Ml Of Medium Research Articles

Polyclonal antibodies inhibit growth of key cellulolytic rumen bacterial species.

Antibodies targeting specific bacterial species could allow for modification of the rumen microbial population to enhance rumen fermentation. However, there is limited knowledge of targeted antibody effects on rumen bacteria. Therefore, our objective was to develop efficacious polyclonal antibodies to inhibit the growth of targeted cellulolytic bacteria from the rumen. Egg-derived, polyclonal antibodies were developed against pure cultures of Ruminococcus albus 7 (anti-RA7), Ruminococcus albus 8 (anti-RA8), and Fibrobacter succinogenes S85 (anti-FS85). Antibodies were added to a cellobiose-containing growth medium for each of the three targeted species. Antibody efficacy was determined via inoculation time (0 h and 4 h) and dose response. Antibody doses included: 0 (CON), 1.3 × 10-4 (LO), 0.013 (MD), and 1.3 (HI) mg antibody per ml of medium. Each targeted species inoculated at 0 h with HI of their respective antibody had decreased (P < 0.01) final optical density and total acetate concentration after a 52 h growth period when compared with CON or LO. Live/dead stains of R. albus 7 and F. succinogenes S85 dosed at 0 h with HI of their respective antibody indicated a decrease (≥ 96%; P < 0.05) in live bacterial cells during the mid-log phase compared with CON or LO. Addition of HI of anti-FS85 at 0 h in F. succinogenes S85 cultures reduced (P < 0.01) total substrate disappearance over 52 h by at least 48% when compared with CON or LO. Cross-reactivity was assessed by adding HI at 0 h to non-targeted bacterial species. Addition of anti-RA8 or anti-RA7 to F. succinogenes S85 cultures did not affect (P ≥ 0.45) total acetate accumulation after 52 h incubation, indicating that antibodies have less of an inhibitory effect on non-target strains. Addition of anti-FS85 to non-cellulolytic strains did not affect (P ≥ 0.89) OD, substrate disappearance, or total VFA concentrations, providing further evidence of specificity against fiber-degrading bacteria. Western blotting with anti-FS85 indicated selective binding to F. succinogenes S85 proteins. Identification by LC-MS/MS of 8 selected protein spots indicated 7 were outer membrane proteins. Overall, polyclonal antibodies were more efficacious at inhibiting the growth of targeted cellulolytic bacteria than non-targeted bacteria. Validated polyclonal antibodies could serve as an effective approach to modify rumen bacterial populations.

Production and characterization of a bioflocculant produced by microorganisms isolated from earthen pond sludge

Bioflocculants are biodegradable polymers produced by microorganisms. The aim of this study was to produce and characterize a bioflocculant by microorganisms isolated from earthen pond sludge. Sludge from the earthen pond was collected from ‘Lapai Gwari’ in Minna, Nigeria. The microorganisms were screened for the potential of producing bioflocculant using kaolin suspension and a selective medium. The isolate with the highest flocculating potential was identified based on its molecular characteristics. The bioflocculant produced was characterized using thermogravimetric analysis (TGA), Fourier-transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). Four bacteria and three fungi were isolated from the sludge. Priestia megaterium (ON184360) was identified as the isolate with the highest flocculant production potential, with 10.67 g of bioflocculant produced from 500 mL of medium, which was confirmed by TGA, FTIR and SEM results. The findings of this study showed that Priestia megaterium is a more efficient alternative for flocculant production.

Intratumoral pan-ErbB targeted CAR-T for head and neck squamous cell carcinoma: interim analysis of the T4 immunotherapy study

BackgroundLocally advanced/recurrent head and neck squamous cell carcinoma (HNSCC) is associated with significant morbidity and mortality. To target upregulated ErbB dimer expression in this cancer, we developed an autologous CD28-based...

A synthetic population-level oscillator in non-microfluidic environments

Synthetic oscillators have become a research hotspot because of their complexity and importance. The construction and stable operation of oscillators in large-scale environments are important and challenging. Here, we introduce a synthetic population-level oscillator in Escherichia coli that operates stably during continuous culture in non-microfluidic environments without the addition of inducers or frequent dilution. Specifically, quorum-sensing components and protease regulating elements are employed, which form delayed negative feedback to trigger oscillation and accomplish the reset of signals through transcriptional and post-translational regulation. We test the circuit in devices with 1 mL, 50 mL, 400 mL of medium, and demonstrate that the circuit could maintain stable population-level oscillations. Finally, we explore potential applications of the circuit in regulating cellular morphology and metabolism. Our work contributes to the design and testing of synthetic biological clocks that function in large populations.

MP01-03 XENO-FREE PEPTIDE-FUNCTIONALIZED BIOINKS IMPROVE EX VIVO CULTURE OF HUMAN TESTICULAR TISSUES

You have accessJournal of UrologyCME1 Apr 2023MP01-03 XENO-FREE PEPTIDE-FUNCTIONALIZED BIOINKS IMPROVE EX VIVO CULTURE OF HUMAN TESTICULAR TISSUES Meghan Robinson and Ryan Flannigan Meghan RobinsonMeghan Robinson More articles by this author and Ryan FlanniganRyan Flannigan More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003212.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The development of human in vitro spermatogenesis (IVS) is a promising regenerative platform for the treatment of male factor infertility. IVS has been demonstrated in rodent models, but despite decades of research remains a challenge in the human sphere. A promising strategy may be to culture fragments of testis tissues in hydrogels. Hydrogels can be functionalized to sequester matrix and signaling peptides in a local manner mimicking tissue niche function. In this study we investigated the use of xeno-free hydrogels functionalized with biomimetic peptide motifs for their ability to improve ex vivo culture of human testicular tissues. METHODS: Human testicular tissues were obtained from hormone-treated gender affirmation patients following informed consent, gently teased apart, cut into small fragments and mixed into bioinks (VitroInk) containing either YIGSR (laminin), COL (collagen), or MMPs (matrix metalloproteinases) peptide motifs, then printed into ∼1 cm3 droplets using a BioX6 extrusion bioprinter (Figure 1A). 1 mL of media was placed on each and they were cultured in 34°C at 5% CO2 and ambient oxygen (21%) for 12 days, with 50% media changes every other day. The media consisted of Human Plasma-Like Medium (HPLM) with 15% CTS™ KnockOut™ SR XenoFree Medium, vitamins, metabolites, antioxidants, growth factors and hormones known to be present in the testis niche. Tissues were analyzed after 12 days by quantitative real-time polymerase chain reaction (qPCR) for gene expression associated with apoptosis, somatic cell function, spermatogonial stem cell maintenance, and spermatogenesis. RESULTS: After 12 days gene expression associated with somatic cell function and spermatogenesis was higher than suspension culture controls in all hydrogel groups (Figure 1B). Compared to freshly harvested testicular tissues, COL- and YIGSR-functionalized hydrogels exhibited similar expression levels of genes associated with somatic cell function, and the highest increased levels of genes associated with early spermatogenesis (Figure 1B). CONCLUSIONS: COL- or YIGSR peptide-functionalized hydrogels sustain testicular tissue function ex vivo in short-term cultures, whereas suspension culture exhibited tissue function loss, suggesting the utility of functionalized hydrogels for human IVS. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e1 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Meghan Robinson More articles by this author Ryan Flannigan More articles by this author Expand All Advertisement PDF downloadLoading ...

Drosophila suzuki: Matsumura (Diptera: Drosophilidae): analysis of experimental conditions for its rearing and study in the laboratory

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is an invasive pest species that uses a wide variety of commercial fruit as a breeding and feeding resource. The damage to the fruit generated by D. suzukii´ oviposition causes considerable economic losses. Therefore, the maintenance of D. suzukii in the lab conditions constitutes a fundamental tool for research proposal. Two of the determining factors affecting the lab rearing conditions are the culture medium and density. In this study, we evaluated the impact of these factors on three components associated with fitness: development time, larval-adult viability and body size, comparing the effects of three culture media: cornmeal (polenta), mashed potato and plum when the flies were reared at three densities: 10, 30 and 50 larvae per 5ml of culture medium. Our results revealed that the flies reared the cornmeal culture medium and at lower densities provide the better conditions for the maintenance of D. suzukii under lab conditions.

OPTIMIZATION OF CULTURE CONDITIONS AND NUTRIENT SOURCES FOR Bacillus sp. ES4 SYNTHESIZES FIBRINOLYTIC ENZYME

Bacillus sp. can biosynthesize fibrinolytic enzymes. Culture conditions and nutrient sources are important factors affecting microbial growth and production of the Fibrinolytic enzyme. Therefore, we investigated the culture conditions of the strain Bacillus sp. ES4, such as temperature, pH, C nutrient source (glucose, sucrose, glycerol, and maltose), N source (yeast extract, meat content, peptone, and tryptone), metal ions (Ca 2+ , Mg 2+ , Mn 2+ , Zn 2+ , K + , Fe 2+ , and Cu 2+ ), and time to obtain enzyme product. The results show that for Bacillus sp. ES4, the best C source was glucose, peptone and yeast extract. Ca2+ and Mg2+ are nutritional factors that greatly influence the biosynthesis of the fibrinolytic enzyme of strain ES4. At the culture conditions of 37 0 C and pH = 6.5, the strains for the best enzyme activity at 24 hours and the enzyme activity of the strain when cultured in a 250 ml flask containing 50 ml of medium for the highest enzyme activity at 450±25. FU/ml.

Fibrin clot and Leukocyte-rich platelet-rich fibrin show similar release kinetics and amount of growth factors: a pilot study

BackgroundIn knee arthroscopic surgery, fibrin clot (FC) and leukocyte-rich platelet-rich fibrin (L-PRF) may be used in augmentation for meniscal repair. Studies have investigated growth factors released from FC and L-PRF; however, it is difficult to compare FC and L-PRF between different studies. Direct comparison of growth factors that may support meniscal healing released from FC and L-PRF may be beneficial in deciding whether to use FC or L-PRF. If no significant difference is seen, the surgeon may decide to use FC which is easier to prepare compared to L-PRF. The purpose of this pilot study is to investigate the release amount and pattern of basic fibroblast growth factor (bFGF), platelet-derived growth factor AB (PDGF-AB), transforming growth factor β1 (TGF-β1), vascular endothelial growth factor (VEGF), and stromal cell-derived factor 1 (SDF-1) from FC and L-PRF.MethodTwenty milliliters (ml) of whole blood was collected from each of the four volunteers. Ten milliliters of whole blood was allocated for preparation of FC and 10 ml for L-PRF. FC and L-PRF were separately placed in 5 ml of culture media. Five milliliters of the culture media was sampled and refilled at 15 min, 1 day, 3 days, 1 week and 2 weeks. The collected culture was used to quantify bFGF, PDGF-AB, TGF-β1, VEGF, and SDF-1 release by Enzyme-linked immune-sorbent assay (ELISA). Mann–Whitney U test was performed to assess significance of differences in amount of each growth factor released between FC and L-PRF. Significance was accepted at P value less than 0.05.ResultsAt two weeks, the cumulative release of TGF-β1 was the highest among all the growth factors in both FC and L-PRF (FC:19,738.21 pg/ml, L-PRF: 16,229.79 pg/ml). PDGF-AB (FC: 2328 pg/ml, L-PRF 1513.57 pg/ml) had the second largest amount, followed by VEGF (FC: 702.06 pg/ml, L-PRF 595.99 pg/ml) and bFGF (FC: 23.48 pg/ml, L-PRF 18.2 pg/ml), which order was also common in both FC and L-PRF. No significant difference in final release amount and pattern was seen between FC and L-PRF.ConclusionThe current pilot study showed that cumulative release amount and release pattern of PDGF-AB, VEGF, TGF-β1, and bFGF did not significantly differ between FC and L-PRF during the two weeks of observation.

In vitro spore germination and gametophyte development of two Cyathea species of South America in response to nutrient media

Abstract Cyathea corcovadensis and Cyathea phalerata are tree ferns native to Brazil, endangered in the state of Rio Grande do Sul. Spore germination and gametophyte development in media with different nutrient formulations and activated charcoal were evaluated, aiming to develop a process for obtaining plants of the two species. Spores were sown in four semi-solid culture media: Meyer, Dyer, MS with 50% and MS with 25% of the original macronutrient concentration. For each medium, 10 replicates were carried out (flasks with 5 mg of spores/30 mL of medium), with and without 1% activated charcoal, respectively. Spore germination and gametophytic development (laminar and cordate stages) were quantified at 30, 60 and 90 days of culture. Cyathea corcovadensis and C. phalerata germinated and developed gametophytes in all media. For both species, the highest percentages of germination and cordate gametophytes (more advanced development stage) were recorded in Meyer medium without activated charcoal, which has higher concentrations of macronutrients and no micronutrients compared to the other evaluated media. We recommend cultivating the plants in Meyer medium for greater gametophytic development and subsequent sporophyte obtention, as a biotechnological tool for C. corcovadensis and C. phalerata conservation and for environmental restoration and enrichment using these tree ferns.

Evalution of Jeevamrutham (Organic Biofertilizer) for It’s use in the Devising an Nutrient Medium for the Isolation of Bacteria

Jeevamrutham is widely used Organic biofertilizer prepared with cow waste, gram flour and Jaggery by fermentation process. The study was aimed at physicochemical and microbiological analysis of jeevamrutham and also to evaluate it’s use as a nutrient medium for isolation of bacteria. The Jeevamrutham prepared and subjected to physicochemical, and microbiological analysis. The filtrate of the Jeevamrutham was studied for its use as a Nutrient (Jeevamrut) medium for the growth of the bacteria. Total solid content of Jeevamrutham was found to be 100 mg/ml. The Standard Plate count (SPC) for Nitrogen fixers, for Phosphate solubilizing bacteria was found to be maximum on 6th day of fermentation. The SPC for Yeast and Molds was found to be maximum on 8th day. The study regarding the evaluation of Jeevamrut medium showed excellent growth of test Gram positive and Gram negative bacteria on Jeevamrut medium. The pigment producers showed same pigmentation pattern on Jeevamrut agar and on conventially used Nutrient agar. The cost of the Jeevamrut medium was calculated per 100 ml of medium and was found to be 60.27% less than that of commercially available media. The Jeevamrut medium is economical, all the ingredients are easily available, cheap and ecofriendly. The Jeevamrut medium can be a best option to use as a Nutrient medium for the cultivation of bacteria in the laboratory

Inhibition of Aflatoxin B1 Synthesis in Aspergillus flavus by Mate (Ilex paraguariensis), Rosemary (Rosmarinus officinalis) and Green Tea (Camellia sinensis) Extracts: Relation with Extract Antioxidant Capacity and Fungal Oxidative Stress Response Modulation

Plant extracts may represent an ecofriendly alternative to chemical fungicides to limit aflatoxin B1 (AFB1) contamination of foods and feeds. Mate (Ilex paraguariensis), rosemary (Romarinus officinalis) and green tea (Camellia sinensis) are well known for their beneficial properties, which are mainly related to their richness in bioactive phenolic compounds. AFB1 production is inhibited, with varying efficiency, by acetone/water extracts from these three plants. At 0.45 µg dry matter (DM)/mL of culture medium, mate and green tea extracts were able to completely inhibit AFB1 production in Aspergillus flavus, and rosemary extract completely blocked AFB1 biosynthesis at 3.6 µg DM/mL of culture medium. The anti-AFB1 capacity of the extracts correlated strongly with their phenolic content, but, surprisingly, no such correlation was evident with their antioxidative ability, which is consistent with the ineffectiveness of these extracts against fungal catalase activity. Anti-AFB1 activity correlated more strongly with the radical scavenging capacity of the extracts. This is consistent with the modulation of SOD induced by mate and green tea in Aspergillus flavus. Finally, rutin, a phenolic compound present in the three plants tested in this work, was shown to inhibit AFB1 synthesis and may be responsible for the anti-mycotoxin effect reported herein.

Production and Optimization of Hyaluronic Acid Extracted from Streptococcus thermophilus Isolates.

Biopolymers, particularly exopolysaccharides produced by microorganisms such as bacteria, yeasts, and algae, have gained popularity in recent years due to their physical, chemical, and functional properties that are widely useful in food, industrial, cosmetic, and pharmaceutical systems. Hyaluronic acid is one type of these polysaccharide. This study investigated the optimal conditions for producing hyaluronic acid from the Streptococcus thermophilus bacterial strain. The isolated Streptococcus thermophilus were cultured on MRS broth, Skim milk, and M17 broth with an addition of 1% lactose. The diagnosed bacterial strains were grown in 100 ml of culture media, placed in volumetric flasks of 250 ml capacity, and incubated at 42˚C for 24 hours, pH 6.8, inoculum volume 1%, and a vibrating incubator at 150 rpm. After the end of the fermentation period, the isolation and purification of HA have performed accordingly: proteins were removed using 1% trichloroacetic acid (TCA), and HA in the supernatant was collected by isopropanol precipitation. The collected HA was dialyzed against ultrapure water and lyophilized. The amount of acid produced was estimated. The results show that the best production of hyaluronic acid was from the S. thermophilus bacterial strain grown on the alternative medium containing whey at a ratio of 450 ml/L and 7.5 g/L yeast extract at 40 ˚C, with a 3% of inoculum volume and 102×108 colony-forming units/ml of bacterial cells, in pH 6.8 and agitation speed of 150 rpm for 18 h, which had the most significant effect on the fermentation process and gave the highest value of HA production of 0.598 g/L and biomass of 6.08 g/L. These results showed the best production method for HA to achieve maximal production yelled.

Multifunctional, stable and low-cost lipopeptide biosurfactant produced by Enterobacter cloacae UCP 1597

Caatinga of Pernambuco is an area with a potential richness of microorganisms that produce biosurfactants, which are considered good candidates to replace synthetic surfactants in industrial applications due to their functional stability and low toxicity. In this context, current study aimed to investigate the biosurfactant production by the bacterium Enterobacter cloacae UCP 1597, isolated from the Caatinga soil. First fermentation was carried out in Erlenmeyer flasks containing 100 ml of medium, according to a 2³ full-factorial design (FFD). The results showed higher reduction in surface tension (28.3 mN/m) in condition 2 of the FFD, where dispersion of 38.46 cm2 of burnt motor oil was also verified. Then, a second fermentation was performed in Fernbach flasks, containing 2 L of selected medium, confirming reduction in surface and interfacial tension to 30.5 and 2.3 mN/m, respectively, as well as excellent emulsifying properties. The critical micellar dilution (CMD) of the crude biosurfactant was determined (70%) and its use in phytotoxicity assay verified the absence of toxicity for cabbage seeds. The biomolecule showed a high yield (13.69 g/L) after extraction with ethyl acetate and anionic and lipopeptide nature. The stability in acid pH, high temperature and salinity, showed an acid-resistant, thermostable, and halotolerant biocompound. Thus, this lipopeptide was shown to be a multifunctional biosurfactant, since it not only has excellent surface-active properties, but it is also a good emulsifier, dispersant, and potent agent to germination of cabbage seeds. Hence, is suggested its promising application in industrial activities or environmental processes under adverse conditions.

Design and Testing of a Disposable Flow Cuvette for Continuous Electroporation of a Bioreactor’s Initial Algae Cultivation

Electroporation is a technique applied both in biomedical and biotechnological fields which uses a high-voltage electric current to temporarily destabilize the plasma membrane of living cells, permitting the introduction of small molecules as well as nucleic acids into the cytosol. Besides viral and chemical transfections, this method is a common way to manipulate living cells. However, the majority of electroporation machines available on the market can only work using batch-based cuvettes treating only a few micrograms of cells. To transform cells in the order of several grams in the quickest possible way, it is necessary to use a continuous-flow method. In this work, we present the design, electric and fluid dynamics simulations, construction and testing of a flow cuvette that can adapt to standard electroporator systems. The flow cuvette connected with a peristaltic pump was able to successfully electroporate 20 mL of medium containing microalgae cells in less than 5 min. Microalgae Scenedesmus almeriensis cells were transfected with a fluorescent siRNA oligo as well as magnetically transformed by introducing magnetic nanoparticles in their cytoplasm. The flow cuvette presented here offers a valid tool for the high-throughput transformation/transfection/transfer of both prokaryotic and eukaryotic organisms, especially suitable for bioreactor cultivation and other industrial biotechnological contexts.

In Vitro Detection of Biochemical Effect in Human CaCo-2 Cell Line after Exposure to a Low Concentration of a Deltamethrin-Based Pesticide

Pesticide residues are chemicals frequently found in food as contaminants. Pesticides may have adverse health effects, particularly when the digestive tract is concerned, as a consequence of food ingestion. Deltamethrin is a pyrethroid pesticide widely used in various fields, such as agriculture, veterinary and in the household, so the ingestion of a small amount of this chemical may occasionally occur. To assess whether exposure to pesticide residues may have a biological effect at the intestinal level, it is primarily necessary to perform in vitro exposure experiments about cell lines models of the intestinal barrier at low concentrations of the chemical. In the present study, CaCo-2 cells were exposed to different concentrations of a Deltamethrin-based commercial pesticide, which was diluted in the cell medium. An MTT viability test indicated that the cytotoxic concentration value of the pesticide inside 1 mL of medium is between 10−6 and 10−5 mL. However, the analysis of Raman spectra found that biochemical changes occur inside cells exposed to a non-cytotoxic concentration of 10−6 mL of the pesticide inside 1 mL of the medium. Such changes involve mainly an increase in the ratio between the amount of lipid with respect to that of the protein components in the cell cytoplasm. The results obtained by Raman micro-spectroscopy were confirmed by fluorescence images obtained by using a fluorophore staining neutral lipids. Overall, the obtained results suggest that Raman micro-spectroscopy can be successfully used to monitor the cellular modifications due to exposure at low concentrations of pesticides, as those values that can be found inside food are residuals.

Effect of sunflower oil (Helianthus annuus) in ruminal fermentation in vitro and emission of gases

Objective: The production of total gas, methane (CH4) and in vitro fermentative variables in diets for sheep with 1 and 2 % sunflower oil were evaluated. Design/Methodology/Approach: Serological vials with 0.5 g of treatment and 50 ml of culture medium were incubated at 39 °C for 24, 48 and 72 h. The production of total gas, CH4, and degradation of dry matter (DEGDM), neutral detergent fiber (DEGNDF), acid (DEGADF), production of volatile fatty acids (VFA) and ammoniacal nitrogen (N-NH3) were estimated. . The experimental design was completely randomized. Results: Total gas production at 48 and 72 h increased (p &lt; 0.05) and at 24 h decreased (p &lt; 0.05) as the oil increased. CH4 production at 24 and 48 h did not present differences (p &gt; 0.05); a linear decrease (p &lt; 0.05) was quantified at 72 h. DEGDM increased (p &lt; 0.05) at 24 and 48 h and decreased (p &lt; 0.05) at 72 h. DEGNDF and DEGADF increased (p &lt; 0.05) at 48 h. Butyric acid content and N-NH3 decreased (p &lt; 0.05) at 48 h. Study Limitation/Implications: An inclusion greater than 2 % of sunflower oil in the diet can reduce the degradability of the food and the microbial protein. Findings/Conclusions: The inclusion of up to 2% of sunflower oil in diets for lambs does not affect the degradation of fibers and is an alternative to reduce the methane emitted into the environment.

PRODUCCIÓN DE BIOGÁS Y CARACTERÍSTICAS FERMENTATIVAS in vitro DE CONSORCIOS BACTERIANOS CELULOLÍTICOS RUMINALES OBTENIDOS DE DIFERENTES FIBRAS

&lt;p&gt;&lt;strong&gt;Background&lt;/strong&gt;. Cellulolytic bacterial consortia (CBC) serve as additives to improve fiber degradation in ruminants, since they improve biogas production and &lt;em&gt;in vitro&lt;/em&gt; fermentation characteristics. &lt;strong&gt;Objective&lt;/strong&gt;. To evaluate the biogas production and fermentative characteristics &lt;em&gt;in vitro &lt;/em&gt;of mulato grass inoculated with ruminal CBC obtained from ground sawdust, ground mulato grass or whole stem as substrates in the selective culture medium. &lt;strong&gt;Methodology&lt;/strong&gt;. The CBC were obtained from ruminal fluid from a Suiz-Bu cow fitted with a ruminal cannula, which was transferred six times in a selective anaerobic medium. The test consisted of preparing sterile biodigesters with 0.5 g of mulatto grass with 63 d of regrowth, 45 mL of culture medium. The inoculation was with 5 mL of a type of CBC. The biodigesters were incubated 72 h at 39 °C. Biogas production was measured at 3, 6, 9, 12, 24, 48 and 72 h. In the culture media, ammonia nitrogen (N-NH&lt;sub&gt;3&lt;/sub&gt;), total bacterial count, cellulase activity, pH, dry matter degradation (DMD) and neutral detergent fiber degradation (NDFD) were determined at 12, 24, 48 y 72 h. In the biogas production a completely random design was used; while in the rest of the variables a completely randomized design was carried out with a 3x4 factorial arrangement, with fiber source and incubation time as factors. &lt;strong&gt;Results&lt;/strong&gt;. The biogas production of the CBC obtained from the ground grass showed higher (p&amp;lt;0.05) accumulated production in all the evaluated times. The CBC obtained from sawdust increased the biogas production by 175% from 24 to 48 h and by 313% from 48 to 72 h. There was an interaction effect (p &amp;lt;0.05) on DMD, NDFD, N-NH&lt;sub&gt;3&lt;/sub&gt;, total bacterial count and cellulase enzymatic activity. The CBC obtained from ground mulatto grass at 72 h increased (p &amp;lt;0.05) the DMD and NDFD. The CBC obtained from sawdust increased DMD by 19.1% and NDFD by 33% from 48 to 72 h. The concentration of bacteria in the observed interactions is within the range of the ruminal ecosystem. The highest (p&amp;lt;0.05) content of N-NH&lt;sub&gt;3&lt;/sub&gt; was determined when the CBC obtained from grass stem with 12, 24 and 72 h of incubation were used. &lt;strong&gt;Implications&lt;/strong&gt;. Using the same fiber source to obtain the CBCs improves the degradation of said fiber. &lt;strong&gt;Conclusion&lt;/strong&gt;. The size and source of fiber are decisive in the type of bacteria that make up the cellulolytic bacterial consortia of ruminal origin under the conditions of this &lt;em&gt;in vitro&lt;/em&gt; test.&lt;/p&gt;

Comparative study of nutrient media for the isolation of mycobacteria

Laboratory diagnostics is of decisive importance in the detection of animal tuberculosis. The proposed nutrient media for isolating of the pathogen are rich in variety. The practical significance of frequently used culture media and their modifications have been studied. Samples of Petragnani, Gerold, Evglevsky, Mordovsky, Pavlovsky, Levenshtein-Jensen, Finn-II and some of their modifications for the primary isolation and subculturing of mycobacteria were studied. Also, in a comparative plan, the Levenshtein-Jensen and Finn-II media were studied in various versions: Levenshtein-Jensen, prescribed and frozen; Levenshtein-Jensen, dry and frozen; Finna-II in the author's version and four modifications, AF-I (glycocol instead of monosodium glutamate), AF-II- (with potato extract), AF-III - (without glycocol) and MF variant (with M. flei extract). For sowing, biomaterial from sick animals, also artificially infected, was used. The studies made it possible to establish the effectiveness of the Levenshtein-Jensen and Finn-II media for the primary isolation of the pathogen. In terms of growth properties, the dry Levenshtein-Jensen medium is superior to the nitive one, but is significantly inferior to Finn's native medium in terms of properties that inhibit foreign microflora. The ability to accelerate the growth of mycobacteria with hydrocarbons was determined by the addition of n-alkanes with a wide range of carbohydrates - from decane to nonadecane (from C10 to C22). A total of 30 different options were tested: by adding liquid paraffin; alcohol extract of M. flei; wax solutions; propolis on hexane; bovine serum and others. The use of n-alkanes as a growth stimulator in the amount of 2-3 drops per 10 ml of nutrient medium (1 drop per 4 ml of medium) made it possible to reduce the cultivation time on 15-20 days and increase the bacterial mass in about 2 times. The reliability of growth stimulation of M. bovis (P&lt;0.05) by n-alkanes on egg media according to the c2 criterion was also confirmed by the results of variation statistics. Weak growth properties in relation to M. avium and a slight stimulation of the growth of M. tuberculosis were revealed.

Influence of conditioned media from glial cell cultures on contractility of uterine in rats of different ages

Background. The physiological regulation of the uterine contractile activity changes with age, which leads to an increased number of prolonged labor and emergency caesarean sections in women giving birth at the age of 35+. One of the modern approaches to correct the function of the reproductive system is the use of from cell cultures. CM from glial cell culture contains neurotrophic factors that play an important role in maintaining the contractile function of the uterus. Current cell culture technologies include cryopreservation.Objective: to research experimentally the effect of CM obtained from intact and cryopreserved cultures of glial cells on the contractile activity of the uterus in rats of different reproductive ages.Materials and methods. The monolayer cell culture was obtained from the dorsal root ganglia of neonatal piglets and cryopreserved in the presence of cryoprotectant dimethyl sulfoxide. CM from native and cryopreserved cultures were collected for 28 days, after which fractions with a molecular weight of &lt; 30 kDa were obtained from them by ultrafiltration. Rats at the age of 6 and 14 months, which corresponds to reproductive age and late reproductive age (LRA), were intraperitoneally injected with 0.2 ml of media from intact (ICM) or cryopreserved (CCM) cultures for 9 days. On the 30th – 32nd day after the end of the administration of CM animals were slaughtered and the uterine contractile activity was determined by the organ bath method, the relative area of myometrium and density of myocytes by histological method, the average area of labeling with specific antibodies to smooth muscle actin by immunohistochemical method. The statistical significance of differences was assessed by the Mann–Whitney test.Results. It was found that spontaneous, OT-, and KCl-induced tension of isometric contraction of the uterus in intact LRA rats decreased by 19, 20, and 14%, respectively, compared with intact reproductive aged animals. After the introduction of ICM and CCM in LRA animals, normalization of isometric contraction parameters was observed. This effect was realized against the background of an increase in the area of the myometrium, the density of myocytes, and actin expression.Conclusions. Intra-abdominal administration of CM from glial cell culture increases the uterine contractile activity in LRA rats. This effect is realized by increasing the relative area of the myometrium, the density of myocytes, and the area of expression of smooth muscle actin. The effect of media from intact and cryopreserved cultures on the contractile activity of the uterus was similar, which makes it possible to use low-temperature culture storage technologies to obtain CM without losing its biological effect.

Determination of Phenolic Compounds in Blue Corn Flour (Zea mays L.) Produced and/or Metabolized by Colletotrichum gloeosporioides in a Fermentation Process

Phenolic compounds are secondary metabolites produced by plants, and their study has been increased in recent years due to their ability to improve human health. The aim of this work was the determination of phenolic compounds presents in blue corn flour before and after a fermentation process, where different proportions were used of blue corn (Zea mays L.) flour and Czapek Dox culture medium (90 mL of culture medium with 10 g of blue corn flour, 80 mL of culture medium with 20 g of blue corn flour and 70 mL of culture medium with 30 g of blue corn flour) and were fermented at 3 different times (20, 25 and 30 days) with the Colletotrichum gloeosporioides fungus. A determination of the phenolic compounds was carried out with five standard solutions, which were cyanidin 3-glucoside (CYA), pelargonidin 3-glucoside (PEL), chlorogenic acid (CLA), quercetin (QRC) and cinnamic acid (CA). The obtained results showed the presence of CA and PEL. The most abundant phenolic compound in the fermented samples was CLA over the naturally occurring compounds in blue corn, which are CYA and PEL. QRC was the phenolic compound with the lowest concentration in blue corn flour samples fermented with Colletotrichum gloeosporioides.