How does a Saccharomyces cerevisiae extract influence the components of isolated rotavirus particles from stool samples collected in a clinical setting from children?

Human Rotavirus (HRV) is the causative pathogen of severe acute enteric infections that cause mortality among children worldwide. This study focuses on developing a new and effective treatment for rotavirus infection using an extract from Saccharomyces cerevisiae, aiming to make this treatment easily accessible to everyone. 15 antigens and 26 antibodies were detected in serum and stool using ELISA. The titers of HRVq1, HRVq2, HRVC1, and HRVC2 on Vero cells were determined to be 1.2x106, 3.0x106, 4.2x106, and 7.5x105 (Plaque forming unit, PFU/ml) four days after infection, respectively. The HRVq1 isolate induced cytopathic effects, i.e., forming multinucleated, rounded, enlarged, and expanding gigantic cells. RT-PCR identified this isolate, and the accession number 2691714 was assigned to GeneBank. The molecular docking analysis revealed that nonstructural proteins (NSPs) NSP1, NSP2, NSP3, NSP4, NSP5, and NSP6 exhibited significant binding with RNA. NSP2 demonstrated the highest binding affinity and the lowest binding energy (-8.9 kcal/mol). This affinity was maintained via hydrophobic interactions and hydrogen bonds spanning in length from 1.12 Å to 3.11 Å. The ADMET and bioactivity predictions indicated that the yeast extract possessed ideal solubility, was nontoxic, and did not cause cancer. The inhibitory constant values predicted for the S. cerevisiae extract in the presence of HRV vital proteins varied from 5.32 to 7.45 mM, indicating its potential as a viable drug candidate. Saccharomyces cerevisiae extract could be utilized as a dietary supplement to combat HRV as an alternative dietary supplement.

Ammi-visnaga extract; a novel phyto-antiviral agent against bovine rotavirus.

The spread of bovine rotavirus has a great impact on animal productivity, milk products, and human public health. Thus, this study aimed to develop a novel, effective and accessible Phyto-antiviral treatment made from methanolic Ammi-visnaga seed extract against rotavirus infection. Rotaviruses were isolated from raw milk and cottage cheese samples randomly collected from Cairo and Qalubia governorates. They were all identified serologically, however, only three of them were both biologically and molecularly confirmed. The methanolic extract derived from Khella seeds (MKSE) was chemically analyzed with mass chromatography. The cellular toxicity of MKSE was tested on Caco-2 cells and its antiviral activity against one of the isolated bovine rotaviruses (BRVM1) was tested by both the cytopathic inhibition assay and the plaque reduction assay. Our results showed that 17.3% of the total collected 150 dairy samples were bovine rotavirus antigen positive. Three representatives of them were phylogenetically identified to be included in group A based on a 379 bp coat protein gene. Visnagin, Benzopyran, Khellin, and Benzenepropanoic acid were the major active components found in the MKSE. The maximum non-toxic concentration of MKSE was 5 µg/mL and the CC50 value was 417 µg/mL. The MKSE exhibited in-vitro antiviral activity against BRVM1 indicated by inhibition of the viral cytopathic effect (SI = 204.5, IP = 98%), causing a 1.5 log decrease in BVRM1 TCID50 and reducing the viral plaques count by the percentage of 93.14% at MNTC (5 ug/ml). In conclusion, our study showed that bovine rotavirus represents a severe health problem that needs attention in Egypt, and it supports using MKSE as a potential natural anti-rotavirus agent.

Isolation and Characterization of Antagonistic Activity of Some Streptomycetes and their Specific Actinophages from Soil.

<b>Background and Objective:</b> Phages specific to actinomycetes are common, active in the soil and gladly detected. Soil streptomycetes are having antibiosis activities against numerous bacteria, fungi and plant viruses. Thus, this study was designed to isolate, purify and characterize some streptomycetes active against some microorganisms from soil followed by isolation of their specific phages. <b>Materials and Methods:</b> Antagonistic activities of these streptomycetes isolates were tested against <i>Bacillus subtilis</i>,<i> Pseudomonas</i> sp., <i>Serratia</i> sp. and <i>Aspergillus niger</i>. To confirm their biological characterization of the streptomycetes isolates under investigation, the 16SrRNA gene was also used. The presence of specific lysate actinophages in the soil samples were tested by spot test technique and then propagated and purified for further characterization. The morphology of the purified actinophages was determined by electron microscopy. <b>Results:</b> The five selected <i>Streptomyces</i> isolates having effective antagonistic activity were biologically and molecularly identified as <i>Streptomyces sclerogranulatus </i>(QQ06), <i>Streptomyces mutabilis </i>(QQ07), <i>Streptomyces heilongjiangensis </i>(QQ08), <i>Streptomyces sparsus </i>(QQ09) and <i>Streptomyces purpurascens </i>(QQ10) strains. Electron micrographs showed the presence of filamentous virus-like particles with lengths of 21.4×928.57, 25×750, 21.4×857.14, 21.4×885.7 and 21.4×857.14 nm specific to <i>Streptomyces</i> strains QQ06, QQ07, QQ08, QQ09 and QQ10, respectively and belong to the family Inoviridae. <b>Conclusion:</b> Phage of Inoviridae was considered as the first time against streptomycetes isolates, therefore, additional and advanced studies should be carried out at the level of molecular characterization.

Expression of DnaK and HtrA genes under high temperatures and their impact on thermotolerance of a Salmonella serotype isolated from tahini product.

BACKGROUND: Salmonella is considered to be the second largest source of infection in food-borne diseases. It is also considered one of the most important dangers particularly in the meat and dairy industry. Therefore, the main objective of our study was to determine the relationship between thermotolerance of a Salmonella serotype and the expression of DnaK and HtrA genes. RESULTS: In this study, expression of the two genes DnaK and HtrA was compared under four different temperatures 37 °C, 42 °C, 50 °C, and 55 °C in two serotypes of Salmonella enterica subsp. enterica. One of them was isolated from tahini product and identified as Salmonella enterica subsp. enterica serovar choleraesuis. This identified serotype was found to be more thermotolerant than the second serotype (Salmonella enterica subsp. enterica serovar typhimurium (ATCC 13311)), which was used as reference. This conclusion was based on D and Z values, which were used to compare thermoresistance ability of the two serotypes under four different temperatures 60 °C, 65 °C, 70 °C, and 75 °C. In addition, the results of qRT-PCR showed that after 43 °C (induction temperature), the relative expression (fold change) of DnaK and HtrA genes increased up to 5 and 47, respectively, comparing to their expression at 37 °C. CONCLUSIONS: Thermotolerance of the identified S. choleraesuis serotype showed significantly high expression levels of DnaK and HtrA genes.