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Microbiological and Physicochemical Research of Thermal Spring and Mountain Spring Waters in the District of Sliven, Bulgaria

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Microbiological and Physicochemical Research of Thermal Spring and Mountain Spring Waters in the District of Sliven, Bulgaria Journal of Advances in Microbiology 20(2): 9-17, 2020; Article no.JAMB.55066 ISSN: 2456-7116 Microbiological and Physicochemical Research of Thermal Spring and Mountain Spring Waters in the District of Sliven, Bulgaria Nedyalka Valcheva1, Ignat Ignatov2* and Georgi Dinkov3 1Department of Biochemistry, Microbiology and Physics, Faculty of Agriculture, Trakia University, Stara Zagora 6000, Bulgaria. 2Scientific Research Center of Medical Biophysics, 32 Nikolai Kopernik St., 1111 Sofia, Bulgaria. 3Idealabs, LLC, 18th Street NW, #700, Washington 20036, USA. Authors’ contributions This work was carried out in collaboration among all authors. Author NV designed the study, performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript. Authors II and GD managed the analyses of the study. Author II managed the literature searches. All authors read and approved the final manuscript. Article Information DOI: 10.9734/JAMB/2020/v20i230213 Editor(s): (1) Hung-Jen Liu, National Chung Hsing University, Taiwan. Reviewers: (1) Schirley Costalonga, Universidade Federal do Espírito Santo, Brazil. (2) R. D. Mavunda, University of Johannesburg, South Africa. (3) Lachezar Pehlivanov, Bulgarian Academy of Sciences, Bulgaria. Complete Peer review History: http://www.sdiarticle4.com/review-history/55066 Received 27 December 2019 Accepted 02 March 2020 Original Research Article Published 05 March 2020 ABSTRACT Defined are the physicochemical properties of four healing non-thermal and thermal spring waters in the area of Sliven District, Bulgaria. The spring waters from the given four water sources are characterized by microbiological indicators and the pathogenic micro-organisms in the samples from the springs water sources mentioned above are determined by the membrane method. It is shown that according to 18 controlled parameters included in the study, the non-thermal healing spring “Hadji Dimitar” in the area of “Hot water” in the town of Shivachevo with water temperature 22.5ºC and the non-thermal spring “Gunchov spring” with water temperature 21.5ºC correspond to all controlled parameters according to Ordinance NO 9/2001, Official State Gazette, issue 30 and decree NO 178/23.07.2004 about the quality of water, intended for drinking purposes. It is established that that thermal healing spring Sliven mineral baths with water temperature 48ºC, _____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected]; Valcheva et al.; JAMB, 20(2): 9-17, 2020; Article no.JAMB.55066 healing spring Banya village with water temperature 37ºС meet the standard requirements. Non- thermal spring "Gunchov Spring", Sliven district with water temperature 21.5ºC does not meet the required microbiological parameters in regards to coli bacteria levels. Keywords: Spring water; physicochemical properties; microbiological indicators. 1. INTRODUCTION microorganisms with valuable properties (enzymes, antibiotics, thermophilic and The causes for that lie in the combination acidophilic stains) are present in mineral and non between hydrological conditions of the continuing – thermal spring waters. tectonic processes in the Earth’s crust [1,2]. By their nature the springs can be separated in cold, This was proved by the results obtained from the warm and hot springs. The first group includes experimental work carried out to determine the the ones with temperature up to 37ºС and this is micro flora of medicinal and spring waters in cold mineral water. The second one ranges Haskovo, Stara Zagora, Burgas [7-14] and Varna between 37ºС and 60ºС and this is warm mineral region [12]. water. The third one with over 60ºС and this is hot mineral water. The hottest mineral spring in There are different studies of physicochemical Bulgaria is the one at Sapareva Banya with and microbiological properties of hot mineral temperature of 101.4ºС. The springing waters waters [16-19] etc. have different mineralogical characteristics. Their content is defined by the ones of the rocks, Isolated bacteria from the healing and spring where the water has been flowing through, and regions have been identified by physiologico- the solubility of the minerals within them (Ignatov, biochemical and well as molecular genetic tests Mosin, 2012). The research with hot mineral and the species include Bacillus subtilis, Bacillus - water from Rupite, Bulgaria with HCO3 -1320- cereus, Bacillus thuringiensis, Bacillus 2+ 3 1488 Ca -29-36 mg/dm shows the possibility of methylotrophicus, Aeromonas hydrophila. origin of life in hot mineral water [1,2]. The research of Bacillus subtilis in hot mineral Several authors [3,4,5] The temperature of the water were performed from Asoodeh, Lagzian water of the source is 76ºС and in the lake is [19] and with one of the authors [2]. There was around 50ºС and is depending with season. study for effects of Bacillus subtilis in heavy There are proofs for cyanobacteria in hot mineral waterr with parameters for study in mineral water water in Rupite [6]. This water contains the [21-23]. The study of Bacillus methylotrophicus following ions and they are structured the first was made in hot water from Tumbarski, et al. living organisms – stromatolites. [24] and Yuan, et al. [25]. For the first time Aeromonas hydrophila was observed in hot In Bulgaria, there are mineral and spring waters, mineral water from Italian team [26]. One of the which are not subjected to physicochemical and authors Valcheva has performed the study with microbiological control by the Regional Health Bacillus cereus and Bacillus thuringiensis [15]. Inspectorate, yet they are the most widely used springs by the population as sources of drinking Strains with high proteolytic, lipolytic and water. Similar springs are located in the territory amylolytic activity have been isolated and of Haskovo District [7-9], Stara Zagora District standardized for further scientific use [8-14]. [10,11], Varna District [12] and Burgas District [13,14]. Antimicrobial activity of the strains of Bacillus sp., was detected against the following saprophytic For many of these sources physicochemical and and pathogenic microorganisms: Penicillium sp., microbiological studies have not been conducted, Fusariummoliniforme, Rhizopus sp., yet they are used for drinking and household Aspergillusniger, Aspergillusoryzae, Aspergillu- needs [15]. sawamori, Mucor sp.и Enterococcus faecalis, in the process of development and growth of the Water is an environment for the development of four Bacillus – Bacillus cereus, Bacillus microorganisms. Studies by many authors, thuringiensis, Bacillus subtilis, Bacillus including our research team, demonstrate that methylotrophicus are the most active strains - 10 Valcheva et al.; JAMB, 20(2): 9-17, 2020; Article no.JAMB.55066 Bacillus methylotrophicusPY5 (Я1), Bacillus hydrocarbonate - calcium - magnesium water. cereus LH1 (Р1), Bacillus cereus WIF15 ( ГИ2) Tested levels of micro-components are within и Bacillus thuringiensis B62 (ХМБ3) [27-31]. normal limits except for arsenic content. If used for drinking water, it is necessary to reduce Pathogenic bacteria exhibit resistance and 4 arsenic within the normal range. The water from retain their vitality in the process of development this source is suitable for the treatment of and interaction between them and the strains of diseases of the musculoskeletal system, the Bacillus sp. at temperature of 37ºС. peripheral nervous system, gastrointestinal diseases, bile-liver diseases and nephro- A relatively low bactericidal effect was urological. demonstrated against the (Gr+) bacterium Enterococcus faecalis. The isolated strains are likelyto have a higher inhibitory ability against (Gr 2.3 Thermal Healing Spring Banya Village -) bacteria compared to (Gr +) bacteria. The village of Banya, with its thermal mineral The yeasts used in the genus Candida exhibit a springs (37ºC), combined with the beautiful simulating effect onthe growth of Bacillus sp. – nature and modern infrastructure, and provides Bacillus methylotrophicus PY5 (Я1), and the perfect conditions for complete recovery! It is Bacillus cereus LH1 (Р1). This indicates that suitable for the treatment of the following types of synergism has occurred between these diseases – allergies, arthritis, lung disease, back microorganisms. pains, depression, and diabetes. The sources from Sliven district, Bulgaria for The established Health Recovery Center research are: combines the most effective recovery package - healthy food, healing gymnastics, many group 2. WATER SPRINGS FOR RESEARCH walks in the unique surroundings, food FROM SLIVEN DISTICT, BULGARIA exclusively of plant origin (vegan), relaxing massages, water treatments with mineral water, 2.1 Sliven Mineral Baths with herbs or different salts, compresses and heat-pads, different saunas, inhalations, The mineral water is hot (48ºC), low-mineralized exercises, juice therapy and herbal teas as 3 (1,986 g/dm ), hydrocarbonate - sulphate - needed. sodium and calcium, slightly fluorine (4.2 mg 3 fluorine per dm ). It contains 27 mg colloidal 2.4 Mountain Spring “Gunchov Spring”, metasilicic acid per liter, neutral reaction (pH 6.8) Karandila Locality, Sliven District due to its CO content (287 mg / dm3).Water from 2 this source is effective in the treatment of The spring is located among the most densely gastrointestinal diseases, liver and bile diseases, forested areas along the Ravna oriver. The diseases of the musculoskeletal system, whole area
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