Intestinal microbiota in human health and disease: the impact of probiotics

The complex communities of microorganisms that colonise the human gastrointestinal tract play an important role in human health. The development of culture-independent molecular techniques has provided new insights in the composition and diversity of the intestinal microbiota. Here, we summarise the present state of the art on the intestinal microbiota with specific attention for the application of high-throughput functional microbiomic approaches to determine the contribution of the intestinal microbiota to human health. Moreover, we review the association between dysbiosis of the microbiota and both intestinal and extra-intestinal diseases. Finally, we discuss the potential of probiotic microorganism to modulate the intestinal microbiota and thereby contribute to health and well-being. The effects of probiotic consumption on the intestinal microbiota are addressed, as well as the development of tailor-made probiotics designed for specific aberrations that are associated with microbial dysbiosis

Agricultural and industrial waste as a substrate for cellulase and amylase production by novel bacterial strain Paenibacillus chitinolyticus CKS1.

Predmet ove disertacije je iskorišćenje otpadnih sirovina, poljoprivrednog i industrijskog porekla, za proizvodnju enzima celulaza i amilaza pomoću novoizolovanog bakterijskog soja Paenibacillus chitinolyticus CKS1. Optimizacija procesa proizvodnje hidrolitiĉkih enzima celulaza i amilaza vršena je korišćenjem komercijalnih supstrata i otpadnih sirovina ivariranjem uticaja pojedinaĉnih parametara primenom metode planiranog eksperimenta i matematiĉko-statistiĉka obrade eksperimentalnih podataka – metode odzivnih površina (RSM). Proces proizvodnje celulalaza (endoglukanaza i egzoglukanaza) je optimizovan na razliĉitim komercijalnim podlogama ali i na otpadnim sirovinama. Soj CKS1 je pokazao predominantnu aktivnost egzoglukanaza (iskazanih kao Avicelazna aktivnost) što je netipiĉno za većinu bakterijskih vrsta. Rezultati su pokazali da se P. chitinolyticus CKS1 moţe koristiti za dobijanje celulazau komercijalnoj podlozi (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,532 U/mL i 1,83 U/mL, pojedinaĉno), ali i na otpadnim sirovinama: otpadnom lekovitom bilju (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,203 U/mL i 1,94 U/mL, pojedinaĉno) i piljevini (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,145 U/mL i 1,75 U/mL, pojedinaĉno). Soj CKS1 je pokazao mogućnost rasta i proizvodnje dve vrste celulaza: endoglukanaza (iskazanih kao karboksimetil celulazna (CMC-azna) aktivnost i egzoglukanaza (iskazanih kao Avicelazna) aktivnost tokom svog rasta na ĉvrstom otpadnom supstratu- repinom rezancu (maksimalna CMC-azna i Avicelazna aktivnost iznosi 3,159 U/g i 4,840 U/g). Metodom odzivnih površina optimizovana je proizvodnja endoglukanaza i egzoklukanaza u podlozi obogaćenoj jeĉmenim mekinjama (maksimalna CMC-aza i Avicelazna aktivnost iznosi 0,405 U/mL i 0,433 U/mL, pojedinaĉno). Dobijena je nešto niţa vrednostAvicelazne aktivnosti, ali optimizovani procesi jasno ukazuju na mogućnost njihovog korišćenja, pošto su u skladu sa principima odrţive proizvodnje i brige o ţivotnoj sredini. P. chitinolyticus CKS1 soj je pokazao i amilolitiĉku aktivnost, netipiĉnu za vrstu kojoj pripada i stoga je metodom odzivnih površina optimizovana proizvodnja β-amilaza na podlozi dobijenoj korišćenjem otpadne vode iz industrije prerade kartonske ambalaţe (0,333 U/mL) i na podlozi baziranoj na melasi i otpadnom repinom rezancu (2,237 U/mL). U cilju što kompletnije analize podobnosti primene dobijenih enzima u industrijskim procesima, izvršena je karakterizacija proizvoda hidrolize nastalih primenom obe klase enzima.UtvrĊeno je da su proizvodi enzimske hidrolize dobijeni pomoću enizima P. chitinolyticus CKS1 vredni biotehnološki proizvodi – glukoza i maltoza. Primenljivost bakterijskih celulaza i amilaza, koje produkuje soj CKS1 se ogleda u mogućnosti hidrolize pamuĉnog materijala, kao celulozne sirovine i jeĉmenih mekinja,kao lignocelulozno-skrobne sirovine, pri ĉemu se oslobaĊa 0,589 mg/mL i 2,74 mg/mL redukujućih šećera pojedinaĉno.Time se otvaraju nove mogućnosti industrijske primene ovih enzima, prevashodno za saharifikaciju skrobnih sirovina i u industriji proizvodnje bioetanola...The subject of this thesis is the utilization of waste materials of agricultural and industrial origin, for the production of enzymes cellulases and amylases using a novel isolated bacterial strain Paenibacillus chitinolyticus CKS1. Optimization of the process of production of hydrolytic enzymes cellulases and amylases was performed using commercial substrates and waste materials by varying the influence of individual parameters but also by employing the method of the planned experiment and mathematical-statistical processing of experimental data - Response Surface Methodology (RSM). The process of producing cellulase (CMC-ase and Avicelase) was optimized using various commercial substrates but also using waste materials.The strain CKS1 showed predominant exoglucanase (Avicelase) activity which is not typical for most bacterial species. Results showed that P.chitinolyticus CKS1 could be used for cellulase (CMC-ase and Avicelase) production, in a liquid medium with comercial substrates (with maximum CMCase and Avicelase activity of 0,532 U/mL and 1,83 U/mLrespectively), but also in a medium with waste substrates: waste medicinal herbs (with maximum CMCase and Avicelase activityof 0,203 U/mL and 1,94 U/mL, respectively) and sawdust (with maximum CMCase and Avicelase activity of 0,145 U/mL and 1,75 U/mL, respectively)..

Agricultural and industrial waste as a substrate for cellulase and amylase production by novel bacterial strain Paenibacillus chitinolyticus CKS1.

Predmet ove disertacije je iskorišćenje otpadnih sirovina, poljoprivrednog i industrijskog porekla, za proizvodnju enzima celulaza i amilaza pomoću novoizolovanog bakterijskog soja Paenibacillus chitinolyticus CKS1. Optimizacija procesa proizvodnje hidrolitiĉkih enzima celulaza i amilaza vršena je korišćenjem komercijalnih supstrata i otpadnih sirovina ivariranjem uticaja pojedinaĉnih parametara primenom metode planiranog eksperimenta i matematiĉko-statistiĉka obrade eksperimentalnih podataka – metode odzivnih površina (RSM). Proces proizvodnje celulalaza (endoglukanaza i egzoglukanaza) je optimizovan na razliĉitim komercijalnim podlogama ali i na otpadnim sirovinama. Soj CKS1 je pokazao predominantnu aktivnost egzoglukanaza (iskazanih kao Avicelazna aktivnost) što je netipiĉno za većinu bakterijskih vrsta. Rezultati su pokazali da se P. chitinolyticus CKS1 moţe koristiti za dobijanje celulazau komercijalnoj podlozi (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,532 U/mL i 1,83 U/mL, pojedinaĉno), ali i na otpadnim sirovinama: otpadnom lekovitom bilju (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,203 U/mL i 1,94 U/mL, pojedinaĉno) i piljevini (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,145 U/mL i 1,75 U/mL, pojedinaĉno). Soj CKS1 je pokazao mogućnost rasta i proizvodnje dve vrste celulaza: endoglukanaza (iskazanih kao karboksimetil celulazna (CMC-azna) aktivnost i egzoglukanaza (iskazanih kao Avicelazna) aktivnost tokom svog rasta na ĉvrstom otpadnom supstratu- repinom rezancu (maksimalna CMC-azna i Avicelazna aktivnost iznosi 3,159 U/g i 4,840 U/g). Metodom odzivnih površina optimizovana je proizvodnja endoglukanaza i egzoklukanaza u podlozi obogaćenoj jeĉmenim mekinjama (maksimalna CMC-aza i Avicelazna aktivnost iznosi 0,405 U/mL i 0,433 U/mL, pojedinaĉno). Dobijena je nešto niţa vrednostAvicelazne aktivnosti, ali optimizovani procesi jasno ukazuju na mogućnost njihovog korišćenja, pošto su u skladu sa principima odrţive proizvodnje i brige o ţivotnoj sredini. P. chitinolyticus CKS1 soj je pokazao i amilolitiĉku aktivnost, netipiĉnu za vrstu kojoj pripada i stoga je metodom odzivnih površina optimizovana proizvodnja β-amilaza na podlozi dobijenoj korišćenjem otpadne vode iz industrije prerade kartonske ambalaţe (0,333 U/mL) i na podlozi baziranoj na melasi i otpadnom repinom rezancu (2,237 U/mL). U cilju što kompletnije analize podobnosti primene dobijenih enzima u industrijskim procesima, izvršena je karakterizacija proizvoda hidrolize nastalih primenom obe klase enzima.UtvrĊeno je da su proizvodi enzimske hidrolize dobijeni pomoću enizima P. chitinolyticus CKS1 vredni biotehnološki proizvodi – glukoza i maltoza. Primenljivost bakterijskih celulaza i amilaza, koje produkuje soj CKS1 se ogleda u mogućnosti hidrolize pamuĉnog materijala, kao celulozne sirovine i jeĉmenih mekinja,kao lignocelulozno-skrobne sirovine, pri ĉemu se oslobaĊa 0,589 mg/mL i 2,74 mg/mL redukujućih šećera pojedinaĉno.Time se otvaraju nove mogućnosti industrijske primene ovih enzima, prevashodno za saharifikaciju skrobnih sirovina i u industriji proizvodnje bioetanola...The subject of this thesis is the utilization of waste materials of agricultural and industrial origin, for the production of enzymes cellulases and amylases using a novel isolated bacterial strain Paenibacillus chitinolyticus CKS1. Optimization of the process of production of hydrolytic enzymes cellulases and amylases was performed using commercial substrates and waste materials by varying the influence of individual parameters but also by employing the method of the planned experiment and mathematical-statistical processing of experimental data - Response Surface Methodology (RSM). The process of producing cellulase (CMC-ase and Avicelase) was optimized using various commercial substrates but also using waste materials.The strain CKS1 showed predominant exoglucanase (Avicelase) activity which is not typical for most bacterial species. Results showed that P.chitinolyticus CKS1 could be used for cellulase (CMC-ase and Avicelase) production, in a liquid medium with comercial substrates (with maximum CMCase and Avicelase activity of 0,532 U/mL and 1,83 U/mLrespectively), but also in a medium with waste substrates: waste medicinal herbs (with maximum CMCase and Avicelase activityof 0,203 U/mL and 1,94 U/mL, respectively) and sawdust (with maximum CMCase and Avicelase activity of 0,145 U/mL and 1,75 U/mL, respectively)..

Agricultural and industrial waste as a substrate for cellulase and amylase production by novel bacterial strain Paenibacillus chitinolyticus CKS1.

Predmet ove disertacije je iskorišćenje otpadnih sirovina, poljoprivrednog i industrijskog porekla, za proizvodnju enzima celulaza i amilaza pomoću novoizolovanog bakterijskog soja Paenibacillus chitinolyticus CKS1. Optimizacija procesa proizvodnje hidrolitiĉkih enzima celulaza i amilaza vršena je korišćenjem komercijalnih supstrata i otpadnih sirovina ivariranjem uticaja pojedinaĉnih parametara primenom metode planiranog eksperimenta i matematiĉko-statistiĉka obrade eksperimentalnih podataka – metode odzivnih površina (RSM). Proces proizvodnje celulalaza (endoglukanaza i egzoglukanaza) je optimizovan na razliĉitim komercijalnim podlogama ali i na otpadnim sirovinama. Soj CKS1 je pokazao predominantnu aktivnost egzoglukanaza (iskazanih kao Avicelazna aktivnost) što je netipiĉno za većinu bakterijskih vrsta. Rezultati su pokazali da se P. chitinolyticus CKS1 moţe koristiti za dobijanje celulazau komercijalnoj podlozi (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,532 U/mL i 1,83 U/mL, pojedinaĉno), ali i na otpadnim sirovinama: otpadnom lekovitom bilju (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,203 U/mL i 1,94 U/mL, pojedinaĉno) i piljevini (maksimalna CMC-azna i Avicelazna aktivnost iznose 0,145 U/mL i 1,75 U/mL, pojedinaĉno). Soj CKS1 je pokazao mogućnost rasta i proizvodnje dve vrste celulaza: endoglukanaza (iskazanih kao karboksimetil celulazna (CMC-azna) aktivnost i egzoglukanaza (iskazanih kao Avicelazna) aktivnost tokom svog rasta na ĉvrstom otpadnom supstratu- repinom rezancu (maksimalna CMC-azna i Avicelazna aktivnost iznosi 3,159 U/g i 4,840 U/g). Metodom odzivnih površina optimizovana je proizvodnja endoglukanaza i egzoklukanaza u podlozi obogaćenoj jeĉmenim mekinjama (maksimalna CMC-aza i Avicelazna aktivnost iznosi 0,405 U/mL i 0,433 U/mL, pojedinaĉno). Dobijena je nešto niţa vrednostAvicelazne aktivnosti, ali optimizovani procesi jasno ukazuju na mogućnost njihovog korišćenja, pošto su u skladu sa principima odrţive proizvodnje i brige o ţivotnoj sredini. P. chitinolyticus CKS1 soj je pokazao i amilolitiĉku aktivnost, netipiĉnu za vrstu kojoj pripada i stoga je metodom odzivnih površina optimizovana proizvodnja β-amilaza na podlozi dobijenoj korišćenjem otpadne vode iz industrije prerade kartonske ambalaţe (0,333 U/mL) i na podlozi baziranoj na melasi i otpadnom repinom rezancu (2,237 U/mL). U cilju što kompletnije analize podobnosti primene dobijenih enzima u industrijskim procesima, izvršena je karakterizacija proizvoda hidrolize nastalih primenom obe klase enzima.UtvrĊeno je da su proizvodi enzimske hidrolize dobijeni pomoću enizima P. chitinolyticus CKS1 vredni biotehnološki proizvodi – glukoza i maltoza. Primenljivost bakterijskih celulaza i amilaza, koje produkuje soj CKS1 se ogleda u mogućnosti hidrolize pamuĉnog materijala, kao celulozne sirovine i jeĉmenih mekinja,kao lignocelulozno-skrobne sirovine, pri ĉemu se oslobaĊa 0,589 mg/mL i 2,74 mg/mL redukujućih šećera pojedinaĉno.Time se otvaraju nove mogućnosti industrijske primene ovih enzima, prevashodno za saharifikaciju skrobnih sirovina i u industriji proizvodnje bioetanola...The subject of this thesis is the utilization of waste materials of agricultural and industrial origin, for the production of enzymes cellulases and amylases using a novel isolated bacterial strain Paenibacillus chitinolyticus CKS1. Optimization of the process of production of hydrolytic enzymes cellulases and amylases was performed using commercial substrates and waste materials by varying the influence of individual parameters but also by employing the method of the planned experiment and mathematical-statistical processing of experimental data - Response Surface Methodology (RSM). The process of producing cellulase (CMC-ase and Avicelase) was optimized using various commercial substrates but also using waste materials.The strain CKS1 showed predominant exoglucanase (Avicelase) activity which is not typical for most bacterial species. Results showed that P.chitinolyticus CKS1 could be used for cellulase (CMC-ase and Avicelase) production, in a liquid medium with comercial substrates (with maximum CMCase and Avicelase activity of 0,532 U/mL and 1,83 U/mLrespectively), but also in a medium with waste substrates: waste medicinal herbs (with maximum CMCase and Avicelase activityof 0,203 U/mL and 1,94 U/mL, respectively) and sawdust (with maximum CMCase and Avicelase activity of 0,145 U/mL and 1,75 U/mL, respectively)..

Antimikrobna aktivnost medicinskih materijala obrađenih srebrom

The main goal of this work was to optimize the method of processing wound dressing products (like gauzes, sanitary pads, cotton wool, compresses, and bandages) with a commercial silver colloidal solution (Koloid doo, Belgrade, Serbia) and then to examine the antimicrobial properties of the obtained items in order to potentially reach the market with new improved wound dressing products. The influence of different silver concentrations used for treatment on antimicrobial activity was investigated only against Escherichia coli. The antimicrobial activity of different types of materials treated with silver solutions of 30 ppm was investigated against Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa, Gram-positive bacteria Staphylococcus aureus, Bacillus subtilis, and Enterococcus faecalis, as well as the fungus Candida albicans. The microbial reduction of the tested materials loaded with a silver solution of 30 ppm (15-20 µg of Ag on 1 g of fabric) against the Gram-negative bacteria E. coli and P. aeruginosa was almost maximal after 2 h of contact (i.e. 95 and 99 %, respectively). In the case of Gram-positive bacteria S. aureus, B. subtilis, and E. faecalis, a longer time is needed to completely eradicate bacteria (over 99 %). Antifungal activity testing against the fungus C. albicans gave moderate antifungal activity results.Cilj ovog rada je ispitivanje antimikrobne efikasnosti pamučnih materijala kao što su gaza, higijenski ulošci, vata, komprese i zavoji obrađeni komercijalnim koloidnim rastvorom srebra (kompanija Koloid doo). Ispitan je uticaj koncentracije koloidnog rastvora srebra kojim je obrađena tkanina na antimikrobnu aktivnost prema Gram-negativnim bakterijama Escherichia coli i Pseudomonas aeruginosa, Gram-pozitivnim bakterijama Staphylococcus aureus, Bacillus subtilis i Enterococcus faecalis i gljivici Candida albicans. Redukcija bakterija postignuta testiranim materijalima obrađenim rastvorom srebra koncentracije 30 ppm (15 do 20 µg Ag na 1 g materijala) prema Gram-negativnim bakterijama E. coli i P. aeruginosa je skoro maksimalna nakon dva sata kontakta, 95 i 99 %, redom. U slučaju Gram-pozitivnih bakterija S. aureus, B. subtilis i E. faecalis, potrebno je duže vreme za potpunu redukciju broja bakterija, osim za B. subtilis, gde je dovoljno dva sata kontakta za maksimalno smanjenje početnog broja bakterija. Antifungalna aktivnost prema gljivici C. albicans je umerena

Decomposition mechanism and kinetics of zinc–isophthalate complex with 2,2’-dipyridylamine as a precursor for obtaining nanosized zinc oxide

Studies related to the synthesis of nanosized ZnO as the antibacterial agent have become an interdisciplinary area gathering chemists, physicists, biologists, and medics. The broad scope of materials based on ZnO resulted in the development of various techniques for its preparation. Considering the dependence of particle shape and size onto physical and chemical properties of ZnO, the synthesis procedure is of major importance. In this work, an unconventional methodology of synthesis is proposed for obtaining nanosized ZnO. Polymeric zinc complex containing 2,2’-dipyridylamine (dipya) and dianion of 1,3-benzenedicarboxylic acid (ipht), [Zn(dipya)(ipht)]n, was used as precursor. Besides the crystal structure of [Zn(dipya)(ipht)]n which was already published [1], the luminescent properties are presented in this work. Also, the amazing antibacterial activity of this precursor prompted us to investigate the relationship between the crystal structure and thermal properties, especially if we bear in mind the lack of similar studies in the literature. Therefore, the mechanism and kinetics of its degradation was investigated under nonisothermal conditions in nitrogen and air atmospheres. Degradation enthalpies, thermodynamic activation parameters, pre-exponential factor, A, and the apparent activation energy, Ea, were determined for each step using Kissinger’s and Ozawa’s equations. The complexity of degradation steps has been analyzed using isoconversional methods. TG/DCS data were collected at four different heating rates: 10, 15, 20 and 25 ºC min –1 , while the formation of nanosized ZnO was confirmed using XRPD and FESEM techniques. The influence of precursor on the crystallite size and morphology of the resulting ZnO along with its antibacterial activity was examined. The obtained results will be discussed and compared

From Agricultural Waste to Biofuel: Enzymatic Potential of a Bacterial Isolate Streptomyces fulvissimus CKS7 for Bioethanol Production

Purpose To avoid a negative environmental and economic impact of agricultural wastes, and following the principles of circular economy, the reuse of agricultural wastes is necessary. For this purpose, isolation of novel microorganisms with potential biotechnological application is recommended. The current researches in bioethanol production are aimed to reduce the production costs using low-cost substrates and in-house produced enzymes by novel isolated microorganisms. In line with this, in this study valorization of these agricultural by-products by novel isolate S. fulvissimus CKS7 to biotechnological value added products was done. Methods Standard microbiological methods were used for the isolation and characterization of strain. Enzymes activities were determinated using DNS method while, the ethanol concentration was determined based on the density of the alcohol distillate at 20 degrees C. Results The maximal enzymatic activities for amylase, cellulases (carboxymethyl cellulase and Avicelase), pectinase and xylanase were achieved using rye bran as a waste substrate for CKS7 growth. Obtained crude bacterial enzymes were used for enzymatic hydrolysis of lignocellulosic materials including horsetail waste, yellow gentian waste, corn stover, cotton material and corona pre-treated cotton material. The maximum yield of reducing sugars was obtained on horsetail waste and corona pre-treated cotton material. Waste brewer's yeast Saccharomyces cerevisiae was successfully used for the production of bioethanol using horsetail waste hydrolysate and corona pre-treated cotton material hydrolysate. Conclusion The obtained results showed that bacterial strain CKS7 has a significant, still unexplored enzymatic potential that could be used to achieve a cleaner, environmental friendly and economically acceptable biofuel production. [GRAPHICS]

Lignocellulosic waste material as substrate for Avicelase production by a new strain of Paenibacillus chitinolyticus CKS1

A novel strain of Paenibacillus chitinolyticus CKS1 was isolated from forest soil and identified as a potent cellulase producer. The strain was able to grow on various commercial substrates including microcrystalline cellulose (Avicel), carboxymethylcellulose (CMC), and cellobiose but also on lignocellulosic waste material such as medicinal herbs waste and sawdust. On all these substrates the strain produced cellulase composed of two subunits (similar to 70 and similar to 45 kDa) that was active on CMC, Avicel and filter paper. The maximal Avicelase activity (1.94 U/ml) was reached in a medium that contained 0.1% (w/v) of medicinal herbs waste, 3 g l(-1) of yeast extract and 5.0 g l(-1) of casein hydrolysate in 0.1 M phosphate buffer pH 7, after 48 h of incubation at 30 degrees C. The Avicelase performed optimally at 80 degrees C and at pH 4.8. Addition of K+ increased the Avicelase activity almost three fold and the enzyme retained 48.39% of the initial activity after 60 min. The product of Avicel and CMC hydrolysis was glucose with traces of other soluble sugars, indicating that the crude cellulase produced on waste material using the novel P. chitinolyticus strain CKS 1 could be used in eco-friendly processes of cellulose bioconversion, such as enzymatic saccharification of lignocellulosic materials in processes performed under acidophilic conditions and high temperatures