3_2015(en)

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ISSN 2410-776X (Online),
ISSN 2410-7751 (Print)

3 2015

Biotechnologia Acta" V. 8, No 3, 2015
https://doi.org/10.15407/biotech8.03.123
Р. 123-128, Bibliography 13, English
Universal Decimal Classification: 579.266

REGULARITIES OF QUANTITATIVE DISTRIBUTION FOR FE(III)-REDUCING BACTERIA IN NATURAL ECOSYSTEMS

Govorukha V. M.¹, Havrylyuk O. A.², Tashyrev O. B.¹

¹Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine, Kyiv
²National Aviation University, Kyiv, Ukraine

The aim of the work was quantitative determination of Fe(III)-reducing bacteria in natural ecosystems of the Antarctic, the Arctic, the Dead and the Black Sea, middle latitude (Ukraine, Abkhazia) and the equatorial zone (Ecuador).

It was used the method of quantitative determination of microorganisms by McCready and the colorimetric method for determination of Fe(II) compounds .

Results. The systemic study of the number of Fe(III)-reducing bacteria of both hemispheres in the ecosystems of six geographic regions was carried out for the first time. High number of Fe(III)-reducing bacteria in natural ecosystems was experimentally shown. The number of Fe(III)-reducing bacteria ranged from 1.1•10² to 2.8•10⁷cells/g of absolutely dry sample.

Conclusions. The presented data showed that Fe(III)-reducing bacteria are an integral part of natural ecosystems and can significantly affect the biogeochemical cycles of iron and carbon compounds transformation.

Key words: quantity of Fe(III)-reducing bacteria, natural ecosystems, biogeochemical cycles of iron.

References

1. Slobodkin A. I. Thermophilic iron-reducing prokaryotes. Dissertation, INMI RAN. Мoskva, Russia, 2008. (In Russian).

2. Lovley D. R. Organic matter mineralization with the reduction of ferric iron: a review. Geomicrobiol. J. 1987, 5(3/4), 375–399.

3. Aristovskaya T. V. Microbiology of the processes of soil formation. Leningrad: Nauka. 1980, 187 p. (In Russian).

4. Becker M., Asch F. Iron toxity in rice-conditions and management concepts. J. Plant Nutr. Soil Sci. 2005, V. 168, P. 558–573.
http://dx.doi.org/10.1002/jpln.200520504

5. Nealson K. H., Myers Ch. R. Microbial reduction of manganese and iron: new approaches to carbon cycling. Applied and Environmental Microbiology. 1992, 58(2), 439–443.

6. Lovley D. R. Dissimilatory metal reduction. Annu. Rev. Microbiol. 1993, 47, 263–290.
http://dx.doi.org/10.1146/annurev.mi.47.100193.001403

7. Shekhovtsova N. V., Verkhovtseva N. V. Ironreducing microorganisms in ultra-deep volcanic rocks. Vestnik OGU. 2011, 12(131), 363–365. (In Russian).

8. Kostka J. E., Wu J., Nealson K. H., Stucki J. W. The impact of structural Fe(III) reduction by bacteria on the surface chemistry of smectite clay minerals. Geochimica et Cosmochimica Acta. 1999, 63(22), 3705—3713.
http://dx.doi.org/10.1016/S0016-7037(99)00199-4

9. Weiss J. V., Emerson D., Backer S. M., Megonigal J. P. Enumeration of Fe(II)-oxidizing and Fe(III)-reducing bacteria in the root zone of wetland plants: Implications for a rhizosphere iron cycle. Biogeochemistry. 2003, V. 64, P. 77–96.
http://dx.doi.org/10.1023/A:1024953027726

10. Sendel Е. Colorimetric methods of trace metals determination. Мosk va: Мir. 1964, 899 p. (In Russian).

11. Kamenskaya E. P., Averyanova E. V. Quantitative determination of micro organisms: guidelines for laboratory work. Biysk: Izd-vо Alt.gosс. tekh. un-ta. 2007, 35 p. (In Russian).

12. Tepper E. Z., Shilnikova V. K., Pereverze ua G. I. Workshop for microbiology. Мoskva: Agropromizdat. 1987, 239 p. (In Russian).

13. Shlegel G. General microbiology. Мoskva: Мir. 1987, 567 p. (In Russian).

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ISSN 2410-7751 (Print)

3 2015

Biotechnologia Acta V. 8, No 3, 2015
ttps://doi.org/10.15407/biotech8.03.116
Р. 116-122, Bibliography 19, Ukrainian
Universal Decimal Classification: 615.222: 582.594.2

BIOTECHNOLOGY OF PRESERVATION OF THE RARE WILD ORCHID OF UKRAINIAN FLORA Ophrys sphegodes subsp. mammosa (Desf.) Soó ex E. Nelson

Sheiko E. A., Kosakivska I. V.

Institute of Botany of the National Academy of Sciences of Ukraine, Kyiv

The aim of the work was to determine the optimal conditions for callusogenesis in (in vitro) cultures of ovules, ovaries and anthers from vanishing wild orchid Ophrys sphegodes subsp. mammosa (Desf.) Soó ex E. Nelson followed by using of the obtained callus tissue to develop micropropagation techniques and conservation ex situ. In this study we used biotechnological (cultivation of explants in vitro) and microscopic (making temporary preparations, light microscopy, cytomorphological callus tissue analysis) methods. Nutrient media with optimal concentrations of growth regulators were selected and calluses derived from the generative organs were obtained (we used for ovaries Murashige–Skoog medium that contains 2.5 mg/l 6 of BAP and 1.5 mg/l of 2,4-D; for ovules — nutrient medium Nitsch and Nitsch containing 2.0 mg/l of 6-benzyladenine and 2.5 mg/l of 2,4-D, for the explants from anther — nutrient medium Nitsch and Nitsch containing 3 mg/l 6 of 6- benzyladenine and 2.5 mg/l of IBA). Cytomorphological analysis reveals the presence of meristematic foci in calluses. It indicates the beginning of the secondary processes of differentiation in the callus tissue. The results may be used in further investigations to obtain O. sphegodes regenerated plants from callus.

Key words: Ophrys sphegodes, in vitro culture, callus, phytohormones.

References

1. Cherevchenko T. M., Lavrentieva A. N., Ivannikov R. V. Biotechnology of tropicaland subtropical plants in vitro. Kyiv: Nauk. Dumka. 2008, 560 p. (In Ukrainian).

2. Moyo M. Plant biotechnology in South Africa: Micropropagation research endeavours, prospects and challenges. South African Journal of Botany. 2011, V. 77, P. 996–1000.

3. Diduh Ya. P. Red Book of Ukraine. Flora. Кyiv: Globalkonsalting. 2009, 900 p. (In Ukrainian).

4. Popovich S. Yu. List of rare biodiversity of reserves and national parks of Ukraine. Fitogenetic fund mikogenetic fund, phytocoenotic fund. Кyiv: Fіtosocіocenter. 2002, 276 p. (In Ukrainian).

5. Vahrusheva L. P., Svolinskiy M. D., Kucher E. N. The new location of Ophrys taurica (Agg.) Nevski in Crimea. Crimea ecosystems, their optimization and security. 2002, V. 12, P. 164–169. (In Russian).

6. Murashige T., Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiology Plant. 1962, V. 15, P. 473–497.

7. Teng W. L., Nicholson L., Teng M. C. Micropropagation of Spathoglottis plicata. Plant Cell Rep. 1997, V. 16, P. 831–835.

8. Kruglova N. N., Seldimirova O. A., Zaycev D. Y., Katasonova A. A. Biotechnological evaluation of explants to obtain regenerated plants of spring wheat in an in vitro culture in order to adaptive selection in the conditions of the Southern Urals. Izv. Chelyab. NC UrO RAN. 2006, 2 (32), 94–98. (In Russian).

9. Teixeira da Silva J. A. Floriculture, Ornamental and Plant Biotechnology: Advances and Topical Issues. Global Science Books. London: UK. 2006, 2506 p.

10. Huan L. V. T., Tanaka M. Callus induction from protocorm-like body segments and plant regeneration in Cymbidium (Orchidaceae). J. Hortic. Sci. Biotechnol. 2004, V. 79, P. 406–410.

11. Teixeira da Silva J. A., Singh N., Tanaka M. Priming biotic factors for optimal protocormlike body and callus induction in hybrid Cymbidium (Orchidaceae), and assessment of cytogenetics stability in regenerated plantlets. Plant Cell Tiss. Org. Cult. 2006, 84 (2), 119–128.

12. Teixeira da Silva J. A., Yam T., Fukai S., Nayak N., Tanaka M. Establishment of optimum nutrient media for in vitro propagation of Cymbidium Sw. (Orchidaceae) using protocorm-like body segments. Prop. Ornamental Plants. 2005, 5 (3), 129–136.

13. Morini S., D’Onofrio C., Bellocchi G., Fisichella M. Effect of 2,4-D and light quality of callus production and differentiation from in vitro quince leaves. Plant Cell Organ Culture Tissue. 2000, V. 63, P. 47–55.
http://dx.doi.org/10.1023/A:1006456919590

14. Tiago S. B., Claudete S. C., Vanildo S., Massuo J. K., Eny I. S. In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae). Acta Botanica Brasilica. 2009, 23 (2), 274–281.

15. Chen J., Chang W. Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae). Plant Sci. 2000, 160 (1), 87–93.
http://dx.doi.org/10.1016/S0168-9452(00)00367-8

16. Huan L. V. T., Takamura T., Tanaka M. Callus formation and plant regeneration from callus through somatic embryo structures in Cymbidium orchid. Plant Sci. 2004, V. 166, P. 1443–1449.
http://dx.doi.org/10.1016/j.plantsci.2004.01.023

17. Yusuf A., Thyagi R. K., Malik S. K. Somatic embryogenesis and plant regeneration from leaf segments of Piper colubrinum. Plant Cell, Tissue and Organ Culture. 2001, V. 65, P. 255–258.
http://dx.doi.org/10.1023/A:1010678609606

18. Gorbunova V. Yu. Androgenesis in vitro in spring wheat. M. S. thesis, Dept. Ecology. Rus. Saint Petersburg. 2000. (In Russian).

19. Barnab?s B. In vitro androgenesis of wheat: from fundamentals to practical application. Euphytica. 2001, V. 119, P. 211–216.
http://dx.doi.org/10.1023/A:1017558825810

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ISSN 2410-7751 (Print)

3 2015

Biotechnologia Acta V. 8, No 3, 2015
https://doi.org/10.15407/biotech8.03.110
Р. 110-115, Bibliography 5, Ukrainian
Universal Decimal Classification: 664.0 637.52

DECREASING OF SODIUM NITRITE CONTENT IN COOKED SAUSAGES USING DENITRIFYING MICROORGANISMS

Bal-Prylypko L. V., Leonova B. I.

National University of Life and Environmental Sciences of Ukraine, Kyiv

The purpose of this work was to study reduction of sodium nitrite in cooked sausages by adding of the optimized amount of denitrifying microorganisms to the bacterial preparation maintaining quality characteristics of the product. To develop biotechnology of boiled sausages «Naturel» we selected bacterial preparation based on nitrite-reducing strains of Staphylococcus carnosus and S. carnosus ssp.utilis. It was used generally accepted and special methods. The content of total pigments and nitrozopigments was determined by a method based on the extraction of meat pigments by aqueous acetone; color stability of final products was evaluated as the difference in optical density of nitroso pigment extracts before and after exposure (40 min) of the sample under the light source; analytical processing of the experimental data was carried out using modern software; quantitative evaluation of color characteristics was performed in the RGB using a multifunctional device Epson Stylus TX400. Mathematical modeling was carried out on the basis of full factorial experiment such as 2², the optimization was performed by Box–Wilson. According to the study, using of the bacterial preparation based on nitrite-reducing strains of Staphylococcus carnosus and S. carnosus ssp. utilis in biotechnology of boiled sausages «Naturel» has a positive effect on the formation of the complex of required color characteristics of final products (for prototypes of sausages the index redness was 1. 61 times higher compared to the control). Degradation of sodium nitrite and formation of nitroso pigments were intensified that improved the stability of color during the storage (the index of color fastness of experimental cooked sausages was higher by 19%). The results of performed investigations illustrate the possibility of production of cooked sausages with a minimized content of synthetic food additives and ingredients.

Key words: boiled sausage, sodium nitrite, denitrifying microorganisms.

References

Details: Hits: 123

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ISSN 2410-7751 (Print)

3 2015

Biotechnologia Acta V. 8, No 3, 2015
https://doi.org/10.15407/biotech8.03.104
Р. 104-109, Bibliography 18, English
Universal Decimal Classification: 577.15 (088.8)

MUCOADHESIVE GEL WITH IMMOBILIZED LYSOZYME: PREPARATION AND PROPERTIES

Dekina S. S., Romanovska I. I., Leonenko І. І., Yegorova A. V.

Bogatsky Physico-Chemical Institute of the National Academy of Sciences of Ukraine, Odesa

The study of non-covalent immobilized lysozyme, as well as physico-chemical and biochemical properties of obtained mucoadhesive gel was the aim of the research. Lysozyme activity was determined by bacteriolytic method (Micrococcus lysodeikticus cells acetone powder was a substrate). Lysozyme immobilization was conducted by the method of entrapment in gel. Enzyme carrier interaction was studied by viscometric, spectrophotometric and spectrofluorimetric methods. Mucoadhesive gel with immobilized lysozyme, possessing antiinflammatory and antimicrobial activities, was prepared. Due to immobilization, protein-polymer complex with the original enzymatic activity was formed. The product is characterized by high mucoadhesive properties, quantitative retaining of protein and bacteriolytic activity, prolonged release of the enzyme, improved biochemical characteristics (extended pH-activity profile, stability in acidic medium and during storage for 2 years), and it is perspective for further studies. The proposed method for lysozyme immobilization in the carboxymethyl cellulose sodium salt gel allows to obtain a stable, highly efficient product, with high adhesive properties for attachment to the mucous membranes, that is promising for use in biomedicine.

Key words: lysozyme, immobilization, mucoadhesive gel.

References

1. State Pharmacopoeia of Ukraine. Supplement 2. DP «Nauk.-exp. farm. center». 1st ed. Kharkiv: RІREG. 2007, 617 p. (In Ukrainian).

2. Pirasa A. M., Maisettab G., Sandreschi S. Preparation, physical-chemical and biological haracterization of chitosan nanoparticles loaded with lysozyme. Int. J. Biol. Macromol. 2014, V. 67, P. 124–131. doi:10.1016/j.ijbiomac.2014.03.016.

3. Takahashi D., Hamada T., Izumi T. Immobilization of lysozyme on poly(N-isopropyl acrylamide)/2-hydroxyethyl methacrylate copolymer core–shell gel beads. Polym. Bull. 2012, 68 (6), 1777–1788. doi:10.1007/s00289-012-0715-0.

4. Anirudhan T. S., Rauf T. A. Lysozyme immobilization via adsorption process using sulphonic acid functionalized silane grafted copolymer. Colloids Surf. B Biointerfaces. 2013, V. 107, P. 1–10. doi:10.1016/j.colsurfb.2013.01.063.

5. Xiao Q., Tao X., Zou H. Comparative study of solid silica nanoparticles and hollow silica nanoparticles for the immobilization of lysozyme. Chem. Eng. J. 2008, 137 (1), 38–44. doi:10.1016/j.cej.2007.09.012.

6. Lu Z., Zhang J., Ma Y. Biomimetic mineralization of calcium carbonate/carboxymethylcellulose microspheres for lysozyme immobilization. Mater. Sci. Eng. C. 2012, 32 (7), 1982–1987. doi:10.1016/j.msec.2012.05.027.

7. Hanu?ov? K., V?penka L., Dobi?? J. Development of antimicrobial packaging materials with immobilized glucose oxidase and lysozyme. Cent. Eur. J. Chem. 2013, 11 (7), 1066–1078. doi:10.2478/s11532-013-0241-4.

8. Kuijpersa A. J., Wachemb P. B., Luyn M. J. In vivo and in vitro release of lysozyme from cross-linked gelatin hydrogels: a model system for the delivery of antibacterial proteins from prosthetic heart valves. J. Contr. Release. 2000, 67 (1–2), 323–336.
doi:10.1016/S0168-3659(00)00221-2.

9. Weia L., Caia C., Lin J. Degradation controllable biomaterials constructed from lysozymeloaded Ca-alginate microparticle/chitosan composites. Polymer. 2011, 52 (22), 5139–5148. doi:10.1016/j.polymer.2011.09.006.

10. Dekina S. S. Romanovskaya I. I., Ovsepyan A. M., Molodaya A. L., Pashkin I. I. Immobilization of lysozyme in polyvinyl alcohol cryogel. Biotechnol. acta. 2014, 7 (3), 69–73.
doi: 10.15407/biotech7.03.069. (In Russian).

11. Levitskiy A. P. Lysozyme instead of antibiotics. Odesa: KP OGT. 2005, 74 p. (In Russian).

12. Hartree E. F. Determination of protein: a modification of the Lowry method, that gives a linear photometric response. Anal. Biochem. 1972, 48 (2), 422–427.

13. Bartenev G. M., Frankel S. J. Polymer physics. Leningrad: Khimiya. 1990, 332 p. (In Russian).

14. Levitsky A. P., Selivanska I. A., Furdychko A. I. Phytogel for dentistry. and others. Patent 78112 Ukraine, MPK (2013.01) А61К 8/30 (2006.01), А61К 8/97 (2006.01), А61Q 11/00. March 11, 2013.

15. Demina N. B., Larionova N. I., Kharenko E. A. Mucoadhesive drug delivery systems: quantitative assessment of interaction between synthetic and natural polymer films and mucosa. Pharmaceut. Chem. J. 2008, 42 (7), 392–399.
doi:10.1007/s11094-008-0132-8.

16. Lapach S. N., Tschubenko A. V., Babich P. N. Statistical methods in biomedical research using Excel. Кyiv: Morion. 2000, 320 p. (In Russian).

17. Rogovin Z. A. Chemistry of cellulose. Moskva: Khimiya. 1972, P. 402–404. (In Russian).

18. Kuzin M. I. Wounds and wound infection. Moskva: Medicina. 1990, 484 p. (In Russian).

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ISSN 2410-7751 (Print)

3 2015

Biotechnologia Acta V. 8, No 3, 2015
https://doi.org/10.15407/biotech8.03.095
Р. 95-103, Bibliography , English
Universal Decimal Classification: 579.864.1:615.331

ANTIBACTERIAL ACTIVITY OF Lactobacillus casei IMV B-7280 IN CASES OF EXPERIMENTAL UROGENITAL STAPHYLOCOCCAL INFE

Babenko L. P.¹, Lazarenko L. M.¹, Demchenko Z. K.¹, Konarbaeva O. A.², Veccio G. Lo³, Spivak M. Ja.¹

¹Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine, Kyiv
²Auezov South Kazakhstan state University, Shymkent, Republic of Kazakhstan
³Stucom Centre of the Studies, Barcelona, Spain

The aim of work was to determine antibacterial activity of Lactobacillus casei ІМV В-7280 probiotic strain on the experimental urogenital tract infection of mice. The influence of intravaginal and/or per os administration of this strain once per day during 7 days on the microflora of vagina, kidneys and intestinal contents of Staphylococcus aureus 8325-4 infected mice was studied.

It was established, that in cases of experimental staphylococcal infection of urogenital tract L. Casei IMV B-7280 had effective antagonistic activity against S. aureus 8325-4 and opportunistic bacteria. After L. casei IMV B-7280 introduction into infected mice reduction or complete elimination of S. aureus 8325-4 in vagina, kidneys and intestinal contents in different periods of observation was established. Under the influence of L. casei ІМВ В-7280 the number of coliform bacteria, streptococci and staphylococci in the vagina was normalized, and fungal flora — decreased even in comparison with intact mice. Normalization of kidneys microflora was also observed. In the intestinal contents of infected mice trea ted with L. casei IMV B-7280 the number of streptococci did not change, staphylococci number decreased, but the number of fungal and coliform flora remained relatively low during the observation period.

L. casei IMV B-7280 probiotic strain is promising to create immunobiotics with antibacterial action, which can be used for the prevention and treatment of urogenital infections caused by opportunistic microorganisms.

Key words: antibacterial activity, immunobiotics, Lactobacillus casei, Staphylococcus aureus.

References

1. Foxman B. Epidemiology of Urinary Tract Infections: incidence, morbidity, and economic costs. Am. J. Med. 2002, V. 113 (Issue 1), P. 5–13.
http://dx.doi.org/10.1016/S0002-9343(02)01054-9

2. Koumans E. H., Sternberg M., Bruce C., McQuillan G., Kendrick J., Sutton M., Markowitz L. E. The prevalence of bacterial vaginosis in the United States, 2001-2004; associations with symptoms, sexual behaviors, and reproductive health. Sex. Transm. Dis. 2007, 34 (11), 864–869.
http://dx.doi.org/10.1097/OLQ.0b013e318074e565

3. Srinivasan S., Fredricks D. N. The human vaginal bacteria biota and bacterial vaginosis. Interdiscip. Perspect. Infect. Dis. 2009, 2008; 2008:750479.
doi: 10.1155/2008/750479.

4. Yakovenko L. F., Sidorik L. L., Romashchenko O. V., Rudenko A. V., Shcherbak M. O., Obukhova I. A., Lazarenko L. M., Spivak M. Ya. The diagnostic significance of indicators of immunity in women with chronic inflammatory diseases of the pelvic organs. Zdorovia Ukrainy. 2012, 3 (7), 15–20. (In Ukrainian).

5. Gupta K., Scholes D., Stamm W. E. Increasing prevalence of antimicrobial resistance among uropathogens causing acute uncomplicated cystitis in women. JAMA. 1999, 281 (8), 736–738.
http://dx.doi.org/10.1001/jama.281.8.736

6. Reid G., Bruce A. W. Urogenital infections in women: can probiotics help? Postgrad. Med. J. 2003,79 (934), 428–432.http://dx.doi.org/10.1136/pmj.79.934.428

7. Marelli G., Papaleo E., Ferrari A. Lactobacilli for prevention of urogenital infections: a review. Eur. Rev. Med. Pharmacol. Sci. 2004, 8 (20), 87–95.

8. Foster L. M., Tompkins T. A., Dahl W. J. A comprehensive post-market review of studies on a probiotic product containing Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011. Benefic. Microb. 2011, 2 (40), 319–334.
http://dx.doi.org/10.3920/BM2011.0032

9. Lazarenko L., Babenko L., Sichel L., Pidgorskyi V., Mokrozub V., Voronkova O., Spivak M. Аntagonistic Action of Lactobacilli and Bifidobacteria in Relation to Staphylococcus aureus and Their Influence on the Immune Response in Cases of Intravaginal Staphylococcosis in Mice. Probiot. Antimicrob. Prot. 2012, 4 (2), 78–89.

10. Kitazawa H., Villena J., Alvarez S. Probiotics: immunobiotics and immunigenenics. USA.: CRC Press. 2014, 402 p.

11. Reid G. The scientific basis for probiotic strains of Lactobacillus. Appl. Environ. Microbiol. 1999, 65 (9), 3763–3766.

12. McGroarty J. A., Reid G. Detection of a Lactobacillus substance that inhibits Escherichia col. Can. J. Microbiol. 1988, 34 (80), 974–978.
http://dx.doi.org/10.1139/m88-171

13. Reid G., Bruce A. W. Selection of lactobacillus strains for urogenital probiotic applications. J. Infect. Dis. 2001, 183 (Suppl 1), 77–80.
http://dx.doi.org/10.1086/318841

14. Osset J., Bartolome R. M., Garcia E., Andreu A. Assessment of the capacity of Lactobacillus to inhibit the growth of uropathogens and block their adhesion to vaginal epithelial cells. J. Infect. Dis. 2001, 183 (30), 485–491.
http://dx.doi.org/10.1086/318070

15. Gardiner G. E., Heinemann C., Bruce A. W., Beuerman D., Reid G. Persistence of Lactobacillus fermentum RC-14 and Lactobacillus rhamnosus GR-1 but not L. rham nosus GG in the human vagina as demonstrated by randomly amplified polymorphic DNA. Clin. Diagn. Lab. Immunol. 2002, 9 (1), 92–96.

16. De Vrese M., Schrezenmeir J. Probiotics, prebiotics, and synbiotics. Adv. Biochem. Eng. Biotechnol. 2008, V. 111, P. 1–66.
doi: 10.1007/10_2008_097.

17. Kovachev S., Dobrevski-Vacheva R. Effect of Lactobacillus casei var rhamnosus (Gynophilus) in restoring the vaginal flora by female patients with bacterial vaginosis-randomized, open clinical trial. Akush. Ginekol. (Sofiia). 2013, V. 52 (Suppl 1), P. 48–53. (In Bulgarian).

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http://dx.doi.org/10.1139/m88-062

19. Reid G., Bruce A. W., Fraser N., Heinemann C., Owen J., Henning B. Oral probiotics can resolve urogenital infections. FEMS Immunol. Med. Microbiol. 2001, 30 (10), 49–52.
http://dx.doi.org/10.1111/j.1574-695X.2001.tb01549.x

20. Falagas M. E., Betsi G. I., Tokas T., Athanasiou S. Probiotics for prevention of recurrent urinary tract infections in women: a review of the evidence from microbiological and clinical studies. Drugs. 2006, 66 (90), 1253–1261.
http://dx.doi.org/10.2165/00003495-200666090-00007

21. Spivak M. Ya., Shinkarenko L. M., Pidgorskyi V. S., Gorchakov D. Yu., Starovoitova S. O., Lazarenko L. M., Timoshok N. O. The strain Lactobacillus casei IMV B-7280 — late interferon inducer and activator of macrophages. Ukraina. Patent N 93133. 10.01.2011. (In Ukrainian).

22. Starovoitova S. A., Lazarenko L. N., Avdieieva L. V., Polishchuk E. I., Timoshok N. O., Shinkarenko L. M., Babenko L. P., Mokrozub V. V., Spivak M. Ya., Cheipesh A. V., Nikolaichuk V. I. Search of Lactobacillus and Bifidobacterium genera strains promising for probiotics creation. Nauk. visnyk Uzhgorod. un-ty (Ser. Biol.). 2009, V. 26, P. 216–219. (In Russian).

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http://dx.doi.org/10.1007/s12602-012-9093-z

26. Z?rate G., Santos V., Nader-Macias M. E. Protective effect of vaginal Lactobacillus paracasei CRL 1289 against urogenital infection produced by Staphylococcus aureus in a mouse animal model. Infect. Dis. Obstet. Gynecol. 2007; 2007: 48358.
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