Phytochemical, antimicrobial and cytotoxicity screening of ethanol extract of Acacia ehrenbergiana Hayne grown in Jazan Region of Saudi Arabia

Purpose: To explore the phytoconstituents of Acacia ehrenbergiana Hayne as well as its biological effects. Methods: Determination of phytoconstituents of ethanol extract of the plant was performed by gas chromatography-mass spectrometry (GC-MS) technique. Antibacterial screening was conducted against the isolates of Gram-positive and Gram-negative microbes while the anti-carcinogenic properties of the ethanol extract on cancerous cells were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) cytotoxicity assay against breast MCF7, ovary cancer A2780 and colon cancer HT29 cells, respectively, in addition to normal MRC5 fibroblast cells. Results: GC-MS analysis identified 15 different phytochemicals in the ethanol extract. The extract exerted significant antimicrobial activity with the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in the range 1.56 - 6.25 and 3.12 – 12.5 mg/L, respectively, against all test bacterial strains. Cytotoxic activity, obtained by MTT assay, was 28.81 ± 0.99, 12.50 ± 2.50, 23.90 ± 0.74 and 50.58 ± 3.24 μg/mL, against the three cancer cell lines and normal fibroblast, respectively. MTT cytotoxicity results was further confirmed by clonogenic survival assay on MCF7 cells. Conclusion: This study highlights the potential interesting ethnopharmacological applications of Acacia ehrenbergiana Hayne to treat drug-resistant pathogens as standardized extract. Keywords: Acacia ehrenbergiana, Phytochemistry, Antimicrobial, Cytotoxicit

Evaluation of specific and non-specific immune response of four vaccines for caseous lymphadenitis in sheep challenged

Background: Caseous lymphadenitis (CLA) is a serious disease affects sheep and goat, caused by Corynebacterium pseudotuberculosis. Due to it is non-treatable disease, so the effective preventive vaccines are considered a significant way to combat the disease. All strains of C. pseudotuberculosis have several virulence factors that associated with their cell invasion, survival, and proliferation such as phospholipase D (PLD), outer lipid coat, and secreted proteases. Aim: The present study was directed to perform a comparative innate and acquired immune response assessment of different four vaccine formulas to evoke protection against induced (CLA) challenge in sheep. Materials and Methods: Negative ELISA (free of CLA) 15 local breed male (Balady) sheep were divided into five groups, each has received a different vaccine while the control has received saline buffer. The first vaccine composed of toxoid PLD alone the second composed of toxoid PLD with bacterin (formalinkilled bacteria), the third vaccine composed of toxoid PLD plus covaccine 8, while the fourth one composed of toxoid PLD plus locally produced polyvalent clostridial vaccine. The specific immune response was evaluated through lymphocyte proliferation assay using ELISA BrdU kit, while the non-specific response was estimated by superoxide anion production and lysozyme activity assays. Results: The study revealed that PLD toxoid could evoke the highest specific immune response, showing a stimulation index (9.12%). On the other hand, combined toxoid PLD with bacterin followed by PLD toxoid showed a significant increase in the non-specific innate immune response. Conclusion: The present study indicated that the toxoid PLD alone vaccine was most efficient and provided innate and acquired immune response in animals against CLA

Serotyping, antibiotic susceptibility, and virulence genes screening of Escherichia coli isolates obtained from diarrheic buffalo calves in Egyptian farms

Aim: In Egypt as in many other countries, river water buffalo (Bubalus bubalis) is considered an important source of high-quality milk and meat supply. The objective of this study was to investigate serotypes, virulence genes, and antibiotic resistance determinants profiles of Escherichia coli isolated from buffalo at some places in Egypt; noticibly, this issue was not discussed in the country yet. Materials and Methods: A number of 58 rectal samples were collected from diarrheic buffalo calves in different regions in Egypt, and bacteriological investigated for E. coli existence. The E. coli isolates were biochemically, serologicaly identified, tested for antibiotic susceptibility, and polymerase chain reaction (PCR) analyzed for the presence of antibiotic resistance determinants and virulence genes. Results: Overall 14 isolates typed as E. coli (24.1%); 6 were belonged to serogroup O78 (10.3%), followed by O125 (4 isolates, 6.9%), then O158 (3 isolates, 5.2%) and one isolate O8 (1.7%), among them, there were 5 E. coli isolates showed a picture of hemolysis (35.7%). The isolates exhibited a high resistance to β lactams over 60%, followed by sulfa (50%) and aminoglucoside (42.8%) group, in the same time the isolates were sensitive to quinolone, trimethoprim-sulfamethoxazole, tetracycline (100%), and cephalosporine groups (71.4%). A multiplex PCR was applied to the 14 E. coli isolates revealed that all were carrying at least one gene, as 10 carried blaTEM (71.4%), 8 Sul1 (57.1%), and 6 aadB (42.8%), and 9 isolates could be considered multidrug resistant (MDR) by an incidence of 64.3%. A PCR survey was stratified for the most important E. coli virulence genes, and showed the presence of Shiga toxins in 9 isolates carried either one or the two Stx genes (64.3%), 5 isolates carried hylA gene (35.7%), and eae in 2 isolates only (14.3%), all isolates carried at least one virulence gene except two (85.7%). Conclusion: The obtained data displayed that in Egypt, buffalo as well as other ruminants could be a potential source of MDR pathogenic E. coli variants which have a public health importance

Incorporation of Plant Extracted Hydroxyapatite and Chitosan Nanoparticles on the Surface of Orthodontic Micro-Implants: An In-Vitro Antibacterial Study

In our study, the structural and morphological applications of hydroxyapatite and chitosan nanoparticles and coated micro-implants were assessed for their ability to combat oral pathogenic bacteria. The hydroxyapatite, as well as chitosan nanoparticles, were synthesized from the Salvadora persica plant. The crystal morphology, phase composition, particle size, and surface functional groups of the nano-samples were analyzed via classical examinations and energy dispersive X-ray analysis. The prepared nanoparticles have been examined for antibacterial activity against four common oral bacterial strains. The antimicrobial effect was also assessed by the Live/Dead BacLight technique in combination with confocal scanning laser microscopy. Titanium micro-implants were coated with regular hydroxyapatite (HAP) and chitosan nanoparticles, and the surface was characterized by scanning electron microscopy. The analysis asserted elemental composition of the prepared nanoparticles and their textural features, metal crystallization, and functional bonds. The antibacterial activity of the nanoparticles was evaluated against oral pathogenic microorganisms by the disc diffusion method, minimum bacterial concentration (MBC), and minimum inhibitory concentration (MIC). Chitosan nanoparticles showed (MICs) of 8 μg mL−1 for (Streptococcus salivarius, Streptococcus mutans and Enterococcus faecalis), and 16 μg mL−1 for Streptococcus sanguinis. HAP nanoparticles showed (MICs) of 16 μg/mL for E. faecalis, and S. sanguis, 8 μg/mL for S. salivarius and finally 4 μg/mL for S. mutans. HAP nanoparticles showed enhanced antibacterial activity and more obvious damage in the bacterial cell membrane than that of synthesized chitosan nanoparticles. The prepared nanoparticles could successfully coat titanium microplates to enhance their efficiency

Bacteriological, molecular and histopathological evaluation of four vaccines’ protective role against caseous lymphadenitis in sheep

The caseous lymphadenitis disease (CLA) due to Corynebacteriumpseudotuberculosis has worldwide distribution and indicates high prevalence in different countries. The bacterium has many pathogenicity factors; firstly, “phospholipase D”, an exotoxin virulence factor that enhances vascular permeability and facilitates bacterial transmission causing endothelial cell damage. The bacterium's exterior lipid cover is the second pathogenicity factor. This protects it from hydrolytic enzymes found in the phagocytes of the host, where the bacteria proliferate and then release after rupturing.  Bacterial proliferation causes CLA abscesses, which are followed by attraction and the formation of an inflammatory response, which increases lymph flow and vascular permeability. The purpose of this study was to look atpostmortem findings, bacterial cultures, and histology of vaccinated sheep to see how well they were protected. On 15 male local sheep bread, four prepared vaccinations against C. pseudotuberculosis biotype 1 were tested. The animals were divided into 5 groups; four of various vaccines used and the fifthwere kept non-vaccinated. A live virulent strain of C. pseudotuberculosis was given to each group. Unvaccinated animals displayed CLA symptoms similar to those seen in spontaneously sick animals.&nbsp

Major Gram-negative bacterial causes isolated from apparent Healthy and diarrheic foals in Egypt, prevalence, identification and antibiotic susceptibility profiles

Diarrhea is one of the most significant illnesses affecting young foals and may be manifested in more than half of foals up to 3 months of age. Numerous infectious causes have been involved but bacterial pathogens are concerned. This study aimed to assess and identify the prime Gram-negative bacteriological etiology of Arabian horse foals’ diarrhea in Egypt and to designate the antibiotic susceptibility profiles of the isolated microorganisms.  Rectal swabs and internal organs were obtained from 216 foals (89 diarrheic and 127 apparently healthy), aged 1 week to 1 year old, reared in Arabian horse farms in Great Cairo, Egypt during a period extended from March 2022 to December 2022. Conventional bacteriological examination was performed using selective media persuaded by routine and advanced biochemical tests. The isolation displayed 648 bacterial isolates; 452 (68.6%) were Gram-negative with the mixed isolation representing about 28%. Escherichia coli constituted the most prevalent; 110 / 452 (24.34%); 65 in apparently healthy foals and 45 among diarrheic ones. The second was Klebsiella pneumoniae (72, 15.92%) at which 50, (15.67%) were found to expose the highest causative agent recovered from diarrheic foals. Imipenem, quinolones, and trimethoprim/sulfamethoxazole were the most effective versus Gram-negative species isolated from diarrheic foals while ampicillin and tetracycline were of no effect. The phenotypic antibiotic susceptibility test revealed a picture of multidrug resistance (MDR) as 111 isolates (28.71%) showed resistance to three or more antibiotics belonging to different groups. The results demonstrated that Klebsiella pneumonia was the highest MDR species (54.16%) followed by Pseudomonas aeruginosa, Salmonella enterica, and Escherichia coli isolates by 43.9, 32.1 and 30.9 % respectively. In conclusion, Gram-negative bacteria constituted the major causative agents of diarrhea in the Arabian foals. The most effective antibacterial drugs were imipenem, quinolones, and trimethoprim/ sulfamethoxazole. The issue of the existence of multidrug resistance isolates should be considered for proper therapy of foal diarrhea

Major Gram-negative bacterial causes isolated from apparent Healthy and diarrheic foals in Egypt, prevalence, identification and antibiotic susceptibility profiles

Diarrhea is one of the most significant illnesses affecting young foals and may be manifested in more than half of foals up to 3 months of age. Numerous infectious causes have been involved but bacterial pathogens are concerned. This study aimed to assess and identify the prime Gram-negative bacteriological etiology of Arabian horse foals’ diarrhea in Egypt and to designate the antibiotic susceptibility profiles of the isolated microorganisms.  Rectal swabs and internal organs were obtained from 216 foals (89 diarrheic and 127 apparently healthy), aged 1 week to 1 year old, reared in Arabian horse farms in Great Cairo, Egypt during a period extended from March 2022 to December 2022. Conventional bacteriological examination was performed using selective media persuaded by routine and advanced biochemical tests. The isolation displayed 648 bacterial isolates; 452 (68.6%) were Gram-negative with the mixed isolation representing about 28%. Escherichia coli constituted the most prevalent; 110 / 452 (24.34%); 65 in apparently healthy foals and 45 among diarrheic ones. The second was Klebsiella pneumoniae (72, 15.92%) at which 50, (15.67%) were found to expose the highest causative agent recovered from diarrheic foals. Imipenem, quinolones, and trimethoprim/sulfamethoxazole were the most effective versus Gram-negative species isolated from diarrheic foals while ampicillin and tetracycline were of no effect. The phenotypic antibiotic susceptibility test revealed a picture of multidrug resistance (MDR) as 111 isolates (28.71%) showed resistance to three or more antibiotics belonging to different groups. The results demonstrated that Klebsiella pneumonia was the highest MDR species (54.16%) followed by Pseudomonas aeruginosa, Salmonella enterica, and Escherichia coli isolates by 43.9, 32.1 and 30.9 % respectively. In conclusion, Gram-negative bacteria constituted the major causative agents of diarrhea in the Arabian foals. The most effective antibacterial drugs were imipenem, quinolones, and trimethoprim/ sulfamethoxazole. The issue of the existence of multidrug resistance isolates should be considered for proper therapy of foal diarrhea

Evaluation of biofilm formation and associated slime encoding determinants in Staphylococcus aureus isolated from clinically diseased pets

Staphylococcus aureus (S.aureus) is an important zoonotic pathogen implemented in various hospital, community as well as livestock infections. Pets as dogs and cats have increased their close social relation with human leading to significant elevation in transmission of zoonoticmultidrug resistant virulent pathogens.Diplex polymerase chain reaction was applied on ten clinical Staphylococcus aureus isolates obtained from wound and nasal samples from dogs and cats for detection of twoslime formation encoding genes;icaA andicaD. The positive genes carrying isolates were encouraged to produce biofilm and evaluated phenotypically by cultivation onto Congo red medium. Quantitative assessment was done using a microtiter plate assay, also the formed biofilm examined by fluorescent and scanning electron microscopy. Four out of tested tenStaphylococcus aureus isolates were found to harbor the two genes separately.The four isolates displayed positive biofilm production onto Congo red medium while only three isolates produced biofilmin sterile polystyrene 96-well microtiter plate. One isolate associated dog infection developed strong biofilm formation which examined byboth fluorescent and scanning electron microscopy.Staphylococcus aureus isolates obtained from diseased dogs and cats can produce biofilm that increasing their virulence and pathogenicity as well as public health concern

Erratum “Phytochemical, antimicrobial and cytotoxicity screening of ethanol extract of Acacia ehrenbergiana Hayne grown in Jazan Region of Saudi Arabia”

No Abstract

Phytochemical, Cytotoxic, and Antimicrobial Evaluation of the Fruits of Miswak Plant, Salvadora persica L.

Salvadora persica L. (Salvadoraceae) is an evergreen shrub growing in the Middle East, Africa, and Southern Asia. It is traditionally known as “miswak” and used as toothbrushes and for the treatment of toothache, gum diseases, boils, chest infection, gonorrhea, headache, spleen troubles, stomachache, and ulcers. To the best of our knowledge, this is the first study aimed at conducting phytochemical, cytotoxic, and antimicrobial investigations of the fruits (berries) of S. persica collected from the Jazan region of Saudi Arabia. Analysis of the ethanol extract of S. persica fruits using GC-MS showed the presence of six esters (20.71%), seven alkanes (15.47%), tetracosamethyl-cyclododecasiloxane (9.91%), eicosamethyl-cyclodecasiloxane (7.27%), and 1-monolinoleoylglycerol (5.17%). The predominant constituents were acetyl dasycarpidan-1-methanol (10.47%), tetracosamethyl-cyclododecasiloxane (9.91%), eicosamethyl-cyclodecasiloxane (7.27%), and 1-monolinoleoylglycerol (5.17%). The petroleum ether extract of the fruits contained mainly eicosamethyl-cyclodecasiloxane (23.81%), 1-monolinoleoylglycerol (11.78%), (Z,Z,Z)-9,12,15-octadecatrienoic acid ethyl ester derivative (10.56%), and tetracosamethyl-cyclododecasiloxane (9.91%). The cytotoxic properties of the ethanol extract were investigated by MTT assay against the breast MCF7, ovary A2780, and colon HT29 cells. The fruit extract of S. persica was selective against the ovarian and colon cancer cells compared to normal fibroblast cells (MRC5) as it showed IC50 values 17.50, 8.35, and 5.12, against MCF7, A2780, and HT29 cells, respectively. Interestingly, the fruit extract was also found to possess selective antimicrobial activity for Streptococcus mutans isolates with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of 3.12 and 6.25 mg/mL, respectively. Interestingly, it was found to be ineffective against other Gram-positive as well as Gram-negative microorganisms. This study provides insight into the bioactive components present in the fruits of the plant that can be utilized for its cytotoxic and antimicrobial properties