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The disinfection of commercial hatching eggs before incubation is a common strategy to reduce potential vertical transmission of bacterial and fungal infections from the eggshell to one-day-old chicks that may prevail in poultry products and eventually reach the end consumer. The present investigation focuses on the parallel testing and application of four different disinfection methods (conventional and alternative) under commercial hatchery conditions against natural eggshell bacterial contamination. Hatching eggs from two ROSS 308 broiler breeder flocks were selected and divided into six different groups: two groups were not disinfected and served as negative controls, and four were independently disinfected following product specifications and protocols. From each group, a sample of 100 hatching eggs was selected for bacterial re-isolation, utilizing a modified shell rinse method. Colony-forming units (cfu) from the shell rinse suspensions were determined and analyzed to establish cfu values for each tested egg. These values were analyzed to determine the bacterial disinfection capacity of the four disinfection methods under commercial hatchery conditions. The tested methods were hydrogen peroxide + alcohol, peracetic acid, low energy electron beam, and the gold standard in practice: formaldehyde. Among these methods, formaldehyde, peracetic acid, and low energy electron beam showed a significant difference when compared to the non-disinfected groups whereas hydrogen peroxide + alcohol did not. The bacterial disinfection capacity of the tested methods was compared as well to the gold standard method formaldehyde fumigation and only low energy electron beam achieved similar disinfection levels as formaldehyde. According to our data, three methods significantly reduce the bacterial load on the eggshell of hatching eggs under commercial hatching conditions, including potential alternative methods such as low energy electron beam that perform similar to the gold standard in practice.

The presence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli on poultry products is an important issue for veterinary and human health due to the zoonotic infection risk for producers and consumers. The present study focuses on testing the efficacy of six different disinfection methods on eggshell samples, aiming to reduce ESBL producing E. coli contamination on the hatching egg. Sterile eggshell cutouts were artificially contaminated with 10.sup.8 cfu/ml CTX-M-1 producing E. coli and used as a carrier model to analyze the efficacy of six disinfection methods. The contaminated samples were separated into two groups; 1) contaminated and disinfected, 2) contaminated and non-disinfected. Six independent disinfection protocols were performed following product specifications and protocols. Each eggshell sample was separately crushed, and the total viable bacterial count was calculated to determine the disinfection efficacy. Five out of six tested methods (formaldehyde gassing, hydrogen peroxide + alcohol spray, essential oils spray, peracetic acid foam, and low energetic electron radiation) demonstrated a reduction or completely eliminated the initial ESBL producing E. coli contamination. One method (essential oils as cold fog) only partly reached the expected efficacy threshold (reduction of >10.sup.2 cfu/ml) and the result differed significantly when compared to the reference method i.e. formaldehyde gassing.

The medicinal potential of marine invertebrates' bioactive components that may act as anti-COVID-19 demonstrated promising results. Ophiocoma dentata , which is common in the Red Sea, is one such source. Therefore, this study aimed to isolate a new compound from the brittle star, Ophiocoma dentata , and evaluate its efficacy as anti-COVID-19 in-silico and in-vitro. Standard procedures were followed in order to assess the isolated compound’s preliminary toxicity and anti-inflammatory properties. Computer virtual screening technology through molecular docking and ADMET studies was conducted as well as a new steroid derivative was isolated for the first time, named 5α-cholesta-4(27), 24-dien-3β, 23 β-diol. Investigation of the Anti-Covid-19 activity of the isolated compound using a Plaque reduction assay revealed 95% inhibition at a concentration of 5 ng/µl (12.48 µM). Moreover, this compound showed an IC 50 of 11,350 ± 1500 ng/ml against the normal fibroblast cells, indicating its safety. Interestingly, this compound exhibited anti-inflammatory activity with an IC 50 of 51.92 ± 0.03 μg/ml compared to a reference drug’s IC 50 of 53.64 ± 0.01 μg/ml, indicating that this compound is a potent anti-inflammatory. In silico data have proved that the isolated compound is a promising viral inhibitor against SARS-CoV2 and is thus recommended as a future nature preventive and curative antiviral drug.

The pressing demand for safe and efficient COVID-19 treatments has intensified interest in Natural products, especially those derived from marine organisms. In this study, a bioactive steroidal compound, 5α-cholesta-9(11)-en-3β,20β-diol, was successfully isolated from the starfish Acanthaster planci . Structural elucidation was achieved using HREIMS, FTIR, and advanced 1D/2D NMR spectroscopy, confirming a molecular formula of C₂₇H₄₆O₂ and characteristic functionalities including hydroxyl and double bond moieties. The compound demonstrated notable anti-SARS-CoV-2 activity, attaining 85% viral inhibition at 5 ng/µl with an IC₅₀ of 5.86 µM, as demonstrated by plaque reduction assays. Molecular docking studies demonstrated significant binding affinities toward key viral targets Mpro, NSP10, and RNA-dependent RNA polymerase with free binding energies of -26.85, -27.59, and − 35.08 kcal/mol, respectively. These affinities surpassed those of their respective co-crystallized reference ligands. In-silico ADMET profiling indicated favorable pharmacokinetic properties, including high BBB penetration, moderate intestinal absorption, and non-hepatotoxicity. Toxicity assessments predicted low carcinogenic risk, a high rat MTD, and minimal ocular and dermal irritancy. Additionally, we developed a predictive web application based on machine learning to estimate IC₅₀ values of SARS-CoV-2 inhibitors, streamlining the drug discovery process. The forecasted values nearly matched the experimental outcomes, demonstrating the model’s reliability and its potential to reduce time, cost, and risk in early-stage drug development. Moreover, machine learning models, particularly XGBoost, demonstrated excellent performance in predicting pIC₅₀ values (RMSE = 0.1357, MAE = 0.1022), supporting the development of a web-based IC₅₀ prediction application. The bioactivity prediction platform ENHPCG further validated the compound’s antiviral potential, estimating an IC₅₀ of 5.95 µM. Overall, these integrated analytical, biological, and computational approaches highlight 5α-cholesta-9(11)-en-3β,20β-diol as a potential SARS-CoV-2 inhibitor and a candidate for further pharmacological development.

The transition to using dual-purpose chickens is an alternative to killing male hatchlings of high performance egg-laying chickens. This study aimed to compare the gastrointestinal tract of a recently developed genetic line of dual purpose male chicken, Lohmann Dual (LD), with that of a broiler line, Ross 308. Eighty birds from each line were grown until they reached an average body weight 2000 g (5 weeks for Ross and 9 for LD birds). Six birds of each line were sampled weekly. Body weight (BW), normalized mass of gastrointestinal segments and relative length of intestine were determined. Histologically the villus height, epithelium height, crypt depth, mucosal enlargement factor and the tunica muscularis thickness were measured in jejunum and ileum. Data were regressed against body weight and genetic line. Jejunal enterocyte microvilli and junctional complexes length were measured. Normalized mass and relative length of the gastrointestinal segments were greater in LD birds than in Ross birds at all ages. After day 7 these decreased steadily over the lifetime of the birds in both genetic lines. The growth curves of the gastrointestinal segments of the LD birds were similar to those of the Ross birds. In birds of the same BW, LD birds had a significantly heavier gizzard, shorter intestine, higher jejunal villi, thicker ileal tunica muscularis and smaller ileal mucosal enlargement factor than were found in Ross birds. The large gizzard in LD chickens presumably increases the degree of food processing and enhances availability of nutrients in the orad part of the intestine leading to a lower nutrient concentration and a smaller absorption surface area in the ileum of the LD compared to the Ross chickens. The anatomical differences between the two lines are important criteria for further selection and should be considered in their feeding management.

The newly recorded Phyllymenia gibesii in the Mediterranean Sea at Alexandria coast of Egypt is regarded as a significant source of bioactive substances and is applied as an antioxidant, anti-inflammatory, and antimicrobial agent. According to the HPLC chromatograms, the acetone extract of P. gibesii comprised ten photosynthetic pigments (chlorophyll-a, chlorophyll-d, α-carotene, β-carotene, phycocyanin, allophycocyanin, antheraxanthin, β-cryptoxanthin, lutein, and violaxanthin). Total carotenoids were the dominant class in the pigments’ profile, achieving a concentration of 257 g/g dry weight. The P. gibbesii extract had a total content of phenols (146.67 mg/g) and a total content of flavonoids (104.40 mg/g). The capacity of all the investigated biological activities augmented with the concentration of the algal extract. The maximal DPPH scavenging capacity was 81.44%, with an inhibitory concentration (IC 50 ) of 9.88 μg/mL. Additionally, the highest ABTS scavenging capacity was 89.62%, recording an IC 50 of 21.77 μg/mL. The hemolytic activity of P. gibbesii attained a maximum capacity of 49.88% with an IC 50 of 100.25 μg/mL. Data also showed the maximum anti-inflammatory effectiveness at 81.25%, with an IC 50 of 99.75 μg/mL. Furthermore, the extract exhibited antimicrobial capacity against all reference strains, particularly at high concentrations (0.1 mg/mL), with the greatest effect on C. albicans and E. coli .

Bioactive compounds were extracted from a locally available brittle star; Ophiocoma dentata , collected from the Red Sea, Egypt. Two new sesquiterpenoids; 8, 11-epoxy-9(15)-himachaladiene-4-ol (O8-ophiocomane) and, 11-epoxy-9(15)-himachaladiene-4-ol (O7-ophiocomane) were isolated and characterized using appropriate techniques. Structure elucidation was estimated via 1D NMR, 2D NMR, FT-IR and mass spectroscopy analyses. The isolated compounds were tested for cytotoxic, antibacterial and antifungal activities. Pure compounds showed a dose dependent reduction in MCF-7 cells viability with LC50 of 103.5 and 59.5 μg/ml for compounds 1 and 2 respectively compared to the chemotherapeutic drug cisplatin (47.4 µg/ml). In vivo experiments showed that O. dentate extract significantly reduced tumor progression and improved hematological parameters and liver functions of tumor-bearing mice when administered either before or after tumor cells’ injection. The most remarkable antimicrobial effects of O. dentate crude extract were against Staphylococcus aureus , Vibrio damsela and Pseudomonas aeruginosa while the pure compounds showed activity against P. aeruginosa alone. Neither the crude extract nor the pure compounds have shown activity against Aeromonas hydrophila . These results indicates that O. dentata extract and newly isolated compounds have shown a promising cytotoxic, antiproliferative and antimicrobial activities that might be useful for pharmaceutical applications.

With respect to the potential natural resources in the marine environment, marine macroalgae or seaweeds are recognized to have health impacts. Two marine algae that are found in the Red Sea, Codium tomentosum (Green algae) and Actinotrichia fragilis (Red algae), were collected. Antibacterial and antioxidant activities of aqueous extracts of these algae were evaluated in vitro. Polyphenols from the extracts were determined using HPLC. Fillet fish was fortified with these algal extracts in an attempt to improve its nutritional value, and sensory evaluation was performed. The antibacterial effect of C. tomentosum extract was found to be superior to that of A. fragilis extract. Total phenolic contents of C. tomentosum and A. fragilis aqueous extract were 32.28 ± 1.63 mg/g and 19.96 ± 1.28 mg/g, respectively, while total flavonoid contents were 4.54 ± 1.48 mg/g and 3.86 ± 1.02 mg/g, respectively. Extract of C. tomentosum demonstrates the highest antioxidant activity, with an IC50 value of 75.32 ± 0.07 μg/mL. The IC50 of L-ascorbic acid as a positive control was 22.71 ± 0.03 μg/mL. The IC50 values for inhibiting proliferation on normal PBMC cells were 33.7 ± 1.02 µg/mL and 51.0 ± 1.14 µg/mL for C. tomentosum and A. fragilis, respectively. The results indicated that both algal aqueous extracts were safe, with low toxicity to normal cells. Interestingly, fillet fish fortified with C. tomentosum extract demonstrated the greatest overall acceptance score. These findings highlight the potential of these seaweed species for cultivation as a sustainable and safe source of therapeutic compounds for treating human and fish diseases, as well as effective food supplements and preservatives instead of chemical ones after performing in vivo assays.

[This corrects the article DOI: 10.1371/journal.pone.0238860.].[This corrects the article DOI: 10.1371/journal.pone.0238860.].

The use of natural compounds is promising in approaches to prevent and treat cancer. The long-term application of most currently employed chemotherapy techniques has toxic side effects. Eugenol, a phenolic phytochemical extracted from certain essential oils, has an anti-cancer effect. The modulation of autophagy can promote either the survival or apoptosis of cancer cells. Triple-negative (MDA-MB-231) and HER2 positive (SK-BR-3) breast cancer cell lines were treated with different doses of eugenol. Apoptosis was detected by a flow-cytometry technique, while autophagy was detected by acridine orange. Real-time PCR and Western blot assays were applied to investigate the effect of eugenol on the gene and protein expression levels of autophagy and apoptotic genes. Treating cells with different concentrations of eugenol significantly inhibited cell proliferation. The protein levels of AKT serine/threonine kinase 1 (AKT), forkhead box O3 (FOXO3a), cyclin dependent kinase inhibitor 1A (p21), cyclin-dependent kinase inhibitor (p27), and Caspase-3 and -9 increased significantly in Eugenol-treated cells. Eugenol also induced autophagy by upregulating the expression levels of microtubule-associated protein 1 light chain 3 (LC3) and downregulating the expression of nucleoporin 62 (NU p62). Eugenol is a promising natural anti-cancer agent against triple-negative and HER2-positive breast cancer. It appears to work by targeting the caspase pathway and by inducing autophagic cell death.