This work aimed to produce and to apply in dry fermented sausage a lactic acid starter culture using a pork plasma culture medium and to evaluate the effect of two levels (0.5% and 1%) of hydroalcoholic extract of marcela (Achyrocline satureioides) in the security and quality of dry fermented sausages. In the first experiment, the microorganism Lactobacillus plantarum was fermented with control of pH, during 36 hours, under continuous agitation of 100 rpm, at a temperature of 37 (± 0.1ºC). After entering in the stationary phase the culture was lyophilizated and applied in dry fermented sausage, evaluating its influence in the microbiological, physico-chemical and sensorial characteristics. For comparative effects, were elaborated treatments without addition of starter culture and with addition of a commercial culture. In the second experiment, dry fermented sausages had been elaborated with 0.5% and 1% of hydroalcoholic extract of marcela and without extract, considered as control. The parameters of pH, water activity, colour and weight loss were evaluated during the manufacture of the dry fermented sausages. During the storage the TBARS values were determined and verified the sensorial acceptance. The microorganism Lactobacillus plantarum had a maximum growth of 9.82 Log UFC.mL-1, after 30 hours of fermentation and it presented 90.05% of survival to the lyophilization. The dry fermented sausages elaborated with the starter culture produced had presented a decrease on pH significantly faster and a lower water activity than the others treatments. Moreover, the microorganism Lactobacillus plantarum improved significantly the flavor of the dry fermented sausages. The marcela extract addition decreased (p<0.05) the TBARS values during the storage of the dry fermented sausages, compared with the control. The treatment with 1% of marcela extract showed a bigger lipid stability than that with 0.5%, however it presented a reduction (p<0.05) in the sensorial acceptance. The two marcela extract levels had not influenced the parameters of pH, water activity, colour and weight loss. Therefore, this study indicated that the pork plasma culture medium is an alternative in the commercial production of lactic acid bacteria and that the starter culture produced provided a dry fermented sausage with greater microbiological security and better sensorial acceptance. The hydroalcoholic extract of marcela decreased the lipid oxidation and the concentration of 0.5% did not modify the sensorial characteristics, being able, therefore, to be used in the elaboration of safer dry fermented sausages to the consumers. / Este trabalho teve por objetivo produzir e aplicar em salame uma cultura starter ácido láctica utilizando um meio de cultura de plasma suíno e avaliar o efeito de dois níveis (0,5% e 1%) de extrato hidro-alcoólico de marcela (Achyrocline satureioides) na segurança e qualidade de salames. No primeiro experimento, o microrganismo Lactobacillus plantarum foi fermentado em um meio de cultura de plasma suíno, com controle de pH, durante 36 horas, sob agitação contínua a 100 rpm e temperatura de 37 (± 0,1ºC). Após entrar na fase estacionária a cultura foi liofilizada e aplicada em salame, avaliando sua influência nas características microbiológicas, físico-químicas e sensoriais. Para efeitos comparativos, foram elaborados tratamentos sem adição de cultura starter e com adição de uma cultura comercial. No segundo experimento, salames foram elaborados com 0,5% e 1% de extrato hidro-alcoólico de marcela e sem extrato, considerados como controle. Os parâmetros de pH, atividade de água, cor e perda de peso foram avaliados durante a fabricação dos salames. Durante o armazenamento os valores de TBARS foram determinados e foi verificada a aceitação sensorial. O microrganismo Lactobacillus plantarum teve um crescimento máximo de 9,82 Log UFC.mL-1, após 30 horas de fermentação e apresentou 90,05% de sobrevivência a liofilização. Os salames elaborados com a cultura starter produzida apresentaram uma queda de pH significativamente mais rápida e uma menor atividade de água que os demais tratamentos. Além disso, o microrganismo Lactobacillus plantarum significativamente melhorou o sabor dos salames. A adição de extrato de marcela diminuiu (p<0,05) os valores de TBARS durante o armazenamento dos salames, comparado ao controle. O tratamento com 1% de extrato de marcela mostrou uma maior estabilidade lipídica do que aquele com 0,5%, porém apresentou uma diminuição (p<0,05) na aceitação sensorial. Os dois níveis de extrato de marcela não influenciaram significativamente os parâmetros de pH, atividade de água, cor e perda de peso. Portanto, este estudo indicou que o meio de cultura de plasma suíno é uma alternativa na produção comercial de bactérias ácido lácticas e que a cultura starter produzida proporcionou salames de maior segurança microbiológica e melhor aceitação sensorial. O extrato hidro-alcoólico de marcela diminuiu a oxidação lipídica e a concentração de 0,5% não alterou as características sensoriais, podendo, portanto, ser utilizada na elaboração de salames mais seguros aos consumidores.

Bakgrund: Med industrialiseringen har de traditionella yoghurtkulturerna med en mångfald av samverkande mjölksyrabakterier fått ge plats åt de mer standardiserade. En gårdskultur framställs genom att obehandlad mjölk spontanfermenteras av gårdens egen bakterieflora och får därigenom en unik karaktär. Bakteriekulturen kan sedan ympas vidare för tillverkning av yoghurt.Syfte: Syftet med studien är framställning och vidareympning samt sensorisk och mikrobiologisk karaktärisering av termofil gårdskultur från obehandlad mjölk. Vidareympningen avser framställning av yoghurt fermenterad av gårdens egen bakterieflora.Metod: Gårdskulturen framställdes och ympades till yoghurt. Yoghurten undersöktes genom mikrobiologisk karaktärisering, antibiotikaresistens och sensorisk profilering samt jämfördes med industriell kultur och en traditionell heirloomkultur.Resultat: Resultatet visade att gårdskulturen skiljde sig både mikrobiologisk och sensoriskt. Gårdskulturen innehöll stammar av enterokocker vilka inte visade på resistens mot analyserade antibiotika.Slutsats: Det är möjligt att framställa en gårdskultur av godtagbar mikrobiologisk och sensorisk kvalitet. Gårdskulturen ger en differentierad mikrobiologisk och sensorisk karaktär i jämförelse med en industriell kultur och en traditionell heirloomkultur Metoden kan vara riskfylld och kulturen bör analyseras med avseende på patogen tillväxt. En unik gårdsyoghurt kan vara en metod för gårdsmejerister att i sitt varumärke bygga på terroir och platsens unicitet. / Background: With industrialization, the traditional yoghurt cultures with a multitude of lactic acid bacteria had to make way for the more standardized. An artisanal farm culture is produced by raw milk spontaneously fermented by the farm's own bacterial flora and thus develops a unique character. The bacterial culture can then be inoculated for the production of yoghurt.Purpose: The pupose of the study is to produce and inoculate as well as sensory and microbiological characterization of a thermophilic artisanal farm culture from raw milk. The inoculation relates to the production of yoghurt fermented by the farm's own bacterial flora.Method: The artisanal farm culture was produced and inoculated into yoghurt which was assessed by microbiological characterization, antibiotic resistance, sensory profiling and then compared with industrial culture and a traditional heirloom culture.Result: The result showed that the artisanal farm culture differed both microbiologically and with regard to sensory paramters. The farm culture contained strains of enterococci which did not show resistance to analyzed antibiotics.Conclusion: It is possible to produce an artisanal farm culture of good microbiological and sensory quality. The artisanal farm culture provides a differentiated microbiological and sensory character in comparison to an industrial culture and a traditional heirloom culture The method may be risky and the culture should be analyzed for pathogens. A unique farm yoghurt can be a method for artisan farm dairies to build their brand based on terroir.

Proposed dissertation thesis deals with wine and artisanal cheeses microbiology. The first part is focused on identification of yeasts isolated from grapes and musts during production of white and red wines. The grape varieties were grown under the integrated and organic farming on Moravian vineyard. Yeasts were identified by ITS-PCR-RFLP method (amplifying internal transcribed spacer ITS: ITS1, ITS2 and 5.8S rDNA) and unknown species were subjected to partial sequencing of ITS rDNA region. In total, 524 isolates were divided into 14 different species belonging to six genus were identified from. The first stages of fermentation process were characterised by predominance of non-saccharomyces species especially H. uvarum. Due to increased ethanol concentration strains of S. cerevisiae prevailed in the later phases of the process. Further, partial aim of this study was to isolate and to apply selected autochthonous S. cerevisiae strains as starter culture during controlled industrial wine fermentation process. Genus Saccharomyces was distinguished from other non-saccharomyces species by ITS-PCR-RFLP. Further, in order to distinguish Saccharomyces genus at the species and the strain level, several molecular methods were applied including PCR-fingerprinting (rep- and RAPD-PCR), species-specific primers (multiplex and touchdown PCR), LSU-DGGE and interdelta PCR. Species-specific primers enabled us to distinguish some species of the Saccharomyces sensu stricto complex. Furthermore, interdelta PCR seems to be useful tool for S. cerevisiae strains identification. Among 120 isolated autochthonous strains belonging to Saccharomyces genus, 45 different strains were identified. Based on its sufficient technological properties (osmo- and ethanol tolerance, low H2S production etc.), S. cerevisiae 1-09 strain isolated from grape berries coming from moravian vineyard was chosen. Strain S. cerevisiae 1-09 was tested in small amount of must and after that also during industrial fermentation of red and white wine production. Based on the results of chemical and sensorial analysis, the strain seems to be suitable for application as the starter culture for winemaking process. The final part of this thesis is focused on quantification and identification of the yeasts isolated from artisanal cheeses and their by-products coming from Western Balkan Countries. Isolated species were identified by ITS-PCR-RFLP, partial sequencing and by physiological tests. Among the 20 yeast species found, D. hansenii, C. zeylanoides and Y. lipolytica were found to be predominant. Moreover, we developed culture-independent, semi-quantitative technique based on construction of ITS-clone library from metagenomic DNA to investigate complex fungal communities associated with artisanal cheeses and their by-products. Novel technique is based on direct extraction of total DNA from the sample. This was compared with culture-dependent ITS-PCR-RFLP and culture-independent LSU-DGGE methods. The results highlighted the discrepancies among these methods. Finally, the divergences among applied methods were confirmed by correlation analysis and by indices of general biodiversity and dominance of species. ITS-clone library approach combines the advantages of cultivation-based analysis and LSU-DGGE with semi-quantification of fungal species without the requirement of their cultivation. This study might open new perspectives in direct and complex analysis of yeasts and moulds in food matrices.

In my thesis, I focused on monitoring of microorganisms in the sample of red grape juice and on the interactions between yeasts, bacteria and filamentous fungi. Three different media were applied for the cultivation of microorganisms; firstly for monitoring of total volume of microorganisms, secondly for yeasts and third time for lactic acid bacteria. The indirect method was used for the determination of the amount of viable cells. This method consists in enumerating of visible macroscopic colonies grown up on agar plates. When the cells grew up, the forms of colonies were analyzed visually and the morphology of microorganisms was detected microscopically. The operating time of enzymes in grape juice in the production of red wine was monitored after application of commercial enzymatic preparation. The enzym action in grape juice was observed on the basis of the process of degradation of high – molecular substrate by enzymes through the use of Ubbelohd´s viscometer. The research findings provided a lot of knowledge about the occurance of microflora in the process of production of red wine. The commercial preparations added to grape juice played a significant role.