Influence of certain carbon and nitrogen sources on antagonistic potentiality of Trichoderma harzianum and Bacillus subtillus against Botrytis allii the incitant of onion neck rot

Gliotoxin Fermentation Agar (GFA) Medium and Nutrient Glucose Agar   (NGA) medium with different carbon and nitrogen sources were used to study the impact of carbon and nitrogen sources on Trichoderma harzianum Rifai and Bacillus subtillus Cohn antagonistic efficiency against growth of Botrytis allii Munn. Results indicated that Trichoderma harzianum gave the highest inhibition % in growth of Botrytis allii when Sucrose was used as a carbon source while the lowest values of inhibition% were appeared by application of Mannitol as a carbon source. Trichoderma harzianum gave the highest inhibition% in growth of B.allii when use Potassium nitrate  as a nitrogen source while the lowest values of inhibition% in growth of B.allii were obtained by application of Beef extract as a nitrogen source. Results showed also that Bacillus subtillus gave the highest inhibition% in growth of B.allii when Mannitol was used as a carbon source while the lowest values of inhibition% in growth of B.allii were appeared by application of Sucrose as a carbon source. Bacillus subtillus gave the highest inhibition% in growth of B.allii when use Glutamic acid as a nitrogen source while the lowest values of inhibition% in growth of B.allii were appeared by application of Tryptophan as a nitrogen source.

MICROPROPAGATION AND EVALUATION OF GENETIC STABILITY OF FOXGLOVE TREE (Paulownia tomentosa)

This reasearch work aimed to fine-tune micropropagation of Paulownia tomentosa in addition to assessing the genetic stability of in vitro raised clones from it. Paulownia tomentosa explants were surface sterilized using clorox (commercial bleach 5.25% sodium hypochlorite) at 10, 20, 25 and 30% + 0.5 g/l mercuric chloride (HgCl2) at different duration times, i.e. 10, 15, 20 and 25 min. In the multiplication stage, shoots were transferred to MS medium at ¾ strength containing BAP and Kin each at (0, 0.5, 1, 2, and 4 mg/l). Whereas, the rooting medium was MS medium at ¾ strength with IBA and NAA treatments each at 0, 0.5, 1, 2, and 4 mg/l. Sterilized explant with 30% Clorox for 20 min recorded highest survival percentage. The treatment of Kin at 4 mg/l gave higher significant shoot length. Whereas BAP application at 2 and 4 mg/l gave highest significant value of both shoot number and leaf number. Both IBA and NAA at 0.5 or 1 mg/l gave highest significant root number/shoot. Whereas, auxin at 4 mg/l gave highest significant root lengths. Young plantlets resulted from in vitro were acclimitized successfully in a mixture of peat moss: perlit (2: 1) by volume that showed 85.93% survival.  The genetic stability of in vitro raised Paulownia tomentosa clones was assessed by using intersimple sequence repeats (ISSRs) markers. All of the three ISSR primers screened, produced clear, reproducible and scorable bands.      The molecular size of Polymerase Chain reaction (PCR) products generated 22 fragments by these ISSR ranged from ≈460 to18660 bp. All banding profiles from micropropagated plants were monomorphic and similar to those of the mother plant, indicating 100% similarity. This confirmed the true to type nature of the in vitro raised clones

IMPLEMENTATION OF HAZARD ANALYSIS CRITICAL CONTROL POINTS (HACCP) PRINCIPLES IN PRODUCTION OF FILLING CREAM

For improving the safety of filling cream, the Hazard Analysis Critical Control Points (HACCP) system was applied during different preparation process. Different biological, chemical, and physical hazards that may exist in every preparing step of filling cream were identified and control measures were used for controlling those identified hazards. Dried milk powder was identified as high potential hazard raw materials contaminated with pathogenic bacteria e.g. S. aureus and Salmonella sp. Whipping step in preparing filling cream was identified as critical control point. For improving the safety of preparing filling cream system, different control measures including, thermal processing (pasteurization), addition of natural preservatives (lemongrass oil and herbs of cinnamon, clove and anise) and reduction of pH (fruit pulps of strawberry, apricots and apple) were established. Different applied control measures could be used for improving the safety of prepared filling cream where different investigated microbiological criteria were reduced to acceptable levels especially pathogenic bacteria. In addition, different HACCP principles as the critical control points, critical limits, monitoring, corrective measures, verifications and records were established. Implementing the HACCP system during preparing filling cream can effectively ensure safety of this product and other food products contained filling cram especially bakeries

Tetracycline resistance phenotypes and genotypes of coagulase-negative staphylococcal isolates from bubaline mastitis in Egypt

Aim: This study was devoted to elucidate the tetracycline resistance of coagulase-negative staphylococci (CNS) derived from normal and subclinical mastitic (SCM) buffaloes' milk in Egypt. Materials and Methods: A total of 81 milk samples from 46 normal buffalo milk samples and 35 SCM buffalo milk samples at private dairy farms of Egypt were used in this study. CNS were identified using phenotypic and molecular methods (polymerase chain reaction [PCR]). CNS isolates were tested for tetracycline resistance using routine methods and multiplex PCR targeting tetracycline (tet) resistance genes followed by sequencing of positive PCR products and phylogenetic analysis. Results: Isolation and identification of 28 (34.5%) CNS from normal and SCM buffaloes' milk, namely, Staphylococcus intermedius (39.2%), Staphylococcus xylosus (25.0%), Staphylococcus epidermidis (10.7%), Staphylococcus hominis (10.7%), and 3.5% to each of Staphylococcus sciuri, Staphylococcus hyicus, Staphylococcus lugdunensis, and Staphylococcus simulans. Using nested PCR, all the 28 CNS isolates revealed positive for 16srRNA gene specific for genus staphylococci and negative for thermonuclease (nuc) gene specific for Staphylococcus aureus species. The presence of tetracycline resistance-encoding genes (tetK, tetL, tetM, and tetO) was detected by multiplex PCR. All isolates were negative for tetL, M, and O genes while 14 (50%) CNS isolates were positive for tetK gene, namely, S. lugdunensis (100%), S. hominis (100%), S. epidermidis (66.6%), S. intermedius (45.4%), and S. xylosus (42.8%). Nucleotide sequencing of tetK gene followed by phylogenetic analysis showed the high homology between our CNS isolates genes of tetracycline resistance with S. aureus isolates including Egyptian ones. This proves the transfer of the tetracycline resistance encoding genes between coagulase-negative and coagulase-positive Staphylococcus spp. Conclusion: CNS isolates have distinguishingly high resistance to tetracycline. Abundant tetracycline usage for mastitis treatment leads to the spread of genetic resistance mechanisms inside CNS strains and among all Staphylococcus spp. Consequently, tetracycline is not effective anymore

ISOLATION AND MOLECULAR IDENTIFICATION OF INDIGENOUS BACTERIAL ISOLATES ABLE TO DEGRADE ORGANOPHOSPHATES

The wide and indiscriminate use of pesticides for pest control in agriculture has inflicted serious harm and problems to humans as well as to the biodiversity. Microbial degradation of pesticides in contaminated soils has been considered advantageous to decontaminate areas that have been polluted by pesticides. Chlorpyrifos and diazinon were the most persistent residues in Egyptian soils. Four bacterial isolates were isolated from organophosphorus insecticides contaminated soils and genetically identified based on DNA sequence of 16s rDNA gene, Cronobacter muytjensii GH10, Achromobacter xylosoxidans GH9OP, Pseudomonas aeruginosa GH2NO8 and Pseudomonas putida GH4SNO/P were able to degrade 92.59%, 97.75%, 91.82%, and 90.78% of diazinon (600mg/l) as compared with 16.99% in control and 93.43%, 78.51%, 93.18% and 95.36% of chlorpyrifos (480mg/l) as compared with 4.28%, in control, respectively after 20 days of incubation

SULPHATED POLYSACCHARIDES (SPS) FROM THE GREEN ALGA ULVA FASCIATA EXTRACT MODULATES LIVER AND KIDNEY FUNCTION IN HIGH FAT DIET-INDUCED HYPERCHOLESTEROLEMIC RATS

Objective: Hypercholesterolemia (HC) was frequently associated with oxidative stress, and release of inflammatory cytokines is to determine the hypolipidemic effects of sulphated polysaccharides from seaweed Ulva fasciata algal extracts through measuring the activities of some parameters related to liver and kidney functions in the serum of hypercholesterolemic rats as compared to normal one.Methods: Different groups of rats were administered a high cholesterol diet. Liver and kidney functions, inflammatory cytokines (TNF-α, CRP, MPO and IL-10), oxidative stress (GSH, MDA and NO), in addition to cell adhesion molecules (ICAM-1 and VCAM-1) were assessed before and after treatment with the algal polysaccharides. In addition, histological examination of liver and kidney were performed to confirm the biochemical findings.Results: The obtained results showed that oxidative stress and inflammatory markers associated with hypercholesterolemia were significantly increased in HC-rats. The histopathological examination of liver and kidney demonstrated severe degeneration with diffuse vacuolar degeneration, necrosis and the presence of fatty droplets. In addition; nephron-histological examination revealed, mild glomerular injury with mild vascular and inflammatory changes. Treatment with the algal sulphated polysaccharides effectively improved these disorders and diminished the formation of fatty liver, as well as renal dysfunction more than the reference drug; fluvastatin. Conclusion: It could be concluded that the consumption of UFP (Ulva fasciata polysaccharides), may be associated with attenuation of inflammatory markers, amelioration of fatty liver and improvement of renal dysfunction, that in turn lead to counteract hypercholesterolemia and its related disorders; such as obesity, and heart disease.Keywords: Non-alcoholic fatty liver disease, Seaweed, Ulva fasciata, Hypercholesterolemia, Hypolipidemic activity, Sulphated polysaccharides (SPs

Biochemical Characterization, Antifungal Activity, and Relative Gene Expression of Two Mentha Essential Oils Controlling Fusarium oxysporum, the Causal Agent of Lycopersicon esculentum Root Rot

Tomato (Lycopersicon esculentum Mill.) is important food in daily human diets. Root rot disease by Fusarium oxysporum caused huge losses in tomato quality and yield annually. The extensive use of synthetic and chemical fungicides has environmental risks and health problems. Recent studies have pointed out the use of medicinal plant essential oils (EOs) and extracts for controlling fungal diseases. In the current research, Mentha spicata and Mentha longifolia EOs were used in different concentrations to control F. oxysporum. Many active compounds are present in these two EOs such as: thymol, adapic acid, menthol and menthyl acetate. These compounds possess antifungal effect through malformation and degradation of the fungal cell wall. The relative expression levels of distinctly upregulated defense-related WRKY genes (WRKY1, WRKY4, WRKY33 and WRKY53) in seedling root were evaluated as a plant-specific transcription factor (TF) group in different response pathways of abiotic stress. Results showed significant expression levels of WRKY, WRKY53, WRKY33, WRKY1 and WRKY4 genes. An upregulation was observed in defense-related genes such as chitinase and defensin in roots by application EOs under pathogen condition. In conclusion, M. spicata and M. longifolia EOs can be used effectively to control this plant pathogen as sustainable and eco-friendly botanical fungicides

Chemical Profile of Cyperus laevigatus and Its Protective Effects against Thioacetamide-Induced Hepatorenal Toxicity in Rats

Cyperus species represent a group of cosmopolitan plants used in folk medicine to treat several diseases. In the current study, the phytochemical profile of Cyperus laevigatus ethanolic extract (CLEE) was assessed using UPLC-QTOF–MS/MS. The protective effect of CLEE at 50 and 100 mg /kg body weight (b.w.) was evaluated on hepatorenal injuries induced by thioacetamide (100 mg/kg) via investigation of the extract’s effects on oxidative stress, inflammatory markers and histopathological changes in the liver and kidney. UPLC-QTOF–MS/MS analysis of CLEE resulted in the identification of 94 compounds, including organic and phenolic acids, flavones, aurones, and fatty acids. CLEE improved the antioxidant status in the liver and kidney, as manifested by enhancement of reduced glutathione (GSH) and coenzyme Q10 (CoQ10), in addition to the reduction in malondialdehyde (MDA), nitric oxide (NO), and 8-hydroxy-2′-deoxyguanosine (8OHdG). Moreover, CLEE positively affected oxidative stress parameters in plasma and thwarted the depletion of hepatorenal ATP content by thioacetamide (TAA). Furthermore, treatment of rats with CLEE alleviated the significant increase in plasma liver enzymes, kidney function parameters, and inflammatory markers. The protective effect of CLEE was confirmed by a histopathological study of the liver and kidney. Our results proposed that CLEE may reduce TAA-hepatorenal toxicity via its antioxidant and anti-inflammatory properties suppressing oxidative stress

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020