Anais eletrônicos do International Symposium on Science and Biotechnology

ANTAGONISTIC ACTIVITY OF ISOLATED PROBIOTICS AGAINST ESCHERICHIA COLI ESBL

Douglas Zarur, Vilmair Zancanaro, Fernanda H. Lorenzon, Emyr Hiago Bellaver — Wed, 20 Dec 2017 00:00:00 -0200

Potentially all microorganisms are able to synthesize several substances in vitro with inhibitory activity for themselves or other microorganisms, producing bactericidal or bacteriostatic effects. The objective of this work was to establish the bacterial growth antagonistic properties of Escherichia coli ESBL by Lactobacillus casei strain Shirota, Bifidobacterium animalis DN173010, isolated of commercial probiotic products, and Kefir grain by the overcoated agar method. Analysis of the results allowed to gauge halos of 7.1 ± 2 mm inhibition of growth by L. casei strain Shirota and 6.3 ± 2 mm by B. animalis DN173010, Kefir grains did not produce inhibition halos against the multiresistant strain. The bacterial growth inhibition activity can be justified by the competition of nutrients from the medium and/or by the secretion of bacteriocins, protein or peptidic compounds ribosomally synthesized in the primary metabolism with antagonistic activity and because proteins are expected to behave differently on certain target microorganisms and culture conditions, and the production of these compounds by the Kefir grains cannot be excluded.

Physicochemical of strawberry pseudofruits of applied zinc oxide nanoparticles applications properties.

Liziane Cássia Carlesso, Gean Luz, Cristiano Reschke Lajus, Luciano Silva — Wed, 20 Dec 2017 00:00:00 -0200

Strawberry cultivation is appreciated in many countries because it has well-defined nutritional and attractive sensory characteristics. This work proposes the application of nanotechnology to the improvement of physico-chemical characteristics in the strawberry crop, with the main objective of analyzing the influence of different treatment doses of fruits with zinc oxide nanoparticles on nutritional performance.The physicochemical analyzes of the pseudofruits from strawberry cultivars of the San Andreas species were carried out at the Chemistry Laboratories, from November 2015 to June 2016 (3x5), factor A (fertilizer: recommended dose of Zn, 50% of the recommended dose in the form of nanoparticles of Zn and 100% of the recommended dose) was used in a randomized complete block design (DBC) In the form of nanoparticles of Zn) and factor B, 2nd Period; 3rd Period; 4th Period and 5th Period, with 7 repetitions. The data collected were subjected to analysis of variance by the F test and the differences between means were compared by the Tukey test (P≤0, 05). The process of treatment of strawberry culture with zinc oxide nanoparticles is efficient for anthocyanin and soluble solids nutrients. The nanoparticles of 100% of the recommended dose demonstrate to be more effective as the zinc oxide in its natural form, to increase the values of soluble solids.

DETERMINATION OF MINIMUM INHIBITORY CONCENTRATION OF KLEBSIELLA PNEUMONIAE KPC FRONT OF THE ESSENTIAL OIL OF MELALEUCA ALTERNIFOLIA

Alessandra Gioppo, Fernanda H. Lorenzon, Emyr Hiago Bellaver — Wed, 20 Dec 2017 00:00:00 -0200

The essential oil of Melaleuca alternifolia (OTT) has antifungal, anti-inflammatory and antimicrobial effects reported in the literature that attributes this last to more than about 70% of terpenes present in its composition, and its mechanism of action consists in the breakdown of the membrane cytoplasmic activity leading to membrane protein damage, cytoplasmic coagulation, changes in electron fluxes and transport mechanisms and above all, the interruption of the proton motive force. In this century, the advent of multidrug-resistant microorganisms to available drugs has raised widespread concern, prompting industry and researchers to opt for alternative mechanisms in the treatment of some bacterial infections. The objective of the study was to evaluate the antibacterial properties of OTT against Klebsiella pneumoniae producing carbapenemase (KPC) and its standard strain K. pneumoniae ATCC 13883 by the Minimal Inhibitory Concentration (MIC) method by macrodilution and Spot-on-the-lawn. stipulated for KPC and K. pneumoniae ATCC to MIC in 0.25% OTT by the two methodologies, proving the antimicrobial activity of the oil and concluding that the mechanism of resistance to carbapenems does not influence the sensitivity to OTT.

FACTORS OF INNOVATION AND COMPETITIVENESS OF BRAZILIAN BIO INDUSTRIES

Wed, 20 Dec 2017 00:00:00 -0200

Innovation is one of the priority elements for the competitive advantage of organizations. Bio-industries are part of a segment of companies that uses biotechnology to add value to their products and services. The identification of key factors of innovation contributes for companies of this niche area to make assertive decision making to enter and remain in an extremely competitive market of large companies and clusters. The methodology of the present study was based on bibliographical research of pertinent Brazilian legislation, the Oslo manual, CAPES database, journals and other publications in the area. In addition, it was considered fundamental elements of the MCTI, EMBRAPII and guidelines of FINEP, FNDCT and FAPESC to subsidize the definition of recent factors that authors cites and approach to define strategies of innovation and competitive positioning of bio-industries. The results contemplate a set of innovation and competitiveness factors within the scope of internal and external performance of this business segment, composed of intellectual capital, R & D & I (Research, Development & Infrastructure), regulatory environment, strategic alliances, costs, human capital, production and control systems, segregated in 53 sub-factors, which allows the identification and subsequent analysis aiming to contribute to the improvement of the respective bio-industries.

REMOVAL OF ORGANIC MATTER FROM WATER DISCARDED BY PAPER AND CELLULOSE MILLS AFTER UV/H2O2 TREATMENT

Wed, 20 Dec 2017 00:00:00 -0200

aper and cellulose mills effluents are composed by several combinations of chemical reagents used in productive stages and of substances originating from raw material, used to produce paper and cellulose pulp. Considering the volume of effluents generated by such industries and the current crisis of water resources, it is blatant the need of an efficient treatment for the discarded water. The advanced oxidative processes constitute an attractive option for treating several effluents. They are capable of promoting degradation and even mineralization of refractory pollutant particles. The current paper looks forward to assessing the performance of the UV/H2O2 process in removing organic matter present in the effluent of a paper mill. The research was carried out in a photochemical reactor with active capacity of 1300 mL. As source of UV radiation, it was used a medium pressure mercury lamp of 125 W. During 120 minutes of UV irradiation, six H2O2 concentrations were testes, varying between 15 and 200 mg.L-1. The results revealed the effectiveness of the process when removing 84% of the CDO and 91% of the BOD5. The performance of the UV/H2O2 process was influenced by the availability of H2O2 and the time of UV irradiation.

ANTIOXIDANT AND PHOTOPROTECTOR EVALUATION OF GRAPE EXTRACTS

Lucas Bertaiolli Andrade, Fernanda Farias Hollas, Claudriana Locatelli — Wed, 20 Dec 2017 00:00:00 -0200

The agroindustry residue contains a large variety of biologically active species that are wasted. The objective of the present study was to evaluate the antioxidant and photoprotective activity of the hydroalcoholic extracts of the Sauvignon Blanc grape (SB) and a mixture of Tannat and Malbec varieties (BTM). The total phenols content was evaluated by the Folin-Ciocalteu methods; total flavonols by AlCl3; antioxidant activity by radical DPPH sequestration, inhibition of the autoxidation of acid β-carotene linoleic system and iron chelation capacity. The photoprotector effect (SPF) was evaluated by scanning spectrophotometer among the wavelengths of 290nm to 320nm with readings every 5nm. The grape extracts BTM and SB presented an index of total phenols of 33.02 and 16.69 mg equivalents of Gallic acid/gram of extract respectively, while the content of flavonoids was 7.16 and 0.46 mg quercetin equivalents/gram of extract. BTM extract was able 98% DPPH sequestration and SB 88.9% in extract concentration of 3200 ᶙg/mL. Iron chelation capacity was greater for the extract from the BTM (95.58%). The results show an SPF 157.22 to BTM extract and 0.413 to SB. In view of the results, it can be stated that MTM bagasse extract has a better biotechnological potential.

TREATMENT OF TEXTILE WASTEWATER EMPLOYING ORGANIC COAGULANT

Wed, 20 Dec 2017 00:00:00 -0200

Among the available treatment methods for textile wastewater, the coagulation/flocculation process is highlighted. Thus, the present study aimed to determine which volume of the tannin coagulant (SETA S5T) presented better results in the treatment of coagulation/flocculation of textile wastewater (company located in the west of Santa Catarina). The trials were carried out at the LEMA (UNOESC Videira), from April to June 2017. During the methodological procedures were tried out the volumes of: 0,5; 1; 1,5 and 2 mL of tannin per liter of sample, using Jar-test equipment. In this the coagulant was initially incorporated into the samples (20 min. - 100 r.p.m), followed by correction of pH (±7) with calcium hydroxide. Finally, 2 mL of cationic polymer was added (20 min. - 40 r.p.m). The efficiency of the process was determined by the analysis of the parameters: Color, Turbidity, COD, Suspended Solids (TSS) and Sedimentable Solids (SS), from Imhoff cone supernatant (1 hour at repose). At the end of the experiments the following percentages of removal were obtained: 0,5 mL: 97,91% for color; 99,34% of turbidity; 41,78% of COD; 77,52% for SST and 56 mg/L SS production. 1 mL: 97,85%; 99,37%; 36,56%, 82% and 86,93. 1,5 mL: 98,2%, 99,3%, 29,83%, 75,82% and 106,01. 2 mL: 98,48%, 99,30%, 23,02%, 72,75% and 124,04. It was concluded from this study that the best tannin volume was 0,5 mL/L.

PHOTOCHEMICAL REDUCTION OF CR(VI) FROM ELECTROPLATING WASTEWATER USING INDUSTRIAL STEEL WASTE AS IRON SOURCE

Wed, 20 Dec 2017 00:00:00 -0200

The use of chromium in different industrial activities, such as electroplating, textile dyeing, leather tanning and metallurgy, results in toxic wastewaters containing chromium species, which must be treated before discharging into receiving waters. In this work, an industrial steel waste (mill scale found on all hot-rolled steel products) was evaluated as iron source to promote the reduction of hexavalent chromium species present in an electroplating wastewater. Mill scale (MS) was characterized by spontaneous magnetization, point of zero charge (pHpzc) and metal leaching. The efficiency of MS was studied in the presence and absence of reducing agents (citric acid and oxalic acid), and its stability and the influence of the homogeneous reaction on the overall efficiency was also evaluated using a batch system. Higher concentrations of MS resulted in an increase in the rate of Cr (VI) reduction. It was observed a negligible effect of citric acid and oxalic acid on Cr(VI) reduction in the absence of MS, however, in the presence of MS particles and organic acids, Cr(VI) reduction is significantly enhanced. It was verified that MS acts only as source of iron for the solution. The results indicate that MS can be used effectively in wastewater treatment, more precisely in Cr (VI) reduction.

PHOTOCHEMICAL UVC/H2O2 OXIDATION SYSTEM AS AN EFFECTIVE METHOD FOR THE DECOLOURISATION OF BIOTREATED PULP AND PAPER INDUSTRY WASTEWATER

Wed, 20 Dec 2017 00:00:00 -0200

Biological oxidation processes have shown good results for the mineralisation of the non-recalcitrant fraction of wastewater from pulp and paper industry. However, they do not always represent a feasible technique for colour removal, requiring a polishing step for decolourisation before the wastewater discharged into a receiving water. In this study, photochemical UVC/H2O2 oxidation system was used as a polishing step in the decolourisation of a wastewater collected after biological oxidation from a pulp and paper industry. The photochemical oxidation assays were performed in a lab-scale photo-reactor whit a quartz tube (Ø = 20mm), filled on UVC lamp (6W), inside to inox tube (Ø = 70mm and L = 285mm). Photochemical reaction rates were determined under different operational conditions: H2O2 dosage (0 - 100 mM) and temperature (15°C , 25°C and 35°C). The results showing that the photochemical UVC/H2O2 oxidation system showed promising results as polishing step for the decolourisation of a bio-treated wastewater from pulp and paper industry. UVC and H2O2 alone showed negligible colour removal, indicating that the hydroxyl radicals generated from hydrogen peroxide photolysis under UVC radiation is the principal reaction mechanism. A higher hydrogen peroxide dosage and wastewater temperature showed a positive influence on the UVC/H2O2 system.

ISOLATION AND MORPHOLOGIC CARACTERIZATION OF STREPTOCOCCUS SP. STRAINS FROM HEALTHY HORSES.

Wed, 20 Dec 2017 00:00:00 -0200

Streptococcus strains are potential producers of hyaluronic acid, and they are present on the nasal cavity, being an etiologic agent of equine strangles. The objective of this work is the identification of b-hemolytic Streptococcus sp. strains isolated from horses. The collect of samples was made between August and October 2017 in properties from Santa Catarina. A questionnaire was carried out to determine the animal health profile. After being collected, the samples were seeded and cultured on 5% sheep blood agar during 48 hours at 37ºC and aerobic/anaerobic conditions. Catalase test and gram staining were carried out in the isolated colonies. During the above-mentioned period 13 samples were collected from healthy horses: 07 females and 06 males, all vaccinated against infectious disease and with strangles historic. None of the isolates obtained was characterized as b-hemolytic, 07 isolates were characterized as Streptococcus α-hemolytic and presented compatible morphology in gram staining. According to another study that isolated 38 samples, the majority of the samples was identified as the cause of equine adenitis. The results obtained from some authors shows the difficult to isolate Streptococcus strains from healthy horses. However, collecting sample from animals that learn the symptoms of strangles will facilitate the isolation of target microorganisms.

PHYTOCHEMICAL PROFILE AND BIOLOGICAL ACTIVITY USING ISABEL GRAPE (VITIS LABRUSCA L.) RESIDUE EXTRACT.

Fernanda Farias Hollas, Lucas Bertaiolli Andrade, Claudriana Locatelli — Wed, 20 Dec 2017 00:00:00 -0200

The grape residual presents high level of active substances composition, characterizing it as a product that the industry could use more efficiently in a better profitable way, in the formulation of beauty goods and food. The goal of this research was to evaluate the phytochemical profile and biological activity of the Isabel Grape residue. In order to obtain the extracts, it was used the method of trituration in different solvents (water, ethanol, methanol, and a hydroalcoholic mixture). Colorimetric tests used showed the phytochemical profile. The technique of Folin-Ciocalteu evaluated the content of total phenol and the method of AlCl3 evaluated the total flavonoids. As for the biological activity, the antioxidant activity was evalueted by the method of DPPH, the inhibition of auto-oxidation of β-carotene/linoleic acid and the ability of metallic ions sequestration. The spectrophotometric reading obtained the photoprotection factor (SPF). The results showed that the hydroalcoholic extract presents a great phenols and flavonoids quantity that could relate with the extract antioxidant and photo protective activity. This extract presented a SPF as 1.87 and a good antioxidant by the used methods. The results showed that the Isabel Grape residue hydroalcoholic extract presents a great biotechnological potential.

TRENDS AND INNOVATIONS IN THE PET FOOD MARKET

Sheila Baroncello, Jane Mary L. N. Gelinski, Vinícius Caliari — Wed, 20 Dec 2017 00:00:00 -0200

Studies show that Brazil in the world ranking is the second country with the largest population of domestic animals, with the front only the US. These animals, which usually occupied a decorative space, were valued as a company and considered an integral part of the family.It is estimated that 59% of Brazilian households have some pets, spending an average of R $ 390,00 on products and services (pharmaceuticals, vaccines, beautification and accessories). In this survey we evaluate the current situation of the pet market, trends and innovations, especially on snacks/foods with high added value. In this research we have compiled national and international circulation periodicals for the last five years (2012-2017). It was found that the world market grew at an annual average of 17%, earning around US $ 1.5 billion since the year 95. The Brazilian market in 2014 reached R $ 16 billion, placing the country in the third position in the ranking world. The food segment is the main representative of the sector, with 67% of revenues. It is verified that the trend of the sector is directed towards health and animal welfare focused on health and practicality. It is generally seen that the market is susceptible to changing societal patterns and demands impose imposed by consumers, and to achieve competitive advantage must constantly be on the side of innovative technologie.

PREVALENCE OF SALMONELLA ENTERICA HEIDELBERG IN THE PRODUCTIVE CHAIN OF BROILERS

Wed, 20 Dec 2017 00:00:00 -0200

Salmonella enterica subsp. enterica Heidelberg is one of the most invasive serovar and deserves attention due to its prevalence in the productive chain of borilers. This research analysed the percentage of S. Heidelberg serotype samples in semi-solid medium (MSRV) and to evaluate the profile of antimicrobial susceptibility. Isolation of samples from avian environment was performed from MSRV method recommended by ISO 6579. Aliquots removed from the MSRV migration zone were inoculated in selective medium agar XLD and VB agar (DIFCO®) with incubation at 37ºC/24h. Were perfomed biochemical and serological analysis of colonies as well molecular analysis based on PCR. A total of 1506 samples were obtained, of which n=376 samples (24.96%) were positive for S. Heidelberg. From isolates, n=1130 did not migrate in MSRV (75.03%). However, n=62 (16.49%) confirmed as being S. Heidelberg. Isolates with migration in MSRV were susceptible to all antimicrobials tested, whereas all at the group without migration (flagellar injury?) showed resistance to Ceftiofur and Neomycin. It is early to affirm the existence of a relation of using antimicrobials in PCB with the flagellar injury of the pathogen. However, monitoring should be performed, due we detect isolates resistant to at least two broad-spectrum antibiotics at the group of isolates with no ability to migrate in MSRV medium.

Keywords: Produtive chain. Salmonella. Antimicrobials. Flagella.

MEASURING THE TAX CHARGE INCIDENTS ON NATIONAL WINE: IMPACT ON THE COMPETITIVENESS OF BIO-INDUSTRIE

Wed, 20 Dec 2017 00:00:00 -0200

The increase in the national tax burden on wine production in heading 2204 of the Tax Incidence Table on Industrialized Products-TIPI, which is the object of this study, is one of the factors of economic loss and competitiveness for small and medium-sized bio-industries. The objective is to evaluate the preventive act, according to the Brazilian legislation, which allows to pay the smallest possible amount of taxes, in order to verify the lower tax burden. It is qualitative, descriptive and analytical research. Viti-winemakers are included in the tax regimes: Real Profit and Presumed Profit. In 2018, Complementary Law nº155 / 2016 will come into force, amending Complementary Law 123/2006, allowing wineries to opt for the Simple National regime. Currently, wines taxed by Presumed Income or Real Profit have an incidence of 54.73% of charges (ICMS, IPI, PIS, COFINS, CPP ..). One of the taxes responsible for increasing the tax burden is the IPI and its change in the calculation method will increase the tax expense by an average of 50%. In the scenario of legal economy the option for the Simple National will imply in tax planning with significant reduction of costs. In this context, in order to guarantee the competitiveness and sustainability of organizations and, taking into account the need to undertake and innovate, tax management will become necessary in bioindustries.

APPLIED COMPOSTING TO THE BIOLOGICAL FERMENTATION OF SOLID WASTE FROM PRODUCTION OF PIG, BOVINE CATTLE AND CHICKENS

Wed, 20 Dec 2017 00:00:00 -0200

The great Catarinense West is economically based in the productive chains of poultry and swine. At the level of the rural producers the activity is intensive, resulting in the high production of waste. This is a weak link in sustainability, given the possibilities of deleterious effects, impacts and externalities on the environment. This study evaluated the technical efficiency of the composting process in the biological stabilization of the solid fraction of swine, poultry and bovine litter. The composting technique - controlled aerobic biological process speeds up the decomposition of organic material. Seven formulations were prepared with the three types of solid material. For each formulation, 6.6m3 of organic material was used, and the fermentation mass was recorded every three days for gas exchange and temperature control. As a result of formulations prepared and submitted to composting, the internal temperature of the mass increased to 40°C on average after ten days. Except for bovine solid waste alone, all formulations had a mean maximum temperature of 54.6°C between thirty to sixty days of fermentation. The biological stability behavior occurred with sixty-eight days in processing. It was concluded that all organic solid waste used for this study can be stabilized by the aerobic fermentation process in solid cultive.

Influence of ethylene, absisic acid or brassinosteroid application over gene expression during strawberry ripening

Luane Bosetto, Ricardo Antonio Ayub, Letícia Reis — Wed, 20 Dec 2017 00:00:00 -0200

Strawberry (Fragaria x ananassa Duch.) is a crop of high commercial value that has been used as a model for non-climacteric fruit ripening studies, aiming the reduction of postharvest losses, due its high perishability. While for the climacteric fruits, ethylene is the main responsible throughout the ripening process, in the non-climacteric fruits, this process is not completely understood. The aim of this work was to evaluate the effects of ethylene (ET), abscisic acid (ABA) or brassinosteroid (BR) application over strawberry ripening, analyzing gene expression on key points of each pathway. The strawberry fruits cv. Camino Real in four ripening stages (green, white, pink and red) were harvested (postharvest) or not (field) and treated with ethephon, abscisic acid, epibrassinolide or none (control), in ethanol 2%. After consecutive steps, the cDNA was obtained, allowing the analysis of relative gene expression by Real Time PCR. The results shown that these regulators play a role over the ripening process, acting each one in a specific point, mainly in white (ET) and pink stages (ABA and BR), which was observed in the field assay. At postharvest, the stress caused by the harvest altered the plant response. Therefore, we concluded that these phytohormones act together in the non-climacteric fruit ripening regulation, however, due its high complexity, more studies are needed.

THE BEST TISSUE TO OBTAIN DNA FROM APPLE TREE USING CTAB EXTRACTION PROTOCOL

Wed, 20 Dec 2017 00:00:00 -0200

The DNA extraction is the key for all molecular biology procedures. In plants, this stage is absolutely affected by the tissue compounds, that are variable between the species. In apple tree, polyphenols are the main compounds that affects DNA quality and integrity. The aim of this work was to apply a protocol for DNA extraction using different apple tree tissues and to determine the best one to obtain high DNA quality. The protocol tested was an adaptation of CTAB, using leaf, bud, stem, apple skin and flesh. The experiment was carried out in a completely randomized design with five replicates, considering each tissue. To determinate the best quality of DNA was used de relation 260/280 nm and 260/230 nm. Quality parameters and DNA concentration were obtained with absorbance spectrophotometer. It was performed variance analysis and Tukey's test, with a significance level at 5 %. The DNA integrity was observed in agarose gel 0.8 %. The highest mean concentration were 374.53 ng/µL in stem and 395.58 ng/µL in bud tissue. The best quality (260/280 ≈ 2.0, 260/230 ≈ 1.8) was obtain with leaf, bud and stem, the same for DNA integrity. The best tissues to obtain DNA from apple tree with high concentration and quality were bud, stem and leaf, suggesting higher efficiency using this protocol.

REUSE OF BYPRODUCTS OF INDUSTRIAL PROCESSES IN NEW PROCESSES

Tiago Borga, Marcio Alexandre Kreusch — Wed, 20 Dec 2017 00:00:00 -0200

MOLECULAR ANALYSIS AND PROFILES OF ANTIBIOTICS RESISTANCE OF SALMONELLA ENTERITIDIS ISOLATES FROM POULTRY CHAIN PRODUCTION OF THE BRAZILIAN SOUTHERN REGION

Francielli Mello Ferri, Jane Mary Lafayette Neves Gelinski, César Milton Baratto — Wed, 20 Dec 2017 00:00:00 -0200

The agroindustrial sector of meats production and derivatives presents importance for the economic development in the Brazil's Southern States. Salmonella Enteritidis was isolated in all sectors of the productive chain of poultry and PCR-based techniques have demonstrated good applicability for identification and epidemiology studies of food pathogens. The aim of this work was to select molecular techniques with greatest potential to detect polymorphism of Salmonella Enteritidis strains, besides to determine the antibiotic resistance profile, and to find correlation between them. Sixty-eight Salmonella Enteritidis strains isolated from the poultry production chain in the southern States of Brazil were analyzed with the PAST program by UPGMA method and the similarity index Euclidean. The molecular technique RAPD, using the 23L primer, presented highest polymorphism, with 10 polymorphic groups and a Discriminatory Power of 0.96. For the resistance test, 75% of the isolates showed multiresistance, moreover the isolates were 100% resistant to erythromycin and 77% resistant to nalidixic acid. In conclusion, the RAPD molecular technique with the primer 23L have good applicability in epidemiological studies and the high incidence of mutiresistance isolates serves as an alert for the indiscriminate use of antibiotics in productions animals.

GRAPE JUICE STABILITY IN BAG-IN-BOX AND GLASS BOTLLES

Wed, 20 Dec 2017 00:00:00 -0200

This work studied the composition and sensory properties of grape juice after packaging under different configurations: bag-in-box (BIB) and glass bottles. The experiments were carried out over a period of 210 days and analysis were performed at initial time (just before packaging) and after 30, 60, 90, 120, 150, 180 e 210 days of storage. The samples of grape juice were analyzed for soluble solids content (ºBrix), pH, titratable acidity measurements (meq), color intensity was determined by the sum of measuring the absorbance of wine at different weave length (420, 520, 620 nm) using a UV–vis. Tint was determined using the formula (Abs420/Abs520). The total anthocyanins were determined by the pH-differential method. Comparing BIB and glass bottles, in both pH and acid showed the same variations, where acidic get lower with the days pass. BIB shown higher brix than bottles. The total anthocyanins decreased from 240 mg/ 100g just before packaging to 120 mg/100 g after 210 days, the same pattern was observed in both packages. Color index was almost constant during 210 days, while tint shown a slight increase during 210 days in both packages. Thus, we concluded that the grape juice stored in BIB have the same properties of grape juice stored in glass boltless and BIB may be used to substitute glass bottles, which are heaviest and expensive than BIB.

USE OF PROBIOTICS IN COLORRETAL CANCER INDUCED BY 1,2-DIMETHYLHYDRAZINE IN ANIMAL MODEL: A REVIEW

Wed, 20 Dec 2017 00:00:00 -0200

Colorectal cancer is one of the most frequent in the Brazilian population. The available forms of treatment are associated with a high risk of complications, thus highlighting the need to elaborate new therapeutic strategies. Intake of probiotics may be a viable therapeutical option for the treatment of colorectal cancer. The goal of the present study was to review the use of probiotics in the treatment of colorectal cancer induced by 1,2-dimethylhydrazine in rats. This is a literature review of articles published between 1999 and 2017 in the pubmed database, with the descriptors: "probiotics", "colorectal cancer" and "dimethylhydrazine". Found 23 articles within the search inclusion criteria. The probiotic strains studied showed differences in type, concentration and mode of intervention. The use of probiotics was mostly isolated or associated with prebiotic fibers. The main benefits were: reduction of incidence and size of tumors, modulation of the microbiota in a positive way, better anti-inflammatory response and lower DNA damage. The need for more studies to provide more complete and delineated data, with mixtures of strains and specific amounts, provides more conclusive results on the real benefits of this type of supplementation in colorectal cancer.

Keywords - Colorectal Cancer. Probiotics. Dimethylhydrazine. Supplementation.

ISOLATION AND CHARACTERIZATION OF MICROORGANISMS Β-GALACTOSIDASE PRODUCERS WITH POTENTIAL BIOTECHNOLOGICAL APPLICATION

Wed, 20 Dec 2017 00:00:00 -0200

Lactose is the main carbohydrate in milk, its degradation is due to the action of β-galactosidase on glucose and galactose. The objective of this work was to isolate new β-galactosidase-producing microorganisms and optimize their production. The isolates tested were obtained from dairy residues using X-gal as indicator for the production of the enzyme. An isolate obtained had the optimum culture conditions through fractional factorial design, analyzing the effect of temperature, concentrations of whey powder, yeast extract and lactose on the production of the enzyme. The enzyme was partially purified and characterized. The results showed the presence of β-galactosidase only in the cell precipitate and among the studied factors the temperature and the concentration of yeast extract had an effect on the production of the enzyme in 14 hours of culture. Cell enzyme solubilization tests demonstrated that sonication of these in phosphate buffer pH 7.0 for 20 minutes with 0.5% TritonX or 0.2% SDS were more efficient for release of the enzyme into the supernatant. The optimum activity of the enzyme obtained was at 45 °C and pH 7.0. Therefore, the V1 microorganism, based on the parameters evaluated to date, has potential for the production of β-galactosidase with interesting characteristics, as well as the potential use of alternative means for production using whey powder.

Keywords - lactase. zero lactose food. lactose intolerance. optimization.

DETERMINATION OF THIOCYANIDE IN HUMAN SALIVA USING A COLORIMETRIC REACTION

Wed, 20 Dec 2017 00:00:00 -0200

The reaction Fe³⁺ + CSN^- <--> D Fe(SCN)²⁺ is very simple and easy to observe, because Fe³⁺ has a slight yellow color, CSN^- is colorless and Fe(SCN)²⁺ are deep blood-red (λmax = 447 nm), the determination of equilibrium constant of this reaction is presented in many laboratory manuals. When Fe³⁺ concentration is much higher than CSN^- concentration, all thiocyanate ions are assumed to be converted to the Fe(SCN)²⁺ complex ions. The same calibration curve can be exploited in the quantitative analysis of SCN^-. In the first part the equilibrium constant was determined . At the second part each student measures his or her saliva thiocyanate ion concentration with visible spectrophotometry. In the first part of this manuscript, the constant of equilibrium was determinate. At the second part, thiocyanide concentrations was determinate in human saliva. This laboratory practice was carried out in UNOESC at the first and the second semester of 2007. The equilibrium constants found were close to that reported in literature 93. The thiocyanide concentrations found in saliva had an average value of 1.51 mM for nonsmoker woman’s.

ISOLATION OF BACILLUS SP. FROM ORGANIC VINEYARD SOIL AND EVALUATION OF ENTOMOPATHOGENIC POTENTIAL AGAINST ANASTREPHA FRATERCULUS LARVAE

Wed, 20 Dec 2017 00:00:00 -0200

Bacillus, insulation, entomopathogenic, AnThe viticulture is a farming activity of great economic importance, during the last decades there was a microbial diversity in these areas, and one of the groups of bacteria with higher diffusion is the Bacillus. In contrast one of the major pests of fruit trees of lump is to Anastrepha fraterculus, their combat occurs through several tracks. The objective of this work was to isolate, and check the activity entomopatogênica Bacillus gram-positive bacterium from soils of organic vineyard in larvae of Anastrepha fraterculus. Were collected 3 soil samples from organic vineyard, and from this were isolated Bacillus gram-positive bacterium with the presence of spores and crystals. After the selection were performed bioassays of pathogenicity in larvae of Anastrepha fraterculus. We used four treatments (T1=water, T2=Isolate BS-10S, T3=Isolate BS-8S and T4=Isolate BS-6S). In each treatment were placed 0.5 ml of growing end of each crop in disposable cups containing 50 mL of artificial diet and 10 larvae of second urges. After 48 hours in B.O.D at 25ºC, were counted the dead larvae. According to the results were obtained eight isolates of Bacillus, and three had similar morphology to gender sphaericus. The bacterium tested, the isolate BS-6S (5,67X109 CFU/mL) was the only one that presented itself to pathogenic larvae of Anastrepha fraterculus.

Keywords - Bacillus, insulation, entomopathogenic, Anastrepha, viticulture.

MOLECULAR MARKERS IN THE DIAGNOSIS OF PERIODONTAL DISEASE

Queila Da Luz Samistraro, Claudriana Locatelli, César Milton Baratto — Wed, 20 Dec 2017 00:00:00 -0200

Periodontal diseases are highly complex and have etiology multifactorial, with the participation of numerous biological mechanisms. This work aims to discuss some scientific research that uses molecular biology methods to evaluate periodontal disease. Several studies in the literature show that these tools can understanding the mechanisms involved in pathogenesis and the identification of new biomarkers of periodontal diseases, which can be used future diagnoses to detect the presence of active disease, to predict progression and evaluate the response to periodontal therapy. Several markers of lipid peroxidation, protein oxidation and DNA damage induced by reactive oxygen species can be measured in saliva or oral mucosa. Clinical studies have shown an association with oral pathologies at least for some of the established salivary markers of oxidative stress. The results showed an important correlation between oxidative stress biomarkers and periodontal disease.

PHARMACOTHERAPEUTIC FOLLOW-UP IN A BASIC HEALTH UNIT IN CAÇADOR - SC: CASE STUDY

Wed, 20 Dec 2017 00:00:00 -0200

Inappropriate use of medications may decrease or exempt the expected therapeutic effect. The objective of this study was to promote pharmacotherapeutic follow-up in a patient with psoriasis under treatment with methotrexate (MTX). The method used was the case study type. The data were collected through consultation, using a structured clinical document and adapted from the Dáder method. During follow-up, misuse of folic acid was identified, which should be administered once a week after administration of MTX, and the patient administered 1 tablet daily until the end of the card. MTX has been shown to interact with hydrochlorothiazide, which may lead to severe leukopenia. The possibility of a severe interaction between MTX and NSAIDs was also observed, a relevant factor since the patient's main complaint was hyperalgesia, a fact that may lead to self-medication and anti-inflammatory use. The data obtained were discussed with the nurse and the doctor of basic health unit, to elaborate the pharmaceutical Intervention. The pharmaceutical intervention allowed correcting the detected problems, as well as stimulating the patient to be able to promote self-care. The importance of the multidisciplinary team in decision-making was perceived during the work.

APPLICATION OF ORGANIC ACIDS AND SALT IN REPLACEMENT OF SODIUM METABISULFITE IN DEHYDRATED CUBE APPLES

Barbara Helen Giordan, Fabiana A. S. M. Soares, Johannes Newton do Prado — Wed, 20 Dec 2017 00:00:00 -0200

Increasingly the population adopts a healthy diet and becomes demanding about the nutritional value of food, which has increased the demand for dehydrated foods as a healthy and practical option for intermediate meals. Enzymatic browning is a phenomenon that induces changes in color, taste and nutritional losses and is related to the enzyme polyphenoloxidase, which uses phenolic compounds as substrates. The objective of this work was to evaluate the application of conservative solutions containing citric acid (AC), ascorbic acid (AA) and sodium chloride (NaCl), replacing sodium metabisulfite in preserving the color of dehydrated apples. Analyzes were performed to evaluate the stability of the apples, through the pH content, soluble solids and sensorial analysis. At the end of the 10 hrs drying process, the final humidity of the apples was 3.58%. The pH levels in all solutions remained similar, decreasing as the acid concentration increased. Soluble solids contents also presented few variations, explained due to the content of soluble solids present naturally in the fruits in natura. According to the analyzes performed, it can be seen that the conservative solution containing ascorbic acid, citric acid and NaCl was effective to control enzymatic browning, serving as a potential substitute for Sodium Metabisulfite.

LEGAL AND NORMATIVE INSTRUMENTS IN CIVIL CONSTRUCTION WASTE MANAGEMENT IN BRAZIL

Scheila Lockstein, Maria Rita Chaves Nogueira, Petrick Anderson Soares — Wed, 20 Dec 2017 00:00:00 -0200

Civil construction waste (CCW) cause to financial and logistical problems and is environmentally degrading. This work describes the legal basis and guidelines governing the management of CCW in Brazil. The Brazilian legal framework in the area began around 1981, with the National Environmental Politics. However, only in 2010 was promulgated the National Politics on Solid Waste (PNRS). In an innovative way, the PNRS establishes an order of priority for waste management, from non-generation to treatment and waste disposal. The National Environmental Council (CONAMA) through resolution 307/02 establishes guidelines, criteria and procedures for the management of CCW. Later, some items and articles of this resolution were changed by the Resolutions 348/04, 431/11, 448/12 and 469/15. On the other hand, the Brazilian Association of Technical Standards through the (NBR) 10004 defines waste and classifies them according to the physical nature, chemical composition and potential risk to the environment. In 2004, it published a set of norms referring to the solid waste of the civil construction stands out NBR 15114, 15115 and 15116. Finally, there is a legal and technical structure that guides and norms the management of CCW in the country, with emphasis on PNRS and CONAMA Resolution 307/02.

CHARACTERIZATION AND USE OF ISABEL'S GRAPE SEED OIL AFTER THE VINIFICATION PROCESS

Wed, 20 Dec 2017 00:00:00 -0200

Vitiviniculture has become an important activity for the growth of small farms. In the region of Santa Catarina there are many wineries, which in turn, generate waste after the vinification process (bagasse). With this objective, we evaluated the oil of the grape seed (Vitis labrusca, variety “Isabel”) after the vinification, for possible use in cosmetic. The physical-chemical characterization of the oil of the grape seed was obtained through the analyzes of yield 7.29%; acidity 0.045mg KOH / g; volatile matter 0.64%; peroxide index 6.63 meq O2; density 0.91 g.cm3; pH 5, retraction index 1.47n and saponification 194%. It was verified that the grape seed oil analyzed, obtained values accepted by the current legislation and, therefore, can be used as raw material for new products. Contributing in the reduction of the environmental impact generated by the wineries and with their possible reutilization, generating increase of income to the producer. With the results achieved, we developed a hand cream with different concentrations (1% and 3% oil) and evaluated its organoleptic characteristics (appearance, color, smell and touch), physical-chemical (pH, centrifuge and spreadability), and sensory (absorption, drying, stickiness, spreadability, consistency, sensation: oily, hydration and softness of the skin).

Keywords: vinification. Reuse of residues. Grape seed oil. Cosmetic.

Financial support: UNOESC.

E-mails - may89_05@hotmail.com; vinicius.caliari@unoesc.edu.br

IN VITRO AND IN VIVO TECHNOLOGICAL VALIDATION OF ISOTHERMAL EQUIPMENT DEVELOPED FOR THE PROCESSING OF COOLED SWINE SEMEN

Wed, 20 Dec 2017 00:00:00 -0200

Agribusiness is a segment that contributes greatly to Brazil's economy. In this segment, the production of animals for animal protein in the food industry stands out. In order to meet the demand of pigs, the artificial insemination-IA with cooled semen is used in its reproduction. In this research, with the aim of increasing the efficiency of the AI process with cooled semen, a new isothermal technology was developed for the processing of swine semen.The experiment was authorized by the CEUA protocol 002/2013 and carried out in the northern plateau of Santa Catarina. Were used ten AG337 males from 16 to 21 months of age and 430 dairy females aged 200 to 230 days. The in vitro test was utilized to measure the efficiency of the equipment. The ejaculate was collected, diluted, stored for 10 days at 15 to 18°C. For the in vivo test, the doses were stored for up to 4 days. For in vitro analysis, plasma membrane integrity tests were used for phase contrast, progressive motility, total motility and morphology, all using the CASA system. It was obtained an average increase of biological quality of 15% compared to the control. The number of animals born increased by 0.5 piglet / delivery compared to the control group. The technology proved to be efficient and adequate to improve the outcome of the artificial insemination process in a Genetic Dissemination Unit.

keywords: Isothermal Technology. Sperm Quality. Artificial Insemination. Pigs.

MICROBIOLOGICAL CHARACTERIZATION OF A BRAZILIAN KEFIR GRAINS MILK

Wed, 20 Dec 2017 00:00:00 -0200

Kefir is a product resulting from the symbiosis between lactic acid bacteria and acetic acid bacteria, presenting probiotic action, such as immune system stimulation, antimicrobial activity against pathogens, intestinal microbiota balance and antitumor action. The present study aimed to the characterization and microbiological identification population the grains to the Brazilian milk kefir. A sample of kefir produced in the Videira/Santa Catarina region was evaluated. For this, ten grams of the sample were inoculated in 30mL of distilled water with 3 grams of skimmed milk powder and incubated for 24 hours at 28ºC. The sample was diluted and plated for isolation on MRS agar, M17 agar, Mycosel agar, YM agar and SD agar. The predominant population was lactic acid bacteria (70.5%), followed by yeasts (29.5%). The former ranged from 109 to 1011 CFU / g and yeasts from 104 to 105 CFU / g. Gram staining and optical microscopy were also performed to identify the prevalence of the microbiota by bacillary cells (short and curvad long) and gram-positive cocci, which grew in association with slightly oval yeast cells. In the catalase test there was a predominance of positive catalase. The result of the presence of distinct microbial populations in kefir shows the need to better explore the microbial composition of the same.

ALTERNATIVE APPROACHES TO UNDERSTANDING THE RELATIONSHIP BETWEEN BASIC SANITATION AND SCHOLAR PROFICIENCY

Fernando Dirceu Matias, Petrick Anderson Soares, Dirceu Scaratti — Wed, 20 Dec 2017 00:00:00 -0200

The basic sanitation has made little progress on the public policy agenda in Brazil. Currently, the lack of a general sewage network reaches 53% of the Brazilian population. The relationship between sanitation access and health is well known, but, the connection between the basic sanitation and other components of the human development index (HDI), such as education, is neglected. The access to school sanitation infrastructure and impacts on scholar development has not been explicitly studied in Brazilian literature. The question that arises is how access to basic sanitation improves scholar proficiency. Researches that are being developed about this subject have been revealed that the precariousness of access to sanitation is directly linked to variations in indicators that evaluate the quality of basic education, such as school attendance rate, primary school dropout rate and age-series distortion. Correlation analysis is the main method used for such analyses, using data from the demographic and school census. To the best of our knowledge, no research using the Data Envelopment Analysis (DEA) tool has been developed to better understand the relationship between access to basic sanitation and basic education in Brazil.

INSIGHTS INTO REDUCTION OF CHROMIUM(VI) BY COAL MINING WASTE

Sabrina Munaro, Laís Bruna Verona, Jean Carlo Santos, Petrick Anderson Soares — Wed, 20 Dec 2017 00:00:00 -0200

MUNARO, Sabrina; VERONA, Laís Bruna; SANTOS, Jean Carlo; SOARES, Petrick Anderson

Abstract

Pyrite (PE) is a major coal mining waste product and natural sulfur mineralc. The dissociation of PE tends to releases dissolved ferrous and other ions, which upon reaction with atmospheric oxygen brings acidity to the natural water bodies. Stringent environmental regulations have called for alternatives to deal with tailing problems of mineral processing. Based on this concept, it is possible to take advantage of the “redox” characteristics of PE to treat wastes, such as Cr(VI) containing waste streams. In this work, PE, an coal mining waste, was evaluated as iron source to promote the reduction of hexavalent chromium. PE was characterized physically-chemically and by x-ray diffraction. The adsorption phenomena on the coal mining waste was evaluated, and it was found that the PE has high adsorption potential. The effect of solution pH and catalyst PE dosage on the reduction rate of chromium(VI) for chromium(III) were studied in a batch system. It was observed that higher concentrations of PE resulted in an increase in the rate of Cr (VI) reduction. A maximum rate value at pH 3 was also observed. PE acts as a source of Fe(II), presents effciency in the reduction of chromium (VI), so it can be an option in the treatment of wastewater.

Pyrite; Reduction Reactions; Hexavalent Chromium
e-mail: smunaro26@gmail.com

ANALYSIS OF ASCORBIC ACID STABILITY TO 20% BY USING SILICONE EMULSIONS DIFFERENT VEHICLES AND PRESERVATIVES

Wed, 20 Dec 2017 00:00:00 -0200

Vitamin C has physiological effects on the skin such as: inhibition of melanogenesis, promotion of collagen synthesis and antioxidant action. The development of cosmetics containing vitamin C in the acid form presents difficulties inherent in their physico-chemical characteristics that prevent the stability of the final product. The objective of this study was to analyze the stability of 20% ascorbic acid in emulsions developed with different preservatives and still using water, propylene glycol or glycerin as vehicle. 5 formulations containing 20% ascorbic acid were prepared in silicone emulsions. The samples were analyzed at 0, 15, 30 and 45 days for organoleptic characteristics and ascorbic acid content. Samples 1 and 5 had a greater reduction in the content, possibly due to the fact that in both the water was used as the total vehicle of the emulsion. Tests 2.3 and 4 had lower content reduction, probably due to the use of a binary system containing only 10% water and glycerine or propylene glycol as the emulsion vehicle. Regarding preservatives, metabisulphite was not found to be effective against the oxidation of vitamin C in the presence of large amounts of water, since the sample 5 used metabisulfite and water as the total vehicle of the formulation and was one of the samples that presented the most decrease in the content of active principle.

PRESENTATION OF THE SECOND INTERNATIONAL SYMPOSIUM ON SCIENCE AND BIOTECHNOLOGY

Jane Lafayette Neves Gelinski — Wed, 20 Dec 2017 00:00:00 -0200

PRESENTATION

Dear Colleagues It was a great pleasure that we conducted the II International Symposium on Science and Biotechnology-II ISSB – Entrepreuneurship and Innovation with support of the Science and Biotechnology Graduate Program from Universidade do Oeste de Santa Catarina, in Videira, Brazil, between the 04th and 110th of november 2017. We led a very good scientific program that met the expectations of professionals and students. The event was planned to be a discussion forum and with the aim of expanding the professional partnerships on biotechnology and innovation. Now we present the Proceedings of II ISSB with abstracts of research in two Areas: Biotechnology Applied to Agroindustry and Health / Environmental Biotechnology. We hope that it can be a good source of scientific knowledge and information.

We hope see all of you at III ISSB!

SCIENTIFIC COMMITTEE II ISSB

November 10, 2017

Videira, SC – Brazil

SHORT COURSE: MICROBIAL FUEL CELLS: NOVEL BIOTECHNOLOGY FOR ENERGY GENERATION

Lucas Ricardo Cardoso Marcon — Wed, 20 Dec 2017 00:00:00 -0200

Microbial fuel cells (MFCs) provide new opportunities for the sustainable production of energy from biodegradable, reduced compounds. MFCs function on different carbohydrates but also on complex substrates present in wastewaters. MFC is equivalent to stacks and batteries, its main components are: anode, cathode, electrodes, and membrane switched protons. In the anode chamber, by means of anaerobic degradation, the organic compounds (present in the sewage) are degraded and electrons are released (which will generate the electric current) and positive ions. The cathode chamber (aerobic) is generated a positive potential, through forced or natural aeration, in this chamber can also be inserted aerobic bacteria to complete the biological treatment of the sewage. Different models of MFC can be used: two cameras, single chamber, and multiple. The MFC can be designed for the evaluation of Biochemical Oxygen Demand (BOD) in a continuous way, besides being able to treat diverse organic compounds, studies also evaluate the possibility of insertion of landfill leachate.

Keywords: Sewage treatment. Electron transfer. Electric potential. Green energy. Renewable energy.

SHORT - COURSE: MOLECULAR BIOLOGY APPLIED TO CLINICAL ANALYZES

Bibiana Paula Dambrós, Francielli Mello Ferri — Wed, 20 Dec 2017 00:00:00 -0200

In recent years, genetics, cellular biology, genome sequencing of pathogens, among other resources, have significantly changed the opportunities for epidemiological investigations, pathogenesis studies, diagnosis and control of microbial diseases. The PCR technique has expanded the possibilities of DNA analysis and has made molecular biology find new applications even in areas outside its traditional field, such as medicine, agriculture and biotechnology. PCR presents a wide range of applications in various branches of scientific research. This reaction allows a particular region of the genome of any organism to be multiplied in millions of copies, which facilitates genetic analysis and allows the development of diagnostic techniques, much more sensitive and more specific than those traditionally used. The mini-course will have a brief presentation on molecular techniques and after the practical part with the following steps: Extraction of bacterial DNA, Quantification of DNA by the absorption spectrum of nucleic acids, Polymerase Chain Reaction (PCR) associated with length polymorphism DNA fragments (PCR_RFLP / ARDRA) terminating with Sample Analysis using the technique of Agarose Gel Electrophoresis.

Keywords: Genetics. Epidemiological. Polymerase Chain Reaction.

SHORT COURSE: STRUCTURAL ANALYSIS OF BIOMOLECULES

Carlos Alexandre Breyer — Wed, 20 Dec 2017 00:00:00 -0200

Understanding the molecular structure of biomolecules has revolutionized biology and medicine, and it is possible to understand at a molecular level the events that govern life. This understanding has enabled a revolution in medicine to develop new drugs based on structural analysis and the molecular understanding of diseases, whether genetic or caused by pathogenic organisms. Nowadays new methodologies have been outstanding in structural biology, a highlight is given to cryo-electron microscopy, a revolutionary methodology that has been helping the classical X-ray crystallography. So much stands out the importance of cryo-electron microscopy that is laureated with the Nobel Prize in Chemistry of the year 2017 to its developers. In the mini-course offered in this event we take the participants a sample of the possibilities of the structural analysis of biomolecules, presenting the main techniques used, as well as analysis programs that allow better understanding of the structures of biomolecules such as nucleic acids and proteins.

eywords: X-ray crystallography. Structural biology. Biomolecules.

CULTIVATION OF EDIBLE AND MEDICINAL MUSHROOMS

Elisandra Minotto — Wed, 20 Dec 2017 00:00:00 -0200

Mushrooms generally belong to Basidiomycetes which harbors numerous species with diversity of metabolites of nutraceutical and therapeutic significance. They are famous healthy food because show high rates of nutrients such as proteins, fibers, carbohydrates, minerals, vitamins, and lipids. When compared with other countries, the edible mushrooms cultivation is considered too small in the Brazil. They are important to development of small properties because can be cultivated in agroindustrial residues generating their economic improvement. Thus, the aim of this short course is to give to the event participants general notions regarding the cultivation of edible and medicinal mushrooms. This theoretical and practical short course aimed to develop knowledge based on lectures and laboratory techniques about the selected theme. Contents showed: diversity of mushrooms and their main morphological and physiological characteristics; nutritional and medicinal properties; biotechnological importance of fungi: methods and culture media for isolating of different mushrooms: Agaricus bisporus, Lentinula edodes and Pelurotus spp.; strain selection of the edible mushrooms, making of spawns and their application to natural substrates, and standardization of cultivation conditions to ensure proper mycelial colonization of the substrates to be followed by different.

eywords: Basidiomycetes. Agaricus bisporus. Health.