GROWTH, YIELD AND YIELD COMPONENT OF INOCULATED CHICKPEA AND FABA BEAN PLANTS AS AFFECTED BY USING METHYLOTROPHIC BACTERIA

Two pot experiments were performed in Giza research station, AgriculturalResearch Center (ARC) using 2 isolates of Pink-Pigmented FacultativelyMethylotrophic bacteria (PPFMs) originated from chickpea and Faba bean. Foliarapplication with PPFM isolates were conjugated with specific rhizobial inoculumand N-fertilization (15 Kg N Fed-1). Nodulation status, nitrogen fixation and growthyield and yield component were recorded. Results clearly indicated that Chickpeawas superior in its response to foliar application with PPFM.C. As it gave higherrecords of number and dry weight of nodules, dry matter and N-content of plants ascompared to Faba bean. A field experiment was also conducted in sandy loam soil atSouth EL-Tahreer province to investigate the effect of foliar application withPPFM.C strain + specific Rhizobia and N-fertilization on nodulation, growth andyield of chickpea legume plants. Results indicated that foliar application withPPFM.C in the presence of specific rhizobial inoculation scored significant increasesin economic turnover of chickpea in the range of 21-32% as compared to Nfertilizationat rate 50 Kg N Fed-1. Foliar application with 5 L Fed-1 in the presenceof 15 Kg N Fed-1 and specific rhizobial inoculation led to an increase of 518 kg fed -1productivity of seed yield , with economic turnover of 2491 L.E

ISOLATION, PURIFICATION AND IDENTIFICATION OF SOME MICROORGANISMS PRODUCE PLANT GROWTH PROMOTING SUBSTANCES (METHYLOTROPHIC BACTERIA)

Recently, the potential economical importance of the methylotrophic bacteria encouraged the isolation of this group. In the present study five Egyptian isolates were obtained from green leaves surface of legume plants named PPFM.C (ChickPea), PPFM.Ph (Common bean), PPFM.F (Faba bean), PPFM.P (Peanut) and PPFM.S (Soybean) ,to study their general characters which belonging to  methylotrophic bacteria. Morphological studies indicate that all isolates were short rods, gram negative and motile. All Physiological studies to the isolates gave the same results except PPFM.F which could not grow in peptone medium. All isolates were sensitive to Kanamycin but they were resistant to Erythromycin. There was a great range in the ability of the isolates to grow on different sodium chloride concentrations indicating that PPFM.Ph grew well in 5 % sodium chloride, and they were able to excrete and produce cytokinin. Molecular biology studies indicated that there was a great similarity between PPFM.C and PPFM.Ph (99.34%). Identification was carried out to the5 isolates, PPFM.F may be related to Methylobacterium mesophilicum, PPFM.P may be related to M. fujisawaense and PPFM.Ph, PPFM.C and PPFM.S were related to M. radiotolerans

Tetherin antagonism by SARS-CoV-2 ORF3a and spike protein enhances virus release

The antiviral restriction factor, tetherin, blocks the release of several different families of enveloped viruses, including the Coronaviridae. Tetherin is an interferon-induced protein that forms parallel homodimers between the host cell and viral particles, linking viruses to the surface of infected cells and inhibiting their release. We demonstrate that SARS-CoV-2 infection causes tetherin downregulation and that tetherin depletion from cells enhances SARS-CoV-2 viral titres. We investigate the potential viral proteins involved in abrogating tetherin function and find that SARS-CoV-2 ORF3a reduces tetherin localisation within biosynthetic organelles where Coronaviruses bud, and increases tetherin localisation to late endocytic organelles via reduced retrograde recycling. We also find that expression of Spike protein causes a reduction in cellular tetherin levels. Our results confirm that tetherin acts as a host restriction factor for SARS-CoV-2 and highlight the multiple distinct mechanisms by which SARS-CoV-2 subverts tetherin function

Establishing SARS-CoV-2 membrane protein-specific antibodies as a valuable serological target via high-content microscopy

The prevalence and strength of serological responses mounted toward SARS-CoV-2 proteins other than nucleocapsid (N) and spike (S), which may be of use as additional serological markers, remains underexplored. Using high-content microscopy to assess antibody responses against full-length StrepTagged SARS-CoV-2 proteins, we found that 85% (166/196) of unvaccinated individuals with RT-PCR confirmed SARS-CoV-2 infections and 74% (31/42) of individuals infected after being vaccinated developed detectable IgG against the structural protein M, which is higher than previous estimates. Compared with N antibodies, M IgG displayed a shallower time-dependent decay and greater specificity. Sensitivity for SARS-CoV-2 seroprevalence was enhanced when N and M IgG detection was combined. These findings indicate that screening for M seroconversion may be a good approach for detecting additional vaccine breakthrough infections and highlight the potential to use HCM as a rapidly deployable method to identify the most immunogenic targets of newly emergent pathogens

Altered subgenomic RNA abundance provides unique insight into SARS-CoV-2 B.1.1.7/Alpha variant infections

B.1.1.7 lineage SARS-CoV-2 is more transmissible, leads to greater clinical severity, and results in modest reductions in antibody neutralization. Subgenomic RNA (sgRNA) is produced by discontinuous transcription of the SARS-CoV-2 genome. Applying our tool (periscope) to ARTIC Network Oxford Nanopore Technologies genomic sequencing data from 4400 SARS-CoV-2 positive clinical samples, we show that normalised sgRNA is significantly increased in B.1.1.7 (alpha) infections (n = 879). This increase is seen over the previous dominant lineage in the UK, B.1.177 (n = 943), which is independent of genomic reads, E cycle threshold and days since symptom onset at sampling. A noncanonical sgRNA which could represent ORF9b is found in 98.4% of B.1.1.7 SARS-CoV-2 infections compared with only 13.8% of other lineages, with a 16-fold increase in median sgRNA abundance. We demonstrate that ORF9b protein levels are increased 6-fold in B.1.1.7 compared to a B lineage virus in vitro. We hypothesise that increased ORF9b in B.1.1.7 is a direct consequence of a triple nucleotide mutation in nucleocapsid (28280:GAT > CAT, D3L) creating a transcription regulatory-like sequence complementary to a region 3’ of the genomic leader. These findings provide a unique insight into the biology of B.1.1.7 and support monitoring of sgRNA profiles to evaluate emerging potential variants of concern

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Altered subgenomic RNA abundance provides unique insight into SARS-CoV-2 B.1.1.7/Alpha variant infections

B.1.1.7 lineage SARS-CoV-2 is more transmissible, leads to greater clinical severity, and results in modest reductions in antibody neutralization. Subgenomic RNA (sgRNA) is produced by discontinuous transcription of the SARS-CoV-2 genome. Applying our tool (periscope) to ARTIC Network Oxford Nanopore Technologies genomic sequencing data from 4400 SARS-CoV-2 positive clinical samples, we show that normalised sgRNA is significantly increased in B.1.1.7 (alpha) infections (n = 879). This increase is seen over the previous dominant lineage in the UK, B.1.177 (n = 943), which is independent of genomic reads, E cycle threshold and days since symptom onset at sampling. A noncanonical sgRNA which could represent ORF9b is found in 98.4% of B.1.1.7 SARS-CoV-2 infections compared with only 13.8% of other lineages, with a 16-fold increase in median sgRNA abundance. We demonstrate that ORF9b protein levels are increased 6-fold in B.1.1.7 compared to a B lineage virus in vitro. We hypothesise that increased ORF9b in B.1.1.7 is a direct consequence of a triple nucleotide mutation in nucleocapsid (28280:GAT > CAT, D3L) creating a transcription regulatory-like sequence complementary to a region 3’ of the genomic leader. These findings provide a unique insight into the biology of B.1.1.7 and support monitoring of sgRNA profiles to evaluate emerging potential variants of concern