Impact of wheat processing on growth, serum biochemistry, and ruminal microbiota in sheep (Ovis aries).

This study investigated the impact of wheat processing methods (wheat flour vs wheat pellets) on the growth performance, serum biochemical parameters, and rumen microbiome composition in sheep. Results indicated that feeding of wheat flour resulted in significantly higher terminal weight and average daily gain (P<0.05) and lower cholesterol and ALP04 levels (P<0.05) in sheep compared to those fed wheat pellets. Analysis of 16s rDNA high-throughput sequencing data revealed significantly higher microbial richness (Chao1 index) in the rumen of sheep fed wheat flour (P<0.05), even though the phylum-level composition dominated by Firmicutes, Bacteroidetes, and Proteobacteria was similar in both groups of sheep. Notably, sheep fed wheat flour were found to have a significantly higher relative abundance of Bacteroidetes (P<0.05). At the genus level, Succinivibrionaceae_UCG-001 and Prevotella_1 were significantly more abundant in the rumen of sheep fed wheat flour (P<0.05). Correlation analysis identified that both terminal weight and average daily gain were positively correlated with ruminal abundance of Bacteroidetes and Prevotella_1, while ALP04 was negatively correlated with the abundance of these taxa. Functional prediction using PICRUSt2 indicated enrichment of pathways related to the ABC-type glycerol-3-phosphate transport system, and periplasmic components in both wheat flour and pellet fed sheep. Overall, these findings suggest that dietary wheat flour modulates rumen microbiota composition, and improves growth performance in sheep.

Aspergillus Species from the Sabkha Marsh: Potential Antimicrobial and Anticancer Agents Revealed Through Molecular and Pharmacological Analysis.

Introduction: This study aimed to investigate the fungal growth and diversity in the Sabkha marsh. The anti-bacterial properties of the isolated fungi were assessed using an agar disk diffusion assay, and the crude extracts were tested for their anticancer activities. Liquid chromatography-mass spectrometry was employed to identify the active compounds of the fungal secondary metabolites. In-silico studies were conducted to predict the toxicity, pharmacokinetic properties, and safety profiles of the identified compounds. Results: The analysis revealed that the isolated fungi belonged to the Aspergillus species, specifically Aspergillus flavus and Aspergillus niger. The crude extract of A. flavus exhibited significant anticancer activity against various cancer cell lines, while the antifungal activities against pathogenic bacteria varied between the two fungi. Liquid chromatography-mass spectrometry analysis identified several compounds in the fungal isolates. In Aspergillus flavus, the compounds included Aflavinine, Dihydro-24-hydroxyaflavinine, Phomaligin A, Hydroxysydonic acid, Gregatin B, Pulvinulin A, Chrysogine, Aspergillic acid, Aflatoxin B1, and Aflatoxin G1. In Aspergillus niger, the compounds identified were atromentin, fonsecin B, firalenone, rubrofusarin, aurasperone E, aurasperone D, aurasperone C, nigerone, and αß-dehydrocurvularin. Conclusion: This study demonstrated promising fungal growth and diversity in the Sabkha marsh, with Aspergillus species being the most prevalent. The fungal crude extract showed anticancer activities against various cancer cell lines, while the antifungal activities against pathogenic bacteria varied between the two fungi. Future research should focus on investigating the antimicrobial activities of these fungi against multidrug-resistant bacteria and exploring the genetic changes in bacteria and cancer cells treated with these fungal extracts. Additionally, it is important to test the anticancer activity of the active compounds separately to determine which one is the active agent against cancer cells. This information can be used in drug development trials.

Phytochemical Analysis of Centaurea calcitrapa L. Aerial Flowering Parts Serial Solvent Extracts and Its Antibacterial and Antioxidant Activities.

To evaluate the phytochemical composition, antibacterial, and antioxidant activity of successive extracts of Centaurea calcitrapa L. (C. calcitrapa) aerial flowering parts, they were assessed in vitro. Using a spectrophotometer, the sample absorbance at 517 nm was used to quantify the scavenging activity. The negative control was DPPH. In the current study, the diffusion using agar wells technique was adapted to measure antimicrobial activity. Phytochemical analysis was performed using the recommended standard procedures. The methanol extract of C. calcitrapa exhibited high levels of total phenolic acids expressed as gallic acid (GA), measured as (97.25 ± 0.73 mg GAE/g) content compared to the chloroform, acetyl acetate, and aqueous extracts (27.42 ± 0.29, 64.25 ± 0.96, and 17.25 ± 0.73 mg GAE/g), respectively. Additionally, the methanol extract had a higher total tannin (27.52 ± 0.53 mg TAE/g) content compared to the chloroform, ethyl acetate, and aqueous extracts (12.02 ± 0.55, 26.01 ± 0.81, and 7.35 ± 0.56 mg TAE/g), respectively, while the aqueous extract contains a lower percentage of flavonoids (141.10 ± 1.31 mg RTE/g) compared to the higher content achieved by the methanol extract (425.93 ± 1.27 mg RTE/g). The hydroxyl groups of the flavonoid and the phenolic compounds found in C. calcitrapa are essentially scavenging free radicals. Radical scavenging activity was highest in the methanol extract (IC50 = 2.82 µg/mL), aqueous extract (IC50 = 8.03 µg/mL), ethyl acetate extract (IC50 = 4.79 µg/mL), and chloroform extract (IC50 = 6.33 µg/mL), as compared to the standard scavenging activity (IC50 = 2.52 µg/mL). The antibacterial properties of C. calcitrapa against Gram-negative bacterial strains Klebsiella pneumoniae, Escherichia coli, Enterobacter aerogenes, and Acinetobacter baumanii, in addition to Gram-positive strains Staphylococcus haemolyticus, Enterococcus faecalis, and Staphylococcus aureus, revealed inhibition zone diameter. The findings of this investigation establish that the aerial flowering parts of C. calcitrapa have substantial antibacterial action against human infections, and the plant can serve as a significant antioxidant that can be employed to prevent and treat severe degenerative diseases brought on by oxidative stress. qPCR showed that C. calcitrapa extracts elevate both SOD1 and SOD2 (cellular oxidation markers) with remarkable folds (1.8-fold for SOD1 and SOD2) with ethyl acetate plant extract against ascorbic acid as a control. This result reflects that C. calcitrapa extracts have remarkable antioxidant activity.

Diversity and antimicrobial susceptibility patterns of clinical and environmental Salmonella enterica serovars in Western Saudi Arabia.

The diverse environmental distribution of Salmonella makes it a global source of human gastrointestinal infections. This study aimed to detect Salmonella spp. and explore their diversity and antimicrobial susceptibility patterns in clinical and environmental samples. Pre-enrichment, selective enrichment, and selective plating techniques were adopted for the Salmonella detection whereas the API 20E test and Vitek Compact 2 system were used to confirm the identity of isolates. Salmonella serovars were subjected to molecular confirmation by 16S rDNA gene sequencing. Disc diffusion method and Vitek 2 Compact system determined the antibiotic susceptibility of Salmonella serovars. Multiple antibiotic resistance index (MARI) was calculated to explore whether Salmonella serovars originate from areas with heavy antibiotic usage. Results depicted low Salmonella prevalence in clinical and environmental samples (3.5%). The main detected serovars included Salmonella Typhimurium, S. enteritidis, S. Infantis, S. Newlands, S. Heidelberg, S. Indian, S. Reading, and S. paratyphi C. All the detected Salmonella serovars (27) exhibited multidrug resistance to three or more antimicrobial classes. The study concludes that the overall Salmonella serovars prevalence was found to be low in environmental and clinical samples of Western Saudi Arabia (Makkah and Jeddah). However, antimicrobial susceptibility patterns of human and environmental Salmonella serovars revealed that all isolates exhibited multidrug-resistance (MDR) patterns to frequently used antibiotics, which might reflect antibiotic overuse in clinical and veterinary medicine. It would be suitable to apply and enforce rules and regulations from the One Health approach, which aim to prevent antibiotic resistance infections, enhance food safety, and improve human and animal health, given that all Salmonella spp. detected in this investigation were exhibiting MDR patterns.

Abundant resistome determinants in rhizosphere soil of the wild plant Abutilon fruticosum.

A metagenomic whole genome shotgun sequencing approach was used for rhizospheric soil micribiome of the wild plant Abutilon fruticosum in order to detect antibiotic resistance genes (ARGs) along with their antibiotic resistance mechanisms and to detect potential risk of these ARGs to human health upon transfer to clinical isolates. The study emphasized the potential risk to human health of such human pathogenic or commensal bacteria, being transferred via food chain or horizontally transferred to human clinical isolates. The top highly abundant rhizospheric soil non-redundant ARGs that are prevalent in bacterial human pathogens or colonizers (commensal) included mtrA, soxR, vanRO, golS, rbpA, kdpE, rpoB2, arr-1, efrA and ileS genes. Human pathogenic/colonizer bacteria existing in this soil rhizosphere included members of genera Mycobacterium, Vibrio, Klebsiella, Stenotrophomonas, Pseudomonas, Nocardia, Salmonella, Escherichia, Citrobacter, Serratia, Shigella, Cronobacter and Bifidobacterium. These bacteria belong to phyla Actinobacteria and Proteobacteria. The most highly abundant resistance mechanisms included antibiotic efflux pump, antibiotic target alteration, antibiotic target protection and antibiotic inactivation. antimicrobial resistance (AMR) families of the resistance mechanism of antibiotic efflux pump included resistance-nodulation-cell division (RND) antibiotic efflux pump (for mtrA, soxR and golS genes), major facilitator superfamily (MFS) antibiotic efflux pump (for soxR gene), the two-component regulatory kdpDE system (for kdpE gene) and ATP-binding cassette (ABC) antibiotic efflux pump (for efrA gene). AMR families of the resistance mechanism of antibiotic target alteration included glycopeptide resistance gene cluster (for vanRO gene), rifamycin-resistant beta-subunit of RNA polymerase (for rpoB2 gene) and antibiotic-resistant isoleucyl-tRNA synthetase (for ileS gene). AMR families of the resistance mechanism of antibiotic target protection included bacterial RNA polymerase-binding protein (for RbpA gene), while those of the resistance mechanism of antibiotic inactivation included rifampin ADP-ribosyltransferase (for arr-1 gene). Better agricultural and food transport practices are required especially for edible plant parts or those used in folkloric medicine.

Clinical Characteristics, Outcomes and Prognostic Factors for Critical Illness in Hospitalized COVID-19 Patients in Saudi Arabia: A Retrospective Cohort Study.

Background: A good understanding of the possible risk factors for coronavirus disease 19 (COVID-19) severity could help clinicians in identifying patients who need prioritized treatment to prevent disease progression and adverse outcome. In the present study, we aimed to correlate clinical and laboratory characteristics of hospitalized COVID-19 patients to disease outcome in Saudi Arabia. Materials and Methods: The present study included 199 COVID-19 patients admitted to King Fahd Specialist Hospital, Buraydah, Qassim, Saudi Arabia, from April to December 2020. Patients were followed-up until discharge either for recovery or death. Demographic data, clinical data and laboratory results were retrieved from electronic patient records. Results: Critical COVID-19 cases showed higher mean of age and higher prevalence of co-morbid conditions. Fifty-five patients died during the observation period. Risk factors for in hospital death for COVID 19 patients were leukocytosis (OR 1.89, 95% CI 1.008-3.548, p = 0.081), lymphocytopenia (OR 2.152, 95% CI 1.079-4.295, p = 0.020), neutrophilia (OR 1.839, 95% CI 0.951-3.55, p = 0.047), thrombocytopenia (OR 2.152, 95% CI 0.852-5.430, p = 0.085), liver injury (OR 2.689, 95% CI 1.373-4.944, p = 0.003), acute kidney injury (OR 1.248, 95% CI 0.631-2.467 p = 0.319), pancreatic injury (OR 1.973, 95% CI 0.939-4.144, p = 0.056) and high D dimer (OR 2.635, 95% CI 0.747-9.287, p = 0.091). Conclusion: Clinical and laboratory data of COVID-19 patients may help understanding the pathogenesis of the disease and subsequently improve of the outcome of patients by determination of the associated risk factors and recognition of high risk group who are more liable for complications and in hospital death. The present study put an eye on some parameters (laboratory and clinical) that should be alarming signs that the patient is at high risk bad prognosis.

Antimicrobial Resistance, Virulence Factor-Encoding Genes, and Biofilm-Forming Ability of Community-Associated Uropathogenic Escherichia coli in Western Saudi Arabia.

To explore the prevalence of multidrug-resistant community-associated uropathogenic Escherichia coli (UPEC) and their virulence factors in Western Saudi Arabia. A total of 1,000 urine samples were examined for the presence of E. coli by selective plating on MacConkey, CLED, and sheep blood agar. Antimicrobial susceptibility patterns were determined using Vitek® 2 Compact (MIC) and the disc diffusion method with Mueller-Hinton agar. Genes encoding virulence factors (kpsMTII, traT, sat, csgA, vat, and iutA) were detected by PCR. The overall prevalence of UTI-associated E. coli was low, and a higher prevalence was detected in samples of female origin. Many of the isolates exhibited resistance to norfloxacin, and 60% of the isolates showed resistance to ampicillin. No resistance to imipenem, meropenem, or ertapenem was detected. In general, half of the isolates showed multiple resistance patterns. UPEC exhibited a weak ability to form biofilms, where no correlation was observed between multidrug resistance and biofilm-forming ability. All uropathogenic E. coli isolates carried the kpsMTII, iutA, traT, and csgA genes, whereas the low number of the isolates harbored the sat and vat genes. The diversity of virulence factors harbored by community-associated UPEC may render them more virulent and further explain the recurrence/relapse cases among community-associated UITs. To the best of our knowledge, this study constitutes the first exploration of virulence, biofilm-forming ability, and its association with multidrug resistance among UPEC isolates in Saudi Arabia. Further investigations are needed to elucidate the epidemiology of community-associated UPEC in Saudi Arabia.

L-Asparaginase activity analysis, ansZ gene identification and anticancer activity of a new Bacillus subtilis isolated from sponges of the Red Sea.

This study describes the isolation of various marine bacteriafrom sponges collected from the Red Sea (Saudi Arabia) andL-asparaginase (anti-cancer enzyme) production from bacterialisolates. The 16S rDNA based phylogenetic analysis revealed thatthe isolate WSA3 was a Bacillus subtilis. Its partial-length genesequence was submitted to GenBank under the accession numberMK072695. The new B. subtilis strain harbored the exact size(1128 bp) of the new L-asparaginase (ansZ) gene as confirmedby PCR and in gel visualization, which was submitted to the NCBIdatabase (accession number MN566442). The molecular weightof partially purified L-asparaginase was determined as 45 kDa bySDS-PAGE. In addition, the enzyme L-asparaginase did not showglutaminase activity which is very important from a medical pointof view. Moreover, 100 µg/mL of the partially purified B. subtilis Lasparaginaseshowed promising anti-cancer activities when testedagainst three cancer cell lines (HCT-116, MCF-7, and HepG2).

Dissecting Streptococcus pyogenes interaction with human.

Streptococcus pyogenes is a species of Gram-positive bacteria. It is also known as Group A Streptococcus (GAS) that causes pathogenesis to humans only. The GAS infection has several manifestations including invasive illness. Current research has linked the molecular modes of GAS virulence with substantial sequencing determinations for the isolation of genomes. These advances help to comprehend the molecular evolution resulting in the pandemic strains. Thus, it is indispensable to reconsider the philosophy that involves GAS pathogenesis. The recent investigations involve studying GAS in the nasopharynx and its capability to cause infection or asymptomatically reside in the host. These advances have been discussed in this article with an emphasis on the natural history of GAS and the evolutionary change in the pandemic strains. In addition, this review describes the unique functions for major pathogenicity determinants to comprehend their physiological effects.

Bacterium Hafnia alvei secretes l-methioninase enzyme: Optimization of the enzyme secretion conditions.

I isolated bacteria from blue cheese in order to find bacterial strains secreting l-methioninase enzyme, and optimized the conditions for the most efficient enzyme secretion. The efficient isolate, identified according to the 16S rRNA gene sequence analysis, was Hafnia alvei belonging to Enterobacteriaceae. I confirmed that the H. alvei strain harbored the methionase gene, mdeA (1194 bp). The environmental (pH, temperature) and nutritional (carbon and nitrogen sources and Mg concentration) factors influencing the l-methioninase production of H. alvei were optimized. The highest yield of l-methioninase enzyme was reached after 48 h of incubation when the acidity of the growing medium was adjusted to pH 7.5 and the temperature was 35 °C. The following concentrations of the supplements increased the l-methioninase yield in the medium: galactose (2.0 g L-1), MgSO4 (0.25 g L-1), l-methionine as an inducer (2.0 g L-1), and l-asparagine as an additional N source (1.5 g L-1). I introduce a bacterial strain of H. alvei that is previously unreported to secrete l-methioninase enzyme and show that a carbon source is a mandatory supplement whereas l-methionine is not a mandatory supplement for l-methioninase enzyme production of H. alvei.

Gold nanoparticles synthesised by flavonoid tricetin as a potential antibacterial nanomedicine to treat respiratory infections causing opportunistic bacterial pathogens.

In this study, flavonoid tricetin was used as a reducing and capping agent for the synthesis of gold nanoparticles (AuNPs). Further, the antibacterial efficacy of the synthesised AuNPs was evaluated against the opportunistic bacterial pathogens that cause respiratory infections. The optimum levels for the synthesis of AuNPs were found to be pH 8, temperature 30 °C, tricetin 125 µM and chloroauric acid 250 µM. The tricetin synthesised AuNPs exhibited in spherical shape with an average size of 12 nm. FT-IR results confirmed that the hydroxyl (OH) and carbonyl (CO) groups of tricetin were mainly participated in the synthesis of AuNPs. The opportunistic bacterial pathogens isolated from immunocompromised patients suffering with different respiratory infections were identified as Staphylococcus aureus, Enterobacter xiangfangensis, Bacillus licheniformis, Escherichia fergusonii, Acinetobacter pittii, Pseudomonas aeruginosa, Aeromonas enteropelogenes and Proteus mirabilis. The antibacterial studies confirmed the broad-spectrum antibacterial activity of AuNPs against the tested Gram-positive and Gram-negative bacteria. The synthesised AuNPs showed high biocompatibility on primary normal human dermal fibroblast (NHDF-c) cells up to 50 µM mL-1. Best of our knowledge, this is the first report on the synthesis of AuNPs using tricetin, which may be a potential antibacterial nanomedicine to treat bacterial infections.