Pathogenic Properties Research Articles

Sequence Analysis of the Malaysian Low Pathogenic Avian Influenza Virus Strain H5N2 from Duck.

The avian influenza viruses (AIV) of the H5 subtype have the ability to mutate from low pathogenic (LPAI) to highly pathogenic (HPAI), which can cause high mortality in poultry. Little is known about the pathogenic switching apart from the mutations at the haemagglutinin cleavage site, which significantly contributes to the virus virulence switching phenomenon. Therefore, this study aimed to compare the molecular markers in the haemagglutinin (HA), neuraminidase (NA), and matrix (M) genes of a locally isolated LPAI AIV strain H5N2 from Malaysia with the reference HPAI strains using bioinformatics approaches, emphasising the pathogenic properties of the viral genes. First, the H5N2 strain A/Duck/Malaysia/8443/2004 was propagated in SPF eggs. The viral presence was verified by haemagglutination assay, RT-PCR, and sequencing. Results showed successful amplifications of HA (1695 bp), NA (1410 bp), and M (1019 bp) genes. The genes were sequenced and the deduced amino acid sequences were analysed computationally using MEGA 11 and NetNGlyc software. Analysis of the HA protein showed the absence of the polybasic cleavage motif, but presence of two amino acid residues that are known to affect pathogenicity. There were also two glycosylation sites (glycosites) compared to the reference HPAI viruses, which had three or more at the HA globular head domain. No NA stalk deletion was detected but the haemadsorbing and active centres of the studied NA protein were relatively similar to the reference HPAI H5N2 isolates of duck but not chicken origins. Six NA glycosites were also identified. Finally, we observed a consistent M1 and M2 amino acid sequences between our LPAI isolate with the other HPAI H5N1 or H5N2 reference proteins. These data demonstrate distinct characteristics of the Malaysian LPAI H5N2, compared to HPAI H5N2 or H5N1 from ducks or chickens, potentially aiding the epidemiological research on genetic dynamics of circulating AIV in poultry.

Prevalence and genetic characterization of Campylobacter from clinical poultry cases in China.

Campylobacter is one of the leading causes of bacterial foodborne diseases globally. Poultry is considered a major reservoir for the transmission of Campylobacter to humans. The occurrence and epidemiology of Campylobacter in healthy poultry have been extensively studied, but the prevalence in diseased poultry has rarely been reported. In this study, a total of 1,563 intestinal samples, 547 gallbladder samples, and 3,729 parenchyma organ samples were collected from diseased chickens, ducks, and geese in Yangzhou Veterinary Hospital between December 2016 and September 2017. The bacterial isolates were analyzed for genotype, resistance, and virulence genes. The overall prevalence of Campylobacter was 17.9%, and a significantly higher prevalence was observed in the diseased chickens (21.8%) than in the diseased ducks (12.1%) and geese (11.9%). Campylobacter was significantly more prevalent in the intestinal tract (39.6%), followed by the gallbladder (11.7%) and parenchyma organ (9.7%). Among 84 whole-genome sequenced Campylobacter isolates, we identified 47 multi-locus sequence typing types, of which ST-51 and ST-825 were the most predominant sequence types (STs) for Campylobacter jejuni and Campylobacter coli, respectively. Phylogenetic analysis showed that Campylobacter isolates from different tissues from the same host tended to cluster together with the same STs, which indicated Campylobacter infection in different tissues in vivo. The antimicrobial resistance genes most frequently identified were gyrA (T86I) (97.6%) and tet (O) (65.5%). Our results revealed the high prevalence of Campylobacter in diseased poultry in China, which will help the farms take the necessary action to develop effective mitigation strategies for reducing Campylobacter infection in poultry.IMPORTANCECampylobacter is a major cause of campylobacteriosis worldwide, and poultry is the main reservoir for its transmission. Campylobacter was generally considered to be a harmless commensal organism in poultry without pathogenic properties. However, it was proposed that a Campylobacter-like organism may be the cause of vibrionic hepatitis, which poses a significant public health risk. The occurrence and epidemiology of Campylobacter in healthy poultry have been studied systematically, but little is known about the epidemiology of Campylobacter isolates from diseased poultry in China. Therefore, this study determined the prevalence and molecular characterization of Campylobacter from diseased chickens, ducks, and geese in Yangzhou Veterinary Hospital between December 2016 and September 2017, which was critical for improving the diagnosis and prevention of Campylobacter infections.

ПЕРСПЕКТИВЫ РАЗРАБОТКИ НОВЫХ ОРИГИНАЛЬНЫХ АНТИСЕПТИЧЕСКИХ СРЕДСТВ НА ОСНОВЕ УСНИНОВОЙ КИСЛОТЫ

Introduction. Environment changing not least because of human activities determines the appearance of modified microorganisms. Many pathogens have enhanced their pathogenic properties. Their increasing drug resistance represents quite a serious problem. Among potentially efficient antiseptic substances, usnic acid and its derivatives are greatly emphasized on. Aim of our study was to conduct a narrative review of the prospects for the practical (pharmaceutical) use of usnic acid as an antiseptic. Research Materials. To perform the literature analysis, we used sources from international databases, such as Web of Science, Scopus, and PubMed, as well as from the domestic library system, eLibrary. Results. Usnic acid is found in large amounts in various lichen species. It has quite a broad spectrum of antimicrobial action. Its anti-inflammatory effects and enhancement of tissue regeneration wasproven both experimentally and clinically. The operation of usnic acid and its derivatives is determined by inhibiting the synthesis of ribonucleic acid and breaking the deoxyribonucleic acid replication. Moreover, it has a cytotoxic effect and destroys the membranes of microorganisms. In pharmaceutical industry, usnic acid is produced in many forms, such as powders, capsules, antiseptic gels, creams, and protective tapes. They are used in combined treatment of infectious diseases, in general therapy, in post-burn wound healing procedures, and in dentistry. We have concluded that lichens are very promising as raw materials for isolating usnic acid. It is necessary to search for new highly efficient methods to manufacture and use it in various farmaceutical forms.

Aerobiology in Poland: Achievements and challenges

Aerobiology is a scientific discipline that focuses on investigating biological particles (bioaerosols) passively transported in the atmosphere, mainly fungal spores and pollen grains. Due to the allergic and pathogenic properties of these particles, aerobiological data is widely used in allergology, plant pathology, and agriculture. In addition, the results of aerobiological monitoring have found applications in biogeography, paleoecology, forestry, veterinary medicine, forensic science, and climate change research, as well as in the assessment of threats to the deterioration of cultural heritage. In this review, the contribution of Polish scientists to the development of worldwide aerobiology is presented. The most important results on the atmospheric dispersion of bioaerosols, spatiotemporal modeling, and novel methods used in aerobiological research are described. Moreover, the cooperation at the national and international levels as part of scientific programs and the perspectives for further development of aerobiology in Poland are discussed.

Differential contagiousness of respiratory disease across the United States

The initial contagiousness of a communicable disease within a given population is quantified by the basic reproduction number, R0. This number depends on both pathogen and population properties. On the basis of compartmental models that reproduce Coronavirus Disease 2019 (COVID-19) surveillance data, we used Bayesian inference and the next-generation matrix approach to estimate region-specific R0 values for 280 of 384 metropolitan statistical areas (MSAs) in the United States (US), which account for 95% of the US population living in urban areas and 82% of the total population. We focused on MSA populations after finding that these populations were more uniformly impacted by COVID-19 than state populations. Our maximum a posteriori (MAP) estimates for R0 range from 1.9 to 7.7 and quantify the relative susceptibilities of regional populations to spread of respiratory diseases. One-Sentence SummaryInitial contagiousness of Coronavirus Disease 2019 varied over a 4-fold range across urban areas of the United States.

Mechanisms of oral microbial biofilm formation in healthy people and patients with chronic generalized periodontitis

Relevance. Oral biofilms are integrated three-dimensional symbiontic microbial structures embedded in the extracellular matrix that form not only on the mucous membrane but also on hard surfaces such as enamel and root of teeth, as well as dentures. Our study correlated biofilms and the onset of such a serious oral disease as chronic generalized periodontitis.Material and Methods. The analysis of national and international literature in PubMed, Google Search and eLIBRARY databases over the past 15 years served as material and methods.Results. There is currently an increased interest in studying the mechanisms of microbial biofilms and the factors affecting this process. Planktonic bacteria are more sensitive to antimicrobials than bacteria in biofilms. Several factors, such as gas signalling molecules, may become an important tool for intermicrobial communication in a biofilm. Biofilm microorganisms are highly resistant to antibacterial drugs. Bacteria release enzymes and toxins that stimulate the body to produce large amounts of specific antibodies and cytokines. However, the immune components are blocked by the biofilm extracellular matrix and cannot enter the biofilm. Immune complexes are formed in the infection foci, damage the body's tissues, and aggravate inflammation.Conclusion. Periodontal diseases are the most common oral diseases in humans, and their onset is closely related to the pathogenic properties of the oral microbiota, which exists in biofilms. The mechanisms of biofilm formation are complex and associated with the expression of various microorganism/matrix pathogenicity factors by oral commensal microorganisms. Further study is necessary to understand the way of biofilm formation neutralization and to find an effective treatment for chronic generalized periodontitis.

Multi-omics analysis of attenuated variant reveals potential evaluation marker of host damaging for SARS-CoV-2 variants.

SARS-CoV-2 continues to threaten human society by generating novel variants via mutation and recombination. The high number of mutations that appeared in emerging variants not only enhanced their immune-escaping ability but also made it difficult to predict the pathogenicity and virulence based on viral nucleotide sequences. Molecular markers for evaluating the pathogenicity of new variants are therefore needed. By comparing host responses to wild-type and variants with attenuated pathogenicity at proteome and metabolome levels, six key molecules on the polyamine biosynthesis pathway including putrescine, SAM, dc-SAM, ODC1, SAMS, and SAMDC were found to be differentially upregulated and associated with pathogenicity of variants. To validate our discovery, human airway organoids were subsequently used which recapitulates SARS-CoV-2 replication in the airway epithelial cells of COVID-19 patients. Using ODC1 as a proof-of-concept, differential activation of polyamine biosynthesis was found to be modulated by the renin-angiotensin system (RAS) and positively associated with ACE2 activity. Further experiments demonstrated that ODC1 expression could be differentially activated upon a panel of SARS-CoV-2 variants of concern (VOCs) and was found to be correlated with each VOCs' pathogenic properties. Particularly, the presented study revealed the discriminative ability of key molecules on polyamine biosynthesis as a predictive marker for virulence evaluation and assessment of SARS-CoV-2 variants in cell or organoid models. Our work, therefore, presented a practical strategy that could be potentially applied as an evaluation tool for the pathogenicity of current and emerging SARS-CoV-2 variants.

Modern views on the problem of antifungal resistance of strains of fungi of the genera Fusarium and Aspergillus in the context of the concept of One Health

the «One Health» concept integrates human, animal and environmental health and emphasizes the need for collaboration between different sectors such as medicine, veterinary medicine, ecology, public health and other sciences to achieve greater understanding and management of common health threats. This concept arose from the realization that many diseases, including infectious diseases, cancer and mental disorders, have common roots related to the state of the ecosystem. For example, the spread of infectious diseases can be caused by climate changes, loss of biodiversity, inappropriate use of antibiotics, xenobiotics, and other factors. In addition, pathogens of animal diseases can also acquire pathogenic properties and pose a threat to public health. «One Health» implements such an approach to health, which instead of the traditional vertical approach is system-oriented, it encourages comprehensive actions to prevent and control disease at different levels. This approach includes comprehensive medical diagnostics, monitoring, research and coordination, as well as strengthening cooperation between scientists, doctors, veterinarians, ecologists and other specialists. Undoubtedly, an important issue in the context of the implementation of the «One Health» concept is the study of the formation of resistance to antifungal drugs in microscopic fungi, including representatives of the genera Fusarium and Aspergillus. The ratio of natural to artificial resistance can be highly variable among different fungal strains, even within the same species or genus. This is due to the genetic diversity of fungi and their ability to mutate and adapt to new conditions. Considering these factors, it is important to constantly monitor the resistance of mycelial fungi to antifungal drugs and detect the emergence of new resistant strains. This will make it possible to use antimicrobials correctly and provide effective treatment.

The Role of Innate Immunity against Mycobacterium Leprae: A Systematic Literature Review

Introduction: Innate immunity is the body's defense system that exists from birth and is the body's first defense against infection. This system involves multiple defense mechanisms that work together to recognize, deter, and respond to pathogens such as bacteria, viruses, and fungi. In the context of Mycobacterium leprae, the role of the innate immune system is very important because this bacterium has unique pathogenic properties and is capable of infecting human body cells in complex ways.
 Methods: The literature search process was carried out on various databases (PubMed, Web of Sciences, EMBASE, Cochrane Libraries, and Google Scholar) regarding the role of innate immunity against Mycobacterium leprae. This study follows the preferred reporting items for systematic reviews and meta-analysis (PRISMA) recommendations.
 Results: One of the main aspects of the role of the innate immune system against Mycobacterium leprae is the recognition of this pathogen by phagocytic cells, such as macrophages. Mycobacterium leprae evades recognition and destruction by these cells in a number of ways, including cell surface modification of the immune response.
 Conclusion: The innate immunity response plays a significant role in the response to Mycobacterium leprae infection. Innate immunity acts as an initial and fast response to mitigate the Mycobacterium leprae infection from getting more massive.

Molecular Identification, Effector genes and Virulence of Isolates of <i>Parastagonospora nodorum</i> from Altai Krai (Russia)

Septoria blotches represent some of the most harmful wheat diseases that encompass all grain-producing countries worldwide. They are caused mainly by fungi of the genera Parastagonospora and Zymoseptoria. The aims of the research were to analyze the nucleotide sequences of two phylogenetically informative DNA loci, ITS and tub2, of Parastagonospora nodorum isolates from the Altai Krai, to detect the presence of Tox1, Tox3, ToxA effector genes and to study the virulence of the isolates. Microscopic analysis of fragments of affected plant tissue was used for primary identification of the Septoria blotch. The causative agent from the studied plant samples was identified as P. nodorum. Fungal colonies cultivated on potato-glucose agar displayed a well-developed light brown velvety mycelium with dark periphery (mixed type) and numerous pycnidia exhibiting high sporulation capacity. Microbiological diagnostics were complemented by molecular genetic studies. Sequencing of ITS and tub2 loci isolates revealed complete genetic identity of all eight studied monoconidial isolates obtained from different samples. PCR-based detection of tree Tox-genes demonstrated that the only Tox3 was present in the geno-types of 80 tested P. nodorum monoconidial isolates. The ToxA and Tox1 genes were not found in the studied isolates. The virulence of P. nodorum isolates was evaluated in laboratory conditions using an isolated wheat leaves assay. Mixture of four isolates from spring wheat and one isolate from winter wheat, were characterized as virulent. When spring and winter wheat cultivars were infected with two isolates obtained from oats, pathogenic but avirulent properties were detected. The isolate from triticale was avirulent to winter wheat cultivars and virulent to spring wheat cultivars.

The influence of bacteriophages on the manifestation of their pathogenic properties by Xanthomonas campestris bacteria

The use of bacteriophages to control the development of different bacterioses, including those caused by bacterium Xanthomonas campestris is one of the most promising areas of development in the field of plant protection. At the same time, the selection of the most active bacteriophages and the determination of parameters of their use remains one of the most urgent tasks of this direction. The use of phage preparations with low initial concentration of phage particles may not only prevent the development of bacterial disease, but also lead to the emergence of phage-resistant bacterial strains. In this paper, the influence of bacteriophages in various concentrations on bacterial strains and the possibility of latter infecting the host plant is analyzed. It was established that the treatment of bacterial suspensions by bacteriophages with a titer of 107-108 PFU/ml excluded the development of disease in plants inoculated with this bacterial suspension. In case of a decrease in the initial titer of bacteriophage, in some cases, symptoms of the development of the disease began to appear, down to the point that after treatment of bacterial suspensions by phages with a titer of 103 PFU/ml, the development of bacterial disease in host plants inoculated with this bacterial suspension was noted in all the studied cases.

New Approach for Studying of Isoforms and High-Homology Proteins in Mammalian Cells.

In mammals, a large number of proteins are expressed as more than one isoform, resulting in the increased diversity of their proteome. Understanding the functions of isoforms is very important, since individual isoforms of the same protein can have oncogenic or pathogenic properties, or serve as disease markers. The high homology of isoforms with ubiquitous expression makes it difficult to study them. In this work, we propose a new approach for the study of protein isoforms in mammalian cells, which makes it possible to individually detect and investigate the functions of an individual isoform. The approach was developed to study the functions of isoforms of the PHF10 protein, a chromatin subunit of the PBAF remodeling complex. We demonstrated the possibility of induced simultaneous suppression of all endogenous PHF10 isoforms and the expression of a single recombinant FLAG-tagged isoform. For this purpose, we created constructs based on the pSLIK plasmid with a cloned cassette containing the recombinant gene of interest and miR30 with the corresponding shRNAs. The doxycycline-induced activation of the cassette allows on and off switching. Using this construct, we achieved the preferential expression of only one recombinant PHF10 isoform with a simultaneously reduced number of all endogenous isoforms. Our approach can be used to study the role of point mutations, the functions of individual domains and important sites, or to individually detect untagged isoforms with knockdown of all endogenous isoforms.

Updates on the Virulence Factors Produced by Multidrug-Resistant Enterobacterales and Strategies to Control Their Infections.

The Enterobacterales order is a massive group of Gram-negative bacteria comprised of pathogenic and nonpathogenic members, including beneficial commensal gut microbiota. The pathogenic members produce several pathogenic or virulence factors that enhance their pathogenic properties and increase the severity of the infection. The members of Enterobacterales can also develop resistance against the common antimicrobial agents, a phenomenon called antimicrobial resistance (AMR). Many pathogenic Enterobacterales members are known to possess antimicrobial resistance. This review discusses the virulence factors, pathogenicity, and infections caused by multidrug-resistant Enterobacterales, especially E. coli and some other bacterial species sharing similarities with the Enterobacterales members. We also discuss both conventional and modern approaches used to combat the infections caused by them. Understanding the virulence factors produced by the pathogenic bacteria will help develop novel strategies and methods to treat infections caused by them.

NMDA receptor inhibitor MK801 alleviated pro-inflammatory polarization of BV-2 microglia cells

Microglia have both protective and pathogenic properties, while polarization plays a decisive role in their functional diversity. Apart from being an energetic organelle, mitochondria possess biological capabilities of signaling and immunity involving mitochondrial dynamics. The N-methyl-D-aspartate (NMDA)-type glutamate receptor displays excitatory neurotransmission, excitatory neurotoxicity and pro-inflammatory properties in a membrane location- and cell context-dependent manner. In this study, we have provided experimental evidence showing that by acting on mitochondrial dynamics, NMDA receptors displayed pro-inflammatory properties, while its non-competitive inhibitor MK801 exhibited anti-inflammatory potential in Lipopolysaccharide (LPS)-challenged BV-2 microglia cells. LPS stimulation increased the protein phosphorylation of cells regarding their NMDA receptor component subunits and Calcium/Calmodulin-dependent Protein Kinase II (CaMKII), along with mobilizing intracellular calcium. Additionally, parallel changes occurred in the activation of Transforming Growth Factor-β (TGF-β)-Activated Kinase 1 (TAK1), NF-κB p65 and NF-κB DNA binding activity, acquisition of pro-inflammatory M1 polarization and expression of pro-inflammatory cytokines. LPS-treated cells further displayed signs of mitochondrial dysfunction with higher expressions of the active form of Dynamin-Related Protein 1 (Drp1), NADPH Oxidase-2 (NOX2) expression and the generation of DCFDA-/MitoSOX-sensitive Reactive Oxygen Species (ROS). NMDA receptor blockade by MK801, along with CaMKII inhibitor KN93, Drp1 inhibitor Mdivi-1 and antioxidant apocynin alleviated LPS-induced pro-inflammatory changes. Other than the reported CaMKII/TAK1/NF-κB axis, our in vitro study revealed the CaMKII/Drp1/ROS/NF-κB axis being an alternative cascade for shaping pro-inflammatory phenotypes of microglia upon LPS stimulation, and MK801 having the potential for inhibiting microglia activation and any associated inflammatory damages.

Transfer of patient’s peripheral blood mononuclear cells (PBMCs) disrupts blood–brain barrier and induces anti-NMDAR encephalitis: a study of novel humanized PBMC mouse model

BackgroundAnti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe autoimmune neuropsychiatric disease. Brain access of anti-NMDAR autoantibody through the blood–brain barrier (BBB) is essential for pathogenesis. Most previous animal models limit the investigation of etiologies of BBB damage in patients.MethodsIn this study, we established a novel humanized mouse model of anti-NMDAR encephalitis by intraperitoneal injection of patients’ peripheral blood mononuclear cells (PBMCs) into BALB/c Rag2−/−Il2rg−/−SirpαNODFlk2−/− mice.ResultsWe found that engraftment of patients’ PBMCs not only produced potent anti-GluN1 autoantibodies, but also disrupted BBB integrity to allow brain access of autoantibodies, resulting in a hyperactive locomotor phenotype, anxiety- and depressive-like behaviors, cognitive deficits, as well as functional changes in corresponding brain regions. Transcriptome analysis suggested an exaggerated immune response and impaired neurotransmission in the mouse model and highlighted Il-1β as a hub gene implicated in pathological changes. We further demonstrated that Il-1β was produced by endothelial cells and disrupted BBB by repressing tight junction proteins. Treatment with Anakinra, an Il-1 receptor antagonist, ameliorated BBB damage and neuropsychiatric behaviors.ConclusionsOur study provided a novel and clinically more relevant humanized mouse model of anti-NMDAR encephalitis and revealed an intrinsic pathogenic property of the patient’s lymphocytes.

REDUCED CARBON FOOTPRINT THROUGH TARGETED RECYCLING

The importance of returning production and consumption waste into circulation is obvious, both in terms of environmental safety and economic feasibility. The cyclicality of resources is one of the main elements of the concept of a circular economy, the expediency of which is generally recognized today. The processing of various waste fractions differs significantly in terms of energy saving and profitability relative to the production of primary material and the complexity of technological processes. Composting is a natural process of decomposition of organic matter. Compost has no phytotoxic and pathogenic properties, it is characterized by the presence of humic acids. Composting involves reducing the volume of waste, which allows solving environmental problems associated with the accumulation of waste, turning them into a safer product used as an organic fertilizer. Composts are used to increase crop productivity, increase soil fertility, add additional nutrients to the soil, increase plant growth, and suppress diseases caused by soil pathogen. Composting is one of the most effective methods for reducing the toxicity of biodegradable organic matter. The decrease in toxicity is associated with the decomposition of organic pollutants, the mineralization of the substrate, and processes that limit their availability.

Evolutionary Formation and Distribution of Puumala Virus Genome Variants, Russia.

We analyzed Puumala virus (PUUV) sequences collected from bank voles from different regions of Russia. Phylogenetic analysis revealed PUUV reassortments in areas with the highest hemorrhagic fever with renal syndrome incidence, indicating reassortment might contribute to pathogenic properties of PUUV. Continued surveillance is needed to assess PUUV pathogenicity in Russia.

ROLE OF MICROBIAL FACTOR IN MALE INFERTILITY PATHOGENESIS. LITERATURE REVIEW

The role of infectious pathology in the genesis of infertility is still being studied. The present review considers possible mechanisms of infertility development associated with the microbial profile of the male reproductive system. It makes an attempt to study the role of pathogenic properties of microorganisms in the development of infertile conditions if an inflammatory process is localized in the male urogenital tract, in particular its effects on the fertility properties of spermatozoa (reduction in their number, inhibition of their motility, changes in their morphology and fertilizing capacity). Besides, the review presents a discussion on the role of reactivity and resistance of an organism in maintaining chronic inflammation in male reproductive organs.

Longitudinal gut microbiome analyses and blooms of pathogenic strains during lupus disease flares

ObjectiveWhereas genetic susceptibility for systemic lupus erythematosus (SLE) has been well explored, the triggers for clinical disease flares remain elusive. To investigate relationships between microbiota community resilience and disease activity,...

Effect of Colchicum ritchii derived nanoparticles on apple ring rot disease without changing the taste through the prevented metabolites changes and increased sialic acid synthesis

ObjectiveIn this current study, we were planned to produce Colchicum ritchii flower extract associated calcium chloride nanoparticle (CRNPs) in a simple organic method and evaluate their apple fruit pathogenic resistance as well as their shelf-life extension properties during storage. MethodsThis CRNPs was confirmed by analysis of various established physical and chemical methods. Moreover, the apple fruit pathogenic resistance as well as their shelf-life extension properties during storage were evaluated by standardizing methods. ResultsOur results showed that the CRNPs were contrived and proven by physical and chemical evaluations. Moreover, the biological observation of CRNPs indicates that they have good pathogenic resistance properties against the Botryosphaeria dothidea without changing the taste through the prevented metabolites changes and increased sialic acid synthesis. The effective dose of pathogenic resistance in CRNPs was 40 µg/mL. The levels of defensive enzymes such as chitinase (CHI), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL), and β-1,3-glucanase (GLU) significantly increased in CRNPs treated fruits. The increasing antioxidant enzymes activities including superoxide dismutase (SOD) and catalase (CAT) were shown in CRNPs treated apple fruit. The CRNPs-treated apple fruit was found to increase in salicylic acid (SA) and SA glucoside (SAG) levels, owing to increased SA synthesis. Moreover, shelf-life extension studies of fresh cut apples were conducted by immersing them in 20 and 40 µg/mL CRNPs as well as a standard solution of 2% calcium chloride for 5 mins. The standard solution of calcium chloride as well the CRNPs were proved as a good preservative and they were preserved the apple fruits shelf-life up to 25 days. ConclusionIn this study, we have concluded that the CRNPs were produced by fusion with Colchicum ritchii flower extract and calcium chloride in a superlative natural method. Our results recommended that the CRNPs is acted as a best source for apple ring rot resistance without changing the taste through the prevented metabolites changes and increased sialic acid synthesis as well as the antioxidant properties. In addition, the CRNPs is superlative source for apple shelf-life extension up to 25 days without any toxicology nature.