Endophytic bacteria isolated from Urtica dioica L.- preliminary screening for enzyme and polyphenols production.

Endophytes, especially those isolated from herbal plants, may act as a reservoir of a variety of secondary metabolites exhibiting biological activity. Some endophytes express the ability to produce the same bioactive compounds as their plant hosts, making them a more sustainable industrial supply of these substances. Urtica dioica L. (common stinging nettle) is a synanthropic plant that is widely used in herbal medicine due to the diversity of bioactive chemicals it contains, e.g., polyphenols, which demonstrate anti-inflammatory, antioxidant, and anti-cancerous capabilities. This study aimed at isolating endophytic bacteria from stinging nettles for their bioactive compounds. The endophytic isolates were identified by both biochemical and molecular methods (16S rRNA) and investigated for enzymes, biosurfactants, and polyphenols production. Each of the isolated bacterial strains was capable of producing biosurfactants and polyphenols. However, three of the isolated endophytes, identified as two strains of Bacillus cereus and one strain of Bacillus mycoides, possessed the greatest capacity to produce biosurfactants and polyphenols. The derivatized extracts from culture liquid showed the 1.633 mol l-1 (9.691 mg l-1) concentration of polyphenol compounds. Therefore, the present study signifies that endophytic B. cereus and B. mycoides isolated from Urtica dioica L. could be a potential source of biosurfactants and polyphenols. However, further study is required to understand the mechanism of the process and achieve efficient polyphenol production by endophytic bacteria.

Microbial Diversity and Bioactive Compounds in Dried Lycium barbarum Fruits (Goji): A Comparative Study.

This study compares the microbial diversity and content of bioactive compounds in dried goji berries available on the Polish market to those of the most highly valued goji berries from the Ningxia region in China. The content of phenols, flavonoids, and carotenoids were determined, as well as the antioxidant capacities of the fruits. The quantitative and qualitative composition of the microbiota inhabiting the fruits was assessed using metagenomics by high-throughput sequencing on the Illumina platform. The highest quality was demonstrated by naturally dried fruits from the Ningxia region. These berries were characterized by a high content of polyphenols and high antioxidant activity, as well as high microbial quality. The lowest antioxidant capacity was shown by goji berries cultivated in Poland. However, they contained a high amount of carotenoids. The highest microbial contamination was found in the goji berries available in Poland (>106 CFU/g), which is important in terms of consumer safety. Despite the widely accepted benefits of consuming goji berries, both the country of cultivation and the preservation method may influence their composition, bioactivity, and microbial quality.

Aronia melanocarpa (Michx.) Elliot, Chaenomeles superba Lindl. and Cornus mas L. Leaf Extracts as Natural Preservatives for Pork Meat Products.

The aim of this study was to investigate the possibility of using Aronia melanocarpa, Chaenomeles superba, and Cornus mas leaf extracts as natural preservatives for pork meat products. Pork sausages were stored in modified atmosphere packaging (MAP) (80% N2 and 20% CO2) at 4 °C for 29 days. The total psychrotrophic counts (TPC) were determined during the storage period, along with the numbers of Enterobacteriaceae and lactic acid bacteria (LAB). The extracts improved the microbial quality of the meat products but to a lesser extent than sodium nitrate (III). They reduced the amounts of Enterobacteriaceae and LAB. The A.melanocarpa leaf extract showed the strongest preservative effect. The bacterial biodiversity of the meat products was investigated based on high-throughput sequencing of the 16S rRNA gene. Two predominant bacteria phyla were identified, Proteobacteria and Firmucutes, mostly consisting of genera Photobacterium, Brochothrix, and Carnobacterium. The extracts also influenced microbial community in sausages decreasing or increasing bacterial relative abundance. The extracts significantly inhibited lipid oxidation and improved the water-holding capacity of the meat, with C. superba extract showing the strongest influence. In addition, A. melanocarpa and C. superba improved the redness (a*) of the sausages. The results of this study show that A. melanocarpa, C. superba, and C. mas leaf extracts can extend the shelf life of meat products stored in MAP at 4 °C.

Antifungal Activity of Polyoxometalate-Ionic Liquids on Historical Brick.

Moulds inhabiting mineral-based materials may cause their biodeterioration, contributing to inestimable losses, especially in the case of cultural heritage objects and architectures. Fungi in mouldy buildings may also pose a threat to human health and constitute the main etiological factor in building related illnesses. In this context, research into novel compounds with antifungal activity is of high importance. The aim of this study was to evaluate the antifungal activity of polyoxometalate-ionic liquids (POM-ILs) and their use in the eradication of moulds from historical brick. In the disc diffusion assay, all the tested POM-ILs inhibited growth of a mixed culture of moulds including Engyodontium album, Cladosporium cladosporioides, Alternaria alternata and Aspergillus fumigatus. These were isolated from the surfaces of historical brick barracks at the Auschwitz II-Birkenau State Museum in Oswiecim, Poland. POM-IL coatings on historical brick samples, under model conditions, showed that two compounds demonstrated very high antifungal activity, completely limiting mould growth and development. The antifungal activity of the POM-ILs appeared to stem from their toxic effects on conidia, as evidenced by environmental scanning transmission electron microscopy observations. The results herein indicated that POM-ILs are promising disinfectant materials for use not only on historical objects, but probably also on other mineral-based materials.

Multistep approach to control microbial fouling of historic building materials by aerial phototrophs.

The scientific multistep approach described herein is a result of two years of research into a control method against microbial fouling and biodeterioration of historic building materials by phototrophs. A series of tests were conducted to select the best antifouling agent for eliminating 'green' coatings and protecting surfaces against biofouling. Of the seven active compounds, two with the best penetration abilities were subjected to a photosynthetic activity inhibition test using confocal microscopy. Of the two, a quaternary ammonium salt (QAC) - didecyldimethylammonium chloride (DDAC) - was found to be the most effective. Ten biocides containing QACs at different concentrations were then tested against 'green' coatings on wood, brick and plaster, with the best four being selected for further research in model conditions. As a result, biocides containing >14% (v v-1) DDAC were found to be successful antifouling agents for protecting historical materials against biodeterioration by phototrophs.

Viability, Enzymatic and Protein Profiles of Pseudomonas aeruginosa Biofilm and Planktonic Cells after Monomeric/Gemini Surfactant Treatment.

This study set out to investigate the biological activity of monomeric surfactants dodecyltrimethylammonium bromide (DTAB) and the next generation gemini surfactant hexamethylene-1,6-bis-(N,N-dimethyl-N-dodecylammonium bromide) (C6) against the environmental strain Pseudomonas aeruginosa PB_1. Minimal inhibitory concentrations (MIC) were determined using the dilution method. The viability of the planktonic cells and biofilm was assessed using the plate count method. Enzymatic profile was determined using the API-ZYM system. Proteins were extracted from the biofilm and planktonic cells and analysed using SDS-PAGE. The MIC of the gemini surfactants was 70 times lower than that of its monomeric analogue. After 4 h of treatment at MIC (0.0145 mM for C6 and 1.013 mM for DTAB), the number of viable planktonic cells was reduce by less than 3 logarithm units. At the concentration ≥MIC, a reduction in the number of viable cells was observed in mature biofilms (p < 0.05). Treatment for 4 h with gemini surfactant at 20 MIC caused complete biofilm eradication. At sub-MIC, the concentration of some enzymes reduced and their protein profiles changed. The results of this study show that due to its superior antibacterial activity, gemini compound C6 can be applied as an effective microbiocide against P. aeruginosa in both planktonic and biofilm forms.

Candida albicans Impairments Induced by Peppermint and Clove Oils at Sub-Inhibitory Concentrations.

Members of Candida species cause significant health problems, inducing various types of superficial and deep-seated mycoses in humans. In order to prevent from Candida sp. development, essential oils are more and more frequently applied, due to their antifungal activity, low toxicity if used appropriately, and biodegrability. The aim of the study was to characterize the early alterations in Candida albicans metabolic properties in relation to proteins and chromosomal DNA profiles, after treatment with peppermint and clove oils at sub-inhibitory concentrations. The yeasts were affected by the oils even at a concentration of 0.0075% v/v, which resulted in changes in colony morphotypes and metabolic activities. Peppermint and clove oils at concentrations ranging from 0.015× MIC (minimal inhibitory concentration) to 0.5× MIC values substantially affected the enzymatic abilities of C. albicans, and these changes were primarily associated with the loss or decrease of activity of all 9 enzymes detected in the untreated yeast. Moreover, 29% isolates showed additional activity of N-acetyl-ß-glucosaminidase and 14% isolates-α-fucosidase in comparison to the yeast grown without essential oils addition. In response to essential oils at 0.25-0.5× MIC, extensive changes in C. albicans whole-cell protein profiles were noted. However, the yeast biochemical profiles were intact with the sole exception of the isolate treated with clove oil at 0.5× MIC. The alterations were not attributed to gross chromosomal rearrangements in C. albicans karyotype. The predominantly observed decrease in protein fractions and the yeast enzymatic activity after treatment with the oils should be considered as a phenotypic response of C. albicans to the essential oils at their sub-inhibitory concentrations and may lead to the reduction of this yeast pathogenicity.

Identification of Carotenoids and Isoprenoid Quinones from Asaia lannensis and Asaia bogorensis.

The aim of the study was to identify and quantitatively assess of carotenoids and isoprenoid quinones biosynthesized by six different strains of acetic acid bacteria, belonging to genus Asaia, that are common beverage-spoiling bacteria in Europe. Bacterial cultures were conducted in a laboratory liquid culture minimal medium with 2% sucrose. Carotenoids and isoprenoid quinones were investigated using UHPLC-DAD-ESI-MS analysis. In general, tested strains of Asaia spp. were able to produce 10 carotenoids and 3 isoprenoid quinones: menaquinone-7, menaquinone-8, and ubiquinone-10. The main identified carotenoids in Asaia lannensis strains were phytofluene, neurosporene, α-carotene, while for Asaia bogorensis, neurosporene, canthaxanthin, and zeaxanthin were noted. What is more, tested Asaia spp. were able to produce myxoxanthophyll, which has so far been identified primarily in cyanobacteria. The results show that A. lannensis are characterized by statistically higher concentrations of produced carotenoids, as well as a greater variety of these compounds. We have noted that carotenoids were not only accumulated by bacterial cells, but also some strains of A. lannensis produced extracellular carotenoids.

Synthesis, Structure and Antimicrobial Properties of Novel Benzalkonium Chloride Analogues with Pyridine Rings.

Quaternary ammonium compounds (QACs) are a group of compounds of great economic significance. They are widely used as emulsifiers, detergents, solubilizers and corrosion inhibitors in household and industrial products. Due to their excellent antimicrobial activity QACs have also gained a special meaning as antimicrobials in hospitals, agriculture and the food industry. The main representatives of the microbiocidal QACs are the benzalkonium chlorides (BACs), which exhibit biocidal activity against most bacteria, fungi, algae and some viruses. However, the misuses of QACs, mainly at sublethal concentrations, can lead to an increasing resistance of microorganisms. One of the ways to avoid this serious problem is the introduction and use of new biocides with modified structures instead of the biocides applied so far. Therefore new BAC analogues P13-P18 with pyridine rings were synthesized. The new compounds were characterized by NMR, FT-IR and ESI-MS methods. PM3 semiempirical calculations of molecular structures and the heats of formation of compounds P13-P18 were also performed. Critical micellization concentrations (CMCs) were determined to characterize the aggregation behavior of the new BAC analogues. The antimicrobial properties of novel QACs were examined by determining their minimal inhibitory concentration (MIC) values against the fungi Aspergillus niger, Candida albicans, Penicillium chrysogenum and bacteria Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. The MIC values of N,N-dimethyl-N-(4-methylpyridyl)-N-alkylammonium chlorides for fungi range from 0.1 to 12 mM and for bacteria, they range from 0.02 to 6 mM.

Prebiotics and age, but not probiotics affect the transformation of 2-amino-3-methyl-3H-imidazo[4,5-f]quinoline (IQ) by fecal microbiota - An in vitro study.

Heterocyclic aromatic amines (HAAs) are carcinogens which are formed in meat cooked using high-temperature methods. The human gastrointestinal (GI) microbiota plays a crucial role in maintaining health in humans of different ages, and especially in the elderly. However, the GI microbiota, whose metabolism and composition changes with age, may also be responsible for the activation of mutagenic substances reaching the colon with diet. Probiotics and prebiotics are promising in terms of reducing the destructive effects of HAAs. The aim of the study was to determine if fecal microbiota derived from the feces of 27 volunteers: infants (up to 18 months), adults (aged 23-39 years), the sub-elderly (aged 64-65 years), and the elderly (aged 76-87 years), and the presence of probiotics or prebiotics, affected the transformation of IQ (2-amino-3-methylimidazo[4,5-f]quinoline) to 7-OH-IQ (2-amino-3,6-dihydro-3-methyl-7H-imidazo[4,5-f]quinoline-7-one). The compounds were identified using LC-MS(n), NMR, and FTIR. Their genotoxicity was compared in the comet assay. Individual strains capable of IQ transformation were also identified. 7-OH-IQ was detected in six persons (two children and four elderly individuals). The degree of IQ conversion ranged from 26% (4-month-old girl) to 94% (81-year-old woman) of the initial quantity. Four Enterococcus isolates: two Enterococcus faecium and two Enterococcus faecalis strains, as well as one Clostridium difficile strain (LOCK 1030, from the culture collection) converted IQ to 7-OH-IQ. The genotoxicity of samples containing 7-OH-IQ was even three times higher (P < 0.05) than those with IQ and was correlated with the degree of IQ conversion and 7-OH-IQ concentration.

Protection of Historical Wood against Microbial Degradation-Selection and Application of Microbiocides.

The aim of this study was to select effective and safe microbiocides for the disinfection and protection of historical wooden surfaces at the former Auschwitz II-Birkenau concentration and extermination camp. We tested seven active compounds against bacteria and moulds, of which didecyldimethylammonium chloride and N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine were effective even at 0.02%-2%. Subsequently, eight microbiocides containing the selected active ingredients were chosen and applied three times on the surface of wood samples colonized by bacteria and moulds. ABM-1 and ABM-2-6% solution; Rocima 101-8%; Preventol R 80-12%; Acticide 706 LV-15% and Boramon-30% were the most effective disinfectants. Under laboratory conditions, ABM-1, Boramon and Rocima 101 ensured antimicrobial protection of new wood samples for six months. In situ, 30% Boramon and 8% Rocima 101 applied by spraying effectively protected the historical wood from bacterial and mould growth for 12 and 3 months, respectively. Colour and luminance of the new wood were not altered after exposure to the biocides. Boramon and Rocima 101, applied by the spraying method, caused no significant change in the colour of the historical wood. Results from this study were used to develop a procedure for the protection of wood in historical buildings against biodeterioration.

Simultaneous Saccharification and Fermentation of Sugar Beet Pulp with Mixed Bacterial Cultures for Lactic Acid and Propylene Glycol Production.

Research into fermentative production of lactic acid from agricultural by-products has recently concentrated on the direct conversion of biomass, whereby pure sugars are replaced with inexpensive feedstock in the process of lactic acid production. In our studies, for the first time, the source of carbon used is sugar beet pulp, generated as a by-product of industrial sugar production. In this paper, we focus on the simultaneous saccharification of lignocellulosic biomass and fermentation of lactic acid, using mixed cultures with complementary assimilation profiles. Lactic acid is one of the primary platform chemicals, and can be used to synthesize a wide variety of useful products, including green propylene glycol. A series of controlled batch fermentations was conducted under various conditions, including pretreatment with enzymatic hydrolysis. Inoculation was performed in two sequential stages, to avoid carbon catabolite repression. Biologically-synthesized lactic acid was catalytically reduced to propylene glycol over 5% Ru/C. The highest lactic acid yield was obtained with mixed cultures. The yield of propylene glycol from the biological lactic acid was similar to that obtained with a water solution of pure lactic acid. Our results show that simultaneous saccharification and fermentation enables generation of lactic acid, suitable for further chemical transformations, from agricultural residues.

Unusual Use of Gd-EOB-DTPA in MRCP in Order to Reveal the Source of Bile Leakage in a Patient with Neuroendocrine Tumor - Case Report.

BACKGROUND: MRCP is the method of choice in diagnosing pathologies of the biliary system. One of them is bile fistulae. They are uncommon but tend to cause many diagnostic problems. The possible way to improve MRCP is using it with intravenous injection of hepatobiliary-specific contrast agents. As it is eliminated via the hepatobiliary system, it can be visualized in the bile ducts and may help to reveal disorders undetected by a standard MRCP. CASE REPORT: We report a case of a 36-year-old woman with leakage in the biliary system which led to creation of a subcutaneous bile reservoir. By means of a regular MRCP protocol it was impossible to reveal any disorders of the biliary system and thus a decision to inject Gd-EOB-DTPA was taken. As a result, a fistula with its opening in the fundus of the gall bladder was revealed. Patient was qualified for treatment with somatostatin analogues in order to stop bile secretion. CONCLUSIONS: The Gd-EOB-DTPA in combination with regular T2-weighted MRCP may be helpful in detecting anomalies of the biliary system. Although a high price of the procedure restricts its accessibility, such advantages as lower risk of complications, lower costs of hospitalization, and less traumatic nature make it a technique that may take precedence over ERCP in ambiguous cases.

Antigenotoxic activity of lactic acid bacteria, prebiotics, and products of their fermentation against selected mutagens.

Dietary components such as lactic acid bacteria (LAB) and prebiotics can modulate the intestinal microbiota and are thought to be involved in the reduction of colorectal cancer risk. The presented study measured, using the comet assay, the antigenotoxic activity of both probiotic and non-probiotic LAB, as well as some prebiotics and the end-products of their fermentation, against fecal water (FW). The production of short chain fatty acids by the bacteria was quantified using HPLC. Seven out of the ten tested viable strains significantly decreased DNA damage induced by FW. The most effective of them were Lactobacillus mucosae 0988 and Bifidobacterium animalis ssp. lactis Bb-12, leading to a 76% and 80% decrease in genotoxicity, respectively. The end-products of fermentation of seven prebiotics by Lactobacillus casei DN 114-001 exhibited the strongest antigenotoxic activity against FW, with fermented inulin reducing genotoxicity by 75%. Among the tested bacteria, this strain produced the highest amounts of butyrate in the process of prebiotic fermentation, and especially from resistant dextrin (4.09 µM/mL). Fermented resistant dextrin improved DNA repair by 78% in cells pre-treated with 6.8 µM methylnitronitrosoguanidine (MNNG). Fermented inulin induced stronger DNA repair in cells pre-treated with mutagens (FW, 25 µM hydrogen peroxide, or MNNG) than non-fermented inulin, and the efficiency of DNA repair after 120 min of incubation decreased by 71%, 50% and 70%, respectively. The different degrees of genotoxicity inhibition observed for the various combinations of bacteria and prebiotics suggest that this effect may be attributable to carbohydrate type, SCFA yield, and the ratio of the end-products of prebiotic fermentation.

Assessment of microbial contamination within working environments of different types of composting plants.

UNLABELLED: The objective of the study was to determine the degree of microbiological contamination, type of microflora, bioaerosol particle size distribution, and concentration of endotoxins in dust in different types of composting plants. In addition, this study provides a list of indicator microorganisms that pose a biological threat in composting facilities, based on their prevalence within the workplace, source of isolation, and health hazards. We undertook microbiological analysis of the air, work surfaces, and compost, and assessed the particle size distribution of bioaerosols using a six-stage Andersen sampler. Endotoxins were determined using gas chromatography-mass spectrometry (GC-MS). Microbial identification was undertaken both microscopically and using biochemical tests. The predominant bacterial and fungal species were identified using 16S rRNA and ITS1/2 analysis, respectively. The number of mesophilic microorganisms in composting plants amounted to 6.9×10(2)-2.5×10(4) CFU/m3 in the air, 2.9×10(2)-3.3×10(3) CFU/100 cm2 on surfaces, and 2.2×10(5)-2.4×10(7) CFU/g in compost. Qualitative analysis revealed 75 microbial strains in composting plants, with filamentous fungi being the largest group of microorganisms, accounting for as many as 38 isolates. The total amount of endotoxins was 0.0062-0.0140 nmol/mg of dust. The dust fraction with aerodynamic particle diameter of 0.65-1.1 µm accounted for 28-39% of bacterial aerosols and 4-13% of fungal aerosols. We propose the following strains as indicators of harmful biological agent contamination: Bacillus cereus, Aspergillus fumigatus, Cladosporium cladosporioides, C. herbarum, Mucor hiemalis, and Rhizopus oryzae for both types of composting plants, and Bacillus pumilus, Mucor fragilis, Penicillium svalbardense, and P. crustosum for green waste composting plants. The biological hazards posed within these plants are due to the presence of potentially pathogenic microorganisms and the inhalation of respirable bioaerosol. Depending on the type of microorganism, these hazards may be aggravated or reduced after cleaning procedures. IMPLICATIONS: This study assessed the microbial contamination in two categories of composting plants: (1) facilities producing substrates for industrial cultivation of button mushrooms, and (2) facilities for processing biodegradable waste. Both workplaces showed potentially pathogenic microorganisms, respirable bioaerosol, and endotoxin. These results are useful to determine the procedures to control harmful biological agents, and to disinfect workplaces in composting plants.

Halophilic microbial communities in deteriorated buildings.

Halophilic microorganisms were traditionally isolated from an aquatic environment. There has been little research conducted into halophiles inhabiting the terrestrial environment in which historic monuments deteriorate. Salt efflorescence deposited on the walls is an observed phenomenon on the surface of historic buildings, and would favour the growth of halophiles. However, some conditions have to be fulfilled in order for efflorescence to occur: (1) the presence of salts, (2) porosity, (3) a source of water. Salt crystallization influences the material structure (cracking, detachment, material loss), but active growth of halophilic microorganisms may be a serious threat to historic materials as well, leading to aesthetical changes such as coloured biofilms, orange to pink or even violet stains. This is why it is important to investigate halophilic microorganisms, taking into consideration both the environmental conditions they need to grow in, material characteristics they inhabit, the mechanisms they possess to cope with osmotic stress, and the methods that should be applied for their identification.

[Amygdalin - structure and clinical significance]. / Amigdalina - budowa i znaczenie kliniczne.

In this publication we described amygdalin. It was isolated for the first time in the 19th century. Amygdalin is called interchangeably vitamin B17 or laetrile. Since more than a hundred years, there has been reports about its unique anticancer properties. We tried to introduce the present knowledge about therapeutic efficacy of laetrile. Most of these studies has been made in the in vitro environment. The lack of appropriate studies forced scientists to examine the positive influence of amygdalin on many diseases like: bladder cancer, prostate cancer, cervical cancer, colon cancer, promyelocytic leukemia, chronic kidney disease, psoriasis and other.

Assessment of biological colonization of historic buildings in the former Auschwitz II-Birkenau concentration camp.

The objective of this study was to assess biological colonization of wooden and brick buildings in the former Auschwitz II-Birkenau concentration camp, and to identify the organisms colonizing the examined buildings. Microbiological analysis did not reveal increased microbial activity, and the total microbial count of the barrack surfaces did not exceed 103 CFU/100 cm2. However, certain symptoms of biodegradation of the buildings were observed. The predominant microflora consisted of bacteria of the genera Bacillus, Sporosarcina, Pseudomonas, Micrococcus, Streptomyces, and Staphylococcus, as well as fungi of the genera Acremonium, Cladosporium, Alternaria, Humicola, Penicillium, and Chaetomium. The microflora patterns varied both in wooden and brick buildings. The structural elements of wooden and brick barracks, and especially of the floors and lower parts of bathroom walls, were infected by cyanobacteria and algae, with the most numerous being cyanobacteria of the genera Scytonema, Chroococcus, Gloeothece, Leptolyngbya, diatoms of the genus Diadesmis, and chlorophytes of the genera Chlorella and Apatococcus. The outer surfaces of the examined buildings were primarily colonized by lichens and bryophytes, with nearly 30 species identified. The dominant species of lichens belonged to the genera Candelariella, Caloplaca, Lecanora, Lecidea, Lepraria, Physcia, and Protoparmeliopsis, and those of bryophytes to the genera Bryum, Ceratodon, Marchantia, and Tortula. The quantity and species diversity of lichens and mosses were much lower in wooden barracks than in brick ones. The external surfaces of those barracks were only affected by Lecanora conizaeoides, Lecanora symmicta, Lepraria cf. incana, and Strangospora pinicola. The study results revealed vast biodiversity among the species colonizing historic buildings. The presence of these groups of organisms, resulting from their natural expansion in the environment, is undesirable, as their excessive growth and spread may lead to progressive biodegradation of buildings. Our assessment of biological contamination will enable the development of a disinfection and conservation plan for the examined buildings.

The evaluation of microbial contamination in the working environment of tanneries.

BACKGROUND: Due to their animal material processing, tannery workers may be exposed to biological agents. The aim of the study was the microbial contamination assessment of tanneries with different production specifications. Health risk was estimated based on particle size distribution. Moreover, indicators of microbial contamination of tanneries were selected. MATERIALS AND METHODS: The studies were conducted in 2 types of tanneries - processing raw hides and producing chrome-tanned leather. Air was sampled with MAS-100 Eco Air Sampler, leathers using RODAC Envirocheck contact plates and swab method, microbial numbers were determined by a culture method. For the bioaerosols size distribution analysis, a six-stage Andersen sampler was used; identification was performed using microscopy and biochemical methods. Microbial contamination was identified by 16S RNA and ITS1/2 rDNA analysis for bacteria and fungi respectively. RESULTS: The microbial number in the air ranged between 1.2 x 10(3) and 3.7 x 10(3) CFU/m3. While on the leather, it ranged between 7.6 x 10(1) and 5.5 x 10(5) CFU/100 cm2. Bacteria dominated in the tanneries (air: 51-92%, leathers: 60-100%). Results indicate that potential health risks arise from the fungal small bioaerosol particles presence (0.65-2.1 microm). Eleven indicator microorganisms were determined: B. pumilus, B. subtilis, B. cereus, C. lubricantis, C. cladosporioides, P. commune, P. echinulatum, P. chrysogenum, P. crustosum C. parapsilosis and C. albidus. CONCLUSIONS: Microbial contamination evaluation in the tanneries showed the increased bacteria and fungi number in the air in relation to the outdoor air, which indicates an occupational inhalation risk to workers. The designated indicators of microbial contamination in the tanneries are associated with their specific and potentially pathogenic working environment.

Differentiation of strains from the Bacillus cereus group by RFLP-PFGE genomic fingerprinting.

Bacillus mycoides, Bacillus pseudomycoides, Bacillus weihenstephanensis, Bacillus anthracis, Bacillus thuringiensis, and Bacillus cereus belong to the B. cereus group. The last three species are characterized by different phenotype features and pathogenicity spectrum, but it has been shown that these species are genetically closely related. The macrorestriction analysis of the genomic DNA with the NotI enzyme was used to generate polymorphism of restriction profiles for 39 food-borne isolates (B. cereus, B. mycoides) and seven reference strains (B. mycoides, B. thuringiensis, B. weihenstephanensis, and B. cereus). The PFGE method was applied to differentiate the examined strains of the B. cereus group. On the basis of the unweighted pair group method with the arithmetic mean method and Dice coefficient, the strains were divided into five clusters (types A-E), and the most numerous group was group A (25 strains). A total of 21 distinct pulsotypes were observed. The RFLP-PFGE analysis was successfully used for the differentiation and characterization of B. cereus and B. mycoides strains isolated from different food products.