The purpose of this investigation was to examine the change in pulmonary ventilation (V_E), ventilatory equivalent of oxygen (VE_O_2) and lactic acid (LA) in relation to oxygen uptake (V_O_2) as predictors of the lactate threshold (LT). Eight healthy male (21.9 ± 3.0 years) subjects conducted three incremental-load tests. In each test the initial work rate consisted of 4 minutes of unloaded pedaling ("0" load) followed by incremental-load work of 360 Kgm • min^-1 at 60 rpm for trial I and trial II, while during trial III the work rate consisted of 540 Kgm • min^-1 of incremental-load work at 90 rpm. Work load was increased every third minute until the subject reached voluntary exhaustion. Blood samples from a forearm vein were collected during trial II (60 rpm) and trial III (90 rpm) and analyzed for lactic acid. In our subjects the measured (x̄ ± SD) lowest VE_O_2 for O_2 in relation to V_O_2 for trial I of 22.9 ± 1.9 occurred at a V_O_2 of 1.27 ± 0.8 L • min^-1. For trial II the VE_O_2 of 22.4 ± 1.3 occurred at a V_O_2 of 1.30 ± 0.09 L • min^-1, while for trial III a VE_O_2 of 24.4 ± 2.5 occurred at a V_O_2 of 1.84 ± 0.15 L • min^-1. The lowest VE_O_2 and onset of LA accumulation as calculated from individual exponential equations relating V_E to V_O_2 yielded V_O_2 values at 1.77 ± 0.18 L • min^-1 and 1.74 ± 0.25 L • min^-1 for trial II, and 1.83 ± 0.19 L • min^-1 and 2.02 ± 0.53 during trial III. Utilizing ln[LA]-ln V_O_2 equations, the LT occurred at a V_O_2 of 1.30 ± 0.07 L • min^-1 during trial II and 1.85 ± 0.12 L • min^-1 during trial III. It was concluded that during the 60 rpm test that the lactate threshold was best predicted by measured lowest VE_O_2 and the plot of the In[LA] to In V_O_2 relationship. The methods used in this study provide a valid and reliable estimate of the lactate threshold and support the use of measured lowest VE_O_2 , a respiratory gas exchange measure, as an indirect measure of the lactate threshold.

Outgrowth of spoilage yeasts and moulds and post-processing acidification can limit the shelf-life of some fermented dairy products including fresh lactic curd cheeses. The possibility of using high pressure processing (HPP) for controlling these problems was investigated in a commercially manufactured fresh lactic curd cheese (pH 4.3-4.4) and fermented milk models (pH 4.3-6.5). The effects of HPP at 300 and 600 MPa on inactivation of glycolytic enzymes of lactic acid bacteria were also evaluated. Fresh cheeses made from pasteurised bovine milk using a commercial Lactococcus starter preparation were treated with high pressures ranging from 200 to 600 MPa (less than or equal to 22°C, 5 min) under vacuum packaging conditions and subsequently stored at 4°C for 8 weeks. Treatment at greater than or equal to 300 MPa substantially reduced the viable count of Lactococcus and effectively prevented the outgrowth of yeasts and moulds for 6 to 8 weeks without adversely affecting the sensory and textural attributes of the product. However, it had no significant effects (p less than 0.01) on variation of titratable acidity during storage. Fermented milk models were prepared by individually growing Lactococcus lactis subsp. lactis C10, Lactococcus lactis subsp. cremoris BK5, Streptococcus thermophilus TS1, Lactobacillus acidophilus 2400 and Lactobacillus delbrueckii subsp. bulgaricus 2517 in UHT skim milk and diluting the resulting fermented milk with UHT skim milk up to pH 6.5. Pressure treatment of the milk models at pH 5.2 resulted in substantial inhibition of post-processing acidification during storage and markedly reduced the viable count of Lactococcus at both 300 and 600 MPa and other bacteria only at 600 MPa. Treatment of the milk model at 600 MPa decreased the viable counts of Candida zeylanoides and Candida lipolytica (wildtype spoilage yeasts of lactic curd cheese, added as challenge cultures) from 105 CFU mL-1 to below the detection limit (log 0 CFU mL-1) at all pH levels tested (pH 4.3-6.5) and effectively controlled their outgrowth for 8 weeks. Treatment of milk model at 300 MPa had a similar effect only on C. zeylanoides. The viable count of C. lipolytica was reduced by 2.6, 2.4 and 2.3 logs by treatment at 300 MPa at pH levels of 4.3, 5.2 and 6.5, respectively, which subsequently recovered by 2.9, 2.8 and 3.2 logs within 3 weeks. Glycolytic enzymes of various starter bacteria showed different responses to pressure treatment. The lactate dehydrogenase in L. lactis subsp. lactis and Lb. acidophilus was quite resistant to pressures up to 600 MPa, but it was almost completely inactivated in S. thermophilus at pressure levels as low as 300 MPa. The â-galactosidase in Lb. acidophilus was more pressure stable than â-galactosidase in S. thermophilus and Phospho-â-galactosidase in L. lactis subsp. lactis. The findings of this study suggests HPP at 300-600 MPa as an effective method for controlling the outgrowth of some spoilage yeasts and moulds in fresh lactic curd cheeses. The results obtained with selected lactic acid bacteria in fermented milk models can be used to assist in establishing HPP operating parameters for development of new generation cultured dairy products, of reduced acidity and extended shelf-life.

Lic.-avh. Stockholm : Karolinska institutet, 2005.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2006. / Härtill 4 uppsatser.

Introdução: A aplicação clínica de biomateriais está se expandindo para diversas especialidades médicas. Dentre os diversos tipos de biomateriais, os classificados como temporários merecem atenção especial, pois são assimilados pelo organismo após exercerem sua função, evitando, assim, procedimento cirúrgico para sua retirada. O copolímero de poli (ácido láctico-co-glicólico) (PLGA) é um tipo de biomaterial temporário, rotineiramente utilizado na medicina na forma de fios de sutura e implantes ortopédicos. A mistura do PLGA com o poli (isopreno) resulta em uma blenda (PLGA / PI), de alta resistência e tenacidade, que foi desenvolvida pelo Laboratório de Biomateriais do Instituto de Engenharia da UFRGS. Entretanto, não existem estudos “in vivo” testando a reação óssea desta blenda. Objetivo: Testar a histotoxicidade da blenda de PLGA / PI em relação ao biopolímero já consagrado PLGA. Método: Foram utilizados 46 ratos machos wistar (Rattusnorvegicus - linhagem albina), divididos em 2 grupos conforme o material implantado (PLGA ou PLGA / PI) na calota craniana, e subdivididos em tempos de morte (15, 30, 60 e 90 dias). Os procedimentos foram realizados na Unidade de Experimentação Animal (UEA) do Hospital de Clínicas de Porto Alegre (HCPA). Após a morte, a calota craniana foi retirada, submetida ao exame histopatológico e aplicado o escore de Dadas e cols (14) modificado. Resultados: A diferença da histotoxicidade dos dois materiais não foi significativa nos períodos 15, 30 e 90 dias, porém foi significativa no período 60 dias. Conclusão: A histotoxicidade do PLGA / PI, ao final do estudo (90 dias), foi semelhante ao PLGA, demonstrando equivalência em longo prazo. O período de 60 dias pós-cirúrgico (grupo da blenda) foi o único em que a histotoxicidade mostrou-se significativamente maior. Mais estudos devem ser feitos para melhorar o entendimento desta variação. / Introduction: Clinical application of biomaterials is expanding to various medical specialties. Among the different types of biomaterials, those classified as temporary deserve special attention because they are assimilated by the body after exercising their function, thereby avoiding surgical procedure for their removal. Co-polymer poly (lactic-co-glycolic acid) (PLGA) is a type of temporary biomaterial, routinely used in medicine as suture threads and orthopedic implants. The mixture of PLGA with poly (isoprene) results in a high-strength and thoughness blend (PLGA / PI), developed by the Biomaterials Laboratory of the Engineering Institute/ UFRGS. However, there are no studies “in vivo” testing the bone reaction of that blend. Objective: To Test histotoxicity of PLGA / PI blend over the already established biopolymer, PLGA. Method: Forty six male Wistar rats (Rattus norvegicus – albino strain), divided into 2 groups according to the material (PLGA or PLGA / PI) implanted in the skull and sub divided into periods of death (15, 30, 60 and 90 days). The procedures were developed in the Animal Experiment Unit (AEU) of Hospital de Clínicas de Porto Alegre (HCPA). After death, the skull was removed, submitted to histopathologic examination and the modified Dadas’ et all score was used (14). Results: The histotoxicity difference of the two materials was not significant in the periods of 15, 30 and 90 days, but it was significant in the period of 60 days. Conclusion: At the end of the study (90 days), the PLGA / PI histotoxicity was similar to PLGA, showing longterm equivalence. The 60-day post-surgical period was the only one in which histotoxicity was significantly higher (blend group). More studies shall be done in in order to better understand that variation.

Introdução: A aplicação clínica de biomateriais está se expandindo para diversas especialidades médicas. Dentre os diversos tipos de biomateriais, os classificados como temporários merecem atenção especial, pois são assimilados pelo organismo após exercerem sua função, evitando, assim, procedimento cirúrgico para sua retirada. O copolímero de poli (ácido láctico-co-glicólico) (PLGA) é um tipo de biomaterial temporário, rotineiramente utilizado na medicina na forma de fios de sutura e implantes ortopédicos. A mistura do PLGA com o poli (isopreno) resulta em uma blenda (PLGA / PI), de alta resistência e tenacidade, que foi desenvolvida pelo Laboratório de Biomateriais do Instituto de Engenharia da UFRGS. Entretanto, não existem estudos “in vivo” testando a reação óssea desta blenda. Objetivo: Testar a histotoxicidade da blenda de PLGA / PI em relação ao biopolímero já consagrado PLGA. Método: Foram utilizados 46 ratos machos wistar (Rattusnorvegicus - linhagem albina), divididos em 2 grupos conforme o material implantado (PLGA ou PLGA / PI) na calota craniana, e subdivididos em tempos de morte (15, 30, 60 e 90 dias). Os procedimentos foram realizados na Unidade de Experimentação Animal (UEA) do Hospital de Clínicas de Porto Alegre (HCPA). Após a morte, a calota craniana foi retirada, submetida ao exame histopatológico e aplicado o escore de Dadas e cols (14) modificado. Resultados: A diferença da histotoxicidade dos dois materiais não foi significativa nos períodos 15, 30 e 90 dias, porém foi significativa no período 60 dias. Conclusão: A histotoxicidade do PLGA / PI, ao final do estudo (90 dias), foi semelhante ao PLGA, demonstrando equivalência em longo prazo. O período de 60 dias pós-cirúrgico (grupo da blenda) foi o único em que a histotoxicidade mostrou-se significativamente maior. Mais estudos devem ser feitos para melhorar o entendimento desta variação. / Introduction: Clinical application of biomaterials is expanding to various medical specialties. Among the different types of biomaterials, those classified as temporary deserve special attention because they are assimilated by the body after exercising their function, thereby avoiding surgical procedure for their removal. Co-polymer poly (lactic-co-glycolic acid) (PLGA) is a type of temporary biomaterial, routinely used in medicine as suture threads and orthopedic implants. The mixture of PLGA with poly (isoprene) results in a high-strength and thoughness blend (PLGA / PI), developed by the Biomaterials Laboratory of the Engineering Institute/ UFRGS. However, there are no studies “in vivo” testing the bone reaction of that blend. Objective: To Test histotoxicity of PLGA / PI blend over the already established biopolymer, PLGA. Method: Forty six male Wistar rats (Rattus norvegicus – albino strain), divided into 2 groups according to the material (PLGA or PLGA / PI) implanted in the skull and sub divided into periods of death (15, 30, 60 and 90 days). The procedures were developed in the Animal Experiment Unit (AEU) of Hospital de Clínicas de Porto Alegre (HCPA). After death, the skull was removed, submitted to histopathologic examination and the modified Dadas’ et all score was used (14). Results: The histotoxicity difference of the two materials was not significant in the periods of 15, 30 and 90 days, but it was significant in the period of 60 days. Conclusion: At the end of the study (90 days), the PLGA / PI histotoxicity was similar to PLGA, showing longterm equivalence. The 60-day post-surgical period was the only one in which histotoxicity was significantly higher (blend group). More studies shall be done in in order to better understand that variation.

Introdução: A aplicação clínica de biomateriais está se expandindo para diversas especialidades médicas. Dentre os diversos tipos de biomateriais, os classificados como temporários merecem atenção especial, pois são assimilados pelo organismo após exercerem sua função, evitando, assim, procedimento cirúrgico para sua retirada. O copolímero de poli (ácido láctico-co-glicólico) (PLGA) é um tipo de biomaterial temporário, rotineiramente utilizado na medicina na forma de fios de sutura e implantes ortopédicos. A mistura do PLGA com o poli (isopreno) resulta em uma blenda (PLGA / PI), de alta resistência e tenacidade, que foi desenvolvida pelo Laboratório de Biomateriais do Instituto de Engenharia da UFRGS. Entretanto, não existem estudos “in vivo” testando a reação óssea desta blenda. Objetivo: Testar a histotoxicidade da blenda de PLGA / PI em relação ao biopolímero já consagrado PLGA. Método: Foram utilizados 46 ratos machos wistar (Rattusnorvegicus - linhagem albina), divididos em 2 grupos conforme o material implantado (PLGA ou PLGA / PI) na calota craniana, e subdivididos em tempos de morte (15, 30, 60 e 90 dias). Os procedimentos foram realizados na Unidade de Experimentação Animal (UEA) do Hospital de Clínicas de Porto Alegre (HCPA). Após a morte, a calota craniana foi retirada, submetida ao exame histopatológico e aplicado o escore de Dadas e cols (14) modificado. Resultados: A diferença da histotoxicidade dos dois materiais não foi significativa nos períodos 15, 30 e 90 dias, porém foi significativa no período 60 dias. Conclusão: A histotoxicidade do PLGA / PI, ao final do estudo (90 dias), foi semelhante ao PLGA, demonstrando equivalência em longo prazo. O período de 60 dias pós-cirúrgico (grupo da blenda) foi o único em que a histotoxicidade mostrou-se significativamente maior. Mais estudos devem ser feitos para melhorar o entendimento desta variação. / Introduction: Clinical application of biomaterials is expanding to various medical specialties. Among the different types of biomaterials, those classified as temporary deserve special attention because they are assimilated by the body after exercising their function, thereby avoiding surgical procedure for their removal. Co-polymer poly (lactic-co-glycolic acid) (PLGA) is a type of temporary biomaterial, routinely used in medicine as suture threads and orthopedic implants. The mixture of PLGA with poly (isoprene) results in a high-strength and thoughness blend (PLGA / PI), developed by the Biomaterials Laboratory of the Engineering Institute/ UFRGS. However, there are no studies “in vivo” testing the bone reaction of that blend. Objective: To Test histotoxicity of PLGA / PI blend over the already established biopolymer, PLGA. Method: Forty six male Wistar rats (Rattus norvegicus – albino strain), divided into 2 groups according to the material (PLGA or PLGA / PI) implanted in the skull and sub divided into periods of death (15, 30, 60 and 90 days). The procedures were developed in the Animal Experiment Unit (AEU) of Hospital de Clínicas de Porto Alegre (HCPA). After death, the skull was removed, submitted to histopathologic examination and the modified Dadas’ et all score was used (14). Results: The histotoxicity difference of the two materials was not significant in the periods of 15, 30 and 90 days, but it was significant in the period of 60 days. Conclusion: At the end of the study (90 days), the PLGA / PI histotoxicity was similar to PLGA, showing longterm equivalence. The 60-day post-surgical period was the only one in which histotoxicity was significantly higher (blend group). More studies shall be done in in order to better understand that variation.

[Truncated abstract] Antimicrobial growth promoters are added to feed to prevent lactic acidosis in ruminant animals by selectively inhibiting rumen bacteria that produce lactic acid. However, recently imposed or impending bans on the use of antimicrobial growth promoters in animal production have lead to a critical need to find practical alternatives that are safe for the animal and consumer and that obtain similar production benefits. I investigated bioactive plants of Australia for their potential to prevent lactic acidosis in ruminants. The unifying hypothesis tested was that plants would be identified that selectively inhibit lactic acid-producing bacteria and consequently protect against lactic acidosis. This hypothesis was tested in a three phase process: phase 1, plant selection and collection; phase 2, a three stage protocol for screening plants and essential oils; phase 3, in vivo experiments and chemical fractionation of the most promising plant. I developed an in vitro bioassay that simulated acidosis by adding glucose to rumen fluid in Bellco tubes and incubating for 5 h (Chapter 4). The pH and gas production were used as indicators of acidosis and fermentation activity. I used this bioassay to screen ninety-five plants (dried and ground material from 79 species) and ten essential oils and included a negative control (oaten chaff) and a positive control (virginiamycin). One plant, Eremophila glabra, produced a similar pH (5.63) to the positive control (5.43) although it inhibited gas production to a moderate extent (P < 0.05). ... Seven serrulatane diterpenes were identified to be the major secondary metabolites in E. glabra. The metabolites were screened using a broth dilution and microtitre spectrophotometry method and were selective against S. bovis at between 320 and 1077 [mu]g/ mL. The serrulatanes from E. glabra were probably responsible for the activity against acidosis that I observed in vitro, because they selectively inhibited lactateproducing bacteria. It is also possible that a synergy between serrulatanes and possibly other metabolites are responsible for the activity observed in vitro. The results from my experiments support the role that bioactive plants may have to replace the antibiotics that are added to livestock feed. Australian plants were identified containing compounds that were active against the bacterial processes responsible for ruminant acidosis. To my knowledge this is the first work undertaken to identify bioactive plants of Australia for their potential to prevent acidosis. I developed in vitro screening bioassays that targeted key indicators of acidosis. These bioassays enabled me to identify 5 plants from the 104 screened that could potentially control acidosis. One of these plants in particular, E. glabra, showed a level of activity in vitro that was comparable to antibiotic protection against acidosis. The exciting in vitro results were not demonstrated in vivo but only one dose level of E. glabra was used, which was based on the in vitro work. In contrast to the in vitro system the rumen is a continuous flow system with greater complexity and it is possible that the concentration of E. glabra that I used in vivo was not optimum. This places importance on future dose response experiments to confirm the efficacy of E. glabra in vivo.

Thesis (MSc (Wine Biotechnology))--University of Stellenbosch, 2007. / Bacteriocins are ribosomally synthesized antimicrobial peptides produced by bacteria and are active against other bacteria, either in the same species (narrow spectrum) or across genera (broad spectrum). The application of bacteriocins during the vinification process might help to prevent the production of undesired compounds by inhibiting the indigenous bacterial microflora and allowing malolactic fermentation to be conducted by a selected bacterial strain. Furthermore, the use of bacteriocins might allow reducing the total sulphur dioxide amount in wine. The purpose of this study was the selection of lactic acid bacteria (LAB) belonging to the genera Oenococcus, Lactobacillus and Pediococcus with the ability to produce bacteriocins, with respective biological activity against undesired indigenous wine LAB and the capability to complete malolactic fermentation. The first objective of this study was the screening of LAB isolated from South African red wines for the production of bacteriocins. Only 27 strains out of 330 wine isolates, belonging to the species Lb. plantarum, Lb. paracasei, Lb. hilgardii and O. oeni, showed activity towards various wine-related and non wine-related indicator strains with the colony-overlay method. It is the first time that bacteriocin activity is reported in O. oeni. The second objective was the detection and identification of known structural bacteriocin genes of Lb. plantarum wine strains. Furthermore, the web server BAGEL was used to in silico analyse putative bacteriocin-encoding genes in the genome of O. oeni and primers were designed to amplify four possible bacteriocin-encoding genes. A PCR-based screening revealed the presence of the plantaricin encoding genes plnA, plnEF, plnJ and plnK in five selected Lb. plantarum strains. Moreover, PCR analysis rendered positive results with all four chosen putative bacteriocin-encoding genes in the eight tested O. oeni strains with antimicrobial activity. The latter genes of O. oeni were heterologously expressed in different Escherichia coli host strains, but no antimicrobial activity could be detected. The third objective of this study was the transformation and expression of the heterologous bacteriocin genes nisin A and pediocin PA-1 in two selected Lb. plantarum strains. To enhance their antimicrobial activity a plasmid containing the nisin A gene was successfully cloned into the two strains. Indeed, an enhanced antimicrobial activity could be detected, but the transformed plasmid was not stable. The fourth objective in this project was the evaluation of bacteriocin production in liquid media. A co-culture experiment with a plantaricin producing Lb. plantarum strain and an Enterococcus faecalis strain as indicator was performed. A complete inhibition of cell growth of Ent. faecalis was observed within 72 hours. The last objective was the evaluation of the impacts of phenolic compounds on the activity of nisin and pediocin. The short term influence of two phenolic acids, two flavan-3-ols, grape tannins and oak tannins on the activity of nisin and pediocin PA-1 was investigated. No influence on the activity was detected. Furthermore, synergistic effects on bacterial growth inhibition were observed. This study confirms the potential use of either bacteriocin additives or bacteriocin-producing LAB in order to control the bacterial microflora during the vinification process.

<p>Pivski trop čini približno 85% od ukupnih sporednih proizvoda proizvodnje piva, i dostupan je po veoma niskim cenama tokom čitave godine. Pivski trop ima veliku perspektivu za primenu u biotehnologiji i proizvodnji visoko vrednih proizvoda. Jedna od veoma ekolo&scaron;ki i ekonomski isplativih alternativa je upotreba pivskog tropa u proizvodnji mlečne kiseline, jer se poslednjih par decenija uočava intenzivan rast potražnje za mlečnom kiselinom. Mlečna kiselina je najvažnija hidroksikarbonska kiselina &scaron;iroko rasprostranjena u prirodi, sa velikom primenom u prehrambenoj, farmaceutskoj, tekstilnoj i hemijskoj industriji i industriji prerade kože.<br />Cilj istraživanja ove doktorske disertacije je ispitivanje primene pivskog tropa u proizvodnji mlečne kiseline. Prvo je izvr&scaron;ena optimizacija enzimske hidrolize pivskog tropa u cilju dobijanja &scaron;to je moguće veće koncentacije redukujućih &scaron;ećera neophodne za mlečno-kiselu fermentaciju. Hidrolizat pivskog tropa je dobijen enzimskom hidrolizom dodatkom komercijalnih enzima za razgradnju skroba i celuloze. Parametri čiji je uticaj na efikasnost enzimske hidrolize ispitanu su: pH vrednost, temperatura hidrolize i količina dodatih enzima. Nakon &scaron;to su određeni najbolji uslovi razgranje pivskog tropa, dobijeni postupak hidrolize je primenjen u proizvodnji hidrolizata pivskog tropa koji je kori&scaron;ćen u mlečno-kiselim fermentacijama.<br />Nakon toga je ispitana mlečno-kisela fermentacija sa dva proizvodna mikoorganizma. Kao proizvodni mikroorganizmi u mlečno-kiselim fermentacijama primenjena su dva soja bakterija mlečne kiseline: Lactobacillus fermentum PL-1 i Lactobacillus rhamnosus ATCC 7469. Ispitan je uticaj dodatka različitih koncentracija ekstrakta kvasca (0,5-5,0%) uz korekciju pH vrednosti tokom fermentacije sa dodatkom kalcijum-karbonata. U zavisnosti od udela L-(+)- i D-(-)-mlečne kiseline koje nastaju tokom fermentacije izabran je proizvodni mikroorganizam koji proizvodi vi&scaron;e L-(+)-mlečne kiseline.<br />U daljim ispitivanjima je ispitan uticaj korekcije pH pomoću natrijum-hidroksida kao i dodatak različitih koncentracija ekstrakta kvasca (0,5-5,0%) i redukujućih &scaron;ećera (2,7; 5,4 i 8,1%) u hidrolizatu pivskog tropa na mlečno-kiselu fermentaciju pomoću odabranog soja bakterija mlečne kiseline. Na osnovu dobijenih rezultata izabrana je najbolja koncentracija redukujućih &scaron;ećera i ekstrakta kvasca koji će se koristiti u daljim istraživanjima.<br />Takođe je ispitana i mogućnost zamene skupog ekstrakta kvasca i glukoze sa obnovljivim sirovinama, kao &scaron;to su pivski kvasac, džibra i bistra džibra.<br />Ispitan je uticaj dodatka različitih koncentracija pivskog kvasca (0,5-5,0%), džibre (5-20%) i bistre džibre (5-50%) pre fermentacije kao i dodatak bistre džibre u dolivnoj fermentaciji, na mlečno-kiselu fermentaciju hidrolizata pivskog tropa.<br />Ispitan je i dolivni postupak fermentacije hidrolizata pivskog tropa dodatkom glukoze, glukoze i ekstrakta kvasca i sladovine. Takođe je ispitana mogućnost izvođenja vi&scaron;e uzastopnih fermentacija sa imobilisanim ćelijama odabranog soja bakterija mlečne kiseline u kalcijum-alginatu.<br />Na osnovu eksperimentalnih rezultata zaključujeno je da je dodatak kalcijum-karbonata imao pozitivan uticaj na proizvodnju mlečne kiseline sa L. fermentum i L. rhamnosus. Sa dodatkom kalcijum-karbonata povećali su se utro&scaron;ak redukujućih &scaron;ećera, koncentracija i prinos mlečne kiseline i vijabilnost ćelija L. fermentum i L. rhamnosus. Ekstrakt kvasca i kalcijum-karbonat su imali značajan uticaj na proizvodnju mlečne kiseline sa L. fermentum i L. rhamnosus. U fermentacijama sa L. fermentum najveći prinos ukupne mlečne kiseline (44%) je postignut sa dodatkom 5,0% ekstrakta kvasca i 2,0% kalcijum-karbonata. U fermentacijama sa L. rhamnosus najveći prinos ukupne mlečne kiseline (98%) i L-(+)-mlečne kiseline (96%) je ostvaren u fermentaciji sa dodatkom 2,0% ekstrakta kvasca i 2,0% kalcijum-karbonata. Na osnovu rezultata odlučeno je da se u daljim ispitivanjima mlečno-kisele fermentacije hidrolizata pivskog tropa kao proizvodni mikoorganizam koristi L. rhamnosus.<br />Primenom natrijum-hidroksida za korekciju pH je skratila fermentaciju za 48 sati a ostvareno je i značajno povećanje zapreminske produktivnosti L-(+)-mlečne kiseline (za 200%, povećanje sa 0,21 na 0,63 g/l&middot;h-1). Korekcija pH u svim daljim istraživanjima je vr&scaron;ena sa dodatkom natrijum-hidroksida.<br />U mlečno-kiselim fermentacijama sa različitim početnim koncentracijama redukujućih &scaron;ećera (2,7; 5,4 i 8,1%) i sa dodatkom različitih koncentracija ekstrakta kvasca (0,5-5,0%), najveći prinos L-(+)-mlečne kiseline i zapreminska produktivnost od 91,29% i 1,69 g/l&middot;h-1, kao i vijabilnost ćelija L. rhamnosus od 9,7&middot;109 CFU/ml ostvareni su u fermentaciji sa početniom koncentracijom redukujućih &scaron;ećera od 5,4% i dodatkom 5,0% ekstrakta kvasca.<br />Na osnovu ostvarenih rezultata u istraživanjima sa dodatkom džibre i dodacima tokom fermentacije kao i u fermentacijama sa imobilisanim ćelijama je kori&scaron;ćen hidrolizat pivskog tropa sa početnom koncentracijom redukujućih &scaron;ećera od 5,4%.<br />U mlečno kiseloj fermentaciji sa dodatkom pivskog kvasca najveći prinos L-(+)-mlečne kiseline (89,01%) i zapreminska produktivnost (0,89 g/l&middot;h-1) L-(+)-mlečne kiseline su ostvareni u fermentaciji sa dodatkom 5,0% pivskog kvasca i korekcijom početne koncentracije redukujućih &scaron;ećera na 5,0%. Na osnovu rezultata utvrđeno je da se može izvr&scaron;iti delimična ili potpuna zamena ekstrakta kvasca pivskim kvascem uz značajno smanjenje cene podloge za mlečno-kiselu fermentaciju, bez značajnog smanjenja efikasnosti mlečno-kisele fermentacije.<br />U mlečno-kiseloj fermentaciji sa dodatkom džibre i bistre džibre najveć koncetracija, prinos i zapreminska produktivnost L-(+)-mlečne kiseline od 31,03 g/l, 86,15% i 0,93 g/l&middot;h-1, ostvareni su u fermentaciji sa dodatkom 50% bistre džibre. Najvi&scaron;a koncentracija, prinos i zapreminska produktivnost L-(+)-mlečne kiseline ostvareni u dolivnoj fermentaciji sa dodatkom glukoze i bistre džibre tokom mlečno-kisele fermentacije su iznosili su 48,02 g/l, 87,82% i 0,96 g/l&middot;h-1.<br />U fermentacijama sa dodatkom nutritijenata tokom mlečno-kisele fermentacije najveća vrednost koncetracije, prinosa i zapreminske produktivnosti L-(+)-mlečne kiseline od 116,08 g/l, 93,32% i 2,04 g/L&middot;h-1, su ostvarene u fermentaciji sa dodatkom glukoze i ekstrakta kvasca tokom fermentacije. Na osnovu rezultata utvrđeno je da se dolivni postupak fermentacije može koristiti u cilju povećanja efikasnosti mlečno-kisele fermentacije.<br />Izvr&scaron;ena je imobilizacija ćelija L. rhamnosus u kalcijum-alginatu uz izuzetno visoku vijabilnost (1010 CFU/ml). Imobilisane ćelije L. rhamnosus su uspe&scaron;no kori&scaron;ćene u tri mlečno-kisele fermentacije. Prinos L-(+)-mlečne kiseline i zapreminska produktivnost su u sve tri fermentacije bili izuzetno visoki, pri čemu su najveći prinos L-(+)-mlečne kiseline i zapreminska produktivnost od 95,2% i 1,76 g/l&middot;h-1, ostvareni u drugoj fermentaciji. Upotrebom imobilisanih ćelija L. rhamnosus je osim povećanja prinosa i zapreminske produktivnosti L-(+)-mlečne kiseline skraćena fermentacija za 12 sati u poređenju sa &scaron;aržnim fermentacijama.</p> / <p>Brewers spent grain represents (BSG) about 85% of the total by-products from brewing process and is available at low price during the whole year. Due to its chemical composition BSG has great potential use in biotechnology and production of high-value products. One of very eco-friendly and economical alternative uses of BSG is in production of lactic acid (LA), since in the last few decades the demand for the LA has significantly risen, mostly because of development of biodegradable lactic polymers, which are eco-friendly and nontoxic.<br />Lactic acid is the most important hydrocarboxylic acid with an asymmetrical carbon atom, widely distributed in nature, and it has shown great potential in fields of food, pharmaceutical, textile, leather and chemical industries.<br />The aim of this doctoral thesis was to investigate the application of BSG in lactic acid production. First, the optimization of enzymatic hydrolysis of BSG was conducted, with the goal to achieve high reducing sugar concentrations, as much as possible, that are necessary on LA fermentation. BSG hydrolysis was conducted by usage of commercial enzymes for degradation of starch and cellulose. Effect of pH value, temperature and enzyme dosage on BSG hydrolysis efficiency was investigated. After the best conditions for BSG hydrolysis were determined, the optimized procedure for BSG hydrolysis was used for the production of BSG hydrolysate that will be used in LA fermentations.<br />After optimization of BSG hydrolysis, LA fermentation by two LA producing microorganisms was investigated. The strains investigated were two LA bacteria strains: Lactobacillus fermentum PL-1 and Lactobacillus rhamnosus ATCC 7469. The effect of yeast extract (0.5; 1.0; 2.0; 3.0; 4.0, and 5.0%) addition in BSG hydrolysate, with the correction of pH value during LA fermentation by the addition of calcium-carbonate, on LA fermentation was investigated. Based on the results achieved for L-(+)- and D-(-)-LA ratio the LAB strains that produced more L-(+)-LA was chosen for further research.<br />In further research the effect of pH correction (with addition of NaOH), yeast extract (0.5, 1.0, 2.0, 3.0, 4.0, and 5.0%) addition and reducing sugar concentration (2.7; 5.4 and 8.1%) in BSG hydrolysate on LA fermentation was investigated. Based on the results achieved the best yeast extract and reducing sugars concentrations was determined and used in further analysis or research. Also the possible replacement of expensive yeast extract and glucose with cheap alternatives, like brewer`s spent grain and stillage was investigated. The effect of brewer`s spent grain (0.5; 1.0; 2.0; 3.0; 4.0, and 5.0%), whole stillage (5, 10, 15 i 20%) and thin stillage (5, 10, 15, 20, 30, 40, 50%) addition before fermentation as well as thin stillage addition in fed-batch fermentation in BSG hydrolysate on LA fermentation were investigated.<br />Also fed-batch fermentation procedure (addition of glucose, glucose and yeast extract and wort during fermentation) was investigated. The possible application of cells immobilized in Ca-alginate for LA fermentation of BSG hydrolysate was also investigated.<br />Based on the results it was concluded that BSG can be successfully utilized as a raw material in production of LA, after optimization of hydrolysis and addition of nitrogen source.<br />According to the results of chemical composition before and after optimized hydrolysis 78.6% of total cellulose was hydrolyzed.<br />Addition of calcium-carbonate had positive effect on LA production by L. fermentum i L. rhamnosus. With the addition of calcium-carbonate reducing sugar utilization, LA yield and concentration and cell viability (both L. fermentum i L. rhamnosus) increased. Addition of calcium-carbonate and yeast extract had a positive effect on LA fermentation by L. fermentum and L. rhamnosus. In LA fermentation by L. fermentum the highest LA yield (44%) was achieved with addition of 5.0% of yeast extract and 2.0% of calcium-carbonate. In L. rhamnosus fermentations the highest total LA yield (98%) and L-(+)-LA yield (96%) was reached when 2.0% of yeast extract and 2.0% of calcium-carbonate were added.<br />Based on the results achieved it was concluded that BSG hydrolysate, with the addition of yeast extract, is a good fermentation media for LA fermentation with L. rhamnosus, and it was decided that L. rhamnosus will be used in further research of LA fermentation on BSG hydrolysate.<br />Addition of NaOH instead of calcium-carbonate for the pH correction shortened the fermentation time by 48 h and increased the L-(+)-LA volumetric productivity (by 200%, from 0.21 to 0.63 g/L&middot;h-1). Based on this results pH correction in further experiments was done by addition of NaOH.<br />In LA fermentation with different reducing sugar (2.7, 5.4 and 8.1%) and yeast extract concentrations (0.5-5.0%), the highest L-(+)-LA yield and volumetric productivity of 91.29%, and 1.69 g/L&middot;h-1, respectively, as well as L. rhamnosus cell viability (9.67 log CFU/mL), were achieved with the reducing sugar content of 5.4% and yeast extract content of 5.0%.<br />Based on this results in further experiment with the addition of stillage, in fed-batch fermentation and fermentation with immobilized cell BSG hydrolysate with 5.4% of reducing sugars and 5.0% yeast extract was used.<br />In fermentation with the addition of brewer&rsquo;s spent yeast the highest L-(+)-LA yield (89.01%) and volumetric productivity (0.89 g/L&middot;h-1) were achieved in the fermentation of BSG hydrolysate with 5.0% of reducing sugar and 5.0% of brewer&rsquo;s yeast. Based on the results achieved it was concluded that yeast extract can be partial or complete replaced by brewer&rsquo;s spent yeast with significant decrease of media cost, without the decrease in LA fermentation efficiency.<br />In fermentation with the addition of thin stillage the highest L-(+)-LA concentration, yield, and volumetric productivity of 31.03 g/L, 86.15%, and 0.93 g/L&middot;h-1, respectively, was obtained in fermentation with the addition of 50% of thin stillage. The highest L-(+)-LA concentration, yield, and volumetric productivity achieved in fed-batch fermentation with the addition of glucose and thin stillage during fermentation, were 48,02 g/L, 87,82% i 0,96 g/L&middot;h-1.<br />In fed-batch fermentation the highest L-(+)-LA concentration, yield, and volumetric productivity of 116.08 g/L, 93.32%, and, 2.04 g/L h-1, respectively, were achieved in fermentation with glucose and yeast extract addition during fermentation. The results showerd that fed-batch fermentation could be used to increase L-(+)-LA fermentation efficiency<br />Immobilization of L. rhamnosus cells with high viability (1010 CFU/mL) in Ca-alginate was conducted. Immobilized cells we successfully utilized in three repeated batch fermentation. L-(+)-LA yield and volumetric productivity were very high in all three batch fermentation, with the highest results achieved (95.20% and 1.76 g/L&middot;h-1, respectively) in second fermentation. Application of immobilized L. rhamnosus cells increased L-(+)-LA yield and volumetric productivity and shortened the fermentation time for 12 h in comparison with batch fermentation.</p>