scholarly journals Influence of siderophores and iron on Mycobacterium bovis isolation from pathological material

2021 ◽  
Vol 7 (1-2) ◽  
pp. 21-26
Author(s):  
A. I. Zavgorodniy ◽  
S. A. Pozmogova ◽  
V. V. Bilushko ◽  
Kalashnyk Kalashnyk ◽  
O. I. Gologurska
Keyword(s):  
Mycobacterium Bovis ◽  
Culture Filtrate ◽  
Nutrient Medium ◽  
Culture Media ◽  
Alcoholic Extract ◽  
Simultaneous Presence ◽  
Growth Properties ◽  
Pathological Material ◽  
Bacterial Mass

The article presents the results of studying the effect of siderophores and iron on the isolation of Mycobacterium bovis from pathological material. It has been established that the simultaneous presence of iron and siderophore from M. phlei in the nutrient medium makes it possible to detect the growth of M. bovis from pathological material 6–8 days earlier; ensures the growth of more colonies and bacterial mass. The presence of heterologous to mycobacteria siderophore (from Nocardia spp.) in the medium reduces the elective (growth) properties of the medium. Siderophores found in the culture filtrate or alcoholic extract of M. phlei can be valuable additives to culture media for the accelerated isolation of M. bovis from pathological material

ВПЛИВ ДЖЕРЕЛ АЗОТНОГО ЖИВЛЕННЯ НА СИНТЕЗ КАРОТИНОЇДІВ ДЕЯКИМИ ШТАМАМИ БАЗИДІОМІЦЕТІВ

2014 ◽  
Vol 4 (01) ◽  
pp. 22
Author(s):  
A. K. Veligodska ◽  
O. V. Fedotov ◽  
A. S. Petreeva
Keyword(s):  
Culture Filtrate ◽  
Nutrient Medium ◽  
Culture Fluid ◽  
Carotenoid Content ◽  
Nitrogen Compounds ◽  
Total Carotenoid ◽  
Carotenoid Accumulation ◽  
Total Carotenoid Content ◽  
Peptone Medium ◽  
Acetone Extracts

<p>The influence of certain nitrogen compounds - components of glucose-peptone medium (GPM) on the accumulation of carotenoids by some strains was investigated by surface cultivating basidiomycetes. The total carotenoid content was set in acetone extracts of mycological material spectrophotometrically and calculated using the Vetshteyn formula.</p> <p>As the nitrogen-containing components used GPM with 9 compounds, such as peptone, DL-valine, L-asparagine, DL-serine, DL-tyrosine, L-proline, L-alanine, urea, NaNO<sub>3</sub>. The effect on the accumulation of specific compounds both in the mycelium and in the culture fluid of carotenoids by culturing certain strains of Basidiomycetes was identified.</p> <p>Adding to standard glucose-peptone medium peptone at 5 g/l causes an increase of carotenoid accumulation by strain <em>L. sulphureus</em> Ls-08, and in a concentration of 4 g/l by strains of <em>F. hepatica </em>Fh-18 and <em>F. fomentarius</em> Ff-1201.</p> <p>In order to increase the accumulation of carotenoids in the mycelium  we suggested to make a standard glucose-peptone medium with proline or valine for cultivating of <em>L. sulphureus</em> Ls- 08 strain; alanine for <em>F. fomentarius</em> Ff-1201 strain; proline, asparagine and serine - for strain Fh-18 of <em>F. hepatica</em>. The results can be implemented in further optimization of the composition of the nutrient medium for culturing strains of Basidiomycetes wich producing carotenoids.</p> <p><em>Keywords: nitrogen-containing substances, Basidiomycetes, mycelium</em><em>,</em><em> culture filtrate, carotenoids</em></p>

RESEARCH OF THE INFLUENCE OF ANTIBACTERIAL AND FUNGICIDAL PREPARATIONS ON FORMATION BIOFILM OF MICROORGANISMS

2021 ◽  
Vol 1 (4) ◽  
pp. 448-458
Author(s):  
Ekaterina M. Lenchenko ◽  
◽  
Dmitriy V. Stepanov ◽  
Dmitriy A. Blumenkrants ◽  
◽  
...  
Keyword(s):  
Culture Media ◽  
Animal Husbandry ◽  
The Body ◽  
Heterogeneous Structure ◽  
Candida Spp ◽  
Functional Patterns ◽  
Frequency Occurrence ◽  
Pathological Material

The results of studies general patterns formation heterogeneous structure biofilms gram-negative and gram-positive bacteria, as well as yeast-like fungi Candida spp. are presented. Рrocesses intercellular communication of various systematic groups microorganisms has common morphological and functional patterns biofilm formation. Heteromorphic structures of biofilms united by the intercellular matrix have been revealed in natural, industrial, and clinical conditions, both in the body of mammals and birds, and in food products, devices and equipment, animal husbandry and food production technologies. Indication in a large number of microcolonies, as well as yeast and micellar phases in isolates from pathological material of animals, was a differential sign in local and systemic pathologies. Under the influence drugs on biofilms microorganisms, a direct correlation was established between morphometric and densitometric indicators, reflecting a decrease in the frequency occurrence clusters and optical density, respectively. Under the bacteriostatic effect of chemotherapeutic and disinfecting drugs, accumulations altered cells of spheroplastic type, capable forming stable and unstable L-forms, were revealed. For detection of viable microorganisms in a heterogeneous population microorganisms in vitro and in vivo, fluorescence microscopy and culture media with growth factors for the repair cell wall of L-forms bacteria are promising.

Detection and characterization of purified antigenic proteins from culture filtrate of Mycobacterium bovis strain AN5

2020 ◽  
Author(s):  
Malihe Honarmand Kashi ◽  
Nader Mosavari ◽  
Mitra Salehi ◽  
Naheed Mojgani
Keyword(s):  
Molecular Weight ◽  
Western Blotting ◽  
Mycobacterium Bovis ◽  
Culture Filtrate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Immunological Methods ◽  
Antigenic Proteins ◽  
Specific Reactions ◽  
Dot Blotting

Background and Objectives: Bovine tuberculosis diagnosis is usually performed by various tests with specific limitations. Mycobacterium bovis culture filtrate contains antigenic proteins that could be used to improve the sensitivity of bovine tuber- culosis diagnosis. The objective of this study was to identify and purify antigenic proteins from culture filtrates of M. bovis strain AN5 for use in immunological assays. Materials and Methods: Secreted proteins were purified from the heat-treated culture filtrate of M. bovis strain AN5. Pro- teins were precipitated with ammonium sulfate, fractionated by Sephadex G50 chromatography. The protein concentrations and the approximate molecular weight were determined by lowry method and 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Immunological methods, including dot-blotting and western blotting, as- sessed the quality of the isolated proteins. Results: The quantity of antigenic proteins in the culture medium was measured at far more than 15% of the amount of pro- teins secreted into medium. Three main chromatographic fractions obtained and showed concentrations of proteins ranging from 14 to 60 μg/μl with molecular weights in the 10 to 180 kDa range. The purified antigens showed positive reactions to the infected cattle serum throughout dot-blotting. Western blotting revealed a total of 15 to 70 kDa molecular weight proteins. Conclusion: Immunoblotting analysis made it possible to detect and recognize novel antigens that are useful for bovine tu- berculosis diagnosis improvement. This is significant since non-specific reactions were not observed when we utilized serum of cattle experimentally infected with M. bovis as a polyclonal antibody.

scholarly journals Mycobacterium tuberculosis Culture Filtrate Proteins plus CpG Oligodeoxynucleotides Confer Protection to Mycobacterium bovis BCG-Primed Mice by Inhibiting Interleukin-4 Secretion

2009 ◽  
Vol 77 (12) ◽  
pp. 5311-5321 ◽  
Author(s):  
Denise Morais da Fonseca ◽  
Celio Lopes Silva ◽  
Pryscilla Fanini Wowk ◽  
Marina Oliveira e Paula ◽  
Simone Gusmão Ramos ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Bovis ◽  
Culture Filtrate ◽  
Vaccine Development ◽  
Interleukin 4 ◽  
Interleukin 17 ◽  
Bcg Vaccination ◽  
Cpg Oligodeoxynucleotides ◽  
Culture Filtrate Proteins ◽  
Ifn Γ

ABSTRACT Culture filtrate proteins (CFP) are potential targets for tuberculosis vaccine development. We previously showed that despite the high level of gamma interferon (IFN-γ) production elicited by homologous immunization with CFP plus CpG oligodeoxynucleotides (CFP/CpG), we did not observe protection when these mice were challenged with Mycobacterium tuberculosis. In order to use the IFN-γ-inducing ability of CFP antigens, in this study we evaluated a prime-boost heterologous immunization based on CFP/CpG to boost Mycobacterium bovis BCG vaccination in order to find an immunization schedule that could induce protection. Heterologous BCG-CFP/CpG immunization provided significant protection against experimental tuberculosis, and this protection was sustained during the late phase of infection and was even better than that conferred by a single BCG immunization. The protection was associated with high levels of antigen-specific IFN-γ and interleukin-17 (IL-17) and low IL-4 production. The deleterious role of IL-4 was confirmed when IL-4 knockout mice vaccinated with CFP/CpG showed consistent protection similar to that elicited by BCG-CFP/CpG heterologous immunization. These findings show that a single dose of CFP/CpG can represent a new strategy to boost the protection conferred by BCG vaccination. Moreover, different immunological parameters, such as IFN-γ and IL-17 and tightly regulated IL-4 secretion, seem to contribute to the efficacy of this tuberculosis vaccine.

scholarly journals Influence of the incubation conditions on culture media to optimize primary isolation of Mycobacterium bovis

2016 ◽  
Vol 37 (5Supl2) ◽  
pp. 3693
Author(s):  
Cássia Yumi Ikuta ◽  
Flávia Morato ◽  
Gisele Oliveira de Souza ◽  
Marcos Bryan Heinemann ◽  
Marcos Amaku ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Bovine Tuberculosis ◽  
Culture Media ◽  
Growth Time ◽  
Solid Culture ◽  
Sodium Pyruvate ◽  
Primary Isolation ◽  
Granulomatous Lesions ◽  
Tuberculosis Diagnosis ◽  
Carbon Dioxide Co2

The isolation of Mycobacterium bovis is critical to a surveillance system for bovine tuberculosis based on detection of lesions in abattoirs. Thus, four solid culture media and three incubation conditions were investigated to elucidate which combination overcomes the others by assessing growth, time to the first appearance of colonies and their number. Ninety-seven samples of granulomatous lesions were submitted to the decontamination procedure by 1-hexadecylpyridinium chloride at 0.75% w/v, and inoculated on two egg-based media, Stonebrink’s (ST) and Löwenstein-Jensen’s with sodium pyruvate (LJp), and two agar-based media, tuberculosis blood agar (B83) and Middlebrook 7H11 medium (7H11). Each medium was incubated at 37°C for 90 days in three incubation conditions: in air, in air containing 10% carbon dioxide (CO2), and in air in slopes closed with burned hydrophobic cotton and subsequently plugged with a cork to create a microaerophilic atmosphere. The colonies appeared faster and in higher number when incubated in air containing 10% CO2 (p < 0.01), independent of media. B83 showed a faster growth and detected more isolates at 30 days of incubation, when compared to ST (0.0178), LJp (p < 0.0001) and 7H11 (p < 0.0001), though there was no difference between B83, ST and LJp at 60 and 90 days of incubation. 7H11 presented the lowest number of isolates (p < 0.0001) and a longer period for the appearance of the first colony (p < 0.001). According to our findings, the concomitant use of ST and B83 media incubated in air containing 10% CO2 increases the isolation of M. bovis in a shorter period of time, which improves bovine tuberculosis diagnosis.

scholarly journals Vaccination of Cattle with Mycobacterium bovis Culture Filtrate Proteins and Interleukin-2 for Protection against Bovine Tuberculosis

2000 ◽  
Vol 68 (10) ◽  
pp. 5809-5815 ◽  
Author(s):  
D. Neil Wedlock ◽  
Bridget Vesosky ◽  
Margot A. Skinner ◽  
Geoffrey W. de Lisle ◽  
Ian M. Orme ◽  
...  
Keyword(s):  
Immune Responses ◽  
Mycobacterium Bovis ◽  
Culture Filtrate ◽  
Lipid A ◽  
Interleukin 2 ◽  
Delayed Type Hypersensitivity ◽  
Control Group ◽  
High Dose ◽  
Monophosphoryl Lipid A ◽  
Culture Filtrate Proteins

ABSTRACT In this study vaccines prepared from culture filtrate proteins (CFP) of Mycobacterium bovis and interleukin-2 (IL-2) were tested in cattle for their capacity to stimulate immune responses and to protect against an intratracheal challenge with virulent M. bovis. Nine groups of cattle were vaccinated with combinations of different doses of CFP and bovine IL-2 mixed with a monophosphoryl lipid A (MPL) adjuvant. An additional group was vaccinated withM. bovis BCG. Immune responses in CFP–IL-2-vaccinated animals differed from those seen in BCG-vaccinated animals by inducing high antigen-specific antibody responses and low levels of gamma interferon and IL-2 released from purified protein derivative-stimulated whole-blood cultures. In a concurrent experiment, additional animals were added to the high-dose CFP–IL-2, MPL control, and BCG groups and these expanded groups of animals were challenged intratracheally with virulent M. bovis. Although the lung lesion scores were significantly lower for both the CFP–IL-2-and BCG-vaccinated groups compared to the MPL control group, the overall level of protection was greatest for the BCG-vaccinated animals. There were more animals with extrathoracic spread of disease in the CFP–IL-2 group than in the other groups. While vaccination of cattle withM. bovis CFP gave an encouraging reduction in tuberculous lesions and did not induce a delayed-type hypersensitivity response to PPD, future CFP vaccines must prevent any extrathoracic spread of disease.

Antigenic characterization of Mycobacterium bovis BCG culture filtrate

1994 ◽  
Vol 41 (4) ◽  
pp. 345-353
Author(s):  
V.K. Gupta ◽  
G.C. Ram ◽  
M.P. Bansal
Keyword(s):  
Mycobacterium Bovis ◽  
Culture Filtrate ◽  
Mycobacterium Bovis Bcg ◽  
Antigenic Characterization

scholarly journals Development of the technology for preparation of enzymatic hydrolysate of waste chick embryos

2021 ◽  
Vol 21 (3) ◽  
pp. 200-205
Author(s):  
Yu. S. Ovsyannikov ◽  
M. S. Dursenev
Keyword(s):  
Nutrient Medium ◽  
Mass Fraction ◽  
Chemical Properties ◽  
Chick Embryos ◽  
Nutrient Media ◽  
Enzymatic Hydrolysate ◽  
E Coli ◽  
Physico Chemical ◽  
Growth Properties ◽  
Solid Nutrient Medium

The development of technologies for preparation of protein nutritional bases for microbiological nutrient media, from production waste of mainly readily available or non-food products, is a promising area in biotechnology. Researchers of Vyatka State Agrotechnological University assume that non-food secondary raw materials, such as waste chick embryos (WCEs) used in the production of anti-influenza products, could be used for these purposes, after removal of the virus-containing allantoic fluid. The aim of the study was to develop a technology for preparation of WCE enzymatic hydrolysate (WCEEH), and to evaluate growth properties of the hydrolysate-based solid nutrient medium, using Escherichia coli M-17 and Pseudomonas alcaligenes IP-1 test strains. Materials and methods: the authors offer methodological approaches to obtaining WCEEH and substantiate hydrolysis parameters. The obtained WCEEH was characterised in terms of physico-chemical properties: pH, amine nitrogen, total nitrogen, sodium chloride, degree of protein cleavage. The growth properties of the hydrolysate-based nutrient medium were studied using E. coli M-17 and Ps. alcaligenes IP-1 test strains. Results: the experiments demonstrated the feasibility of performing enzymatic hydrolysis of WCEs, and assessed physico-chemical properties of the prepared WCEEH batches. The study demonstrated the possibility of using the prepared hydrolysate as a component of solid nutrient media for growing the selected test strains. Conclusions: the study substantiated the optimal technological parameters for WCE enzymatic hydrolysis: pH (7.6 ± 0.3), duration (48 ± 2 h), temperature (49 ± 1) °C. The loading of hydrolysis components was optimised: mass fraction of the substrate—500 g/L, mass fraction of the hydrolysing agent—100 g/L. The physico-chemical properties of WCEEH make it suitable for preparation of microbiological media; the hydrolysate-based solid nutrient medium consistently ensures the growth of E. coli M-17 and Ps. alcaligenes IP-1 test strains with standard properties. The growth properties of the experimental medium are comparable to those of the meat-peptone broth-based nutrient medium.

scholarly journals An innovative modification of the nutrient medium formulation for the isolation and differentiation of enterobacteria

2021 ◽  
Vol 37 ◽  
pp. 00063
Author(s):  
Vladimir Ermakov ◽  
Nikolay Titov
Keyword(s):  
Hydrogen Sulfide ◽  
Nutrient Medium ◽  
Culture Media ◽  
Laboratory Conditions ◽  
Environmental Objects ◽  
Single Section ◽  
Single Compartment ◽  
Petri Dishes ◽  
Ferment Lactose ◽  
Medium Formulation

In laboratory conditions, the modified nutrient differential diagnostic medium Drigalski agar with lactose is recommended to be used for cultivation (isolation) and differentiation of enterobacteria of the Enterobacteriaceae family. The differentiation of enterobacteria on a modified medium is carried out according to their ability to ferment lactose, mannitol, glucose, sucrose, gelatin and form hydrogen sulfide. The environment can also be used for conducting sanitary and microbiological studies of environmental objects. The environment can be used to perform the ONPG test. In the course of preparing the modified Drigalski agar for operation in laboratory conditions, we recommend using the medium according to one of the options: when one of the carbohydrates is added, the medium is poured into single-section reusable or disposable petri dishes; when two carbohydrates are added, the medium is poured into two-section reusable or disposable petri dishes; when a complex of carbohydrates lactose + mannitol and glucose + sucrose is added, the medium is poured into two-section reusable or disposable petri dishes; when using all four drives separately, the medium is dispensed into single-compartment reusable or disposable petri dishes. We recommend preparing modified Drigalski agar at a plant for the production of ready-made culture media with filling the medium in two-section petri dishes (complex of carbohydrates lactose + mannitol in one section, glucose + sucrose in another section), or with filling the medium in four-section petri dishes (with each carbohydrate in separate section).

scholarly journals Prolactin modulate proinflammatory inmune response induced by culture filtrate protein from Mycobacterium bovis in THP1 cells through different signaling pathways

2013 ◽  
Vol 4 ◽  
Author(s):  
Mart�nez-Neri Priscila ◽  
L�pez-Rinc�n Gonzalo ◽  
Mu�oz-Valle Jos� ◽  
L�pez Pulido Edgar ◽  
Estrada-Ch�vez Ciro ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Mycobacterium Bovis ◽  
Culture Filtrate ◽  
Culture Filtrate Protein
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