Eurasian Journal of Applied Biotechnology

OBTAINING ASEPTIC CULTURE IN VITRO OF RHODIOLA SEMENOWII L. FROM VARIOUS TYPES OF EXPLANTS

2024-10-07T12:24:07+05:00

In vitro cultivation of callus cells allows for the stable production of secondary metabolites, being an alternative to the use of medicinal plant biomass from natural populations. Standardization of cultivation protocols and technological processes for the synthesis of secondary metabolites in vitro conditions determines significant interest in the use of tissue culture methods (PTC – Plant Tissue Culture). The aim of the study was to obtain an aseptic culture of Rhodiola semenowii in vitro from various types of explants, as well as the effect of the hormonal composition of the nutrient medium on the induction of callusogenesis in the culture of isolated organs of R. semenowii. As a result of the research, the optimal mode of obtaining R. semenovii aseptic cultures from various explants was determined. It has been shown that the use of 70% EtOH in combination with 0.01% HgCl₂ or 20% NaClO makes it possible to obtain aseptic cultures from any type of R. semenovii explants. It has been established that the type of explant is R. semenovii influenced both the frequency of callus formation and the beginning of its formation. The least optimal types of R. semenovii explants are stem and rhizome segments, the frequency of callusogenesis varied from 5.8% to 75.6%. The induction of callusogenesis also depended on the hormonal composition of the nutrient medium: the highest percentage of callus formation in all types of explants was on media containing 0.5mg L⁻¹ thidiazuron (TDZ) + 1.0 mg L⁻¹1-naphthaleneacetic acid (NAA) and 0.5mg L⁻¹2,4D + 2.0 mg L⁻¹ 6-Benzylaminopurine (BAP).

INFLUENCE OF ALTERNARIA SPP. FUNGI METABOLITES ON DEVELOPMENT OF WHEAT SPROUTS

2024-10-24T12:28:55+05:00

The study purpose was in vitro investigation of phytotoxic activity of culture broth filtrates (CBFs) of five Alternaria Spp. fungi isolates on development of wheat sprouts. Fungi isolates were received from wheat seeds cultivated under conditions of Northern Kazakhstan. According to the study findings, culture filtrates of all the isolates studied varied with regard to their influence on wheat sprout development. Stimulant effects on seed germinating power were revealed with 14- and 21-day CBFs of A. infectoria (7/4), 14-day CBF of A. brassicae (7/2) and 21-day CBF of A. porri (8/1). It was found that long-term cultivation of fungi on a culture medium contributed to media pH modification towards alkalization due to production of metabolites of various chemical nature, including toxins, which could trigger growth inhibition of radicles and sprouts. However, the CBFs were predominantly low-toxic or non-toxic. Under effect of the 14-day CBF, significant positive correlation was identified between pH values and sprout lengths (0,86±0,25), while negative correlation was found between pH values and germinating power (-0,75±0,33). Change of pH value on the 21^st day significantly affected radicle length (-0,92±0,20). Based on the findings it can be concluded that the 14-day CBF of all the Alternaria fungi isolates studied was most toxic for wheat sprouts.

IDENTIFICATION OF SECONDARY METABOLITES BY GAS CHROMATOGRAPHY WITH MASS SPECTROSCOPY IN CALLUS TISSUES OF CISTANCHE DESERTICOLA Y.C. MA

2024-10-16T18:26:04+05:00

The aim of this study is to analyze the types of secondary metabolite compounds found in cistanche (Cistanche deserticola) plants. Methanolic extracts of callus tissues and shoots (stolon) of cistanche were obtained by lyophilic drying and grinding to powdered state using 80% methanol. Secondary metabolites within cystanche callus were first identified by GC-MS (gas chromatography-mass spectroscopy). The identification results of the methanolic extracts revealed various types of compounds including fatty acids (hexadecanoic acid, methyl ester); phenolic compounds and their derivatives (phenol, 2,4-bis(1,1-dimethylethyl)-, 2-methoxy-4-vinylphenol, homovanillyl alcohol, phenol, 2,6-dimethoxy;) sterols (γ-sitosterol); sugars, glycosides and other carbohydrates (ethyl α-D-glucopyranoside, sucrose), glycols and their derivatives (tetraethylene glycol, diethylene glycol monodododecyl ether), etc. Some of the detected compounds may be precursors or by-products of phenylethanoid glycosides biosynthesis. We also found a correlation between callus color and its chemical composition of secondary metabolites. The data can be used for further optimization of C. deserticola cultivation methods to increase the content of target secondary metabolites.

POTENTIAL FOR ACCELERATED REPLICATION OF FRAXINUS SOGDIANA USING IN VITRO CULTURE AND IDENTIFICATION OF A RELIC TREE USING DNA-BARCODING

2024-10-29T09:57:28+05:00

Four species of Fraxinus including F. angustifolia subsp. syriaca (Boiss.) Yalt., F. sogdiana Bunge, F. pennsylvanica Marshall and F. americana L. grow in Kazakhstan. The study aimed to determine the conditions for awakening endogenous dormancy in vitro of F. sogdiana. In the control group without vernalization, the sterile culture yield at room temperature was 30%. However, with cold treatment, this yield increased to 60%, which is twice as high. It was observed that on MS medium without the addition of hormones, 22 microshoots were obtained from unwounded seeds, whereas 39 microshoots were obtained from wounded seeds during microclonal propagation.Phylogenetic relationships and species identification were analyzed using three DNA barcoding markers, comparing F. sogdiana with species from the NCBI database. rbcL and ITS markers did not show satisfactory interspecific genetic variability among F. species. Notably, matK gene sequences effectively differentiated F. sogdiana from other species. These results provide fundamental information that is valuable for future biotechnological and molecular studies.

ASSESSMENT OF BIOLOGICAL RISKS IN INFECTIOUS DISEASES TO ENSURE BIOLOGICAL SAFETY

2024-09-17T10:49:16+05:00

The territory of the Republic of Kazakhstan is unfavorable for a number of infectious diseases of animals and humans that pose an enzootic biological threat. Such nosological units include anthrax, blackleg, rabies, leukemia, brucellosis, tuberculosis, pasteurellosis, sheep pox, camel pox, CCHF, highly pathogenic avian influenza, foot-and-mouth disease, tick-borne encephalitis, influenza A and B, infectious hepatitis, Newcastle disease, seasonal flu, coronavirus infection COVID-19, etc., some of which affect only animals or humans, and some affect both animals and humans. While some of these diseases are natural focal and pose a constant biological threat, others appear from time to time, penetrating from neighboring countries or develop as a result of movement from the wild - reservoirs and hidden carriers of pathogens. A number of exotic diseases pose a real threat of penetration from outside the country due to interstate movements of people, animals, feed and animal products.

ORAL VACCINE: AS A MEANS OF PREVENTING RABIES IN WILDLIFE

2024-11-20T12:57:04+05:00

Rabies is a zoonotic infection of viral etiology that poses a danger to the health of both animals and humans. The disease is widespread everywhere, with the exception of some European countries, island states, Australia and Antarctica, which have managed to eliminate rabies in their territories. The causative agent of the infection is a representative of the family Rhabdoviridae, affecting all living organisms including vertebrates, invertebrates, plants and fish. A particular danger to humans is the genus Lyssavirus, which in most reported cases enters the body through contact of damaged skin with the saliva of an infected animal. The incubation time depends on the distance of the infection site to the brain. Affecting the nervous system and reaching the brain, the rabies virus leads to paralysis followed by imminent death. The main reservoirs for the virus are wild animals, in particular carnivores, as well as bats. One of the most effective ways to prevent transmission of the virus is to monitor and carry out preventive measures, including vaccination. The purpose of this review article is to show by global experience how oral vaccines have contributed to a significant reduction in the spread of rabies among wild carnivorous animals, which ultimately influenced the dynamics of morbidity among domestic animals and humans.

DESIGNING A PCR TEST SYSTEM FOR THE GENETIC DIAGNOSIS OF RICKETTSIOSES

2024-11-25T11:27:56+05:00

To ensure the biological safety of the country, it is necessary to develop priority areas of biotechnology, create advanced means of detecting pathogenic biological agents. Rickettsia bacteria, one of the most widespread zoonotic pathogens worldwide, continues to be a serious problem for public health. One of the solutions to the problems related to the situation of the effectiveness of rickettsiosis monitoring is the widespread introduction into clinical practice of modern diagnostic methods based on nucleic acid amplification (MANC) methods, in particular PCR. The paper presents the test results of experimental series of rickettsia detection kits.

Purpose. Development and construction of experimental series of PCR test systems for the detection of rickettsioses, conducting control tests to work out optimal conditions in order to achieve maximum sensitivity and specificity of the drug.

ANTIVIRAL IL-33-STUMULATED GROUP 2 INNATE LYMPHOID CELLS (CD-90 AND CD-117) FROM MOUSE LUNGS

2024-10-02T08:01:55+05:00

This article represents not only a significant scientific value, but also a great teaching and instruction value for researchers and motivated students to improve the skill spectrum in flow cytometry. This article provides step-by- step procedure description and results based on in vivo studies on murine lung tissue, cell of interest isolation with dual protocols for flow cytometry method as well as IL-33 impact in dynamics. The newly described innate lymphoid cells of group 2 (ILC2) play an important role in type 2 immune reactions, epithelial repair in mucosal tissue and metabolic homeostasis. ILC2 releases large amounts of type 2 cytokines such as interleukin 4 (IL-4), IL-5 and IL-13, which stimulate type 2 immunity, such as protection against worms. However, without strict regulation, the level of ILC2 can cause undesirable type 2 immune pathologies, including allergic inflammation of the respiratory tract, hypersensitivity of the respiratory tract and atopic dermatitis. Viral infections of the respiratory tract, which are typical triggers of type 1 immune reactions, often lead to type 2 pulmonary immune pathologies, such as asthma and its exacerbations. Interestingly, pulmonary virus infections induce the release of IL-33 with subsequent induction of ILC2-mediated type 2 pulmonary immunopathology, which is independent of the adaptive immune system.

NEW BACTERIOLYTIC ENZYME PREPARATION N-ACETYL-D-GLUCOSAMINIDASE

2024-12-04T18:46:51+05:00

A new nag1 gene encoding N-acetyl-D-glucosaminidase from Streptomyces kursanovii was cloned. The soluble form of N-acetyl-D-glucosaminidase was obtained after improving its expression in E. coli ArcticExpressTM (DE3) cells. The homogeneous form of the enzyme was isolated by affinity chromatography, and its bacteriolytic ability against cells of the Gram-positive bacterium Micrococcus lysodeikticus was studied. It was demonstrated that the new enzyme's bacteriolytic capacity surpasses that of chicken lysozyme.

THE EFFECT OF THE IODINE-CONTAINING COMPOUNDS ON GENE EXPRESSION OF PATHOGENIC MICROORGANISMS

2024-12-04T16:05:11+05:00

The global increase in multidrug resistance highlights the need for new, effective antimicrobial agents. Iodine-containing complexes, due to their broad-spectrum antimicrobial properties, low likelihood of resistance development, and stability under various conditions, represent a promising avenue for the development of innovative therapeutic approaches.

The aim of this study was to collect and analyze research assessing the impact of iodine-containing complexes with organic ligands on the gene expression of certain pathogenic strains of microorganisms with multidrug resistance. Bioinformatics approaches were employed to investigate gene expression, protein interactions, and metabolic pathways, utilizing tools like STRING, KEGG, Reactome, and UniProt. Models of gene and proteome interaction analysis were also used to interpret the data.

Key findings include the suppression of central metabolic cycles, such as the tricarboxylic acid cycle (TCA), increased glycolytic activity, a shift in metabolism toward anaerobic respiration, and activation of oxidative stress defense systems. The study also highlights changes in gene expression associated with membrane remodeling, DNA repair, regulation of virulence factors, and nutrient transport. These adaptive responses reflect the multifaceted antimicrobial action of iodine-containing complexes, which target key cellular components and processes, making them highly effective against multidrug-resistant pathogens.

Iodine-containing complexes exhibit high antimicrobial potential due to their ability to disrupt essential bacterial life processes, impairing metabolism, structure, and adaptive mechanisms. Further research into the molecular mechanisms of iodine’s action and its effects on pathogen virulence could open new avenues for addressing the challenge of antibiotic resistance.