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Characterisation of a thermotolerant yeast, Kluyveromyces marxianus CBS712
7-13Views:140Fermentation at high temperature with application of thermotolerant microorganisms is a technological advantage in bioethanol production. Among the yeasts, K. marxianus has outstanding thermotolarance. The industrial application of the IMB3 strain occurs usually at 45C. The final aim of our project is the genetic modification of the K. marxianus CBS712 strain in order to achieve ethanol production at higher temperature than the currently applied. This requires the characterization of the CBS712 strain, with special attention to the determination of the temperature limit of its growth and the amount of the ethanol produced. The temperature limit of growth was 48C in YPD medium. Elevation of the temperature above 45C led to an exponential drop of the cell viability. Ethanol production was tested in shaking flasks, in MYFM medium, under oxigene limited conditions, applying variable concentrations of glucose (12–20%) and different temperatures (45–47 ºC). Preliminary results have revealed that the elevation of glucose concentration increased the amount of ethanol produced. The amount of ethanol (appr. 5%)+ produced at the highest glucose concentration was not different at the tested temperatures (45, 46 and 47 ºC). The observation indicates the potential in raising the thermotolerance of the strain.
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Effect of cadmium and zinc contamination on the population dynamics of soil microorgani
73-77Views:124Changes in the population dynamics of microorganisms in a soil artificially contaminated with various doses of cadmium and zinc was examined from a quantitative point of view, under laboratory circumstances. The research was based on a chernozem soil originating from the area of a long-term microelement contamination model experiment (Nagyhörcsökpuszta, Hungary), which was carried out during 1991 in the Experimental Site of the Institute of Soil Science and Agricultural Chemistry, Centre for Agricultural Researche Hungarian Academy of Sciences, Budapest, Hungary. According to the amount of bacteria, microscopic fungi and nitrifying bacteria, it can be stated that the effect of contamination can be observed even in the perspective of nearly two decades. In more cases significant changes in the number of soil bacteria and microscopic fungi could be observed, and the nitrification activity increased in case of both microelements. Therefore the further research of changes in microbial activity of these soils can provide novel scientific results.
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Testing the antimicrobial activity of essential oils
71-74Views:151The vapor phase of some essential oils proved to have antimicrobial activity. Utilization of the vapor phase of Eos is presently understood as one of the possible alternatives to synthetic food preservatives which could be used in the future. However, testing the vapor phase of EOs against microorganisms causing food-borne diseases (e.g. Salmonella enteritidis or Staphylococcus aureus) or food spoilage is relatively new. Consequently, due to the large number of known EOs, research on their antimicrobial activity is still largely in the phase of in vitro rather than in vivo testing. Moreover, no standard and reliable method for fast screening of a wide range of samples exists. Thus, the aim of this study is to show results concerning tests of the antimicrobial activity of EOs against S. enteritidis or S. aureus, which were conducted by two modifications of the disc volatilization method we developed. The lately developed method has the potential to become widely used for fast screening of EO antimicrobial activity in the vapor phase.
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Essential oil extraction from herbs and their use in the food industry
59-74Views:383Essential oil extraction of wild caraway and thyme was performed using a classical (HD) and microwave hydro-distillation (MWHD) and a laboratory supercritical fluid extraction (SFE) with a carbon dioxide as solvent. Our experiments demonstrated that the extraction yield of the essential oil performed in same conditions was influenced by the location of growth area; the maximum extraction yield of 10 ml 100 g-1 caraway was obtained from dried seeds collected from Csíkmadaras. This quantity far exceeded the yield of the Újtusnád samples. In the case of wild caraway (Carum carvi L.), the extraction method influenced thecomposition of the essential oil (carvone/limonene ratio), the highest limonene content being achieved by classical hydro-distillation. In the case of thyme, this effect was not detected, the thymol/carvacrol ratio was independent from the given extraction method. The obtained thyme essential oil possesses high antimicrobial activity demonstrated by agar diffusion test. The thyme extract provides a good protection against microorganisms collected on the surface of fresh vegetables following bacterial stains: Citrobacter portucalensis, Pseudomonas hunanensis, Pseudomonas baetica, Pseudomonas parafulva, Bacillus mojavensis and Enterobacter cloacae. Protective effect was also detected on the vegetable surface of by chitosan-based edible film coating during a 6-day-long storage period at a temperature of 4 °C. The caraway essential oil used as soft cheese seasoning with a direct, dilution-free method, proved to be unsuitable because the uneven distribution and confer a strong, unpleasant taste to the product in comparison with the ground wild caraway seed-dressed cheese.
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The effect of keeping technology on the microbiological status of raw milk
67-75Views:136The importance of the quality of raw milk increased after Hungary had joined to the EU. On delivery of raw milk, the microbiological quality, especially total plate count of the milk is very important. Twenty-two farms (7 large, 4 medium-sized, and 11 small farms) were included in the study. We considered the different farm size, keeping- and milking circumstances during the selection of farms. The examined large farms use loose housing system (cubicle, deep litter) and milking parlour. Most of them use preand post-milking disinfection. In the medium-sized farms, loose,
deep litter and tie-stall housing system, as well as milking parlour, pipeline milking and bucket milking occurred. All of them use preand post-milking disinfection. Small farms use tie-stall housing system, bucket milking and udder preparation by water. Unfortunately, they do not use pre- or post-milking disinfection. In the large and medium-sized farms mainly Holstein Friesian, in the small farms Hungarian Simmental breeds can be found.
The aim of our research was to examine the microbiological status of the raw milk produced in dairy farms (total plate count, coliform count, Escherichia coli count, Staphylococcus aureus count, psychrotroph bacteria count, furthermore yeast and mold count); sources of the contamination; connection between the microbiological quality of produced milk and housing-, milking technologies of farms; furthermore the hygienic circumstances of milking and milk handling of the farms, by the examination of coliform bacteria and Escherichia coli contamination.
During the examination of the connection between the different farm sizes, various housing- and milking forms and the microbiological characteristics we observed similar tendencies in the case of total plate count, coliform count, yeast and molds count, furthermore psychrotroph bacteria count. The value of these parameters was significantly higher in small farms, and infarms which use tie-stall housing forms, bucket milking, udder preparation with water, and which do not use pre- and post-milking disinfection.
The results showed that besides cooling, the milking procedure and the type of udder preparation had the largest effect on the total plate count. Statistical analysis shows that in medium and small farms the combination of pipeline milking – tie stall housing system – disinfectant preparation of the udder; in large farms the combination of milking parlour – loose cubicle housing system – dry preparation of the udder are the most appropriate in the aspect of the total plate count. We experienced that in farms where the hygienic instructions are not followed – and therefore
equipment used during the milking and handling of milk is very contaminated – or rather the separation of mastitic cows’ milk is not appropriate, different microorganisms may contaminate the produced milk. -
Saccharomyces cerevisiae growth kinetics study dairy byproduct
169-172Views:243By guess, annual volume of milk whey is 185–190 million tons and this volume probably will increase next years. Whey has significant biochemical oxygen demand due to its high organic matter content so whey as sewage is one of the most pollutant by-products in the food industry. Apart from environmental pollution, benefit of several whey constituents for human health is another reason to utilize whey. Corn and potato, as well as the processing of milk in the food industry in large quantities of by-products generated by low cost, substantial quantities of starch and lactic acid, which are due to high biological oxygen demand are considered as hazardous waste. Some of them are destroyed sewage storage tanks, and those products are excellent substrates for the growth of microorganisms could be. The traditional nutrient solution optimization methods are solution and time-consuming and are not able to determine the real optimum because of the interaction of factors involved.
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Usage of microbiological products in the protection of the sunflower
36-40Views:85White mould is the most important desease of sunflower. We are not able to grow sunflower on the same area for 5-6 years, because of sclerotia. One of the protection methods is if we destroy the sclerotia in the soil with hyperparasite microorganisms, so we can reduce the comeback time of sunflower.
We carried out our farm size researches through 3 years, with the most important sunflower hybrids of Hungary in the area of Vásárhelyi Róna Kft. We can conclude that the microbiological products (Koni, Trifender, Mico’sol) effect positively on the yield of sunflower hybrids, but the weather (moisture) can significantly modify this effect.
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Identification of the Slovak traditional cheese “Parenica” microflora
227-239Views:266Numerous studies have demonstrated the higher accuracy, faster time-to-results and lower costs provided by MALDI Biotyper systems compared to classical methods. In this study, the culturable population of total count of bacteria, enterococci, coliforms bacteria, lactic acid bacteria (LAB) and microscopic fungi and yeasts from cow’s dairy products was identified using the MALDI-TOF MS Biotyper. Altogether, 50 samples of the Slovak cheese “Parenica” were examined. Total numbers of bacteria were cultured on Plate count agar at 37 °C for 24–48 h, aerobically; enterococci were cultured on Enterococcus selective agar at 37 °C for 24–48 h, aerobically; coliforms bacteria were cultured on Violet Red Bile lactose agar at 37 °C for 24–48 h, aerobically. The LAB were cultured on MRS (Main Rogosa agar), MSE and APT agar at 30 °C in microaerophilic conditions. The microscopic fungi and yeasts were cultured on Malt extract agar at 25 °C for 5 days, aerobically. Isolated strains (total 669) were subjected to identification by the MALDI-TOF MS. Among total count the identified bacteria mostly were Acinetobacter baumannii, Bacillus cereus, Micrococcus luteus and Staphylococcus warneri. Escherichia coli and Enterobacter cloacae were the most abundant coliform bacteria representatives identified. Coliform bacteria included Citrobacter, Hafnia and Klebsiella. Altogether three genera belonged to the LAB – Lactobacillus, Lactococcus and Leuconostoc were identified with Lactococcus lactis, Lactobacillus plantarum, Lactobacillus coryniformis, L. fructivorans and Leuconostoc mesenteroides were considered as the dominated LAB species in dairy products. Among yeasts, Kluyveromyces lactis, Candida zeylanoides and Yarrowia lipolytica were among the most isolated.
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Bacteria in the milk of sheep with or without mastitis- mini Review
47-52Views:373From a nutritional point of view, sheep milk is more valuable than cow and goat milk and the interest for sheep milk is increasing in many countries. However, sheep milk is easily contaminated during milking, handling, and transport and it is an ideal medium for bacterial propagation. Consequently, sheep milk spoils quite quickly. The proper, clean handling of milk is not only of sanitarian interest, but it also serves the farmers’ interests, because contaminated milk may not be distributed, and is unsuitable for producing good quality products. Following this technological trend, this review addresses the bacterial composition of sheep milk with and without mastitis. Even though sheep milk contains a lot of bacteria, this review article highlighted total plate count, Enterobacteriaceae, coliform, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Campylobacter, Salmonella spp. and Streptococcus spp. Mastitis in sheep is a vital cause of mortality, reduction in milk production and early culling. The reported risk factors for mastitis in sheep were age, a case of mastitis, breed, husbandry systems, and location. The main priority should be implementation of programs to minimize human pathogenic bacteria and mastitis in raw ewe milk.
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Effects of some herbicides on the microbiological characteristics of soil nitrogen cycle under maize plantation
93-100Views:89Nitrogen is a key element for the living organisms and influence not only for the quantity but for the quality of the yield, considerable. Availability of nitrogen from the soil is influenced by several microbiological processes of the Nitrogen-cycle. Among the intensive agricultural production the herbicide application cannot be omitted more information needs therefore about the inhibitor effect of herbicides on the different microorganisms.
An experiment was set up on calcareous chernozem soil under maize culture. Effect of four different herbicides (Acenit, Frontier, Merlin, and Wing) was investigated. The effect of herbicides was measured to four microbiological parameters of the Nitrogencycle (abundance of nitrifying bacteria, nitrate solubilisation, biomass nitrogen and urease enzyme activity). There were singledouble- and five times of recommended doses of herbicides applied for two onsecutive vegetation periods.
From the results of the different doses of herbicides, the following can be stated:
– The Acenit has a stimulating effect on nitrifying bacteria in general. The Frontier and Merlin also influenced the quantity of nitrifyers, however in certain cases decreased in another cases increased the number of bacteria.
– The double doses and five times doses of herbicides was found to be increasing the nitrate content of soil, -especially in 2006.
– The quantity of microbial biomass nitrogen increased in the 60% of treatments and decreased in the 40% of the treatments.
– Except of the result of Wing in 2006 and Merlin in 2005, the effect of simple dose herbicides was the smallest on the urease enzyme activity. According to the results the effect of Merlin was positive; the effect of Wing was negative on the soil enzyme’s activity.
Regarding the application of four different herbicides in three different doses on the microbiological parameters of soil (at two consecutive years-) in 39% of the treatments has resulted a significant inhibitory effect, 28% of the treatments, however have significant stimulating effect on the parameters measured. More than 50% of the inhibitory effect was measured in case of the Wing, at more than 50% of the Frontier the microbiological parameters have not changed. -
Definition antioxidant activity of selenium-enriched food sprout, as well as their microbiological analysis
25-30Views:255In this present study, we prepared selenium-enriched food sprouts, where the antioxidant capacity was analysed, we also determined their microbiological status. We took into account the fact, we choose micronutrients to our treatment, that selenium can be delivered to the body by a small amount with the most widely consumed food.
We focused during our research to determine that the increasing concentrations of selenium treatment, in which we used sprouts, knowing fully well that it has an impact on aboriginal antioxidant capacity of sprouts, which is mainly due to high vitamin content of sprouts.
Furthermore, we think it is important to make microbiological analysis, because germination conditions, for example temperature, pH, all this will create an ideal environment for the growth of microorganisms. So we had goal to determine, how the used selenium concentration affect the total plate count, coliform bacteria count and Staphylococcus aureus count of sprouts.
We determined the aboriginal water-soluble and lipid-soluble antioxidant capacity of sprout with the PHOTOCHEM chemiluminometer and we applied pour plate technique for the mapping of the mycrobiological state of sprouts.
Experimental results are evaluated, we state that increasing concentrations of selenite or selenate treatment, is primarily water-soluble antioxidant capacity of sprouts was affected. The water-soluble antioxidant capacity of wheat sprout was much higher than the measured values in pea sprout, this may be linked to what we measured. That is much higher ascorbic acid content in case of wheat sprout, which is well known as one of the most important antioxidant properties compounds of wheat sprout.
We conclude from the results of the microbiological, that the highest concentrations of selenite or selenate treatment has a relative significant anti-microbial effect in case of wheat sprouts. Coliform and total plate count showed no clear decreasing tendency, although the values of treatments in both cases obtained were below the control values.
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Energy production systems of phototroph microorganisms (classification of photobioreactors)
35-39Views:72In the field of alternative energy sources there is an argue in the comparison of its effects on the benefits and disadvantages to the economics and the environment. New studies are born which are in contradiction with each other. The demand for bioenergy feedstock is growing rapidly however there are the environmental problems caused by the extending energy crop plantations. There is such a significant need for land to grow traditional energy crops on (rape, soy, palm-oil, sugarcane, etc.) that the food purpose agricultural capacity could be in danger. Probably the extensively
growing energy crops play a role in the very high prices of food. In some countries like China for example laws prohibit the use of food based crops such as corn for energie production. In the case of corn based ethanol production the cost only for the feedstock itself is over 60% of the whole preparation costs which significantly effects the entire economy of the energy productions process.
The microalgaes however have a huge biotechnological potential and their production is notably cheaper then the traditionaly grown food crops growing expenses. They play a significant role as feedstocks in todays industrial production in such fields as comestible production, cosmetics, pharmaceutics and biotechnology especially in biofule production. In the field of economy the major aspects here are also the technological designs
and the construction. For the future industrial production the closed type systems seem to have more advantages compared to the open, pond-type systems. For high value material production the more innovative and more easily developed closed photobioreactors are the profitable regardless the vast techological designs used in the construction. -
Microbiological quality of bulk tank raw milk from two dairy farms in Hajdú-Bihar County, Hungary
105-112Views:327Two main channels have been identified to be responsible for microbiological contamination of raw milk and milk products. Firstly, contamination has occurred due to udder infection from the cow or the blood which harbours most bacteria that come in contact with the raw milk. Secondly, via external factors (may include faeces, skin, contaminated water, environment etc.) which are associated with the operation of milking. There is direct contact with the milk and/or surfaces before, during or after the milking, posing public health risk and economic decline. The aim of this study was to examine the bacteriological quality of bulk tank raw milk samples collected from two different size dairy farms (Farm 1 and Farm 2) of different housing forms (cubicle loose and deep litter) in Hajdú-Bihar County, Hungary in July, 2017. Three samples were taken from each farm, and the total plate count, coliform count, Escherichia coli count, Staphylococcus aureus count, and yeast and mould count were determined in them.
The results clearly showed low level of all measured bacteria group load in Farm 1 samples in comparison to Farm 2 with the exception of coagulase-negative Staphylococcus (CNS) which represented high level in general, indicating significant difference (P<0.05). The mean value of total plate count in Farm 2 samples was higher (1.0 × 105 CFU/mL) than Farm 1 samples (2.8 × 104 CFU/mL). There was a significant difference (P<0.05) in mean count of coliforms in raw milk samples between Farm 1 and Farm 2. Similarly, results of E. coli were significantly different (P<0.05) with mean count of 1.44 × 102 CFU/mL and 2.02 × 103 CFU/mL for Farm 1 and Farm 2 respectively.
Results of Staphylococcus aureus also showed significant difference (P<0.05) with mean count of 9.7 × 101 CFU/mL for Farm 1 and 6.28 × 102 CFU/mL for Farm 2. The mean of mould count recorded was 1.07 × 102 CFU/mL and 4.93 × 102 CFU/mL for Farm 1 and Farm 2 respectively. The recorded mean of yeast count was 1.68 × 103 CFU/mL and 3.41 × 103 CFU/mL for Farm 1 and Farm 2 respectively; however, both farms showed no significant difference (P>0.05) in terms of mean of mould and yeast count. Although Farm 2 produced six times lower milk quantity than Farm 1, the measured microbial parameters were high. Both farms’ microbiological numbers were higher above the permitted limit values as stated by Regulation (EC) No 853/2004, Hungarian Ministry of Health (MoH) 4/1998 (XI. 11.).
This could be an indication of non-conformance to effective GMP, ineffective pre–milking disinfection or udder preparation, poor handling and storage practice, time and temperature abuse and inadequate Food Safety Management System Implementation. Therefore, our recommendation is as follows; establish control measures for pre- and postharvest activities involved in the milking process which would be an effective approach to reduce contamination of the raw milk by pathogenic microorganisms from these farms, strict sanitation regime and hygiene protocol be employed and applied to cows, all equipment, contact surfaces and minimize handling of the milk prior, during and after milking. This will also serve as scientific information to the producers for continual improvement in their operations.
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Búzaminták fungicid szermaradvány ellenőrzése és gombafertőzöttségének kimutatása
45-51Views:88To protect crops with chemicals against different microorganisms are very important because residues of toxins could pollute infected grains. This could have the meaning of serious danger to the human and animal health. Similarly important is knowledge of details about presence or absence of any practically applied protective compounds. Experimental studies were performed on fungicide treated cereals of the Cereal Research Non-Profit Company, Szeged. Chromatographic methods were used (GC and some GC-MS) to find low level fungicide residues, microwave-assisted isolation of ergosterol was followed by HPLC to detect possible fungal infections and a more detailed microbiological analysis completed the work. Concerning the dry year 2000, no fungicides were found in the samples (by November), the ergosterol level was found to be less than 8 ppm, meaning good quality, coupled with an extremely low level of fungal infections. This was confirmed by direct microbiological testing. In comparison with the microbiological results from 1999, a negative difference can be recognized, most probably due to the rainy weather throughout the year.
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Alternatives of sewage sludge use in the crop production
83-87Views:118The produced plants reduce the greenhouse effect because they fix CO2 that contributes to the causing of the greenhouse effect with about 50%. The production of fertilizers is not only a costly process but it needs a considerable energy at the same time. Nowadays, the reduction of the proportion of the fertilizer is significant. One of the reasons of this is that during the production such by-products are produced in a big quantity in which the necessary vegetal nutrients can be found in a considerable measure these enrich the organic matter of soil. The latter is essential condition for the microorganisms in the soil, without which the sustainable plant cultivation can not be achieved. Besides high prices of artificial fertilizers the utilization of the wastes is economically justified. Finally the other reason for the reduction of a usage of artificial fertilizer is that the wrong use of the fertilizer may cause environmental pollutions. I examined the cultivation application of the sewage sludge in laboratory circumstances during my work.
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The effect of bentonite on the quantity change of soil microorganisms, the CO2-production and the activity of saccharase enzyme
287-293Views:96We examined the impact of bentonite – the perspective improving material of sandy soils – and treatments of livestock manure composted with bentonite on sandy soils, within the framework of a small-plot experiment.
The adjustment of the experiment was made on the Experiment Site of the Nyíregyháza Research Centre of the University of Debrecen, Centre of Agricultural Sciences (UD CAS). We collected soil samples from parcels treated with increasing bentonite doses (5, 10, 15, 20 t/ha) on the one hand, and from the parcels treated with livestock manure composted with increasing doses of bentonite, on the other.
We performed laboratory research in the soil microbiology laboratory of the Soil Science Faculty of UD CAS DAS, during which we determined the total number of bacteria, the quantity of microscopic fungi, the number of cellulose-decomposing bacteria, the CO2-production of the soil and the activity of saccharase enzyme.
During the evaluation of the examinations, we made a statistical analysis using SPSS 9.0. We determined the average of measurements, the standard deviation of controls, the standard deviation, the significance value and we also performed a correlation analysis.
Concerning the impacts of bentonite treatment and the treatment of livestock manure composted with bentonite on the examined microbiological features of sandy soil, we can summarize the following:
• Our results prove that the microbiologic activity of the soil has increased owing to the impact of bentonite treatments regarding total number of bacteria and the quantity of microscopic fungi. ”Pure bentonite” treatments – although not significantly in every case – increased these values, but larger doses decreased them. The treatments of livestock manure composted with bentonite resulted in a larger increase regarding both parameters.
• The number of cellulose-decomposing bacteria was increased by the low doses of both series, and was decreased by the higher doses. Higher bentonite doses decreased it in a higher – significant – degree than those of treatments of livestock manure composted with bentonite, whose low dose caused salient number of bacteria.
• Regarding the carbon-dioxide formation, we have experienced an increase even in the case of low dose treatments (nevertheless, the increase did not prove to be significant), but – similarly to the quantitative changes in the number of cellulose-decomposing bacteria – the large doses of both series of treatment decreased the CO2-production of the soil.
• Bentonite also increased the activity of saccharase enzyme significantly. We learned that”pure bentonite” treatments increased the activity of the enzyme to a higher degree than composted treatments. Moreover, it can be stated that the treatments of larger doses of both bentonite and livestock manure composted with bentonite have decreased the enzyme activity – not significantly, though.
• Based on the correlation analysis, it can be stated that as an impact of the treatments, the microbiological activity of the soil has also increased with the increase of the number of soil microbes, as in both treatment series we have experienced a tight positive correlation (r=0.81-0.82) between the change of total number of bacteria and the CO2-production of soil. In both treatments, there was a medium correlation between the total number of fungi and soil respiration (r=0.63-0.63). Furthermore, it can be stated that the usage of organic manure had a positive effect on the activity of cellulose-decomposing bacteria, as in this treatment series there was a positive correlation (r=0.65) between this physiological group and CO2-production. Both treatments prosperously impacted the activity of saccharase enzyme, because there was a medium correlation (r=0.62-0.64) between the activity of the enzyme and soil respiration.