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Acne vulgaris, a prevalent skin condition, arises from an imbalance in skin flora, fostering bacterial overgrowth. Addressing this issue, clindamycin molecularly imprinted polymeric nanoparticles (Clin-MIP) loaded onto polyurethane nanofiber scaffolds were developed for acne treatment. Clin-MIP was synthesized via precipitation polymerization using methacrylic acid (MAA), ethylene glycol dimethacrylate (EGDMA), and azoisobutyronitrile (AIBN) as functional monomers, crosslinkers, and free-radical initiators, respectively. MIP characterization utilized Fourier-transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM) before being incorporated into polyurethane nanofibers through electrospinning. Further analysis involved FTIR, scanning electron microscopy (SEM), in vitro release studies, and an ex vivo study. Clin-MIP showed strong antibacterial activity against S. aureus, with inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of 0.39 and 6.25 µg/mL, respectively. It significantly dropped the bacterial count from 1 × 108 to 39 × 101 CFU/mL in vivo and has bactericidal activity within 180 min of incubation in vitro. The pharmacodynamic and histopathology studies revealed a significant decrease in infected animal skin inflammation, epidermal hypertrophy, and congestion upon treatment with Clin-MIP polyurethane nanofiber and reduced pro-inflammatory cytokines (NLRP3, TNF-α, IL-1ß, and IL-6) conducive to acne healing. Consequently, the recently created Clin-MIP polyurethane nanofibrous scaffold. This innovative approach offers insight into creating materials with several uses for treating infectious wounds caused by acne.
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Trehalose serves as a crucial osmolyte and plays a significant role in stress tolerance. The influence of exogenously added trehalose (1 and 5 mM) in alleviating the chromium (Cr; 0.5 mM) stress-induced decline in growth, photosynthesis, mineral uptake, antioxidant system and nitrate reductase activity in Vigna radiata was studied. Chromium (Cr) significantly declined shoot height (39.33%), shoot fresh weight (35.54%), shoot dry weight (36.79%), total chlorophylls (50.70%), carotenoids (29.96%), photosynthesis (33.97%), net intercellular CO2 (26.86%), transpiration rate (36.77%), the content of N (35.04%), P (35.77%), K (31.33%), S (23.91%), Mg (32.74%), and Ca (29.67%). However, the application of trehalose considerably alleviated the decline. Application of trehalose at both concentrations significantly reduced hydrogen peroxide accumulation, lipid peroxidation and electrolyte leakage, which were increased due to Cr stress. Application of trehalose significantly mitigated the Cr-induced oxidative damage by up-regulating the activity of reactive oxygen species (ROS) scavenging enzymes, including superoxide dismutase (182.03%), catalase (125.40%), ascorbate peroxidase (72.86%), and glutathione reductase (68.39%). Besides this, applied trehalose proved effective in enhancing ascorbate (24.29%) and reducing glutathione content (34.40%). In addition, also alleviated the decline in ascorbate by Cr stress to significant levels. The activity of nitrate reductase enhanced significantly (28.52%) due to trehalose activity and declined due to Cr stress (34.15%). Exogenous application of trehalose significantly improved the content of osmolytes, including proline, glycine betaine, sugars and total phenols under normal and Cr stress conditions. Furthermore, Trehalose significantly increased the content of key mineral elements and alleviated the decline induced by Cr to considerable levels.
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Cromo , Estresse Oxidativo , Fotossíntese , Espécies Reativas de Oxigênio , Trealose , Vigna , Trealose/metabolismo , Trealose/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Fotossíntese/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Vigna/efeitos dos fármacos , Vigna/crescimento & desenvolvimento , Vigna/metabolismo , Minerais/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Clorofila/metabolismo , Antioxidantes/metabolismoRESUMO
Background: Diabetes Mellitus is a multisystem chronic pandemic, wound inflammation, and healing are still major issues for diabetic patients who may suffer from ulcers, gangrene, and other wounds from uncontrolled chronic hyperglycemia. Marshmallows or Althaea officinalis (A.O.) contain bioactive compounds such as flavonoids and phenolics that support wound healing via antioxidant, anti-inflammatory, and antibacterial properties. Our study aimed to develop a combination of eco-friendly formulations of green synthesis of ZnO-NPs by Althaea officinalis extract and further incorporate them into 2% chitosan (CS) gel. Method and Results: First, develop eco-friendly green Zinc Oxide Nanoparticles (ZnO-NPs) and incorporate them into a 2% chitosan (CS) gel. In-vitro study performed by UV-visible spectrum analysis showed a sharp peak at 390 nm, and Energy-dispersive X-ray (EDX) spectrometry showed a peak of zinc and oxygen. Besides, Fourier transforms infrared (FTIR) was used to qualitatively validate biosynthesized ZnO-NPs, and transmission electron microscope (TEM) showed spherical nanoparticles with mean sizes of 76 nm and Zeta potential +30mV. The antibacterial potential of A.O.-ZnO-NPs-Cs was examined by the diffusion agar method against Gram-positive (Staphylococcus aureus and Bacillus subtilis) and Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa). Based on the zone of inhibition and minimal inhibitory indices (MIC). In addition, an in-silico study investigated the binding affinity of A.O. major components to the expected biological targets that may aid wound healing. Althaea Officinalis, A.O-ZnO-NPs group showed reduced downregulation of IL-6, IL-1ß, and TNF-α and increased IL-10 levels compared to the control group signaling pathway expression levels confirming the improved anti-inflammatory effect of the self-assembly method. In-vivo study and histopathological analysis revealed the superiority of the nanoparticles in reducing signs of inflammation and wound incision in rat models. Conclusion: These biocompatible green zinc oxide nanoparticles, by using Althaea Officinalis chitosan gel ensure an excellent new therapeutic approach for quickening diabetic wound healing.
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Althaea , Quitosana , Diabetes Mellitus , Nanopartículas Metálicas , Óxido de Zinco , Humanos , Animais , Ratos , Óxido de Zinco/química , Quitosana/química , Althaea/metabolismo , Interleucina-6 , Fator de Necrose Tumoral alfa , Nanopartículas Metálicas/química , Antibacterianos/farmacologia , Antibacterianos/química , Cicatrização , Anti-Inflamatórios/farmacologia , Inflamação , Flores , Testes de Sensibilidade Microbiana , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
Pseudomonas aeruginosa (P. aeruginosa) is an ESKAPE pathogen that can quickly develop resistance to most antibiotics. This bacterium is a zoonotic pathogen that can be found in humans, animals, foods, and environmental samples, making it a One-Health concern. P. aeruginosa threatens the poultry industry in Egypt, leading to significant economic losses. However, the investigation of this bacterium using NGS technology is nearly non-existent in Egypt. In this study, 38 isolates obtained from broiler farms of the Delta region were phenotypically investigated, and their genomes were characterized using whole genome sequencing (WGS). The study found that 100% of the isolates were resistant to fosfomycin and harbored the fosA gene. They were also resistant to trimethoprim/sulfamethoxazole, although only one isolate harbored the sul1 gene. Non-susceptibility (resistant, susceptible with increased dose) of colistin was observed in all isolates. WGS analysis revealed a high level of diversity between isolates, and MLST analysis allocated the 38â¯P. aeruginosa isolates into 11 distinct sequence types. The most predominant sequence type was ST267, found in 13 isolates, followed by ST1395 in 8 isolates. The isolates were susceptible to almost all tested antibiotics carrying only few different antimicrobial resistance (AMR) genes. Various AMR genes that confer resistance mainly to ß-lactam, aminoglycoside, sulfonamide, and phenicol compounds were identified. Additionally, several virulence associated genes were found without any significant differences in number and distribution among isolates. The majority of the virulence genes was identified in almost all isolates. The fact that P. aeruginosa, which harbors several AMR and virulence-associated factors, is present in poultry farms is alarming and threatens public health. The misuse of antimicrobial compounds in poultry farms plays a significant role in resistance development. Thus, increasing awareness and implementing strict veterinary regulations to guide the use of veterinary antibiotics is required to reduce health and environmental risks. Further studies from a One-Health perspective using WGS are necessary to trace the potential transmission routes of resistance between animals and humans and clarify resistance mechanisms.
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Aves Domésticas , Infecções por Pseudomonas , Humanos , Animais , Aves Domésticas/genética , Pseudomonas aeruginosa/genética , Virulência/genética , Fazendas , Tipagem de Sequências Multilocus/veterinária , Egito/epidemiologia , Galinhas/microbiologia , Antibacterianos/farmacologia , Sequenciamento Completo do Genoma/veterinária , Infecções por Pseudomonas/epidemiologia , Infecções por Pseudomonas/veterinária , Fatores de Virulência/genéticaRESUMO
Fungal diseases have always been a major problem for cantaloupe crops; however, synthetic fungicides are hazardous to humans and the environment. Consequently, a feasible alternative to fungicides without side effects could be by using bio agents and naturally occurring plants with antibacterial potential. This study has achieved a novel procedure for managing wilt and root rot diseases by potentially using Trichoderma sp. culture filtrates in consortium with plant extract of Calotropis procera, Rhizoctonia solani, Fusarium oxysporum, and Pythium ultimum, which were isolated from infected cantaloupe roots with identified root rot symptoms. The antagonistic activity of four Trichoderma isolates and analysis of antibiotics and filtrate enzymes of the most active Trichoderma isolate were determined as well as phytochemical analysis of C. procera plant extract using HPLC-UV. The obtained results showed that all Trichoderma isolates considerably lowered the radial growth of P. ultimum, R. solani, and F. oxysporum in varying degrees. The scanning electron micrographs illustrate the mycoparasitic nature of Trichoderma sp. on F. oxysporum. The phytochemical analysis of C. procera indicated that phenolic contents were the major compounds found in extracts, such as vanillin (46.79%), chlorogenic acid (30.24%), gallic acid (8.06%), and daidzein (3.45%) but including only a low amount of the flavonoid compounds rutin, naringenin, and hesperetin. The Pot experiment's findings showed that cantaloupe was best protected against wilting and root rot diseases when it was treated with both Trichoderma sp. culture filtrates (10%) and C. procera extract of (15 mg/mL), both alone and in combination. This study demonstrates that the application of bio agent Trichoderma spp. filtrate with C. procera phenol extract appears useful for controlling wilting and root rot disease in cantaloupe. This innovative approach could be used as an alternative to chemical fungicide for the control of wilting and rot root diseases.
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Calotropis , Cucumis melo , Porcelana Dentária , Fungicidas Industriais , Ligas Metalo-Cerâmicas , Titânio , Trichoderma , Humanos , Polifenóis , Fenóis/farmacologia , Antibacterianos , Compostos Fitoquímicos , Extratos Vegetais/farmacologiaRESUMO
This work aimed to characterize S. aureus isolates from the eyes of healthy and clinically affected equines in the Kafrelsheikh Governorate, Egypt. A total of 110 animals were examined for the presence of S. aureus, which was isolated from 33 animals with ophthalmic lesions and 77 healthy animals. We also investigated the antimicrobial resistance profile, oxacillin resistance mechanism, and the major virulence factors implicated in many studies of the ocular pathology of pathogenic S. aureus. The association between S. aureus eye infections and potential risk factors was also investigated. The frequency of S. aureus isolates from clinically affected equine eyes was significantly higher than in clinically healthy equids. A significant association was found between the frequency of S. aureus isolation from clinically affected equine eyes and risk factors including age and season but not with sex or breed factors. Antimicrobial resistance to common antibiotics used to treat equine eyes was also tested. Overall, the isolates showed the highest sensitivity to sulfamethoxazole (100%) and the highest resistance to cephalosporin (90.67%) and oxacillin (90.48%). PCR was used to demonstrate that mecA was present in 100% of oxacillin- and ß-lactam-resistant S. aureus strains. The virulence factor genes Spa (x region), nuc, and hlg were identified in 62.5%, 100%, and 56%, of isolates, respectively, from clinically affected equines eyes. The severity of the eye lesions increased in the presence of γ-toxin-positive S. aureus. The phylogenetic tree of the Spa (x region) gene indicated a relationship with human reference strains isolated from Egypt as well as isolates from equines in Iran and Japan. This study provides insight into the prevalence, potential risk factors, clinical pictures, zoonotic potential, antimicrobial resistance, and ß-lactam resistance mechanism of S. aureus strains that cause eye infection in equines from Egypt.
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BACKGROUND AND OBJECTIVE: Bacteriological isolation and identification of Mycoplasma species is difficult and time-consuming, therefore, molecular identification of Mycoplasma using PCR targeting specific genes is considered a specific and sensitive method for identification. The aim of current study was to isolate, characterize Mycoplasma infection in dromedary camels in Saudi Arabia. MATERIALS AND METHODS: Nasal swabs were randomly collected from 93 camels and tested for Mycoplasma by sequencing of their 16S rRNA genes using universal primers. RESULTS: The 93 samples, 24 were positive for Mycoplasma. However, no positive results were obtained using species-specific primers for Mycoplasma arginine, M. bovis or M. mycoides subsp. mycoides, thus, 16S rDNA sequencing methods and semi-nested PCR were employed. Sequences were matched to those in GenBank and phylogenetic analysis was performed. Mycoplasma edwardii (77-84% similarity with Mycoplasma edwardii ATCC 23462) and one isolate of Mycoplasma yeastsii (100% similarity with M. yeastsii GM274B) were identified. Further, some Mycoplasma species were identified as previously uncultured. The incidence of Mycoplasma infection in camels in Taif city, Saudi Arabia, was approximately 26%. CONCLUSION: This study provides insights into the accuracy and efficiency of PCR and universal primers for the detection and identification of Mycoplasma, thereby circumventing conventional culturing methods that require several days to complete and exhibit low accuracy.
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Camelus/microbiologia , DNA Bacteriano/genética , Infecções por Mycoplasma/veterinária , Mycoplasma/genética , Reação em Cadeia da Polimerase/veterinária , Sistema Respiratório/microbiologia , Infecções Respiratórias/veterinária , Ribotipagem/veterinária , Animais , Mycoplasma/classificação , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/microbiologia , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/microbiologia , Arábia SauditaRESUMO
Diagnosis and treatment of ocular fungal infection in equine seems very challenging for owners and clinicians. The present study aimed to identify and characterize fungal species isolated from the eyes of clinically healthy and diseased equines (N = 100) from Dakahlia Governorate, Egypt. The work also involved morphological and molecular characterization of the major fungal species. In addition, correlations between the occurrence of isolated fungi and some of the potential risk factors were also investigated. Interestingly, the prevalence rate of ocular mycosis in all examined equines in the study was 28% and there were major clinical signs associated with ocular fungal infection. Moreover, the identified fungal species included Aspergillus flavus, A. fumigatus, A. niger, Penicillium spp., Mucor spp., and Alternari spp. with a corresponding prevalence rate of 63.9%, 27.8%, 15.3%, 18.1%, 13.9%, and 4.2%, respectively, in healthy equine eyes, while their prevalence in diseased equine eyes was 57.1%, 32.1%, 21.4%, 7.1%, 3.6%, and 0%. Furthermore, a statistical significant association (p < 0.05) was found between the frequency of isolation of A. fumigatus and Penicillium and several risk factors (breed, sex, and ground type), while the remaining risk factors and occurrence of fungi were not statistically correlated. A subset of the Aspergillus species samples positive by polymerase chain reaction (PCR) were sequenced and their phylogenetic analysis identified three species of Aspergillus. Taken together, our study provides novel data related to the occurrence of ocular mycosis in equine in Egypt. Given the zoonotic potential of some identified fungi, our data may be helpful for implementation of novel diagnostic and therapeutic strategies for combating this sight-threatening infection in equine.
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Chicken Salmonella enterica serovars are enteric bacteria associated with massive public health risks and economic losses. There is a widespread antimicrobial resistance among S. enterica serotypes, and innovative solutions to antibiotic resistance are needed. We aimed to use probiotics to reduce antibiotic resistance and identify the major probiotic players that modify the early interactions between S. enterica and host cells. One-day-old cobb broiler chicks were challenged with S. typhimurium after oral inoculation with different probiotic strains for 3 days. The adherence of different probiotic strains to Caco-2 intestinal epithelial cells was studied in vitro. Lactobacillus (Lacticaseibacillus) casei ATTC334 and Bifidobacterium breve JCM1192 strains attached to Caco-2 cells stronger than B. infantis BL2416. L. casei ATTC334 and B. breve JCM1192 reduced S. typhimurium recovery from the cecal tonsils by competitive exclusion mechanism. Although B. infantis BL2416 bound poorly to Caco-2 epithelial cells, it reduced S. typhimurium recovery and increased IFN-γ and TNF-α production. L. casei ATTC334, B. breve JCM1192 and B. infantis BL2416 improved body weight gain and the food conversion rate in S. typhimurium-infected broilers. B. longum Ncc2785 neither attached to epithelial cells nor induced IFN-γ and TNF-α release and consequently did not prevent S. typhimurium colonization in broiler chickens. In conclusion, probiotics prevented the intestinal colonization of S. typhimurium in infected chickens by competitive exclusion or cytokine production mechanisms.
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Inflammation is critical for infection control and acts as an arsenal defense mechanism against invading microbes through activation of the host immune system. It works via its inflammasome components to sense the dangerous invading microorganism and send messages to the immune system to destroy them. To date, the function of bovine macrophage inflammasome and its relationship with actin has not been identified. This study aimed to investigate the activation of bovine inflammasome by phase one flagellin from Salmonella typhimurium and its interaction with actin. Bovine monocyte-derived macrophages were prepared and challenged with S. typhimurium SL1344 phase one flagellin. The results demonstrated the relationship between the flagellin-based activation of inflammasome and actin rearrangement. The flagellin-based activation of inflammasome promoted the activation and co-localization of F-actin and the inflammasome complex. Actin was remodeled to different degrees according to the stage of inflammasome activation. The actin redistribution varied from polymerization to filopodia, while at the stage of pyroptotic cell death, actin was broken down and interacted with activated inflammasome complexes. In conclusion, flagellin-dependent inflammasome activation and actin localization to the inflammasome at the stage of pyroptotic cell death may be of importance for appropriate immune responses, pending further studies to explore the exact cross-linking between the inflammasome complex and actin.
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BACKGROUND: Aspergillus sojae has been an important filamentous fungus in Biotechnology due to its use in diverse fermentative processes for the production of various food products. Furthermore, this fungus is a common expression system for the production of enzymes and other metabolites. The availability of molecular genetic tools to explore its biology is thus of big interest. In this study, an Agrobacterium tumefaciens-mediated transformation (ATMT) system for A. sojae was developed and its applicability evaluated. RESULTS: The donor plasmid named pRM-eGFP was constructed for ATMT of A. sojae. This plasmid contains the ble and egfp genes in its transfer DNA element (T-DNA) to confer phleomycin resistance and express the enhanced green fluorescent protein (EGFP) in A. sojae, respectively. Agrobacterium tumefaciens (LBA4404) harboring the donor plasmid and A. sojae (ATCC 20235) were co-cultured under diverse conditions to achieve ATMT. The maximum number of transformed fungi was obtained after three days of co-culturing at 28°C, and selection with 50 µg/ml phleomycin. Polymerase chain reaction (PCR), fluorescence microscopy and Western Blot analysis for EGFP expression confirmed successful genomic integration of the T-DNA element in A. sojae. The T-DNA was mitotically stable in approximately 40% of the fungal transformants after four generations of sub-culturing under phleomycin pressure. CONCLUSION: We successfully established a new ATMT protocol for A. sojae. This transformation system should enable further protein expression studies on this filamentous fungus.