Thesis (Ph. D.)--University of Wisconsin--Madison, 1964. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 75-81).

Bibliography: leaves 200-214. Three structurally related compounds, 2-acetyltetrahydropyridine (ACTPY), 2-ethyltetrahydropyridine (ETPY) and N-heterocycle, 2-acetyl-1-pyrroline (ACPY), were quantified and found to be unique components of mousy wines. 35 lactic acid bacteria (LAB) were screened for the ability to produce mousy off-flavour. In addition to Lactobacillus brevis and L. cellobiosus, a diversity of LAB species, particularly heterofermentative Lactobacillus spp. and Oenococcus oeni exhibited this ability in a range of ethanolic and wine-based media. The substrates and metabolism of mousy compound formation by LAB were also investigated. A pathway for the formation of ACPY and ACTPY by heterofermentative LAB was proposed.

Enterobacter sakazakii has been associated with powdered infant formula outbreaks which caused high mortality rate illnesses in infants in recent years. Current research was mainly focused on searching for natural antimicrobial agents which may be incorporated into baby foods to control this emerging pathogen. Yogurt and muscadine products were used in this study. The antimicrobial effects of yogurt were evaluated on agar plates and in a simulated gastrointestinal model. In the agar spot tests, diluted yogurt sample containing lactic acid bacteria at 106 CFU/mL was antagonistic toward E. sakazakii. However, the antimicrobial effect of yogurt on E. sakazakii in the simulated GI model was not noted. Certain numbers of tested E. sakazakii and lactic acid bacteria in yogurt were able to survive the acidic gastric condition and recovered in the intestinal model. By measuring the viable E. sakazakii cells in liquid cultures, the strong antimicrobial activities of muscadine juices and muscadine seed extracts were demonstrated. Within two hours, all inoculated E. sakazakii at 106 CFU/mL were decreased to non-detectable level. Juice and seed extract from dark-skinned muscadine demonstrated stronger antimicrobial activities than those form white-skinned muscadine. The characteristics of muscadine juices and seed extracts were also analyzed. The high phenolics and organic acid contents, such as ellagic, gallic, tannic, and tartaric acids in muscadine were correlated to the inhibitory effect observed.

Please read the abstract in the section 00front of this document / Dissertation (MSc Agric (Food Science))--University of Pretoria, 1999. / Food Science / unrestricted

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Lactic acid bacteria (LAB) are common inhabitants of the human gastro-intestinal tract (GIT). Some LAB, especially lactobacilli, are well known for their application in fermented foods and probiotic properties. These microorganisms exert many beneficial effects on human health, such as digestion and assimilation of food and preventing pathogens colonising the GIT. Furthermore, some selected probiotic strains are believed to perform a critical role in the treatment of gastro-intestinal disorders, lactose intolerance and in the stimulation of the immune system. Despite the ever increasing consumer interest in probiotic LAB, the mechanisms by which they exert their beneficial effects and the activities of probiotics in the GIT often remain poorly understood. Understanding survival mechanisms of LAB in the GIT, especially the interaction between LAB and pathogens, would be facilitated by the direct in vivo monitoring of these processes. Using the mCherry fluorescence gene, we successfully constructed Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA reporter strains. With this study we showed that fluorescence imaging can be used to detect Lb. plantarum 423 and Ent. mundtii ST4SA in the GIT of mice. The two species colonized the cecum and colon for at least 24 h after one oral administration. To our knowledge, this is the first report on fluorescence imaging of LAB expressing mCherry in a mouse model. Using a bioluminescence marker system, we evaluated the impact of Lb. plantarum 423 and Ent. mundtii ST4SA on orally acquired Listeria monocytogenes infection and the ability of the probiotics to compete with the pathogen in the GIT of mice. Challenging Lb. plantarum 423 and Ent. mundtii ST4SA that were already established in the GIT of mice with L. monocytogenes EGDe had no effect on the survival and persistence of the probiotic strains. We demonstrated that the colonization of mice with Lb. plantarum 423 and Ent. mundtii ST4SA, or a combination of the strains, protected the animals against colonization of the GIT by L. monocytogenes EGDe. Enterococcus mundtii proved more effective than Lb. plantarum 423 in reducing the number of L. monocytogenes EGDe in the mouse model. / AFRIKAANSE OPSOMMING: Melksuurbakterieë (MSB) kom algemeen in die mens se spysverteringkanaal (SVK) voor. Verskeie MSB, veral lactobacilli, is bekend vir hul gebruik in gefermenteerde voedsel en as probiotika. Die bakterieë het baie eienskappe wat die mens se gesondheid kan bevoordeel, insluitend vertering en assimilasie van voedsel en voorkoming van kolonisering van die SVK deur patogeniese bakterieë. Sekere probiotiese rasse speel ook ʼn belangrike rol in die behandeling van SVK versteurings, laktose intoleransie en die stimulering van die immuun stelsel. Alhoewel die belangstelling in probiotiese bakterieë toeneem, is daar min inligting bekend oor die meganismes wat MSB gebruik om hulle voordelige eienskappe in die SVK uit te voer. Die oorlewing van MSB in die SVK, veral die interaksies tussen MSB en patogene, kan met behulp van ʼn in vivo moniteringsisteem bestudeer word. Deur die mCherry fluorisensie geen in Lactobacillus plantarum 423 en Enterococcus mundtii ST4SA te kloneer, het ons daarin geslaag om ʼn effektiewe verklikker sisteem te ontwikkel en kon die voorkoms en migrasie van die twee spesies in die SVK van muise bestudeer word. Lactobacillus plantarum 423 en Ent. mundtii ST4SA het veral die blindederm en kolon gekoloniseer. Beide rasse het na ʼn enkele mondelingse toediening vir ten minste 24 h in die SVK oorleef. Sover ons kennis strek, is hierdie die eerste verslag van fluoriserende MSB wat met behulp van die mCherry geenproduk in die SVK bestudeer is. Deur gebruik te maak van ʼn bioliggewende verklikker sisteem, het ons die vermoë van Lb. plantarum 423 en Ent. mundtii ST4SA om met Listeria monocytogenes in die SVK te kompeteer, bestudeer. Listeria monocytogenes het geen invloed gehad op die kolonisering van Lb. plantarum 423 en Ent. mundtii ST4SA nie. Deur die muise vooraf met Lb. plantarum 423 en Ent. mundtii ST4SA te koloniseer (in kombinasie of met net een van die twee rasse), kon ons daarin slaag om kolonisering van die SVK met L. monocytogenes te voorkom. In die muis model wat in hierdie studie gebruik is, was Ent. mundtii ST4SA meer effektief as Lb. plantarum 423 in die verlaging van Listeria selgetalle.

Thesis (MScEng (Process Engineering))--University of Stellenbosch, 2007. / Lactic acid bacteria (LAB) are producers of bacteriocins, ribosomally synthesized antimicrobial peptides. Bacteriocins are secreted into the surrounding environment where they inhibit growth of other bacteria competing for the same nutrients in a particular environment, usually closely related strains. Some of the bacteriocin-sensitive bacteria include food spoilers and - pathogens, which makes bacteriocins potential natural food preservatives. The need for more natural preservation techniques in the food industry is high: Consumers prefer ready-toeat, minimally processed foods containing no chemical preservatives, but at the same time food spoilage and food-related illnesses are areas of big concern. The antibacterial and antiviral properties of some bacteriocins have also made them suitable for controlling bacterial infections, e.g. as part of pharmaceutical ointments. The increasing rate of resistance against antibiotics by micro-organisms has created a market for alternative treatments for infections. Commercial bacteriocin manufacturing proceeds in controlled fermentations or by extraction from plant material. Enterococcus mundtii ST4SA produces a bacteriocin, bacST4SA, with properties giving it potential for use as a food preservative or as part of a pharmaceutical product. In this study, production of bacST4SA by fermentation of low-cost food-grade growth media, sugarcane molasses, corn steep liquor (CSL) and cheese whey, was considered to increase the economic viability of production for food application. Furthermore, individual de Man Rogosa and Sharpe (MRS) medium components, pH and fed-batch fermentation were evaluated to improve bacST4SA activity...

Thesis (PhD)--Stellenbosch University, 2008. / ENGLISH ABSTRACT: Research on probiotics has increased over the past years, which led to commercialization of a number of probiotic supplements and functional foods. In vitro assays such as tolerance to acid and bile, adhesion to mucus and epithelial cells, antimicrobial activity and antibiotic resistance tests are performed to screen lactic acid bacteria for probiotic properties. Enterococcus mundtii ST4SA produces an antimicrobial peptide (peptide ST4SA) with activity against Gram-positive and Gram-negative bacteria. Lactobacillus plantarum 423 produces plantaricin 423, a typical class II bacteriocin, active against a number of Gram-positive bacteria. A gastro-intestinal model (GIM) simulating the gastro-intestinal tract (GIT) of infants, was developed to study the survival of E. mundtii ST4SA and L. plantarum 423 and evaluate them as possible probiotics. Growth of the two strains in the GIM was compared to the growth of commercially available probiotics. Infant milk formulations were used as growth medium. Changes in pH, the addition of bile salt and pancreatic juice, and intestinal flow rates were controlled by peristaltic pumps linked to a computer with specifically designed software. Strain ST4SA was sensitive to low pH and high concentrations of bile salts. Growth of strain ST4SA was repressed in the first part of the GIM, however, the cells recovered in the ileum. Strain 423 was also sensitive to acidic conditions. However, the cells withstood the presence of bile and pancreatin in the first part of the GIT. Neither of the two strains displayed bile salt hydrolase (BSH) activity. Both strains were resistant to amoxicillin, ampicillin, chloramphenicol, cefadroxil, roxithromycin, meloxicam, doxycycline, erythromycin, novobiocin, rifampicin, tetracyclin, bacitracin, oflaxacin and cephazolin, anti-inflammatory drugs Na+- diklofenak and ibuprofen, and painkillers codeine terprim hydrate aminobenzoic acid, metamizole aspirin and paracetamol. Strain 423 was resistant to ciprofloxacin. Genes encoding cytolysin, non-cytolysin β-hemolysin and cell aggregation substances were detected on the genome of strain ST4SA but they were not expressed. L. plantarum 423 does not contain genes encoding gelatinase, cell aggregation, enterococcus surface protein, hemolysin, non-cytolysin β- hemolysin and enterococcus endocarditis antigen. Both strains inhibited the growth of Listeria monocytogenes ScottA in the GIM. Survival of the strains improved when used in combination and compared well with the survival of commercially available probiotics. Adhesion to epithelial cells is an important prerequisite for bacterial colonization in the GIT. The adhesion of E. mundtii ST4SA and L. plantarum 423 was studied using Caco-2 (human colon carcinoma epithelial) cells. Both strains revealed good adhesion compared to other probiotic strains. No correlation was found between hydrophobicity, auto-aggregation and adhesion to Caco-2 cells. Antibiotics and anti-inflammatory medicaments had a negative effect on adhesion. Different combinations of proteins were involved in the adhesion of E. mundtii ST4SA and L. plantarum 423 to Caco-2 cells. E. mundtii ST4SA, L. plantarum 423 and L. monocytogenes ScottA were stained with fluorescent dyes to visualize adhesion to Caco-2 cells. Adhesion of L. monocytogenes ScottA to Caco-2 cells was not reduced in the presence of strains ST4SA and 423. Cell-free culture supernatants of both strains inhibited the invasion of L. monocytogenes ScottA. The cell structure of Caco-2 cells changed in the presence of L. monocytogenes ScottA. Strains ST4SA and 423 protected Caco-2 cells from deforming. / AFRIKAANSE OPSOMMING: Navorsing op probiotika het die afgelope tyd drasties toegeneem en aanleiding gegee tot die kommersialisering van ‘n groot hoeveelheid probiotiese supplemente en funksionele voedselsoorte. In vitro studies, soos bv. weerstand teen suur en gal, vashegting aan mukus en epiteelselle, antimikrobiese aktiwiteit en weerstand teen antibiotika word uitgevoer om te bepaal of melksuurbakteriëe aan probiotiese standaarde voldoen. Enterococcus mundtii ST4SA produseer ’n peptied met antimikrobiese werking teen Grampositiewe en Gram-negatiewe bakteriëe. Lactobacillus plantarum 423 produseer ‘n tipiese klas II bakteriosien, plantarisien 423, met aktiwiteit teen sekere Gram-positiewe bakteriëe. ’n Gastro-intestinale model (GIM) wat die spysverteringskanaal (SVK) van babas simuleer, is ontwikkel om die oorlewing van E. mundtii ST4SA en L. plantarum 423 te bepaal en hul eienskappe met dié van kommersiële probiotiese stamme te vergelyk. Babamelk formules is as groeimedium gebruik. Verandering in pH, byvoeging van galsoute en pankreassappe, en intestinale vloei is met behulp van peristaltiese pompe gereguleer wat seine vanaf ‘n spesiaal ontwikkelde rekenaarprogram ontvang. E. mundtii ST4SA was sensitief vir lae pH en hoë galsoutkonsentrasies en groei is in die eerste deel van die GIM onderdruk. Selgetalle het wel in die ileum herstel. Stam 423 was ook sensitief vir lae pH, maar het die galsout- en pankreatienvlakke in die laer deel van die SVK weerstaan. Geen galsout-hidrolase aktiwiteit is by enige van die twee stamme gevind nie. Beide stamme het weerstand getoon teen amoksillien, ampisillien, chloramfenikol, cefadroksiel, roksitromisien, meloksikam, doksisiklien, eritromisien, novobiosien, rifampisien, tetrasiklien, basitrasien, oflaksasien, kefazolien, die anti-inflammatoriese medikamente Na+-diklofenak en ibuprofen, en die pynstillers kodeïenterprimhidraataminobensoësuur, metamisoolaspirien en parasetamol. L. plantarum 423 was bestand teen ciprofloksasien. Gene wat kodeer vir sitolisien, nie-sitolisien β-hemolisien III en sel-aggregasie is op die genoom van E. mundtii ST4SA gevind, maar word nie uitgedruk nie. L. plantarum 423 besit nie die gene wat vir gelatinase, selaggregasie substansies, enterokokkus selwandproteïen, hemolise, nie-sitolisien β-hemolisien en enterokokkus endokarditis antigeen kodeer nie. Albei stamme inhibeer die groei van Listeria monocytogenes ScottA in die GIM. Die twee stamme in kombinasie het tot beter oorlewing in die GIM gelei. Stamme ST4SA en 423 vergelyk goed met kommersieël beskikbare probiotika. Vashegting van probiotiese stamme aan epiteelselle is belangrik vir kolonisering in die SVK. Vashegting van E. mundtii ST4SA en L. plantarum 423 is bestudeer deur van Caco-2 (kolon epiteel) selle van die mens gebruik te maak. Die aanhegting van beide stamme aan Caco-2 selle het goed vergelyk met kommersieël beskikbare probiotiese stamme. Geen korrelasie is gevind tussen hidrofobisiteit, aggregasie en vashegting aan Caco-2 selle nie. Antibiotika en antiinflammatoriese medikamente het ‘n negatiewe effek op vashegting gehad. Verskillende kombinasies van proteïene is betrokke in die vashegting van E. mundtii ST4SA en L. plantarum 423 aan Caco-2 selle. E. mundtii ST4SA, L. plantarum 423 en L. monocytogenes ScottA is met fluoreserende kleurstowwe gemerk om vashegting aan Caco-2 selle te monitor. Vashegting van L. monocytogenes ScottA aan Caco-2 selle is nie deur die teenwoordigheid van stamme ST4SA en 423 beïnvloed nie. Sel-vrye kultuursupernatante van beide stamme het die binnedring van L. monocytogenes ScottA verhoed. Die selstruktuur van Caco-2 selle het in die teenwoordigheid van L. monocytogenes ScottA van vorm verander. E. mundtii ST4SA en L. plantarum 423 het die Caco-2 selle teen vervorming beskerm.

Several lactic acid bacteria (LAB) of the Lactococcus, Lactobacillus, Leuconostoc and Pediococcus genera were screened for inhibition of food-borne pathogens and spoilage microorganisms in raw milk and dairy products. Listeria monocytogenes was killed by Lactococcus lactis subsp. lactis and Pediococcus pentosaceus due to their production of bacteriocin-type inhibitors. Staphylococcus aureus was not able to grow in raw milk at temperatures below 5°C even without LAB being present. Gram negative Salmonella enteritidis. Salmonella typhimurium and Escherichia coli, along with spoilage bacteria of the genus Pseudomonas were dramatically inhibited by a Lactobacillus species, designated AS-1, in raw and pasteurized milk as well as in cottage cheese. However, other LAB were not able to inhibit these organisms. Lactobacillus AS-1, did not produce hydrogen peroxide but carbon dioxide was produced. The AS-1 strain was a gram positive coccobacillus, catalase and oxidase negative and produced DL-lactic acid. It deaminated arginine and grew over a temperature range of 5°C to 45°C. It was also able to ferment glucose, galactose, fructose and lactose in addition to 17 other carbohydrates. High numbers (107 CFU/ml) of AS-1 were required to obtain complete inhibition of gram negative bacteria. A selective medium (ASLM) for Listeria monocytogenes was developed to follow the fate of this particular pathogen in association with LAB in raw milk; other selective media were not able to inhibit the growth of background flora of raw milk. ASLM was superior to four other media in allowing only the growth of the target pathogen. For the Lactococcus genus, a selective and differential agar medium (Alsan) was formulated to selectively allow growth of Lactococcus spp. and to differentiate between Lactococcus lactis subsp. lactis and the biovariety diacetylactis, based on citrate utilization. / Graduation date: 1993

潰瘍性結腸炎 (UC) 是一種原因不明的炎症性腸道疾病，治療原則是減輕炎症，但UC的病因有多種，一般消炎藥物如柳氮磺胺吡啶 (sulfasalazine) 等治療都是單靶向並有嚴重副作用，故副作用低、多靶向藥物是必要的。 / Cathelicidin 是一種抗菌抗炎的肽。事實上，鼠的 cathelicidin (mCRAMP) 直腸給藥能緩解小鼠 UC。為了提高療效及方便給藥，mCRAMP 編碼被導入乳酸乳球菌中。乳酸乳球菌是一種能抵抗胃酸的乳酸益生菌，因此口服亦能生產及傳送 cathelcidin 到大腸。 / 小鼠用含3% 葡聚醣硫酸鈉 (DSS) 的水7天以誘導UC。小鼠隨機分為十組，各接受每日一次的口服製劑：(1) 水，(2) DSS，(3, 4) DSS + 10¹° cfu 有或沒有 nisin 誘導的乳酸乳球菌，(5-8) DSS + 10⁸ 或 10¹° cfu有 (N4I) 或沒有 nisin 誘導的 mCRAMP 編碼乳酸乳球菌，(9) DSS + 0.5% 羧甲基纖維素鈉 (CMC-Na) 及 (10) DSS + 600 mg/kg懸浮於0.5% CMC-Na 的 sulfasalazine。 / 研究對 UC 預防效果時，小鼠同時接受 DSS 及治療。所有益生菌製劑中，只有 N4I 能降低中性粒細胞浸潤、脂質過氧化和炎症細胞因子表達，同時保護腸隱窩及黏膜分泌層結構，減少細胞凋亡及腸道菌群。相比之下，sulfasalazine 能抑制炎症但不能阻止結腸結構損傷。 / 進一步研究治療效果時，小鼠在炎症形成後接受四天治療。N4I 能促進結腸黏膜恢復，改善結腸和黏液分泌層的結構。這些作用可能通過刺激細胞增殖和抑制凋亡造成。相對地，sulfasalazine 對結腸組織重組沒有影響。 / 為了研究 mCRAMP 直接消炎作用，小鼠巨噬細胞 RAW 264.7 被脂磷壁酸和脂多醣刺激以模仿 UC 時細菌引起的炎症。mCRAMP 能減輕腫瘤壞死因子-α分泌及IκBα磷酸化並抑制核因子-κB (NF-κB) 活化，炎症酶如誘導型一氧化氮合酶和環氧合酶-2的表達也減少了。mCRAMP可能直接抑制細菌毒素與受體結合和/或直接抑制 NF-κB 產生消炎作用。 / 在研究 mCRAMP 修復黏膜的作用中，證實 mCRAMP 通過 G 蛋白偶聯受體依賴途徑和間接激活表皮生長因子受體、激活下游絲裂原活化蛋白激酶而促進細胞遷移、加速癒合。 / 總括而言，本研究首次顯示mCRAMP編碼乳酸乳球菌對 UC 有保護和治療作用，其抗炎、抗菌及促進黏膜修復作用來自乳酸乳球菌分泌的mCRAMP。多靶向的mCRAMP編碼乳酸乳球菌具有很大潛力，是一種比標準藥物 sulfasalazine 更好的治療結腸炎製劑。 / Ulcerative colitis (UC) is an idiopathic inflammatory bowel disease (IBD). The mainstay of drug treatment is to relieve inflammation. However the aetiology of UC is multi-factorial while most of the anti-inflammatory drugs, such as sulfasalazine, aim at single target with severe side effects. Therefore, a multi-targeted drug with low systemic toxicity is warranted. / Cathelicidin, a host defense peptide, shows anti-microbial and anti-inflammatory effects. Indeed intra-rectal administration of mouse cathelicidin (mCRAMP) alleviated murine colitis. To improve therapeutic efficacy and reduce inconvenience of administration, Lactococcus lactis (L. lactis) was constructed to encode cathelicidin. L. lactis is a lactic acid probiotic which could resist gastric acid and be able to produce and deliver cathelicidin to the colon when given orally. / Murine colitis was induced by 3% dextran sulphate sodium (DSS) given in drinking water for 7 days. Mice were given intragastrically with the following preparations once daily: (1) water, (2) DSS, (3, 4) DSS + 10¹° cfu L. lactis with or without nisin induction, (5-8) DSS + 10⁸ or 10¹° cfu mCRAMP-encoding L. lactis with (N4I) or without nisin induction, (9) DSS + 0.5% sodium carboxymethylcellulose (CMC-Na) and (10) DSS + 600 mg/kg sulfasalazine suspended in 0.5 % CMC-Na. / To study the preventive effects, mice received the above treatments together with DSS administration. N4I but not the other probiotic preparations suppressed inflammation by reducing neutrophil infiltration, lipid peroxidation and inflammatory cytokines expressions. Crypt structure and mucus-secreting layer were conserved together with the reduction of apoptosis and intestinal microbiota. In contrast, sulfasalazine could only suppress inflammation but not the destruction of colonic structure. / To further examine the therapeutic effects, mice received treatments for 4 consecutive days after the inflammation formation. Similarly, only N4I promoted colonic mucosal recovery and preserved colon structure and mucus-secreting layer. These actions are likely mediated through cell proliferation stimulation and apoptosis suppression. Again, sulfasalazine had no effects on colon tissue reconstitution. / The direct anti-inflammatory action of mCRAMP was also studied. Mouse macrophage RAW 264.7 cells were stimulated by lipoteichoic acid and lipopolysaccharide to mimic bacteria-induced inflammation during UC. mCRAMP prevented tumour necrosis factor-α secretion and IκBα phosphorylation followed by nuclear factor-κB (NF-κB) suppression. The inflammatory enzymes including inducible nitric oxide synthase and cyclooxygenase-2 were also reduced. It was postulated that mCRAMP might directly interact with the bacterial toxins to reduce receptor complex binding and/or reduce NF-κB suppression in macrophages. / The repairing action of mCRAMP on mucosal damage was studied in mouse colon cells. mCRAMP incubation reduced the wound size by promoting cell migration through the G-protein coupled receptor and epidermal growth factor receptor transactivation followed by the mitogen-activated protein kinases activation. / In conclusion, the present study demonstrates for the first time the protective and therapeutic roles of mCRAMP-encoding L. lactis in UC. It was the mCRAMP secreted from the probiotic to produce both anti-inflammatory and anti-bacterial actions and further promote mucosal repair. mCRAMP-encoding L. lactis is a multi-targeted agent for IBD. It has a great potential to be a new therapeutic agent better than sulfasalazine for the treatment of UC. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Wong, Ching Man. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 227-250). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.i / Acknowledgement --- p.v / Table of Content --- p.vi / List of Abbreviations --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Ulcerative Colitis --- p.1 / Chapter 1.1.1 --- Overview --- p.1 / Chapter 1.1.2 --- Epidemiology --- p.2 / Chapter 1.1.3 --- Diagnosis --- p.2 / Chapter 1.1.3.1 --- Clinical Presentation --- p.2 / Chapter 1.1.3.2 --- Apparative Diagnostics --- p.3 / Chapter 1.1.3.3 --- Innovative Diagnostics in IBD --- p.4 / Chapter 1.1.4 --- Etiopathogenesis --- p.4 / Chapter 1.1.4.1 --- Genetic Predisposition --- p.4 / Chapter 1.1.4.2 --- Environmental Factors --- p.5 / Chapter 1.1.4.2.1 --- Life Style --- p.5 / Chapter 1.1.4.2.1.1 --- Smoking --- p.5 / Chapter 1.1.4.2.1.2 --- Diet --- p.6 / Chapter 1.1.4.2.1.3 --- Hygiene --- p.6 / Chapter 1.1.4.2.1.4 --- Psychological Stress --- p.6 / Chapter 1.1.4.2.1.5 --- Appendectomy --- p.7 / Chapter 1.1.4.2.2 --- Colonic Mucus --- p.7 / Chapter 1.1.4.2.3 --- Non-steroidal Anti-inflammatory Drugs (NSAIDs) --- p.7 / Chapter 1.1.4.3 --- Alteration of Intestinal Microbiota --- p.8 / Chapter 1.1.4.4 --- Immune Factors --- p.10 / Chapter 1.1.5 --- Existing Treatments --- p.10 / Chapter 1.1.5.1 --- 5-Aminosalicyclic Acid --- p.11 / Chapter 1.1.5.2 --- Corticosteroids --- p.12 / Chapter 1.1.5.3 --- Immunomodulators --- p.12 / Chapter 1.1.5.4 --- Surgical Management --- p.13 / Chapter 1.1.6 --- Emerging Treatments --- p.14 / Chapter 1.1.6.1 --- Antibiotics --- p.14 / Chapter 1.1.6.2 --- Probiotics --- p.14 / Chapter 1.1.6.3 --- Nicotine Patches --- p.14 / Chapter 1.1.6.4 --- Butyrate --- p.14 / Chapter 1.1.6.5 --- Biological Therapies --- p.15 / Chapter 1.1.7 --- Risk of Colorectal Cancer --- p.17 / Chapter 1.2 --- Cathelicidin --- p.17 / Chapter 1.2.1 --- Cathelicidin Family --- p.17 / Chapter 1.2.2 --- Actions and Possible Mechanisms --- p.19 / Chapter 1.2.3 --- Cathelicidin in Ulcerative Colitis --- p.20 / Chapter 1.3 --- Probiotics --- p.21 / Chapter 1.3.1 --- Lactic Acid Bacteria --- p.21 / Chapter 1.3.2 --- Definition of Probiotics --- p.22 / Chapter 1.3.3 --- Possible Mechanisms of Action of Probiotics --- p.23 / Chapter 1.3.3.1 --- Mucous Layer --- p.23 / Chapter 1.3.3.2 --- Host Cell Antimicrobial Peptides --- p.25 / Chapter 1.3.3.3 --- Probiotic Antimicrobial Factors --- p.25 / Chapter 1.3.3.4 --- Epithelial Adherence --- p.27 / Chapter 1.3.4 --- Recent Findings in Ulcerative Colitis Treatment --- p.28 / Chapter 1.4 --- Lactococcus lactis --- p.29 / Chapter 1.4.1 --- Overview --- p.29 / Chapter 1.4.2 --- Gene Expression System --- p.30 / Chapter 1.4.3 --- Nisin-Inducible Controlled Gene Expression (NICE) System --- p.31 / Chapter 1.4.4 --- Recent Studies of Treating Ulcerative Colitis with L. lactis and Recombinant L. lactis --- p.32 / Chapter 1.4.5 --- Safety Concern of the Use of Probiotics and Transgenic Probiotics --- p.33 / Chapter 1.5 --- Aims --- p.35 / Chapter Chapter 2 --- Material and Methodology --- p.36 / Chapter 2.1 --- General Materials --- p.36 / Chapter 2.1.1 --- Chemicals --- p.36 / Chapter 2.1.2 --- Antibodies and Commercial Kits --- p.41 / Chapter 2.1.3 --- Bacteria --- p.43 / Chapter 2.1.4 --- Animals --- p.43 / Chapter 2.1.5 --- Cell Lines --- p.44 / Chapter 2.1.5.1 --- Mouse Colonic Epithelial Cells --- p.44 / Chapter 2.1.5.2 --- Mouse Macrophages --- p.44 / Chapter 2.2 --- Experimental Designs --- p.45 / Chapter 2.2.1 --- Construction of mCRAMP-Encoding Lactococcus lactis --- p.45 / Chapter 2.2.1.1 --- Enumeration of L. lactis --- p.45 / Chapter 2.2.1.2 --- Bacteriostatic Effect of mCRAMP on L. lactis --- p.46 / Chapter 2.2.1.3 --- Construction of mCRAMP-Encoding L. lactis --- p.46 / Chapter 2.2.1.4 --- Detection of mCRAMP Production by Western Immunoblotting --- p.50 / Chapter 2.2.2 --- In vivo Studies --- p.52 / Chapter 2.2.2.1 --- Survival of mCRAMP-Encoding L. lactis in Murine Colon --- p.52 / Chapter 2.2.2.2 --- Toxicity of mCRAMP-Encoding L. lactis --- p.53 / Chapter 2.2.2.3 --- Determination of mCRAMP Expression in Colon Tissue --- p.53 / Chapter 2.2.2.4 --- Induction of Colitis --- p.54 / Chapter 2.2.2.5 --- Probiotic and Sulfasalazine Treatment --- p.54 / Chapter 2.2.2.6 --- Clinical Symptoms --- p.56 / Chapter 2.2.2.7 --- Morphological Analysis --- p.56 / Chapter 2.2.2.7.1 --- Haematoxylin-Eosin (H&E) Staining --- p.56 / Chapter 2.2.2.7.2 --- Periodic Acid-Schiff (PAS) Staining --- p.58 / Chapter 2.2.2.8 --- Assessment of Apoptosis and Proliferation by Immunohistochemistry --- p.60 / Chapter 2.2.2.8.1 --- Determination of Cell Apoptosis by Terminal Deoxynucleotidyl Transferase dUTP Nick-end Labeling --- p.60 / Chapter 2.2.2.8.2 --- Determination of Cell Proliferation by Proliferating Cell Nuclear Antigen (PCNA) Staining --- p.61 / Chapter 2.2.2.9 --- Determination of the Degree of Inflammation --- p.63 / Chapter 2.2.2.9.1 --- Colonic Myeloperoxidase (MPO) Activity --- p.63 / Chapter 2.2.2.9.2 --- Colonic Malondialdehyde (MDA) Level --- p.63 / Chapter 2.2.2.10 --- Fecal Microbiota Count --- p.64 / Chapter 2.2.2.11 --- mRNA Expression of Inflammatory Cytokines --- p.64 / Chapter 2.2.3 --- In vitro Studies --- p.66 / Chapter 2.2.3.1 --- Determination of Anti-inflammatory Effects of mCRAMP --- p.66 / Chapter 2.2.3.1.1 --- Cell Viability --- p.66 / Chapter 2.2.3.1.2 --- Determination of TNF-α Secretion under Stimulation of LTA and LPS --- p.66 / Chapter 2.2.3.1.3 --- Effects of mCRAMP on TNF-α Secretion Under Stimulation of LTA or LPS --- p.67 / Chapter 2.2.3.1.4 --- Effects of Pertussis Toxin (PTX) on the Inhibition of TNF-α Secretion by mCRAMP --- p.67 / Chapter 2.2.3.1.5 --- Nuclear Factor-κB (NF-κB) Luciferase Reporter Gene Assay in RAW 264.7 cells --- p.68 / Chapter 2.2.3.1.6 --- Determination of IκBα Expression and Phosphorylation by Western Immunoblotting --- p.69 / Chapter 2.2.3.1.7 --- Determination of Inducible Nitric Oxide Synthases (iNOS) and Cyclooxygenase-2 (COX-2) Expression by Western Immunoblotting --- p.70 / Chapter 2.2.3.2 --- Determination of Wound Healing Effects of mCRAMP --- p.72 / Chapter 2.2.3.2.1 --- Cell Viability --- p.72 / Chapter 2.2.3.2.2 --- Cell Migration --- p.74 / Chapter 2.2.3.2.3 --- Determination of Epidermal Growth Factor Receptor (EGFR), Extracellular Signal-Regulated Protein Kinase (ERK1/2) and p38 Expression and Phosphorylation by Western Immunoblotting --- p.76 / Chapter 2.3 --- Statistical Analysis --- p.76 / Chapter Chapter 3 --- Result --- p.77 / Chapter 3.1 --- Protective Effects of Cathelicidin-Encoding Lactococcus lactis in Murine Ulcerative Colitis --- p.77 / Chapter 3.1.1 --- Introduction --- p.77 / Chapter 3.1.2 --- Results --- p.79 / Chapter 3.1.2.1 --- Survival of mCRAMP-Encoding L. latis in Murine Colon --- p.79 / Chapter 3.1.2.2 --- Detection of mCRAMP-Encoded by L. lactis in vivo --- p.81 / Chapter 3.1.2.3 --- Toxicity of mCRAMP-Encoding L. lactis --- p.84 / Chapter 3.1.2.4 --- Clinical Symptoms --- p.86 / Chapter 3.1.2.5 --- Histology Evaluation --- p.89 / Chapter 3.1.2.6 --- Apoptosis --- p.93 / Chapter 3.1.2.7 --- Determination of mCRAMP Expression in Colon Tissue --- p.96 / Chapter 3.1.2.8 --- Determination of the Degree of Inflammation --- p.101 / Chapter 3.1.2.9 --- Faecal Microbiota Populations --- p.105 / Chapter 3.1.3 --- Discussion --- p.108 / Chapter 3.2 --- Therapeutic Effects of Cathelicidin-Encoding Lactococcus lactis in Murine Ulcerative Colitis --- p.113 / Chapter 3.2.1 --- Introduction --- p.113 / Chapter 3.2.2 --- Results --- p.115 / Chapter 3.2.2.1 --- Clinical Symptoms --- p.115 / Chapter 3.2.2.2 --- Histology Evaluation --- p.117 / Chapter 3.2.2.3 --- Cell Death and Proliferation in Colitis --- p.123 / Chapter 3.2.2.4 --- Determination of mCRAMP Expression in Colon Tissues --- p.127 / Chapter 3.2.2.5 --- Determination of the Degree of Inflammation --- p.130 / Chapter 3.2.2.6 --- Faecal Microbiota Populations --- p.133 / Chapter 3.2.3 --- Discussion --- p.135 / Chapter 3.3 --- Mechanistic Study of the Anti-inflammatory Effects of mCRAMP in Mouse Macrophages --- p.139 / Chapter 3.3.1 --- Introduction --- p.139 / Chapter 3.3.2 --- Results --- p.145 / Chapter 3.3.2.1 --- Viability of Macrophages --- p.145 / Chapter 3.3.2.2 --- Effects of LTA and LPS on Tumour Necrosis Factor-α (TNF-α) Release from Macrophages --- p.150 / Chapter 3.3.2.3 --- The Inhibition of TNF-α Secretion by mCRAMP --- p.153 / Chapter 3.3.2.4 --- Inhibition of TNF-α Secretion by mCRAMP Independent to GPCR Stimulation --- p.158 / Chapter 3.3.2.5 --- Activation of NF-κB Through Detection of Luciferase Activity --- p.163 / Chapter 3.3.2.6 --- Determination of the Expression and Phosphorylation of IκBα by Western Immunoblotting --- p.166 / Chapter 3.3.2.7 --- Determination of iNOS and COX-2 Expression by Western Immunoblotting --- p.169 / Chapter 3.3.2.8 --- The Suppression of iNOS and COX-2 Expression by mCRAMP was Independent to GPCR and P2X₇ Signalling --- p.183 / Chapter 3.3.3 --- Discussion --- p.188 / Chapter 3.4 --- Mechanistic Study on Wound Healing Effect of mCRAMP in Mouse Colon Epithelial Cells --- p.193 / Chapter 3.4.1 --- Introduction --- p.193 / Chapter 3.4.2 --- Results --- p.196 / Chapter 3.4.2.1 --- Cell Viability --- p.196 / Chapter 3.4.2.1.1 --- MTT Assay --- p.196 / Chapter 3.4.2.1.2 --- BrdU Incorporation --- p.200 / Chapter 3.4.2.2 --- Cell Migration --- p.202 / Chapter 3.4.2.3 --- Determination of Epidermal Growth Factor Receptor (EGFR), Extracellular Signal-Regulated Protein Kinase (ERK1/2) and p38 Expression and Phosphorylation by Western Immunoblotting --- p.210 / Chapter 3.4.3 --- Discussion --- p.215 / Chapter 4 Discussion and Future Perspectives --- p.219 / Publications --- p.224 / References --- p.227

A large amount of cheese is lost every year due to mold contamination. Biopreservation, which is the use of biological entities (microbes) and their metabolites to suppress microbial spoilage instead of chemical preservatives has lately gained increasing interest. Lactic acid bacteria (LAB) have the potential for use in biopreservation, because they are safe to consume and naturally exist in many foods. In this study, fifteen strains of lactobacilli isolated from dairy products, vegetables, and fermented pickles were tested by agar overlay assay for their anti-mold activity. Six strains grown on MRS agar showed strong inhibitory activity against a target mold (Penicillium sp. at 105 spores/ml) isolated from the surface of Cheddar cheese. The isolates were identified by biochemical tests using API CHL50 strips. Five strains were identified as Lactobacillus plantarum, and one strain as Pediococcus pentasaceus. Well-diffusion method was used to demonstrate anti-mold activity in concentrated cell-free supernatants. Supernatants from all strains showed inhibition of the target mold (indicator). The anti-mold compound(s) produced by all the strains was heat-resistant (100o C for 15 min). Supernatants from 5 strains retained the anti-mold activity when the pH was adjusted to 6.8 ± 0.2, while one strain DC2 isolated from cheese lost its anti-mold activity at that pH. Temperature of incubation of cultures affected anti-mold activity. The optimum was 37o C. Very little or no inhibition was noted when cultures were incubated at either 10 or 55 °C. A preliminary study of applying anti-mold lactobacilli in Cheddar cheese was completed. Anti-mold LAB was added to the cheese milk as an adjunct to give 105 cfu/ml. After 1-week and 1-month ripening, mold (10~20spores) was added on to the surface, and the cheese was wrapped loosely. The appearance of the mold on cheese surface was monitored. Mold was not present on the 1-week old cheese “NB in milk” until the 6th day after the control cheese (made without strain NB) showed signs of mold. The 1-month old cheese “NB in milk ” extended the shelf life 17 days longer than the control cheese.