No abstract

Single recessive mutations of the methylotrophic yeast Pichia methanolica acs1, acs2, acs3 and icl1 affecting acetyl-CoA synthetase and isocitrate lyase, and growth on ethanol as sole carbon and energy source, caused a defect in autophagic peroxisome degradation during exposure of methanol-grown cells to ethanol. As a control, a mutation in mdd1, which resulted in a defect of the 'malic' enzyme and also prevented ethanol utilization, did not prevent peroxisome degradation. Peroxisome degradation in glucose medium was unimpaired in all strains tested. Addition of ethanol to methanol-grown cells of acs1, acs2, acs3 and icl1 mutants led to an increase in average vacuole size. Thickening of peroxisomal membranes and tight contacts between groups of peroxisomes and vacuoles were rarely observed. These processes proceeded much more slowly than in wild-type or mdd1 mutant cells incubated under similar conditions. No peroxisomal remnants were observed inside vacuoles in the cells of acs1, acs2, acs3 and icl1 mutants after prolonged cultivation in ethanol medium. We hypothesize that the acs and icl mutants are defective in synthesis of the true effector-presumably glyoxylate-of peroxisome degradation in ethanol medium. Lack of the effector suspends peroxisome degradation at an early stage, namely signal transduction or peroxisome/vacuole recognition. Finally, these defects in peroxisome degradation resulted in mutant cells retaining high levels of alcohol oxidase which further led to increased levels of acetaldehyde accumulation upon incubation of mutant cells with ethanol.

New strains of sulfate-reducing bacteria were obtained from human intestine. These bacteria are vibrio-like (strains SRB Vib-1, SRB Vib-2, SRB Vib-3) and short rod-like (strains SRB Rod-4 and SRB Rod-5) forms. Growth of these strains of bacteria, usa ge of sulfates by bacteria and their production of hydrogen sulfide were studied. The strains of sulfate-reducing bacteria SRB Vib-1, SRB Vib-2, SRB Vib-3 more intensively accumulate biomass compared with strains SRB Rod-4 і SRB Rod-5. The SRB Vib-1 and SRB Vib-2 strains grow most intensively. Accumulation of bacterial biomass (3.8 g/l) is the highest on the third day of cultivation, and after that a stationary growth phase began. The SRB Vib-3 strain of bacteria accumulates 2.89 gram per liter of biomass on the third day of bacteria cultivation. Maximal biomass of SRB Rod-4 and SRB Rod-5 bacteria strains ranged from 2.59 to 3.25 gram per liter on the eighth day of cultivation. The obtained sulfate-reducing bacteria intensively use sulfate ions and produce hydrogen sulfide. The SRB Vib-1, SRB Vib-2, SRB Vib-3 strains produce from 2.99 mM to 3.12 mM of hydrogen sulfide. The rod-shaped strains of sulfate-reducing bacteria use sulfate ions and produce hydrogen sulfide less intensively in the presence of sulfates.

The toxicity of metal ions to microorganisms, in particular at high concentrations, is one of the main impediments to their usage in remediation technologies. The purpose of this work is to analyze the possibility of usage by bacteria of the Desulfuromonas genus, isolated by us from Yavorivske Lake, of ferrum (ІІІ) and manganese (IV) ions at concentrations in the medium of 1,74–10,41 mM as electron acceptors of anaerobic respiration to assesss resistance of sulphur reducing bacteria strains to heavy metal compounds. Cells of Desulfuromonas acetoxidans ІМV V-7384, Desulfuromonas sp. Yavor-5 and Desulfuromonas sp. Yavor-7 were cultivated for 10 days at 30 °C under anaerobic conditions in Kravtsov-Sorokin’s medium without sulphate ions, sulphur, with cysteine as the sulphur source (0.2 g/l) and sodium lactate or citrate as the electron donor (17.86 g/l), in which were added sterile 1 M solutions of C6H5O7Fe and C4H4O4 (control) and also weights of MnO2 to their terminal concentrations 1.74, 3.47, 5.21, 6.94, 10.41 mM. Biomass was determined by the turbidimetric method. In the culture liquid the presence of Fe3+ and Mn4+ were qualitatively determined, and the content of Fe2+ in reaction with о-phenanthroline was determined quantitatively. It was established that sulphur reducing bacteria used with different intensity ferrum (ІІІ) and manganese (IV) ions as electron acceptors during the process of anaerobic respiration at concentrations of 1.74–10.41 mM C6H5O7Fe and MnO2 in the medium, which demonstrated the important role of the investigated microorganisms in reductive detoxication of natural and technogenic media from oxidized forms of transitional heavy metals. An insignificant difference in biomass accumulation during usage of 5.21–10.41 mM ferrum (ІІІ) ions and fumarate is caused by toxicity of the metal ions to cells since the high redox potential of the Fe(III)/Fe(ІІ) pair with increase in concentrations of electron acceptors in the medium did not lead to increase in the biomass accumulation level. The greatest biomass of the bacteria accumulated on the 8–10th days in the medium with the lowest concentration of C6H5O7Fe – 1.74 mM (up to 2.77 g/l), and the lowest biomass – with highest concentration – 10.41 mM (up to 2.41 g/l). After 10 days of cultivation the bacteria of all strains had fully used the ferrum (ІІІ) ions present in the medium. A biomass yield almost twice as low was revealed after manganese (IV) oxide was used by bacteria compared with its use of ferrum (ІІІ) citrate and fumarate at all studied concentrations of electron acceptors in the medium. The highest biomass of bacteria accumulated in the medium with the lowest MnO2 content – 1.74 mM (up to 1.35 g/l), and the lowest biomass in the medium with the highest content – 10.41 mM (up to 1.15 g/l). After 10 days of cultivation bacteria of all strains had not fully restored the manganese (IV) ions present in the medium. The greatest biomass compared with other strains after growth in medium with different C6H5O7Fe and MnO2 contents was accumulated by the strain Desulfuromonas sp. Yavor-7. Since sulphur reducing bacteria strains proved to be resistant to Fe3+ and Mn4+ high concentrations (up to10.41 mM) they can be successfully used in technologies of environmenal remediation from sulphur and heavy metal compounds.

Moroz, O. M., Hnatush, S. O., Tarabas, O. V., Bohoslavets, C. I., Yavorska, G. V., & Borsukevych, B. M. (2018). Sulfidogenic activity of sulfate and sulfur reducing bacteria under the influence of metal compounds. Biosystems Diversity, 26(1), 3-10.Due to their high content in natural environments, heavy metals exhibit toxic effects on living organisms, which leads to a decrease in the biological diversity and productivity of ecosystems. In niches with low oxidation reducing potential, sulfate and sulfur reducing bacteria carry out the reducing transformation of oxidized sulfur compounds with the formation of significant amounts of hydrogen sulfide. H 2 S produced by bacteria interacts with metal ions, precipitating them in the form of sulfides. The aim of this work was to investigate the influence of lead, cuprum (II), iron (II) and manganese (II) salts on the production of hydrogen sulfide by bacteria of the Desulfovibrio and Desulfuromonas genera, isolated from Yavorivske Lake, and to evaluate the efficiency of their use for purifying media, enriched with organic compounds, from hydrogen sulfide and heavy metals. The content of heavy metal ions in the water of Yavorivske Lake was determined by the spectrophotometric method. The bacteria were grown for 10 days at 30 °C in the Kravtsov-Sorokin medium under anaerobic conditions. To study the influence of metal ions on bacteria growth and their H 2 S production, cells were incubated with metal salts (0.5-4.0 mM), washed and grown in media with SO 4 2or S 0 . To determine the level of metal ions binding by H 2 S, produced by bacteria, cells were grown in media with metal compounds (0.5-4.0 mM), SO 4 2or S 0 . Biomass was determined by turbidimetric method. In the cultural liquid the content of H 2 S was determined quantitatively by spectrophotometric method, and qualitatively by the presence of metal cations. The content of metal sulfides in the growth medium was determined by weight method. Sulfate and sulfur-reducing bacteria were resistant to 2.0 mM Pb(NO 3 ) 2 , 2.5 mM CuCl 2 , 2.5 mM FeCl 2 × 4H 2 O and 2.0 mM MnCl 2 × 4H 2 O, therefore they are promising for the development of biotechnologies for the purification of water resources contaminated by sulfur and metal compounds. When present in a medium with sulfates or sulfur of 1.0-1.5 mM lead, cuprum (II), iron (II) or manganese (II) ions, they almost completely bind with the H 2 S produced by bacteria in the form of insoluble sulfides, which confirms the negative results of qualitative reactions to their presence in the cultural liquid.

This study considers the expediency of combined duck meat of regional origin containing other types of raw materials with high protein value in meatcontaining sausages. The model for the studying of feasibility of combining duck meat of regional origin with other types of raw materials was a recipe for smoked sausage containing duck meat and pork, which also including pork fat and spices. The developed formulations contained meat of Peking duck, pork hearts, pork fat, soybean isolates, dry demineralized whey, chicken skin, and soluble dietary fiber. In the model samples of smoked sausage qualitative characteristics, indicators of microbiological safety, nutritional and biological value have been studied according to standard methods. The analysis of the results of the organoleptic evaluation confirms that the combination of Peking duck meat with raw materials of different origins containing protein enhances the appearance of the products, the taste and smell, the texture and the color. The study of the microbiological parameters of the sausages developed has indicated that the number of mesophilic aerobic and optional anaerobic microorganisms in all the tested samples was within the normal limits. The study of the number of bacteria of E. coli group has not detected these microorganisms in any of the samples. It has been proved that the developed variants of meat-containing smoked sausages contained high protein according to the analysis of the nutritional value of the products, low fat concentration and low-calorie foods. It has been confirmed that the duck meat can be combined with protein ingredients of different origins to increase the protein content in the developed products by 29.98 - 31.69%, to reduce the mass fraction of fat by 31.18-35.64%. The high biological value for the content of essential amino acids and the biological efficiency for the content of fatty acids of the ω-6 and ω-3 families have been confirmed. It has been determined that when Peking duck meat is combined with non-meat types of protein-containing raw materials (pork hearts, soybean isolate, dry demineralized whey) as a part of meat-containing smoked sausage, it allows producing nutritious products with high quality indicators.

Technogenic reservoirs mainly contain several possible electron acceptors of anaerobic respiration, many of which are dangerous to the environment. The succession of their reduction (and thus detoxification) by sulfur reducing bacteria is not yet sufficiently studied. We investigated the influence of ferrum (III) citrate, present in the cultivation medium, on the reduction of sulfur, nitrate and nitrite ions by sulfur reducing bacteria Desulfuromonas acetoxidans IMV B-7384, Desulfuromonas sp. Yavor-5 and Desulfuromonas sp. Yavor-7, isolated from Yavorivske Lake. It was established that ferrum (III) citrate inhibits the biomass accumulation and hydrogen sulfide production by bacteria of Desulfuromonas sp. after simultaneous addition to the medium of 3.47 mM S0 and 1.74–10.41 mM ferrum (III) citrate, as compared with growth and hydrogen sulfide production by bacteria in the medium with only sulfur. In the medium with the same initial content (3.47 mM) S0 and ferrum (III) citrate bacteria produced ferrum (II) ions at concentrations 3.5–3.9 times higher than that of hydrogen sulfide. Ferrum (III) citrate inhibits the biomass accumulation, the nitrate or nitrite ions reduction and the ammonium ions production by bacteria of Desulfuromonas sp. after simultaneous addition to the medium of 3.47 mM NaNO3 or NaNO2 and 1.74–10.41 mM ferrum (III) citrate. In the medium with the same initial content (3.47 mM) NaNO3 and ferrum (III) citrate, bacteria produced ammonium ions at concentrations in 1.1 times higher than that of ferrum (II) ions. In the medium with the same initial content (3.47 mM) NaNO2 and ferrum (III) citrate, bacteria reduced 1.5–1.6 times more ferrum (III) than nitrite ions with production of ferrum (II) ions at concentrations 1.7 times higher than that of ammonium ions. The process of nitrate reduction carried out by bacteria of Desulfuromonas genus was less sensitive to the negative influence of ferrum (III) citrate, compared to the process of nitrite ions reduction. When the reduction of nitrate ions by bacteria in the presence of 1.74–10.41 mM ferrum (III) citrate decreased by 1.4–2.2 times, then the reduction of nitrite ions decreased by 1.8–3.2 times compared to their reduction in media with only NaNO3 or NaNO2, respectively. Although the reduction of ferrum (III) by cells in media with 3.47 mM S0, NaNO3 or NaNO2 and 1.74–10.41 mM ferrum (III) citrate decreased by 1.6–2.7, 1.6–2.7 and 1.1–2.2 times, respectively, compared to the reduction in medium with only ferrum (III) citrate, the investigated strains of bacteria were resistant to high concentrations of trivalent ferrum compounds and can therefore can be used in technologies of complex purification of environments polluted by heavy metal and nitrogen compounds.

Sulfate reducing bacteria, capable to reductive transformation of different nature pollutants, used in biotechnologies of purification of sewage, contaminated by carbon, sulfur, nitrogen and metal compounds. H2S formed by them sediment metals to form of insoluble sulfides. Number of metals can be used by these microorganisms as electron acceptors during anaerobic respiration. Because under the influence of metal compounds observed slowing of bacteria metabolism, selection isolated from technologically modified ecotops resistant to pollutions strains is important task to create a new biotechnologies of purification. That’s why the purpose of this work was to study the influence of potassium dichromate, present in medium, on reduction of sulfate and nitrate ions by sulfate reducing bacteria Desulfovibrio desulfuricans IMV K-6, Desulfovibrio sp. Yav-6 and Desulfovibrio sp. Yav-8, isolated from Yavorivske Lake, to estimate the efficiency of possible usage of these bacteria in technologies of complex purification of environment from dangerous pollutants. Bacteria were cultivated in modified Kravtsov-Sorokin medium without SO42- and FeCl2×4H2O for 10 days. To study the influence of K2Cr2O7 on usage by bacteria SO42- or NO3- cells were seeded to media with Na2SO4×10H2O or NaNO3 and K2Cr2O7 at concentrations of 1.74 mM for total content of electron acceptors in medium 3.47 mM (concentration of SO42- in medium of standard composition). Cells were also seeded to media with 3.47 mM Na2SO4×10H2O, NaNO3 or K2Cr2O7 to investigate their growth in media with SO42-, NO3- or Cr2O72- as sole electron acceptor (control). Biomass was determined by turbidymetric method, content of sulfate, nitrate, dichromate, chromium (III) ions, hydrogen sulfide or ammonia ions in cultural liquid – by spectrophotometric method. It was found that K2Cr2O7 inhibits growth (2.2 and 1.3 times) and level of reduction by bacteria sulfate or nitrate ions (4.2 and 3.0 times, respectively) at simultaneous addition into cultivation medium of 1.74 mM SO42- or NO3- and 1.74 mM Cr2O72-, compared with growth and level of reduction of sulfate or nitrate ions in medium only with SO42- or NO3- as sole electron acceptor. Revealed that during cultivation of bacteria in presence of equimolar amount of SO42- or NO3- and Cr2O72-, last used by bacteria faster, content of Cr3+ during whole period of bacteria cultivation exceeded content H2S or NH4+. K2Cr2O7 in medium has most negative influence on dissimilatory reduction by bacteria SO42- than NO3-, since level of nitrate ions reduction by cells in medium with NO3- and Cr2O72- was a half times higher than level of sulfate ions reduction by it in medium with SO42- and Cr2O72-. The ability of bacteria Desulfovibrio sp. to priority reduction of Cr2O72- and after their exhaustion − NO3- and SO42- in the processes of anaerobic respiration can be used in technologies of complex purification of environment from toxic compounds.