Thesis (M.Tech.: Biotechnology)- Dept. of Biotechnology, Durban University of Technology, 2007 xv, 127 leaves / The human gut microbiota play a major role in host health, and attempts are being made to manipulate the composition of the gut microbiota-increase the composition of bacterial groups, such as lactobacilli and bifidobacteria that are perceived as exerting health promoting properties. These bacteria defined as food supplements (probiotics) beneficially affect the host by improving the intestinal microbial balance, and have been used to change the composition of the colonic microbiota. However, such changes may be transient, and the implantation of exogenous bacteria therefore becomes limited. In contrast, prebiotics are naturally occurring carbohydrates that are classified as non-digestible oligosaccharides present in edible plants. These carbohydrates enter the colon as intact compounds, elicit systemic physiological functions and act as fermentable substrates for colonic microflora-influencing the species composition and metabolic characteristics of intestinal microflora providing important health attributes. Currently, a widely marketed prebiotic, inulin is extracted from plants of the family Asteraceae. There are many unexploited plants that are regularly consumed and that may have a prebiotic effect or can have very high levels of inulin which could make them commercially viable. In this study, we investigated prebiotic compounds, especially inulin from locally growing, non-commercialised leafy plants. The aqueous extracts of 22 plants from the families Asparagaceae, Alliaceae, Asteraceae, Solanaceae, Cucurbitaceae, Amaranthaceae, Acanthaceae, Polygonaceae, Portulaceae, Fabaceae, Chenopodiaceae, Pedaliaceae and Apiaceae from Kwa-Zulu Natal were investigated for a prebiotic effect using a modified batch-culture technique with Lactobacillus bulgaricus, Lactobacillus lactis, Lactobacillus reuteri and Bifidobacterium longum, four common probiotics and the inulin content of the plants was determined using high performance liquid chromatography. Of the 22 plants studied, Solanum nigrum, Amaranthus spinosus, Amaranthus hybridus, Asystasia gangetica, Senna occidentalis, Cerathoteca triloba, Asparagus sprengeri, Tulbaghia violacea, Sonchus oleraceus and Taraxacum officinale exhibited a prebiotic effect. The prebiotic effect of the Taraxacum officinale, Sonchus oleraceus and Asparagus sprengeri extracts on L. lactis and L. reuteri was higher than or equivalent to inulin-a commercial prebiotic. In this study, Sonchus oleraceus exhibited the best prebiotic effect-was the only plant to stimulate all the probiotics including B. longum. Of all the plants analysed, Asparagus sprengeri tuber contained the highest amount of inulin (3.55%).

Thesis (PhD (Microbiology))--Stellenbosch University, 2008. / One of the key selection criteria for potential probiotics is the ability to adhere and colonise the host gastrointestinal tract (GIT). Probiotics compete for receptor sites at the host intestinal surface, preventing the colonisation of pathogens, thereby protecting the host from infection. In addition, several important intestinal functions are mediated by the binding of probiotics to host tissue. However, the molecular mechanisms and genotypic characterization of adhesive elements have not received as much attention as other aspects of probiotic research. The present study aims to contribute to this area of research. The first part of the study focused on monitoring the expression of mucus adhesion genes mub, mapA, adhesion-like factor EF-Tu and bacteriocin gene plaA of Lactobacillus plantarum 423, as well as mub, surface layer protein (slp) and EF-Tu of Lactobacillus acidophilus ATCC 4356 when grown in the presence of mucin, bile, pancreatin and at low pH. Real time PCR was used. mub, mapA and EF-Tu of strain 423 were up-regulated in the presence of mucus and expression increased under increasing concentrations of mucus. Expression of mapA was up-regulated under normal gut conditions (0.3%, w/v, bile; 0.3%, w/v, pancreatin; pH 6.5) and at higher levels of bile (1.0%, w/v) and pancreatin (1.0%, w/v). Expression of mub was downregulated in the presence of bile and pancreatin at pH 6.5, whilst the expression of EFTu and plaA remained unchanged. At pH 4.0, the expression of mub and mapA remained unchanged, whilst EF-Tu and plaA were up-regulated. Expression of mapA was down-regulated in the presence of 0.1% (w/v) cysteine, suggesting that the gene is regulated by a mechanism of transcription attenuation that involves cysteine. In the case of L. acidophilus ATCC 4356, none of the genes were up-regulated under increasing concentrations of mucin, whilst only slp and EF-Tu were up-regulated under normal and stressful gut conditions in vitro. In the second part of the study, male Wistar rats were used to evaluate which section of the gastrointestinal tract are colonised by L. plantarum 423 and Enterococcus mundtii ST4SA and determine the effect of adhesion. Fluorescent in situ hybridization (FISH) incorporating strain specific oilgonucleotide probes indicated strong fluorescent signals for L. plantarum 423 along the intestinal lining of the ileum and the cecum. L. plantarum 423 did not colonise the colon as indicated by real timePCR. Fluorescent signals were recorded for E. mundtii ST4SA across the epithelial barrier of cecum and colonic tissue, suggesting that translocation took place. Real time PCR revealed highest cell numbers of strain ST4SA in the cecum and the colon. Haemotoxylin eosin staining of rat tissue revealed no change in morphology or any toxic effects induced upon adhesion of the strains. 16S rDNA PCR and denaturing gradient gel electrophoresis (DGGE) revealed a decrease in enterobacterial species whilst the lactic acid bacterial content remained unchanged. Strains 423 and ST4SA agglutinated yeast cells in vitro, indicating the possible presence of mannose receptors. It is well known that these receptors play a crucial role in the elimination of type 1 fimbriated strains of E. coli. It is thus safe to speculate that mannose receptors may have played a role in diminishing the enterobacterial content in the gut. The third part of the study encompassed characterization of cell surface proteins of L. plantarum 423 and their role in adhesion to Caco-2 cell lines. The strain lacks the typical surface layer protein whilst a multifunctional “intracellular” protein, elongation factor Tu (EF-Tu) and glycolytic enzymes glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and triosephosphate isomerase (TPI) were detected. Removal of surface proteins reduced adherence of strain 423 to Caco-2 cell lines by 40%, suggesting that these proteins play a role in adhesion. The ability of strain 423 to competitively adhere, exclude and displace Clostridium sporogenes LMG 13570 and Enterococcus faecalis LMG 13566 from Caco-2 cell lines, was studied. Adhesion of C. sporogenes LMG 13570 and E. faecalis LMG 13566 was inhibited by 70% and 90%, respectively. Strain 423 excluded C. sporogenes LMG 13570 from Caco-2 cells by 73% and displaced the pathogen by 80%. E. faecalis LMG 13566 was excluded by 60% and displaced from Caco-2 cells by 90%. Despite removal of the surface proteins, L. plantarum 423 was still capable of competitively adhering to Caco-2 cells and reduced adherence of C. sporogenes LMG 13570 by 50% and E. faecalis LMG 13566 by 70%.

The aim of the project is to investigate how a more viscous, gel like formulation can be made of BioGaia´s Prodentis Drops, which is a probiotic product for oral use. The two different strains of Lactobacillus reuteri that are used in the product, together called L. reuteri Prodentis, have been clinically proven to be effective in treatment of gingivitis and caries formation. The existing product is a highly liquid oil suspension that has been described as too runny and difficult to get into tooth pockets and between teeth. Therefore a gel formulation would be preferred. Pre-trials were excecuted to see what combinations and quantities of ingredients could work. Three different formulations using 3 %, 5 % and 7 % beeswax as thickening agent proceeded to another round of trials, as well as one formulation where the original oil was exchanged for hydrogenated rapeseed oil. In the beeswax formulations fumed silicon dioxide was being used as well. Three different analyses were executed, considering bacterial survival, viscosity and phase separation of gels. The bacterial survival proved to be acceptable in all samples even after 14 days of incubation in 37oC. The formulation with 7 % beeswax was the most viscous one, followed by 5 % beeswax, 3 % beeswax and the formulation with hydrogenated oil, respectively. Phase separation could be seen in the hydrogenated oil formulation already after seven days and even more so after 14 days. There were also signs of separation in the formulation with 3 % beeswax after 14 days. It was concluded that in further development of the Prodentis Drops it is recommendable to proceed with the 5 % beeswax formulation.

Nous avons déjà démontré que les probiotiques réduisaient l'apoptose observée dans le système limbique après un infarctus du myocarde (IM), suggérant un rôle anti-dépresseur potentiel des probiotiques. Cette étude a été conçue pour déterminer si les probiotiques pouvaient atténuer le comportement dépressif observé après un infarctus du myocarde. Un autre objectif visait à démontrer qu’une altération de la barrière intestinale pourrait avoir lieu lors d’un IM et que les probiotiques pourraient empêcher cette altération de la perméabilité intestinale. Méthodes: Des rats mâles Sprague-Dawley ont reçu des probiotiques (1 milliard de cellules bactériennes vivantes de Lactobacillus helveticus R0052 et Bifidobacterium longum R0175) ou le véhicule tous les jours en dilution dans leur eau, débutant 1 semaine avant l'induction d'une ischémie myocardique. Un infarctus a ensuite été induit chez la moitié des rats, par l'occlusion de l'artère coronaire antérieure gauche (40 minutes) suivie d'une reperfusion. Les rats témoins, l'autre moitié de la cohorte, ont été soumis à la même procédure sans occlusion coronarienne. Une semaine après l'infarctus, les animaux ont été resoumis au traitement préalable jusqu'au moment du sacrifice. Le comportement dépressif a été évalué par trois tests soit: l'interaction sociale, le test de nage forcée et le test d'évitement passif. Résultats: Les résultats obtenus indiquent qu'en absence d'infarctus, les probiotiques n'ont pas d'effet significatif. Toutefois, en dépit de taille similaire IM, des rats traités avec des probiotiques, ont démontré davantage d'interactions sociales et une meilleure performance dans le test de nage forcée et d'évitement passif, comparativement à des rats du groupe IM sans probiotique (p<0,05). Conclusion: Les probiotiques atténuent le comportement dépressif observé après infarctus du myocarde par un mécanisme qui pourrait impliquer l'intégrité des intestins. / We have already shown that probiotics reduce apoptosis in the limbic system after a myocardial infarct, suggesting a potential anti-depressive role of probiotics. This study was conceived to determine if probiotics could lessen the depressive behaviour observed after a myocardial infarction (MI). We tested the hypothesis that probiotics achieve their central effects through changes in the intestinal barrier. Methods: Male Sprague-Dawley rats received probiotics (1 billion live bacteria of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175) or placebo every day dissolved in drinking water , beginning one week before induction of myocardial ischemia. An infarct has been induced on half of the cohort, by the occlusion left anterior coronary artery (40 minutes) followed by reperfusion. The other half was subjected to the same procedure without coronary occlusion. One week after the infarction, the animals received probiotic treatment until sacrifice after 2 weeks. The depressive behaviour was evaluated by three tests: social interaction, forced swim and foot shock. Results: The obtained results indicate that in the absence of infarction, probiotics have no significant effect. Even though they are of the same MI gravity, the rats treated with the probiotics have shown more social interaction and have a better performance in the forced swim and foot shock tests compared to the untreated MI rats (p<0,05). Conclusion: Probiotics diminish the depressive behaviour observed after a myocardial infarction by a mechanism which could implicate the integrity of the intestine barrier.

De nombreuses pathologies découlent de déséquilibres du microbiote intestinal. Ainsi, des stratégies visent à les prévenir en restaurant cet écosystème par l'apport de probiotiques, de prébiotiques et de synbiotiques. Des résultats prometteurs nécessitant d'être validés rationnellement, nous nous sommes proposés d'établir un premier crible de sélection de bactéries lactiques et de glucooligosaccharides (GOS) pour des activités anti-inflammatoires et anti-prolifératives in vitro. Les probiotiques ont inhibé des pathogènes tout en se révélant résistants aux conditions du tube digestif, tandis que deux GOS ont conduit à leur croissance sélective. De plus, les probiotiques ont réduit la réponse inflammatoire de cellules intestinales et la prolifération de cellules cancéreuses en association avec un GOS. Nous avons donc retenu des produits potentiellement actifs contre les maladies inflammatoires chroniques de l'intestin et du cancer colorectal, par un crible devant être validé in vivo. / Some pathologies are induced by intestinal microbiota disorders. Thus, some strategies aim torestore this ecosystem through probiotics, prebiotics and synbiotics. Promising results need tobe rationally validated, so we aimed to establish the screening first step of lactic acid bacteriaand glucooligosaccharides (GOS) against anti-inflammatory and anti-proliferative activity invitro. Probiotics inhibited pathogens and were resistant to digestive tract conditions whileGOS promoted specifically their growth. Moreover, probiotics reduced inflammatoryresponse of intestinal cells and proliferation of cancer cells when combined with GOS. Thus,we selected compounds potentially efficient against inflammatory bowel diseases andcolorectal cancer, through a screen that need to be validated in vivo

Submitted in complete fulfillment for the Degree of Doctor of Philosophy in Biotechnology, Durban University of Technology, Durban, South Africa, 2016. / Colon cancer is the third most common cancer worldwide with a high morbidity and mortality rate. Therapies are less effective during metastasis, therefore prevention and earlier detection is key to reducing the risk of colon cancer. Increased dietary fibre and probiotic intake is known to lower the risk of colon cancer. Probiotics are defined as “live microorganisms which when administered orally in an adequate amount confer a health benefit on the host”. The International Dairy Federation recommends a viable minimum level of 6–7 log10cfu/g in a probiotic product being consumed. Different biopolymer matrices have been used for encapsulation of probiotics; however, loss of viability is still a major challenge. Citrus pectin is a dietary fibre polysaccharide broken down into smaller fragments to form modified citrus pectin (MCP). The unique bioactivity of MCP against carcinogenesisis is linked to its sugar β-galactose inhibiting the cell signalling protein marker, galectin-3 (gal-3), which is intimately involved in endothelial cell morphogenesis. The vascular endothelial growth factor (VEGF) signalling, which invariably drives angiogenesis can be activated when gal-3 binds to integrins. The bioactivity and uptake of MCP may be improved through a novel approach if conjoined with a supplement for example probiotic. Therefore, the synergistic inhibitory effect of modified citrus pectin alginate (MCPA) probiotic microbeads on gal-3 and VEGF in an azoxymethane (AOM) induced colon carcinogenesis Balb/c mouse model was investigated. A microencapsulation process was used to produce a MCPA microbead containing probiotic, Lactobacillus acidophilus ATCC 4356. Efficiency of the microbead was evaluated in vitro (simulated conditions) and in vivo (Balb/c mouse model). Genomic identification of faecal lactobacilli samples from the treated mice was analyzed. Optimization of AOM dose-time with 10 and 15 mg/kg AOM intraperitoneal (ip) administered to Balb/c mice for 2 and 4 weeks were performed. The optimal AOM dose was initiated prior to intake of MCPA, AP (alginate calcium) probiotic microbeads and MCP in Balb/c mice for 16 weeks; samples were analyzed for colon histopathology and immunohistochemistry. The MCPA probiotic microbeads significantly enhanced the viability of L. acidophilus ATCC 4356 compared to the AP microbeads in vitro (p< 0.05). Exposure of the MCPA probiotic microbeads to 3 h of simulated gastric juice (SGJ) resulted in 82.7% survival of L. acidophilus ATCC 4356. Also, the faecal lactobacilli count in the MCPA probiotic treated mice significantly increased after 28 days by 10.2% compared to the AP probiotic, MCP treated and control mice (p< 0.0001). A total of 4DNA encoding 16S rRNA gene closest to the genera namely Lactobacillus, Bacillus, Enterococcus and Bifidobacterium were identified from faecal samples of the colon cancer-induced Balb/c mice. Azoxymethane at 15 mg/kg for 4 weeks induced optimal gal-3 and VEGF immunoexpression. Furthermore, MCPA probiotic treatment significantly reduced gal-3 immunoexpression in the colon of AOM induced cancer Balb/c mice compared to the control mice (p< 0.0001). The immunoexpresion of VEGF in the MCPA and AP probiotic treated groups was weakly positive and significantly reduced when compared to the control group (p<0.05), while the MCP treated group was barely positive (p< 0.001). Modified citrus pectin alginate is a novel effective means of oral delivery of bacterial cells and bioactive compounds. It has a good biodegradability, inexpensive, non-toxic, proven efficiency, and stability at low temperatures warranting its use as a drug carrier by pharmaceuticals. Modified citrus pectin alginate probiotic microbeads increase bioactivity and chemoprevention against colon pre-cancerous lesions and adenocarcinoma through inhibition of gal-3 and VEGF in the mouse model. Modified citrus pectin alginate can be used in probiotic therapy, which may improve the prevention of colon cancer. / D

Probiotics are live microorganisms which confer a health benefit to the host. The literature strongly supports the benefits of probiotic therapy in preterm infant populations, specifically in the prevention of necrotizing enterocolitis (NEC) and mortality. To this end, probiotics are routinely given to premature infants in several European and Asian countries. However, in spite of the current evidence and neonatal feeding practices elsewhere, probiotic supplements are rarely prescribed in nurseries in North America. Furthermore, there is little or no literature on factors which affect clinical decision-making regarding probiotic supplementation. The study implemented a cross-sectional descriptive survey. The purpose of this study was to: (i) describe current practices involving probiotic supplementation of preterm infant enteral feeds; and (ii) identify factors that affect willingness of health care professionals to support the use of probiotics. Probiotic use was examined in Neonatal Intensive Care Units (NICUs) in Canada and the United States using two cross-sectional internet-based surveys. Survey #1 focused on current practices and targeted neonatologists who serve as clinical directors or department heads. The results were analysed using descriptive statistics. Survey #2 addressed factors that affect probiotic supplementation of preterm infant feedings, and had two versions: the first version targeted physicians and nurse practitioners whereas version 2 targeted neonatal nurses. The development of Survey #2 was guided by the Theoretical Domain Framework which evaluates factors which may affect the willingness of Health Care Professionals to support the use of probiotics in neonatal practice. The results of survey #2 were analysed using Chi-Square, Fisher’s Exact Test, and One-Way ANOVA. The results of the study indicated that only a small proportion of NICUs are administering probiotics to preterm infants and practices vary. The most significant factors influencing clinical decision-making regarding probiotic supplementation were knowledge about probiotics and the evidence, perceptions about the evidence and safety of probiotics, and knowledge about probiotics and clinical guidelines. Improving knowledge about probiotics, addressing safety issues of probiotics products, expanding the evidence base, and developing clinical guidelines may contribute to increased use of probiotics in NICUs. / October 2016

Objective To describe the use of Lactobacillus by hospitalized patients and to examine its relationship with various Clostridium difficile infection (CDI) related outcomes. Methods The characteristics of Lactobacillus users and non-users and the initiation of Lactobacillus with respect to initiation of antibiotic therapy and CDI treatment were described using national hospital discharge database. The relationships between Lactobacillus use and post-CDI length of stay, mortality, switch of CDI therapy, and readmission were analyzed. Results Lactobacillus users and non-users were different in most characteristics. Metronidazole and fluoroquinolones were the most frequently used antibiotics by Lactobacillus users. They were mainly CDI cases, used multiple antibiotics, extremely ill, and started Lactobacillus five or more days after initiation of antibiotics or CDI treatment. Lactobacillus use was associated with increased length of stay and switching of CDI therapy. Conclusions The true association between Lactobacillus use and CDI remains unclear. This study provides foundation for future research.

Além das características tecnológicas das bactérias lácticas para o processamento de alimentos fermentados, novas propriedades são requeridas, como a produção de bacteriocinas. Vários desses compostos protéicos possuem atividade antimicrobiana direcionada à patógenos e apresentam potencial de uso como bioconservadores naturais de alimentos. O objetivo desse trabalho foi avaliar a viabilidade de aplicação de cinco culturas lácticas produtoras de bacteriocinas como bioconservantes em alimentos. Avaliaram-se as propriedades de segurança e probióticas dessas culturas e determinaram-se os parâmetros de produção e purificação parcial das bacteriocinas, bem como os efeitos de fatores limitantes de crescimento (cloreto de sódio e nitrito). Os resultados demonstraram que essas linhagens não possuem atividade de enzimas indicadoras de patogenicidade (termonuclease, -hemolisinas e gelatinases). A tolerância às condições ácidas variou dependendo da linhagem. Lactobacillus plantarum CTC 368 e Enterococcus avium CTC 469 apresentaram maior tolerância a valores de pH 2,0 e 3,0. Lactococcus lactis subsp. cremoris CTC 204 e Enterococcus avium CTC 483 também apresentaram 100% de sobrevivência a pH 3,0 e, mesmo com uma diminuição da tolerância a pH 2,0, a sobrevivência foi cerca de 80%. Com relação à susceptibilidade aos sais de bile, observou-se que a viabilidade variou entre as linhagens em função da concentração e tempo de incubação. Enterococcus avium CTC 469 e Lactococcus lactis subsp. hordinae CTC 484 foram as mais tolerantes e apresentaram sobrevivência de 57% e 58%, respectivamente. As linhagens mostraram excelente sobrevivência tanto a baixos valores de pH, quanto às concentrações de sais biliares indicando potencial probiótico. As características de crescimento e de produção de bacteriocinas foram semelhantes para todas as linhagens e meios avaliados. A viabilidade aumentou durante o período de incubação e a produção de bacteriocinas mostrou uma cinética de metabólito primário. A suplementação do meio MRS com 1,0% de NaCl e 0,1% de NaNO2, não afetou o crescimento de Lc. lactis subsp. cremoris CTC 204, mas promoveu a redução da atividade da bacteriocina de 42,5% e 17,5%, respectivamente. O crescimento de Lactococcus lactis subsp. hordinae CTC 484 foi afetado com a adição de NaCl e NaNO2 ao meio APT, mas ainda reteve 50% da atividade bacteriocigênica. O espectro de atividade das bacteriocinas variou de acordo com a susceptibilidade da linhagem indicadora, a concentração ou atividade da bacteriocina, a viabilidade e/ou presença das células produtoras e os procedimentos de extração das bacteriocinas. Os resultados revelaram que as bacteriocinas de Lactococcus lactis subsp. cremoris CTC 204 e Lactococcus. lactis subsp. hordinae CTC 484 promoveram um efeito bactericida contra Staphylococcus aureus. Concluiu-se que essas duas culturas produtoras de bacteriocinas apresentam propriedades para utilização como bioconservantes em alimentos e que estudos mais aprofundados, relacionados com a purificação da bacteriocina e avaliação de seu uso em alimentos, devem ser realizados. / In addition to the technological characteristics of lactic bacteria for the processing of fermented foods, other properties need to be developed and explored, such as bacteriocin production. Several of these protein-based compounds have antimicrobial activities targeted towards pathogens and have a significant potential for use as natural biopreservatives in processed foods. The objective of this study was to evaluate the viability of using five bacteriocin-producing lactic cultures as biopreservatives in foods. The safety and probiotic properties of these cultures were evaluated, followed by the determination of bacteriocin production and purification parameters and the effect of growth-limiting factors (sodium chloride and nitrite). The results demonstrated that these strains do not show activity of marker enzymes of pathogenicity (thermonuclease, - hemolysins and gelatinases). Tolerance to acidic conditions varied depending on the specific strain. Lactobacillus plantarum CTC 368 and Enterococcus avium CTC 469 exhibited greater tolerance to pH values 2,0 and 3,0. Lactococcus lactis subsp. cremoris CTC 204 and En. avium CTC 483 also presented 100% survival at pH 3,0, and even with reduced tolerance at pH 2,0, the survival rate was 80%. Variation in susceptibility to bile salts was observed between the strains as a function of concentration and incubation time. En. avium CTC 469 and Lactococcus lactis subsp. hordinae CTC 484 were found to be the most tolerant, exhibiting survival rates of 57% and 58%, respectively. The strains demonstrated excellent survival not only at low pH values, but also at high bile salt concentrations, thereby indicating probiotic potential. The growth and bacteriocin-producing characteristics were similar for all the strains and culture media evaluated. Bacterial multiplication increased during the incubation period, while bacteriocin production displayed primary metabolite kinetics. The activity spectrum of the bacteriocins varied with the susceptibility of the indicator strain, the concentration or activity of the bacteriocin, the viability and/or presence of bacteriocinproducing cells and the bacteriocin extraction procedures. The results reveal that the bacteriocins produced by Lc. lactis subsp. cremoris CTC 204 and Lc. lactis subsp. hordinae CTC 484 exerted a bactericidal effect against Staphylococcus aureus. Supplementation of the MRS medium with 1,0% NaCl and 0,1% NaNO2, did not affect the growth of Lc. lactis subsp. cremoris CTC 204, but did reduce by 42,5% and 17,5% the activity of the bacteriocin, respectively. Growth of Lc. lactis subsp. hordinae CTC 484 was affected by the addition of NaCl and NaNO2 to the APT medium, but even so still exerted 50% of its bacteriocinogenic activity. It was concluded that Lc. lactis subsp. cremoris CTC 204 and Lc. lactis subsp. hordinae CTC 484 exhibited adequate properties for use as biopreservatives in foods and that more extensive studies related to the purification of bacteriocins and assessment of their use in foods should be conducted.

Foram conduzidos dois experimentos no confinamento experimental do Departamento de Zootecnia da ESALQ/USP com o objetivo de estudar os efeitos de diferentes aditivos em dietas para bovinos terminados em confinamento. As dietas continham 78,1% de milho moído fino, 12% feno de Tifton-85, 6% melaço de cana, 2,5% mineral, 1,4% uréia e os respectivos aditivos. No experimento 1 foram utilizados 134 tourinhos da raça Nelore com 24 meses de idade e 330 kg de peso corporal médio inicial, alocados em 24 baias por 102 dias. Os tratamentos foram: 1) Controle (sem aditivo), 2) Monensina (30 ppm), 3) Monensina (20 ppm) + Virginiamicina (15 ppm), 4) Monensina (30 ppm) + Virginiamicina (15 ppm), 5) Virginiamicina (17 ppm), 6) Salinomicina (13 ppm) + Virginiamicina (15 ppm). No experimento 2 foram utilizados 93 tourinhos da raça Nelore com 24 meses de idade e 320 kg de peso corporal médio inicial, alocados em 16 baias por 109 dias. Os tratamentos foram: 1) Controle (sem aditivo), 2) Monensina (30 ppm), 3) Levedura (Saccharomyces cerevisiae; 109 UFC/g) 10g/animal/dia, 4) Combinação dos microrganismos probióticos Bifidobacterium bifidum (3,33 x 106 UFC/g), Lactobacillus acidophilus (3,33 x 106 UFC/g), Lactobacillus plantarum (1,66 x 106 UFC/g), Enterococcus faecium (1,66 x 106 UFC/g) e Saccharomyces cerevisiae (3,33 x 105 UFC/g) FLORAFORT® na dosagem de 3g/animal/dia. No experimento 1, a suplementação com monensina sódica exclusiva e em combinação com virginiamicina reduziu (P<0,1) a ingestão de MS (9,89; 9,20; 9,29; 8,98; 9,76; 9,85). O ganho de peso diário dos animais (1,33; 1,33; 1,39; 1,44; 1,45; 1,49) não foi afetado pelos tratamentos (P>0,05). Em comparação com o tratamento controle, a suplementação com Monensina (30 ppm) + Virginiamicina (15 ppm) aumentou a eficiência alimentar dos animais (P<0,05). O uso de aditivos antibióticos foi efetivo (P<0,05) para aumentar os valores de energia líquida de ganho das dietas (1,05; 1,14; 1,17; 1,26; 1,15; 1,16). No experimento 2, a monensina sódica reduziu a ingestão de MS (P<0,1) em comparação com os demais tratamentos (9,35; 8,25; 9,33; 9,20). O ganho de peso diário, a eficiência alimentar e a densidade energética das dietas não foram afetados pela monensina sódica e pelos probióticos testados (P>0,05). / Two trials were conducted at the ESALQ/USP Animal Science Department experimental feedlot to evaluate the effect of different feed additives in feedlot finished cattle. The animals were fed with 78.1% ground corn, 12% Tifton hay, 6% sugarcane molasses, 2.5% mineral, 1.4% urea and the respective additives. The first trial had 134 Nellore bulls with an average of 24 months and 330 Kg of initial body weight, allocated in 24 pens for 102 days. Treatments were: 1) Control (without additive), 2) Monensin (30 ppm), 3) Monensin (20 ppm) + Virginiamycin (15 ppm), 4) Monensin (30 ppm) + Virginiamycin (15 ppm), 5) Virginiamicyn (17 ppm), 6) Salinomicyn (13 ppm) + Virginiamicyn (15 ppm). The second trial had 93 Nellore bulls with an average of 24 months and 320 Kg of initial body weight, allocated in 16 pens for 109 days. Treatments were: 1) Control (without additive), 2) Monensin (30 ppm), 3) Yeast (Saccharomyces cerevisiae) 10g/animal/day 109 UFC/g, 4) Combination of probiotic microorganisms: Bifidobacterium bifidum (3.33 x 106 UFC/g), Lactobacillus acidophilus (3.33 x 106 UFC/g), Lactobacillus plantarum (1.66 x 106 UFC/g), Enterococcus faecium (1.66 x 106 UFC/g) and Saccharomyces cerevisiae (3.33 x 105 UFC/g) FLORAFORT® 3g/animal/day. In the first trial, the results showed that monensin alone or in combination with virginiamycin decreased (P<0.1) dry matter intake (9.89; 9.20; 9.29; 8.98; 9.76; 9.85). The average daily gain(1.33; 1.33; 1.39; 1.44; 1.45; 1.49) was not affected by the treatments (P>0.05). In comparison to the control diet, the monensin (30 ppm) + virginiamycin (15 ppm) supplementation increased feed efficiency (P<0.05). The use of antibiotic additives was effective (P<0.05) to increase the diet energy values (1.05; 1.14; 1.17; 1.26; 1.15; 1.16). In the second trial, monensin decreased dry matter intake (P<0.1) in comparison to the other treatments (9.35; 8.25; 9.33; 9.20). Average daily gain, feed efficiency and diet energy density were not affected by monensin and probiotics (P>0.05).