodology was optimized regarding the amount of crude enzyme extract, incubation time in the presence of the substrates, optimal pH, reuse of the biosensor, and storage time. Samples of green tea and kombucha were purchased in local markets in the city of Goiânia-GO, Brazil. High performance liquid chromatography (HPLC) and Folin-Ciocalteu spectrophotometric techniques were performed for the comparison of the analytical methods employed. In addition, two calibration curves were made, one for catechin with a linear range from 1 to 60 µM (I = -0.152 * (catechin) - 1.846), with a detection limit of 0.12 µM and a quantification limit of 0.38 µM and one for gallic acid with a linear range from 3 to 60 µM (I = -0.0415 * (gallic acid) - 0.0572), with a detection limit of 0.14 µM and a quantification limit of 0.42 µM. The proposed biosensor was efficient in the determination of phenolic compounds in green tea.

Análises de matérias estranhas e microbiológicas de chás industrializados comercializados em Pouso Alegre MG / Analysis of extraneous materials and microbiological from industrialized teas marketed in Pouso Alegre MG

O objetivo deste estudo foi de avaliar as condições higiênico-sanitárias de quatro marcas de chás industrializados comercializados na cidade de Pouso Alegre - MG utilizando parâmetros de presença de matérias estranhas e análises microbiológicas. Foram coletadas no período de setembro a outubro de 2017, amostras de chás de camomila, erva-doce, hortelã e chá misto que contém em sua formulação essas mesmas ervas. Foram utilizados os métodos da Association of Official Analytical Chemists International (AOAC) de acordo com a RDC n°14 para capturar as sujidades leves e nas análises microbiológicas, foram utilizados métodos da International Organization for Standardization (ISO) de acordo com a RDC n° 12 para avaliar coliformes totais e termotolerantes, fungos e Salmonella spp. Nas pesquisas de matérias estranhas somente três marcas de camomila foram aprovadas, estando o restante dos chás em desacordo com a legislação vigente por estarem acima do limite de tolerância aceitável. Na avaliação de coliformes e Salmonella sp. todas as amostras foram aprovadas porém, houve um crescimento considerável de fungos filamentosos nestas amostras...(AU)

The aim of this study was to evaluate the hygienic-sanitary conditions of four brands of industrialized teas marketed in the city of Pouso Alegre - MG using parameters of presence of foreign matter and microbiological analyzes. Were collected in the period from September to October 2017, samples of chamomile teas, fennel, mint and mixed tea that contains in its formulation these same herbs. Were used the methods of the International Association of Analytical Chemists (AOAC) according to RDC n° 14 to capture light dirt and in the microbiological analyzes, were used the methods of the International Organization for Standardization (ISO) according to RDC nº 12 to evaluate total and thermotolerant coliforms, fungi and Salmonella spp. In the research of extraneous materials, only three brands of chamomile were approved, the rest of the teas being in disagreement with the current legislation because they are above the acceptable tolerance limit. In the evaluation of coliforms and Salmonella sp. all samples were approved, however, there was a considerable growth of filamentous fungi in these samples…(AU)

Análises de matérias estranhas e microbiológicas de chás industrializados comercializados em Pouso Alegre MG / Analysis of extraneous materials and microbiological from industrialized teas marketed in Pouso Alegre MG

O objetivo deste estudo foi de avaliar as condições higiênico-sanitárias de quatro marcas de chás industrializados comercializados na cidade de Pouso Alegre - MG utilizando parâmetros de presença de matérias estranhas e análises microbiológicas. Foram coletadas no período de setembro a outubro de 2017, amostras de chás de camomila, erva-doce, hortelã e chá misto que contém em sua formulação essas mesmas ervas. Foram utilizados os métodos da Association of Official Analytical Chemists International (AOAC) de acordo com a RDC n°14 para capturar as sujidades leves e nas análises microbiológicas, foram utilizados métodos da International Organization for Standardization (ISO) de acordo com a RDC n° 12 para avaliar coliformes totais e termotolerantes, fungos e Salmonella spp. Nas pesquisas de matérias estranhas somente três marcas de camomila foram aprovadas, estando o restante dos chás em desacordo com a legislação vigente por estarem acima do limite de tolerância aceitável. Na avaliação de coliformes e Salmonella sp. todas as amostras foram aprovadas porém, houve um crescimento considerável de fungos filamentosos nestas amostras...

The aim of this study was to evaluate the hygienic-sanitary conditions of four brands of industrialized teas marketed in the city of Pouso Alegre - MG using parameters of presence of foreign matter and microbiological analyzes. Were collected in the period from September to October 2017, samples of chamomile teas, fennel, mint and mixed tea that contains in its formulation these same herbs. Were used the methods of the International Association of Analytical Chemists (AOAC) according to RDC n° 14 to capture light dirt and in the microbiological analyzes, were used the methods of the International Organization for Standardization (ISO) according to RDC nº 12 to evaluate total and thermotolerant coliforms, fungi and Salmonella spp. In the research of extraneous materials, only three brands of chamomile were approved, the rest of the teas being in disagreement with the current legislation because they are above the acceptable tolerance limit. In the evaluation of coliforms and Salmonella sp. all samples were approved, however, there was a considerable growth of filamentous fungi in these samples

Candida theae sp. nov., a new anamorphic beverage-associated member of the Lodderomyces clade.

Four strains representing a novel yeast species belonging to the genus Candida were independently isolated in Taiwan and Ecuador. Two strains (G17(T) and G31) were isolated in Taiwan, by pellet precipitation from plastic-bottled tea drinks produced in Indonesia, while two additional strains (CLQCA 10-049 and CLQCA 10-062) were recovered from ancient chicha fermentation vessels found in tombs in Quito, Ecuador. These four strains were morphologically, and phylogenetically identical to each other. No sexual reproduction was observed on common sporulation media. Large-subunit (LSU) rRNA gene sequence analysis revealed the four strains to belong to the Lodderomyces clade, closely related to members of the Candida parapsilosis species complex. The four strains, which have identical LSU D1/D2 sequences, differ from their closest phylogenetic neighbors, Candida orthopsilosis and Candida parapsilosis, by 6-9 nt substitutions, respectively. Physiologically, the four strains are similar to Candida parapsilosis, although they can be distinguished from their closest relative by the assimilation of arbutin, nitrite, and creatine. The Indonesian and Ecuadorian strain sets can also be distinguished from one another based on ITS sequencing, differing by 4 substitutions in ITS1 and 1 single nucleotide indel in ITS2. Collectively, the results indicate that the four strains represent a previously unrecognized species of Candida. The name Candida theae sp. nov. is proposed to accommodate these strains, with G-17(T) (BCRC 23242(T)=CBS 12239(T)=ATCC MYA-4746(T)) designated as the type strain.

Crescimento de fungos potencialmente produtores de micotoxinas em ervas utilizadas para chá coletadas em feiras livres do Distrito Federal / Growth of fungi potentially mycotoxin-producing used in herbal tea collected from fairs of the Distrito Federal

O objetivo deste trabalho foi avaliar a presença, isolar e identificar fungos filamentosos, produtores de micotoxinas Aspergillus sp., Penicillium sp. e Fusarium sp em amostras de ervas utilizadas em chá comercializadas em feiras livres do Distrito Federal...

Crescimento de fungos potencialmente produtores de micotoxinas em ervas utilizadas para chá coletadas em feiras livres do Distrito Federal / Growth of fungi potentially mycotoxin-producing used in herbal tea collected from fairs of the Distrito Federal

O objetivo deste trabalho foi avaliar a presença, isolar e identificar fungos filamentosos, produtores demicotoxinas Aspergillus sp., Penicillium sp. e Fusarium sp em amostras de ervas utilizadas em chá comercializadas em feiras livres do Distrito Federal. Foram analisadas 20 amostras de cinco ervas utilizadas para chás, Aroeira (Schinus molle L.), Barbatimão (Stryphnodendron rotundifolium Mart), Chapéu-de-couro (Echinodorus macrophyllus), e Espinheira- santa (Maytenus ilicifolia), comercializadas em feiras livres da cidade de Taguatinga e Ceilândia- DF, no período de Abril e Maio de 2007. Foi realizada a contagem de bolores e leveduras e posterior verificação da presença de fungos filamentososnas amostras. Do total de amostras estudadas, 80% estão fora do limite permitido pela legislaçãobrasileira (5xl03 UFC/g). As maiores contagens por tipo de erva foram: Espinheira Santa (549x103 UFC/g), Barbatimão (210, 4xl03 UFC/g), Aroeira (200,6x103 UFC/g) e Chapéu de Couro (l74,4xl03 UFC/g). Em 50% das ervas estudadas foi verificada a presença de fungos filamentosos e os gêneros e espécies identificados foram: Aspergillus jlavus,Aspergillus nidulans, Aspergillus niger, Aspergillus fumigatus, Fusarium. e Penicillium. Os gêneros fúngicos encontrados são produtores de micotoxinas e indicam possível contaminação tóxica das amostras analisadas. Os resultados comprovam a má qualidade das amostras de chás naturais comercializadas em bancas de feira livre e exposição do consumidor ao risco real de utilização desses chás ao consumo, evidenciando-se a necessidade de adoção de programa de fiscalização, vigilância e controle de qualidade do material vegetal, disponibilizado para comercialização como bebida. (AU)

The purpose of this study was to assess the presence, isolate and identify filamentous fungi, produce mycotoxinsAspergillus sp., Penicillium sp. and Fusarium sp in samples of herbs traded atmarketed at fairs free of the Federal District, used in tea. The present study analyzed 20 samples of five herbs used for tea, Aroeira (Schinus molte L.), Barbatimão (Stryphnodendron rotundifolium Mart), Chapéu de couro (Echinodorus macrophyllus), and Espinheira - santa (Maytenus ilicifolia), marketed at[airsfree of city of Taguatinga and Ceilãndia- DF, in the period of April and May 2007. It held the count of molds and yeasts and subsequent verification of the presence of filamentous fungi in the samples. Of the total samples studied, 80% are outside the limit permitted by Brazilian law (5x103 CFU/g). The largest counts by type of grass were: Espinheira Santa (549x103 UFC/g), Barbatimão (210.4 xI03 UFC/g), Aroeira (200.6 x103 CFU/g) and Hat of leather (174.4 x103 CFU/g). In 50% of herbs studied verified the presence of filamentous fungi and genera and species were identified: Aspergillus flavus, Aspergillus nidulans, Aspergillus niger, Aspergillus fumigatus, Fusarium and Penicillium. Genera fungicos found are producersof mycotoxins and indicate possible toxic contamination of the samples analyzed. The results show thepoor quality of samples of natural teas sold in stalls of free and fair exposure of consumers to the real riskof the use of tea consumption, highlighting the need for adoption of the program of supervision, monitoring and quality control of the material plant, available for marketing as drink. (AU)

Determinação da qualidade microbiológica e físico-química de chás de Cymbopogon citratus (D. C) Stapf (capim-limão) / Evaluation of the microbiological and physical-chemical qualities of lemongrass tea

Em função do grande volume de produção e comercialização do capim-limão, estima-se que uma contaminação microbiológica deste produto possa representar importante risco para a saúde pública. Esta pesquisa visou avaliar marcas de chá de capim-limão, em relação às qualidades microbiológica e físico-química, tendo como base a legislação brasileira do Ministério da Saúde. Nenhuma das amostras apresentou Salmonella sp, porém em 50% delas, houve presença de coliformes a 35ºC. Em três amostras, evidenciaram-se coliformes a 45ºC, com presença de Escherichia coli em uma destas. 81,25% das amostras apresentaram bolores e leveduras, porém sem relação direta com os teores de umidade observados. Dentre os fungos presentes, identificou-se Aspergillus niger, no entanto sem produção de aflatoxinas. Todos infusos analisados negativaram a contaminação evidenciada nos testes com o produto seco. Os resultados indicaram possíveis falhas nos procedimentos pós-colheita e de comercialização. A implementação do Sistema APPCC (HACCP) poderia substancialmente minimizar esta contaminação.

Considering the great volume of lemongrass production and commercialization, it is estimated that a potential microbiological contamination of this product could represent an important hazard to public health. This study was performed to evaluate different brands of lemongrass tea, regarding their microbiological and physical-chemical quality, based on the legislation of the Brazilian Health Ministry. No traces of Salmonella sp were found, but in 50% of samples, the presence of coliforms was detected at 35ºC. Three samples presented coliforms at 45ºC, with Escherichia coli found in one of them. 81.25% of samples presented molds and yeasts. Among the fungi present, Aspergillus niger was identified, but aflatoxins were absent. None of the infusions analyzed presented the contamination evidenced in the dry product. The results obtained may be indicative of failure in procedures of postharvest and commercialization. The implementation of a hazard analysis and critical control point system (HACCP) could reduce this contamination substantially.

Manufacture of a beverage from cheese whey using a "tea fungus" fermentation.

Kombucha is a sour beverage reported to have potential health effects prepared from the fermentation of black tea and sugar with a "tea fungus", a symbiotic culture of acetic acid bacteria and yeasts. Although black tea is the preferred substrate for Kombucha fermentation, other beverages have also been tested as substrates with fair results. Cheese whey is a by-product with a good amount of fermentable lactose that has been used before in the production of beverages, so the objective of this study was to test three types of whey (fresh sweet, fresh acid and reconstituted sweet) in the elaboration of a fermented beverage using a kombucha culture as inoculum. The isolation and identification of bacteria and yeasts from the fermented tea and wheys was done along with the study of the rates of change in sugar consumption, acid production and pH decrease. Several species of acetic acid bacteria (Acetobacter aceti subsp. aceti, Gluconobacter oxydans subsp. industrius, subsp. oxydans and Gluconoacetobacter xylinus) were isolated from the different kombuchas along with the yeasts Saccharomyces cerevisiae, Kluyveromyces marxianus, and Brettanomyces bruxelensis. The main metabolic products in the fermented wheys included ethanol, lactic and acetic acids. A good growth was obtained in both sweet wheys in which a pH of 3.3 and a total acid content (mainly lactic and acetic acids) of 0.07 mol/l was reached after 96 h. The sweet whey fermented beverages contained a relatively low lactose concentration (< 12 g/l). The final ethanol content was low (5 g/l) in all the fermented wheys. The whey products were strongly sour and salty non sparkling beverages.