Search results for: Staphylococcus sp

Authors: Kumudini Apsara Munasinghe, Devin Gregory Lissau, Ryan Thomas Wade

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Fresh fruits and vegetables are rich in vitamins, minerals, and fibers and help maintain a healthy weight over high-calorie food. Eating fruits and vegetables protects us from free radicals produced by metabolic reactions and safeguards us from cardiovascular disease and cancer. However, there has been an increased concern about foodborne diseases tied to contaminated farmers’ market produce. In addition, very little information is available about the contribution of eating raw fruits and vegetables to human exposure to antibiotic-resistant bacteria. This research aims to identify bacteria isolated from farmers’ market fruits and vegetables and understand their antibiotic resistance. Vegetables and fruits were collected from farmers’ markets around Frostburg and Cumberland areas in Maryland and transported to the microbiology lab at Frostburg State University for the isolation of bacteria. Bacteria were extracted from tomatoes, cucumber, strawberry, and lettuce using Tryptic soy broth overnight at 37°C, and Tryptic Soy agar was used for the streak plate technique to isolate bacteria. Pure cultures were used to identify bacteria using biochemical reactions after conducting Gram staining technique. The research used many biochemical reactions, including Mannitol Salt agar, MacConkey agar, and Eosin Methylene blue agar, for identification. Antibiotic sensitivity was tested for many different types of antibiotics, including amoxicillin, penicillin, tetracycline, ampicillin, and erythromycin. Most prevalent bacteria in the isolates were Staphylococcus, Bacillus, Micrococcus, Enterococcus, Enterobacter, Citrobacter, and other bacteria from the family Enterobacteriaceae. The data obtained from this research will be useful to educate and train farmers and individuals involved in post-harvest processes such as transportation and selling in farmers’ markets. Further results for bacterial antibiotic resistance will be obtained, and unculturable bacteria will be identified by next-generation DNA sequencing.

Keywords: antibiotic resistance, farmers markets, fruits, bacteria, vegetables

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Authors: Said Abdirasak Abidrahman, Ibrahim Mohamed

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Background: Urinary Tract Infections (UTIs) are the commonest infections described among diabetes mellitus patients. More often, empirical antimicrobial therapy is initiated before the laboratory results are made available with minimal treatment success. The knowledge of the etiology and antibiotic susceptibility patterns of the organisms causing urinary tract infections among diabetes mellitus patients remains scarce, despite its vitality. This study sought to determine the prevalence, bacteria species, and drug susceptibility patterns of common causes of urinary tract infections among diabetes mellitus patients attending Bosaso health centers. Materials and methods: We conducted a cross-sectional study involving adult diabetic patients at Bosaso health centers between the months of May and July 2020. Laboratory assay of mid-stream urine samples was done to isolate bacteria causes of UTIs. These were biochemically identified using Gram stain, Kligler iron agar (KIA), Indole test, citrate, urea, coagulase, catalase, motility agar, and lysine iron agar. Their antibiotic susceptibility pattern for the isolated organisms was made for Ampicillin 10μg, Ciprofloxacin 5μg, Cotrimoxazole 25μg, Gentamycin 10μg, Ceftriaxone 10μg, and determined using the Kirby Bauer Disc Diffusion method. Results: Of 177 participants, 69 (39.0%) were males and 108 (61.0%) were females. Their mean age was 33.1 years (range; 18-67 years). Of these, 14.7% (26/177) of the samples revealed significant growth (>= 105 CFU/mL) giving a prevalence of 14.9 % (95% CI: 10.6 to 16.3). The organisms isolated were Escherichia coli -50% (N=13), Klebsiella pneumonia 30.8% (N=8), Staphylococcus aureus 15.4% (N=4), and unidentified organism 3.8% (N=1), and these were associated with such socio-demographic factors like history of catheterization and sexual activity. Antibiotic susceptibility to the commonly used agents for treating UTIs indicated higher sensitivity to Gentamicin and Ceftriaxone.

Keywords: antimicrobials, bacteria, urinary tract infections, diabetes

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Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

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Authors: Sodamade A., Bolaji K. A.

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Problems associated with protein malnutrition are still prevalent in third-world countries, leading to the constant search for plants that could serve as nutrients and medicinal purposes. Huckleberry is one of the plants that has been proven useful locally in the treatment of numerous ailments and diseases. A fresh sample of the plant (Vaccinium pavifolium) was collected from a vegetable garden situated near the Erelu dam of the Emmanuel Alayande College of Education Campus, Oyo. The sample was authenticated at the Forestry Research Institute of Nigeria (FRIN) Ibadan. The leaves of the plant were plucked and processed for leaf protein concentrates before proximate composition, mineral analysis phytochemical and antimicrobial properties were determined using a standard method of analysis. The results of proximate constituents showed; moisture content; 9.89±0.051g/100g, Ash; 3.23±0.12g/100g, crude fat; 3.96±0.11g/100g and 61.27±0.56g/100g of Nitrogen free extractive. The mineral analysis of the sample showed; Mg; 0.081±0.00mg/100g, Ca; 42.30±0.05mg/100g, Na; 27.57±0.09mg/100g, K; 6.81±0.01mg/100g, P; 8.90±0.03mg/100g, Fe; 0.51±0.00mg/100g, Zn; 0.021±0.00mg/100g, Cd; 0.04±0.04mg/100g, Pb; 0.002±0.00mg/100g, Cr; 0.041±0.00mg/100g Cadmium and Mercury were not detected in the sample. The result of phytochemical analysis of leaf protein concentrates of the Huckleberry showed the presence of Alkaloid, Saponin, Flavonoid, Tanin, Coumarin, Steroids, Terpenoids, Cardiac glycosides, Glycosides, Quinones, Anthocyanin, phytosterols, and phenols. Ethanolic extracts of the Vaccinium parvifolium L. leaf protein concentrates showed that it contains bioactive compounds that are capable of combating the following microorganisms; Staphylococcus aureus, Streptococcus pyogenes, Streptococcus faecalis, Pseudomonas aeruginosa, Klebisialae pneumonia and Proteus mirabilis. The results of the analysis of Vaccinium parvifolium L. leaf protein concentrates showed that the sample contains valuable nutrient and mineral constituents, and phytochemical compounds that could make the sample useful for medicinal activities.

Keywords: leaf protein concentrates, vaccinium parvifolium, nutritional characteristics, mineral composition, antimicrobial activity

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Authors: Stéfani T. A. Dantas, Laura T. S. Takume, Bruna F. Rossi, Érika R. Bonsaglia, Ivana G. Castilho, José C. F. Pantoja, Ary Fernandes Júnior, Juliano L. Gonçalves, Marcos V. Santos, Rinaldo A. Mota, Vera L. M. Rall

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Staphylococcus aureus is one of the main pathogens causing contagious bovine mastitis, being more frequently isolated from subclinical form, although the clinical form also occurs. Clinical mastitis cause visual signs, such as swelling, fever, hardening of the mammary gland, or any change in the characteristics of the milk. Considering the subclinical type, there are no visible signs in the animal nor changes in the milk. S. aureus has many important virulence factors for the establishment of its pathogenicity in animals, such as enterotoxins, which are also responsible for foodborne poisoning. Our objective is to perform a comparative analysis between 103 isolates of S. aureus, obtained from the milk of cows with clinical mastitis and 103 more, from subclinical type, in relation to the presence of these enterotoxins and verify if their presence plays an important role in the signs of illness. We will investigate all enterotoxins described till now, such as sea-see, seg-sez, sel26, sel 27, se01, and se02 (This study was approved by the Sao Paulo State University Animal Use Ethics Committee, No. 0136/2017). For the PCR assay, we used Illustra Bacteria Mini Spin Kit for bacterial DNA. At this moment, we have already tested sea-see, seg-ser, sew, and sex, and the results have already been submitted to Fisher Exact Probability Test or Chi-square Test. Considering the isolates obtained from clinical mastitis, the most frequent enterotoxins were selw (99%), selx (78%) and selh (50.5%), and sec, see, sej, sell, selp,and ser were absent. Among the subclinics, selw (82.5%) selm (15.5%) and selx (14.6%) were the most frequent, and sea-see, seg, sei-sel, sem-ser were absent. We have already observed statistically significant differences for seb, seg, seh, sei, selo, selu, selw and selx. Other interesting results were the low number of genes in each isolate from subclinical mastitis [0 genes: 14 (13.6%); 1 gene: 55 (53.4%); 2 genes: 33 (32%) or 3: 1 (0.97%)] compared to clinical isolates [1 gene: 5 (4.9%); 2 genes: 29 (28.1%); 3 genes: 38 (36.9%); 4 genes: 14 (13.6%); 5 genes: 5 (4.9%); 6 genes: 4 (3.9%); 7 genes: 5 (4.9%); 8 genes: 2 (1.9%) and 9 genes: 1 (1%)]. Based on these results, we can conclude that enterotoxins indeed play an important role in clinical signs in cattle with mastitis.

Keywords: mastitis, S. aureus, PCR, staphylococcal enterotoxin

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Authors: Naila Safdar, Faria Riasat, Azra Yasmin

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Lychee is an emerging fruit crop in Pakistan. Two famous cultivars of lychee, Gola and Surakhi, were collected from Khanpur Orchard, Pakistan and their whole fruit (including peel, pulp and seed) was investigated for pomological features and therapeutic activities. Both cultivars differ in shape and size with Gola having large size (3.27cm length, 2.36cm width) and more flesh to seed ratio (8.65g). FTIR spectroscopy and phytochemical tests confirmed presence of different bioactive compounds like phenol, flavonoids, quinones, anthraquinones, tannins, glycosides, and alkaloids, in both lychee fruits. Atomic absorption spectroscopy indicated an increased amount of potassium, magnesium, sodium, iron, and calcium in Gola and Surakhi fruits. Small amount of trace metals, zinc and copper, were also detected in lychee fruit, while heavy metals lead, mercury, and nickel were absent. These two lychee cultivars were also screened for antitumor activity by Potato disc assay with maximum antitumor activity shown by aqueous extract of Surakhi seed (77%) followed by aqueous extract of Gola pulp (74%). Antimicrobial activity of fruit parts was checked by agar well diffusion method against six bacterial strains Enterococcus faecium, Enterococcus faecalis, Staphylococcus aureus, Bacillus subtilis, Bacillus sp. MB083, and Bacillus sp. MB141. Highest antimicrobial activity was shown by methanolic extract of Gola pulp (27mm ± 0.70) and seed (19.5mm ± 0.712) against Enterococcus faecalis. DPPH scavenging assay revealed highest antioxidant activity by aqueous extract of Gola peel (98.10%) followed by n-hexane extract of Surakhi peel (97.73%). Results obtained by reducing power assay also corroborated with the results of DPPH scavenging activity.

Keywords: antimicrobial evaluation, antitumor assay, gola, phytoconstituents, reactive oxygen species, Surakhi

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Authors: Fatma Koksal Cakirlar, Murat Gunaydin, Nevri̇ye Gonullu, Nuri Kiraz

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During the last few decades, the increasing prevalence of methicillin resistant-CoNS isolates has become a common problem worldwide. Macrolide-lincosamide-streptogramin B (MLSB) antibiotics are effectively used for the treatment of CoNS infections. However, resistance to MLSB antibiotics is prevalent among staphylococci. The aim of this study is to determine species distribution and the incidence of inducible clindamycin resistance in CoNS isolates caused nosocomial bacteremia in our hospital. Between January 2014 and October 2015, a total of 484 coagulase-negative CoNS isolates were isolated from blood samples of patients with true bacteremia who were hospitalized in intensive care units and in other departments of Istanbul University Cerrahpasa Medical Hospital. Blood cultures were analyzed with the BACTEC 9120 system (Becton Dickinson, USA). The identification and antimicrobial resistance of isolates were determined by Phoenix automated system (BD Diagnostic Systems, Sparks, MD). Inducible clindamycin resistance was detected using D-test. The species distribution was as follows: Staphylococcus epidermidis 211 (43%), S. hominis 154 (32%), S. haemolyticus 69 (14%), S. capitis 28 (6%), S. saprophyticus 11 (2%), S. warnerii 7 (1%), S. schleiferi 5 (1%) and S. lugdunensis 1 (0.2%). Resistance to methicillin was detected in 74.6% of CoNS isolates. Methicillin resistance was highest in S.hemoliticus isolates (89%). Resistance rates of CoNS strains to the antibacterial agents, respectively, were as follows: ampicillin 77%, gentamicin 20%, erythromycin 71%, clindamycin 22%, trimethoprim-sulfamethoxazole 45%, ciprofloxacin 52%, tetracycline 34%, rifampicin 20%, daptomycin 0.2% and linezolid 0.2%. None of the strains were resistant to vancomycin and teicoplanin. Fifteen (3%) CoNS isolates were D-test positive, inducible MLSB resistance type (iMLSB-phenotype), 94 (19%) were constitutively resistant (cMLSB -phenotype), and 237 (46,76%) isolates were found D-test negative, indicating truly clindamycin-susceptible MS phenotype (M-phenotype resistance). The incidence of iMLSB-phenotypes was higher in S. epidermidis isolates (4,7%) compared to other CoNS isolates.

Keywords: bacteremia, inducible MLSB resistance phenotype, methicillin-resistant, staphylococci

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Authors: Ahilya Singh, Brad Carte, Ramesh Subramani, William Aalbersberg

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A total of 37 actinomycete strains were purified from 25 Solomon Islands marine sediments using four different types of isolation media. Among them, 54% of the strains had obligate requirement of seawater for growth. The ethyl acetate extract of 100 ml fermentation product of each strain was screened for antimicrobial activity against multidrug resistant human pathogens and cytotoxic activity against brine shrimps. A total of 67% of the ethyl acetate extracts showed antimicrobial and/or cytotoxic activities. A strain F-1915 was selected for isolation and evaluation of bioactive compound(s) based on its bioactive properties and chemical profile analysis using the LC-MS. The strain F-1915 was identified to have 96% sequence similarity to Streptomyces violaceusniger on the basis of 16S rDNA sequences using BLAST analysis. The 16S rDNA revealed that the strain F-1915 is a new member of MAR4 clade of actinomycetes. The MAR4 clade is an interesting clade of actinomycetes known for the production of pharmaceutically important hybrid isoprenoid compounds. The ethyl acetate extract of the fermentation product of this strain was purified by silica gel column chromatography and afforded the isolation of one bioactive pure compound. Based on the 1D and 2D NMR spectral data of compound 1 it was identified as a new mono-brominated phenazinone, Marinophenazimycin A, a structure which has already been studied by external collaborators at Scripps Institution of Oceanography but is yet to be published. Compound 1 displayed significant antimicrobial activity against drug resistant human pathogens. The minimum inhibitory concentration (MIC) of compound 1 was against Methicillin Resistant Staphylococcus aureus (MRSA) was about 1.9 μg/ml and MIC recorded against Amphotericin Resistant Candida albicans (ARCA) was about 0.24 μg/ml. The bioactivity of compound 1 against ARCA was found to be better than the standard antifungal agent amphotericin B. Compound 1 however did not show any cytotoxic activity against brine shrimps.

Keywords: actinomycetes, antimicrobial activity, brominated phenazine, MAR4 clade, marine natural products, multidrug resistent, 1D and 2D NMR

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Authors: M. G. Shehata, S. A. El Sohaimy, Marwa M. Abu-Serie, Nourhan M. Abd El-Aziz

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Probiotic strains can potentially be used as bio-preservatives and functional food supplement. Eight lactic acid bacteria strains (LAB) Lactobacillus brevis NRRL B-4527; Streptococcus thermophilus BLM 58; Pediococcusacidilactici ATCC 8042; Lactobacillus rhamnosus CCUG 1452; Lactobacillus curvatus ATCC 51436; Lactococcuslactis sub sp. lactisDSM 20481; Lactobacillus plantarum DMSZ 20079 and Lactobacillus plantarumTF103 were selected to screen the antioxidant, anticancer potential and probiotic properties. LAB strains exhibited good probiotic, antioxidant properties and showed antagonistic activity against food-borne pathogenic (Bacillus subtilis DB 100 host; Candida albicans ATCCMYA-2876; Clostridium botulinum ATCC 3584; Escherichia coli BA 12296; Klebsiellapneumoniae ATCC12296; Salmonella senftenberg ATCC 8400 and Staphylococcus aureus NCTC 10788). Further, in vitro probiotic properties of eight strains displayed excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, in vitro adhesion ability for HT-29 cell line. The antioxidant effect of intracellular and cell-free extract of lactic acid bacteria strains was evaluated by various antioxidant assays, namely, resistance to hydrogen peroxide, DPPH radical scavenging, ABTS radical scavenging, and hydroxyl radical scavenging (HRS). The results showed that intracellular and cell-free supernatant of S. Thermophilus BLM 58, L. lactissubsp.lactis DSM 20481, P. acidilactici ATCC 8042, L. brevis NRRL B-4527 strains possess excellent antioxidant capacity. The intracellular of S. Thermophilus BLM 58 and P. acidilactici ATCC 8042 also showed excellent anticancer activity against Caco-2, MCF-7, HepG-2, and PC-3. Antioxidative property of selected lactic acid bacteria strains would be useful in the functional food manufacturing industry. They could beneficially affect the consumer by providing dietary source of antioxidants.

Keywords: anticancer activity, antioxidant activity, functional food, lactic acid bacteria, probiotic

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Authors: Ebere Christian Ugochukwu, Okafor Josephine, Oyawoye Tomisin

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The occurrence of multidrug resistance patterns that have been developed by bacteria has made it difficult to properly treat infections using standard clinical medications. Hence, the use of herbs as an alternative source of therapy is considered cheap and easily accessible to locals. This research explored the antimicrobial effects of aqueous and ethanolic extracts obtained from Chrysophyllum albidum (commonly called ‘Agbalumo’ in southwest Nigeria and ‘Udara’ in the eastern and southern parts of Nigeria) plant parts (leaves, roots and seeds) against bacteria isolated from urinary catheter tips. The following isolates were obtained; Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Proteus mirabilis, and Klebsiella aerogenes. The agar well diffusion method was used. The average percentages of antimicrobial resistance of the isolates to gentamycin were 45.5% for P. aeruginosa, 42.1% for E. coli, 46.9% for K. aerogenes, and ˃90% for other isolates. Qualitative phytochemical screening of the plant parts extracts was done using chemical test for the screening and identification of bioactive chemical constituents. The ethanolic extract mixtures (leaf, root and seed) had the greatest effect on all the isolates, with inhibition zones (IZs) ranging from 8-26 mm and MICs ranging from <16-32 mg/ml. The Potencies of the C. albidum extracts based on the IZ and MIC values were greater in the extract mixtures, followed by those in the roots. Phytochemical screening revealed that all the extracts contained phenol except for the seeds while tannins were present in all the extracts except the leaves. The activity of the ethanolic extracts of each part at high and low concentrations was greater than that of the aqueous extracts at the same concentrations (p<0.05). The acute toxicity results showed that the LD50 of the extracts was ˃5000 mg/body weight, indicating no toxicity. The antibacterial activities of the extract mixtures and roots on the isolates confirmed the use of C. albidum in folk medicine for the treatment of CAUTIs, hence indicating its antibacterial potential for use in novel antibiotic production.

Keywords: antimicrobials, susceptibility, minimum inhibitory concentration, extracts

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Authors: A. Heshmati, M. Y Alikhani, M. T. Godarzi, M. R. Sadeghimanesh

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Herbal essential oil and extracts are a good source of natural antioxidants and antimicrobial compounds. Hypericum is one of the potential sources of these compounds. In this study, the antioxidant and antimicrobial activity of essential oil and aqueous, methanol, ethanol, ethyl acetate and acetone extract of Hypericum scabrum was assessed. Flowers of Hypericum scabrum were collected from the surrounding mountains of Hamadan province and after drying in the shade, the essential oil of the plant was extracted by Clevenger and water, methanol, ethanol, ethyl acetate and acetone extract was obtained by maceration method. Essential oil compounds were identified using the GC-Mass. The Folin-Ciocalteau and aluminum chloride (AlCl₃) colorimetric method was used to measure the amount of phenolic acid and flavonoids, respectively. Antioxidant activity was evaluated using DPPH and FRAP. The minimum inhibitory concentration (MIC) and the minimum bacterial/fungicide concentration (MBC/MFC) of essential oil and extracts were evaluated against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella typhimurium, Aspergillus flavus and Candida albicans. The essential oil yield of was 0.35%, the lowest and highest extract yield was related to ethyl acetate and water extract. The most component of essential oil was α-Pinene (46.35%). The methanol extracts had the highest phenolic acid (95.65 ± 4.72 µg galic acid equivalent/g dry plant) and flavonoids (25.39 ± 2.73 µg quercetin equivalent/g dry plant). The percentage of DPPH radical inhibition showed positive correlation with concentrations of essential oil or extract. The methanol and ethanol extract had the highest DDPH radical inhibitory. Essential oil and extracts of Hypericum had antimicrobial activity against the microorganisms studied in this research. The MIC and MBC values for essential oils were in the range of 25-25.6 and 25-50 μg/mL, respectively. For the extracts, these values were 1.5625-100 and 3.125-100 μg/mL, respectively. Methanol extracts had the highest antimicrobial activity. Essential oil and extract of Hypericum scabrum, especially methanol extract, have proper antimicrobial and antioxidant activity, and it can be used to control the oxidation and inhibit the growth of pathogenic and spoilage microorganisms. In addition, it can be used as a substitute for synthetic antioxidant and antimicrobial compounds.

Keywords: antimicrobial, antioxidant, extract, hypericum

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Authors: Anyimc, C. J. Aneke, J. O. Orji, O. Nworie, U. C. C. Egbule

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The microbial examination and antimicrobial susceptibility profile of microorganism isolated from the salt mining site in Ebonyi state were evaluated in the present study using a standard microbiological technique. A total of 300 samples were randomly collected in three sample groups (A, B, and C) of 100 each. Isolation, Identification and characterization of organization present on the soil samples were determined by culturing, gram-staining and biochemical technique. The result showed the following organisms were isolated with their frequency as follow: Bacillus species (37.3%) and Staphylococcus species(23.5%) had the highest frequency in the whole Sample group A and B while Klebsiella specie (15.7%), Pseudomonas species(13.7%), and Erwinia species (9.8%) had the least. Rhizopus species (42.0%) and Aspergillus species (26.0%) were the highest fungi isolated, followed by Penicillum species (20.0%) while Mucor species (4.0%), and Fusarium species (8.0%) recorded the least. Sample group C showed high microbial population of all the microbial isolates when compared to sample group A and B. Disc diffusion method was used to determine the susceptibility of isolated bacteria to various antibiotics (oxfloxacin, pefloxacin, ciprorex, augumentin, gentamycin, ciproflox, septrin, ampicillin), while agar well diffusion method was used to determine the susceptibility of isolated fungi to some antifungal drugs (metronidazole, ketoconazole, itraconazole fluconazole). The antibacterial activity of the antibiotics used showed that ciproflux has the best inhibitory effect on all the test bacteria. Ketoconazole showed the highest inhibitory effect on the fungal isolates, followed by itraconazole, while metronidazole and fluconazole showed the least inhibitory effect on the entire test fungal isolates. Hence, the multiple drug resistance of most isolates to appropriate drugs of choice are of great public health concern and cells for periodic monitoring of antibiograms to detect possible changing patterns. Microbes isolated in the salt mining site can also be used as a source of gene(s) that can increase salt tolerance in different crop species through genetic engineering.

Keywords: microorganisms, antibacterial, antifungal, resistance, salt mining site, Ebonyi State

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Authors: Ali Ghorbanipour

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The aim of this study is to investigate the drug resistance pattern and the value of the MIC (minimum inhibitory concentration)method to reduce the impact of infectious diseases and the slow development of resistance. Method: The study was conducted on clinical specimens collected between 2018 to 2021. identification of isolates and antibiotic susceptibility testing were performed using conventional biochemical tests. Antibiotic resistance was determined using kibry-Bauer disk diffusion and MIC by E-test methods comparative with microdilution plate elisa method. Results were interpreted according to CLSI. Results: Out of 249600 different clinical specimens, 18720 different pathogenic bacteria by overall detection ratio 7.7% were detected. Among pathogen bacterial were Gram negative bacteria (70%,n=13000) and Gram positive bacteria(30%,n=5720).Medically relevant gram-negative bacteria include a multitude of species such as E.coli , Klebsiella .spp , Pseudomonas .aeroginosa , Acinetobacter .spp , Enterobacterspp ,and gram positive bacteria Staphylococcus.spp , Enterococcus .spp , Streptococcus .spp was isolated . Conclusion: Our results highlighted that the resistance ratio among Gram Negative bacteria and Gram positive bacteria with different infection is high it suggest constant screening and follow-up programs for the detection of antibiotic resistance and the value of MIC drug susceptibility reporting that provide a new way to the usage of resistant antibiotic in combination with other antibiotics or accurate weight of antibiotics that inhibit or kill bacteria. Evaluation of wrong medication in the expansion of resistance and side effects of over usage antibiotics are goals. Ali ghorbanipour presently working as a supervision at the microbiology department of Massoud medical laboratory. Iran. Earlier, he worked as head department of pulmonary infection in firoozgarhospital, Iran. He received master degree in 2012 from Fergusson College. His research prime objective is a biologic wound dressing .to his credit, he has Published10 articles in various international congresses by presenting posters.

Keywords: antimicrobial profile, MIC & MBC Method, microplate antimicrobial assay, E-test

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Authors: Aditi Taunk, George Iskander, Kitty Ka Kit Ho, Mark Willcox, Naresh Kumar

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Bacterial infections on biomaterial implants and medical devices accounts for 60-70% of all hospital acquired infections (HAIs). Treatment or removal of these infected devices results in high patient mortality and morbidity along with increased hospital expenses. In addition, with no effective strategies currently available and rapid development of antibacterial resistance has made device-related infections extremely difficult to treat. Therefore, in this project we have developed biomaterial surfaces using antibacterial compounds that inhibit biofilm formation by interfering with the bacterial communication mechanism known as quorum sensing (QS). This study focuses on covalent attachment of potent quorum sensing (QS) inhibiting compounds, halogenated furanones (FUs) and dihydropyrrol-2-ones (DHPs), onto glass surfaces. The FUs were attached by photoactivating the azide groups on the surface, and the acid functionalized DHPs were immobilized on amine surface via EDC/NHS coupling. The modified surfaces were tested in vitro against pathogenic organisms such as Staphylococcus aureus and Pseudomonas aeruginosa using confocal laser scanning microscopy (CLSM). Successful attachment of compounds on the substrates was confirmed by X-ray photoelectron spectroscopy (XPS) and contact angle measurements. The antibacterial efficacy was assessed, and significant reduction in bacterial adhesion and biofilm formation was observed on the FU and DHP coated surfaces. The activity of the coating was dependent upon the type of substituent present on the phenyl group of the DHP compound. For example, the ortho-fluorophenyl DHP (DHP-2) exhibited 79% reduction in bacterial adhesion against S. aureus and para-fluorophenyl DHP (DHP-3) exhibited 70% reduction against P. aeruginosa. The results were found to be comparable to DHP coated surfaces prepared in earlier study via Michael addition reaction. FUs and DHPs were able to retain their in vitro antibacterial efficacy after covalent attachment via azide chemistry. This approach is a promising strategy to develop efficient antibacterial biomaterials to reduce device related infections.

Keywords: antibacterial biomaterials, biomedical device-related infections, quorum sensing, surface functionalization

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Authors: Luz Hurtado, Rodrigo Rojas, Jaime Romero, Christopher Concha

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The culture of the marine fish red cusk eel Genypterus chilensis is currently considered a priority for Chilean aquaculture which is a Chilean native species of high gastronomic demand and market value. The microbiota was analyzed in terms of diversity and structure using massive Illumina sequencing. The analysis of alpha diversity was performed in samples of G. chilensis larvae of 6, 18 and 32 dph (days post-hatching) and it was observed that there were significant differences (P = 0.05) between the days of culture for the Chao1 index, being the larvae of 18 dph the one with the highest index followed by the larvae of 6 dph, The lowest value for this index was presented in larvae of 32 dph. There were no significant differences in larvae between the days of culture for the Shannon (P=0.0857) and Simpson (P=0.0714) indices. In general, the larvae of G. chilensis have high rates of diversity. When analyzing the beta diversity, a differentiation between the bacterial communities is observed depending on the day of the culture of the larvae. Considering the PCoA elaborated from the unweighted UniFrac statistic, the explained variance was 46.2% (PC1 29.2% and PC2 17.0%) and in the case of the PCoA elaborated with the weighted UniFrac statistic; the explained variance was 65.5% (PC1 41.8% and PC2 23.7%) these differences were significant based on the Permanova statistical analysis (P= 0.002 and 0.037 respectively). When analyzing the taxonomic composition of the microbiota of the larvae in the different days of culture it was observed that at the phyla level the most abundant in the larvae of 6 dph were Proteobacteria (57%) Verrucomicrobia (24%) and Firmicutes (14%), for the larvae of 18 dph the predominant phyla were Proteobacteria (90%), Dependientiae (5%), Actinobacteria (2%) and Plactomyces (2%), for the larvae of 32 dph the phyla that presented the highest relative abundance were Proteobacteria (57%), Firmicutes (29%), Verrucomicrobia (5%) and Actinobacteria (5%), when comparing the larvae between the days it was observed that the phylum Proteobacteria was the most abundant in the samples of larvae of 6, 18 and 32 dph being the larvae of 18 dph those that present the highest relative abundance, the larvae of 6 dph were those that presented the highest relative abundance for the phylum Verrucomicrobia and in the larvae of 32 dph was observed greater abundance of the phylum Firmicutes compared to the other days of larval culture. At the level of genera, those with the highest relative abundance in larvae of 6 dph were Rubritalea (30%), Psychrobacter (28%), staphylococcus (17%) and Ralstonia (10%), for the larvae of 18 dph the genera with the highest abundance were Psychrobacter (47%), Litoreibacter (13%), Nautella (9%) and Cohesibacter (8%), for the larvae of 32 dph the most abundant genera were Alloiococcus (25%), Dialister (14%), Neptunomonas (13%) and Piscirickettsia (11%). When observing the taxonomic composition of the larvae between the days of larval culture, it is observed that there are differences between them.

Keywords: microbiota, diversity, G. Chilensis, larvae

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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People with diabetes mellitus are more susceptible to developing infections, as high blood sugar levels can weaken the patient's immune system defences. People with diabetes are more adversely affected when they get an infection than someone without the disease, because you have weakened immune defences in diabetes. People who have minimally elevated blood sugar levels experience worse outcomes with infections. Diabetic patients in hospitals do not necessarily have a higher mortality rate due to infections, but they do face longer hospitalisation and recovery times. A study was done in a tertiary care unit in eastern India. Patients with type 2 diabetes mellitus infection were recruited in the study. A total of 520 cases of Type 2 Diabetes Mellitus were recorded out of which 200 infectious cases was included in the study. All subjects underwent detailed history & clinical examination. Microbiological samples were collected from respective site of the infection for microbial culture and antibiotic sensitivity test. Out of the 200 infectious cases urinary tract infection(UTI) was found in majority of the cases followed by diabetic foot ulcer (DFU), respiratory tract infection(RTI) and sepsis. It was observed that Escherichia coli was the most commonest pathogen isolated from UTI cases and Staphylococcus aureus was predominant in foot ulcers followed by other organisms. Klebsiella pneumonia was the major organism isolated from RTI and Enterobacter aerogenes was commonly observed in patients with sepsis. Isolated bacteria showed differential sensitivity pattern against commonly used antibiotics. The majority of the isolates were resistant to several antibiotics that are usually prescribed on an empirical basis. These observations are important, especially for patient management and the development of antibiotic treatment guidelines. It is recommended that diabetic patients receive pneumococcal and influenza vaccine annually to reduce morbidity and mortality. Appropriate usage of antibiotics based on local antibiogram pattern can certainly help the clinician in reducing the burden of infections.

Keywords: antimicrobial resistance, diabetic foot ulcer, respiratory tract infection, urinary tract infection

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Authors: Holm Zerbe, Laura Macias, Hans-Joachim Schuberth, Wolfram Petzl

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Mastitis is among the most important production diseases in cows. It accounts for large parts of antimicrobial drug use in the dairy industry worldwide. Due to the imminent normative to reduce the use of antimicrobial drugs in livestock, new ways for therapy and prophylaxis of mastitis are needed. Recently epigenetic regulation of inflammation by chromatin modifications has increasingly drawn attention. Currently, some epigenetic modifiers have already been approved for the use in humans, however little is known about their actions in the bovine system. The aim of our study was to investigate whether three selected epigenetic modifiers (Vitamin D3, SAHA and S2101) influence the initial immune response towards mastitis pathogens in bovine udder tissue in vitro. Tissue explants of the teat cistern and udder parenchyma were collected from 21 cows and were incubated for 36 hours in the absence and presence of epigenetic modifiers. Additionally, the tissue was stimulated with heat-inactivated particles of Escherichia coli and Staphylococcus aureus, which are regarded as two of the most important mastitis pathogens. After incubation, the explants were tested by RT-qPCR for transcript abundances of immune-related candidate genes. Gene expression was validated in culture supernatants by an AlphaLISA assay. Furthermore, the culture supernatants were analyzed for their chemotactic capacity through a chemotaxis assay. Statistical analysis of data was performed with the program ‘R’ version 3.2.3. Vitamin D3 had no effect on the immune response of udder tissue in vitro after stimulation with mastitis pathogens. The epigenetic modifiers SAHA and S2101 however significantly blocked the pathogen-induced upregulation of CXCL8, TNFα, S100A9 and LAP (P < 0.05). The regulation of IL10 was not affected by treatment with SAHA and S2101. Transcript abundances for CXCL8 were reflected by IL8 contents and chemotactic activity in culture supernatants. In conclusion, these data show the potential of epigenetic modifiers (SAHA and S2101) to block overshooting inflammation in the udder. Thus epigenetic modifiers may serve in future as immune modulators for the treatment and/or prophylaxis of clinical mastitis. (Funded by Deutsche Forschungsgemeinschaft PE 1495/2-1).

Keywords: mastitis, cattle, epigenetics, immunomodulation

Procedia PDF Downloads 235

Authors: Rawia M. Khalil, Ghada Abd-Elbary, Mona Basha, Ghada E. A. Awad, Hadeer A. Elhashemy

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Background: Bacterial infections of the eye are the clinical conditions responsible for ocular morbidity and blindness. Conjunctivitis is an inflammation of the conjunctiva, due to Staphylococcus aureus. Lomefloxacin HCl (LXN) is a third generation flouroquinolone antibiotic with a broad spectrum against wide range of bacteria and very effective against Staph infections especially in conjunctiva (conjunctivitis). The present study aims to develop and evaluate novel ocular proniosomal gels of Lomefloxacin Hcl (LXN); in order to improve its ocular bioavailability for the management of bacterial conjunctivitis. Materials and methods: Proniosomes were prepared by coacervation phase separation method using different types of nonionic surfactants (Span 60,40,20,Tween 20,40,60,80,Brij 35,98,72) solely and as mixtures with Span® 60. The formed gels were characterized for entrapment efficiency, vesicle size and in vitro drug release. The optimum proniosomal gel; P-LXN 7 were characterized for pH measurement, transmission electron microscopy (TEM) and differential scanning calorimetry (DSC) as well as Stability study and microbiological evaluation .The results revealed that only Span 60 was able to form stable LXN proniosomal gel when used individually while the other nonionic surfactants formed gels only in combination with Span 60 at different ratios. The optimum proniosomal gel; P-LXN 7 (Span60:Tween60, 9:1) appeared as spherical shaped vesicles having high entrapment efficiency (>80 %), appropriate vesicle size (187 nm) as well as controlled drug release over 12h. DSC confirmed the amorphous nature and the uniformity of LXN inclusion within the vesicles. Physical stability study did not show any significant changes in appearance or entrapment efficiency or vesicle size after storage for 3 months at 4°C. Ocular irritancy test revealed that P-LXN 7 was safe, well tolerable and suitable for ocular delivery. In vivo antibacterial activity of P-LXN 7 evaluated using the susceptibility test and topical therapy of induced ocular conjunctivitis confirmed the enhanced antibacterial therapeutic efficacy of the LXN-proniosomal gel compared to the commercially available LXN eye drops; Orchacin®. Conclusions: Our results suggest that proniosomal gels could provide a promising carrier of LXN for efficient ocular treatment of bacterial conjunctivitis.

Keywords: bacterial conjunctivitis, lomefloxacin HCl, ocular drug delivery, proniosomes

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Authors: Mohammed Sabbah, Asmaa Al-Asmar, Doaa Abu-Hani, Fuad Al-Rimawi

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Plant extracts are an increasingly popular natural product with a wide range of potential applications in food, industrial, and health care industries. They are rich in polyphenolic compounds and flavonoids, which have been demonstrated to possess a variety of beneficial properties, including antimicrobial and antioxidant activity. Plant extracts have been found to possess antimicrobial activity against a variety of foodborne pathogens and can be used as a natural preservative to extend the shelf life of food products. They have also strong antioxidant activity, which can reduce the formation of free radicals and oxidation of food components. Recently there is an increase interest in bio-based polymers to be used as innovative “bioplastics” for industrial exploitation e.g. packaging materials for food products. Additionally, incorporation of active compounds (e.g. antioxidants and antimicrobials) in bio-polymer materials is of particular interest since such active polymers can be used as active packaging materials (with antimicrobial and antioxidant activity). In this work, different plant extracts have been characterized for their phenolic compounds, flavonoids content, antioxidant activity (both as free radical scavenging ability and reducing ability), and antimicrobial activity against gram positive and negative bacteria (Escherichia coli; Staphylococcus aureus, and Pseudomonas aeruginosa) as well as antifungal activities (against yeast, mold and Botrytis cinera/a plant pathogen). Results showed that many extracts are rich with polyphenolic compounds and flavonoids and have strong antioxidant activities, and rich with phytochemicals (e.g. rutin, quercetin, oleuropein, tyrosol and hydroxytyrosol). Some extracts showed antibacterial activity against both gram positive and negative bacteria as well as antifungal activities and can work, therefore, as preservatives for food or pharmaceutical industries. As an application, two extracts were used as additive to pectin-based packaging film, and results showed that the addition of these extracts significantly improve their functionality as antimicrobial and antioxidant activity. These biomaterials, therefore can be used in food packaging materials to extend the shelf life of food products.

Keywords: plant extracts, antioxidants, flavonoids, bioplastic, edible biofilm, packaging materials

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Authors: Michail Milas, Aikaterini Dafni Tegiou, Nickolas Rigopoulos, Eustathios Giaouris, Zaharias Loannou

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Nanotechnology, a relatively new scientific field, addresses the manipulation of nanoscale materials and devices, which are governed by unique properties, and is applied in a wide range of industries, including food packaging. The incorporation of nanoparticles into polymer matrices used for food packaging is a field that is highly researched today. One such combination is silver nanoparticles with polypropylene. In the present study, the synthesis of the silver nanoparticles was carried out by a natural method. In particular, a ripe banana peel extract was used. This method is superior to others as it stands out for its environmental friendliness, high efficiency and low-cost requirement. In particular, a 1.75 mM AgNO₃ silver nitrate solution was used, as well as a BPE concentration of 1.7% v/v, an incubation period of 48 hours at 70°C and a pH of 4.3 and after its preparation, the polypropylene films were soaked in it. For the PP films, random PP spheres were melted at 170-190°C into molds with 0.8cm diameter. This polymer was chosen as it is suitable for plastic parts and reusable plastic containers of various types that are intended to come into contact with food without compromising its quality and safety. The antimicrobial test against Escherichia coli DFSNB1 and Staphylococcus epidermidis DFSNB4 was performed on the films. It appeared that the films with silver nanoparticles had a reduction, at least 100 times, compared to those without silver nanoparticles, in both strains. The limit of detection is the lower limit of the vertical error lines in the presence of nanoparticles, which is 3.11. The main reasons that led to the adsorption of nanoparticles are the porous nature of polypropylene and the adsorption capacity of nanoparticles on the surface of the films due to hydrophobic-hydrophilic forces. The most significant parameters that contributed to the results of the experiment include the following: the stage of ripening of the banana during the preparation of the plant extract, the temperature and residence time of the nanoparticle solution in the oven, the residence time of the polypropylene films in the nanoparticle solution, the number of nanoparticles inoculated on the films and, finally, the time these stayed in the refrigerator so that they could dry and be ready for antimicrobial treatment.

Keywords: antimicrobial control, banana peel extract, E. coli, natural synthesis, microbe, plant extract, polypropylene films, S.epidermidis, silver nano, random pp

Procedia PDF Downloads 176

Authors: Pampita Chakraborty, Sukumar Mukherjee

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Diabetes Mellitus (DM) is commonly encountered metabolic disorder in clinical practice. An estimated 25 percent of DM patients develop foot problems. Foot ulceration and infection are one of the major causes of morbidity, hospitalization or even amputation. Objective: To isolate and identify bacterial pathogens in Diabetic Foot Ulcer (DFU) and to observe its antimicrobial sensitivity pattern. Methodology: A prospective study was conducted for a period of 9 months at the Department of Microbiology, GD Hospital & Diabetes Institute, Kolkata. 75 DFU patients were recruited in the study. Specimens for microbiological studies obtained from ulcer base were examined as gram stained smear and was cultured aerobically on Nutrient agar, Blood agar and MacConkey agar plates. Antimicrobial sensitivity test was performed by disc diffusion techniques according to CLSI guidelines. Result: In this study out of 75cases, 73% (55/75) were male and 27% (20/75) were females with mean (SD) age of 51.11(±10) years. Out of 75 pus cultures, 63(84%) showed growth of microorganism making total of 81 bacterial isolates with 71.42% of monomicrobial infection and 28.57% of polymicrobial infection. Out of 81 isolates 53(65.43%) were gram negative and 21(25.92%) were gram positive. E.coli was relatively common isolate 21(26%) followed by Staphylococcus aureus 15(18.5%), Klebsiella pneumonia 14(17.28%), Pseudomonas aeruginosa 12 (14.81%), Proteus spp. 3 (3.70%), and Enterococcus faecalis 6 (7.40%). 75% of Gram-negative microorganism were extended Beta-lactamase enzyme (ESBL) producer and around 20 % of Klebsiella and Proteus spp. were carbapenemase enzyme producer. Among Gram positive, around 50% of S.aureus was MRSA, sensitive only to Vancomycin, Teicoplanin & Linezolid. Conclusion: More prevalence of monomicrobial gram-negative bacteria than gram-positive bacteria in DFU was observed. This study emphasizes that Beta-Lactam group of antibiotics should not be the empirical treatment of choice for Gram-negative isolates; instead alternatives like Carbapenems, Amikacin could be a better option. On the other hand, Vancomycin and Linezolid are preferred for most of the infection with gram-positive aerobes. Continuous surveillance of resistant bacteria is required for empiric therapy.

Keywords: antibiotic resistant, antimicrobial susceptibility, diabetic foot ulcer, surveillance

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Authors: Ade O. Oyewole, Sunday O. Okoh, Ruth O. Ishola, Adenike D. Odusote, Chima C. Igwe, Gloria N. Elemo, Anthony I. Okoh

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Azadirachta indica (Neem tree) also referred to as an all-purpose tree is used in a wide range of medical preparations in tropical and subtropical countries for prevention and management of various livestock, crops products and human diseases. In Nigeria however, the potentials of this plant have not been fully exploited thus it causes an environmental nuisance during the fruiting season. With a rise in the demand for herbal personal care products globally extracts from different parts of the neem plant were used as the bio-active ingredients in the formulation of personal care products. In this study, formulated neem soap, body cream, lotion, toothpaste and shampoo are analyzed to determine their antibacterial, antifungal, and toxicity properties. The efficacies of these products for management of infectious diseases, both oral and dermal, were also investigated in vitro. Oil from the neem seeds obtained using a mechanical press and acetone extracts of both the neem bark and leaves obtained by the maceration method were used in the formulation and production of the neem personal care products. The antimicrobial and toxicity properties of these products were investigated by agar diffusion, and haemolytic methods respectively. The five neem products (NPs) exhibited strong antibacterial activities against four multi–drug resistant pathogenic and three none pathogenic bacterial strains (Escherichia coli (180), Listeria ivanovii, Staphylococcus aureus, Enterobacter cloacae, Vibro spp., Streptococcus uberis, Mycobacterium smegmatis), except the neem lotion with insignificant activity against E. coli and S. aureus. The minimum inhibitory concentration (MIC) range was between 0.20-0.40 mg/ mL. The 5 NPs demonstrated moderate activity against three clinical dermatophytes isolates (Tinea corporis, Tinea capitis, and Tinea cruiz) as well as one fungal strain (Candida albican) with the MIC ranging between 0.30 - 0.50 mg/ mL and 0.550 mg/mL respectively. The soap and shampoo were the most active against test bacteria and fungi. The haemolytic analysis results on the 5 NPs indicated none toxicity at 0.50 mg/ mL in sheep red blood cells (SRBC).

Keywords: antimicrobial, Azadirachta indica, multi–drug resistant pathogenic bacteria, personal care products

Procedia PDF Downloads 270

Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

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Authors: Fatma A. Mohamed, Hend M. Ahmed

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This research explores the antimicrobial effects of dielectric barrier discharge (DBD) oxygen plasma treatment combined with ZnO NPs on the cotton fabric, focusing on various treatment durations (5, 10, 15, 20, and 30 minutes) and discharge powers (15.5–17.35 watts) at flow rate 0.5 l/min. After treatment with oxygen plasma and ZnO NPs, the fabric was printed with green tea (Camellia sinensis) at five different concentrations. The study evaluated the treatment's effectiveness by analyzing surface wettability, specifically through wet-out time and hydrophilicity, as well as measuring contact angles. To investigate the chemical changes on the fabric's surface, attenuated total reflectance–Fourier transform infrared (ATR-FTIR) spectroscopy was employed to identify the functional groups formed as a result of the plasma treatment. This comprehensive approach aims to understand how DBD oxygen plasma treatment and ZnO nanoparticles change cotton fabric properties and enhance its antimicrobial potential, paving the way for innovative applications in textiles. In addition to the chemical analysis, the surface morphology of the O₂ plasma/ZnO NPs-treated cotton fabric was examined using scanning electron microscopy (SEM). FTIR analysis revealed an increase in polar functional groups (-COOH, -OH, and -C≡O) on the fabric's surface, contributing to enhanced hydrophilicity and functionality. The antimicrobial properties were evaluated using qualitative and quantitative methods, including agar plate assays and modified Hoenstein tests against Staphylococcus aureus and Escherichia coli. The results indicated a significant improvement in antimicrobial effectiveness for the cotton fabric treated with plasma and coated with natural extracts, maintaining this efficacy even after four washing cycles. This research demonstrates that utilizing oxygen DBD plasma/ZnO NPs treatment, combined with the absorption of tea and tulsi leaf extracts, presents a promising strategy for developing natural antimicrobial textiles. This approach is particularly relevant given the increasing medical and healthcare demands for effective antimicrobial materials. Overall, the method not only enhances the absorption of plant extracts but also significantly boosts antimicrobial efficacy, offering valuable insights for future textile applications.

Keywords: cotton, ZnO NPs, green tea leaf, antimicrobial avtivity, DBD oxygen plasma

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Authors: Kichan Lee, Kwang-Ho Choi, Mi-Hye Hwang, Young Min Son, Bang-Hun Hyun, Byeong Yeal Jung

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Recently, worldwide food safety authorities have been strengthening food hygiene in order to curb foodborne illness outbreaks. The hygiene status of Korean slaughterhouses has been monitored annually by Animal and Plant Quarantine Agency and provincial governments through foodborne pathogens investigation using slaughtered pig and cattle meats. This study presented the prevalence of food-borne pathogens from 2014 to 2016 in Korean slaughterhouses. Sampling, microbiological examinations, and analysis of results were performed in accordance with ‘Processing Standards and Ingredient Specifications for Livestock Products’. In total, swab samples from 337 pig carcasses (100 samples in 2014, 135 samples in 2015, 102 samples in 2016) and 319 cattle carcasses (100 samples in 2014, 119 samples in 2015, 100 samples in 2016) from twenty slaughterhouses were examined for Listeria monocytogenes, Campylobacter jejuni, Campylobacter coli, Salmonella spp., Staphylococcus aureus, Clostridium perfringens, Yersinia enterocolitica, Escherichia coli O157:H7 and non-O157 enterohemorrhagic E. coli (EHEC, serotypes O26, O45, O103, O104, O111, O121, O128 and O145) as foodborne pathogens. The samples were analyzed using cultural and PCR-based methods. Foodborne pathogens were isolated in 78 (23.1%) out of 337 pig samples. In 2014, S. aureus (n=17) was predominant, followed by Y. enterocolitica (n=7), C. perfringens (n=2) and L. monocytogenes (n=2). In 2015, C. coli (n=14) was the most prevalent, followed by L. monocytogenes (n=4), S. aureus (n=3), and C. perfringens (n=2). In 2016, S. aureus (n=16) was the most prevalent, followed by C. coli (n=13), L. monocytogenes (n=2) and C. perfringens (n=1). In case of cattle carcasses, foodborne bacteria were detected in 41 (12.9%) out of 319 samples. In 2014, S. aureus (n=16) was the most prevalent, followed by Y. enterocolitica (n=3), C. perfringens (n=3) and L. monocytogenes (n=2). In 2015, L. monocytogenes was isolated from 4 samples, S. aureus from three, C. perfringens, Y. enterocolitica and Salmonella spp. from one, respectively. In 2016, L. monocytogenes (n=6) was the most prevalent, followed by C. perfringens (n=3) C. jejuni (n=1), respectively. It was found that 10 carcass samples (4 cattle and 6 pigs) were contaminated with two bacterial pathogen tested. Interestingly, foodborne pathogens were more detected from pig carcasses than cattle carcasses. Although S. aureus was predominantly detected in this study, other foodborne pathogens were also isolated in slaughtered meats. Results of this study alerted the risk of foodborne pathogen infection for humans from slaughtered meats. Therefore, the authors insisted that it was important to enhance hygiene level of slaughterhouses according to Hazard Analysis and Critical Control Point.

Keywords: carcass, cattle, foodborne, Korea, pathogen, pig

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Authors: Maryam Beiranvand

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Multi-drug resistance in various microorganisms has increased globally in many healthcare facilities. Less effective antimicrobial activity of drug therapies for infection control becomes trouble. Since 1980, no new type of antimicrobial drug has been identified, even though combinations of antibiotic drugs have been discovered almost every decade. Between 1981 and 2006, over 70% of novel pharmaceuticals and chemical agents came from natural sources. Microorganisms have yielded almost 22,000 natural compounds. The identification of antimicrobial components from endophytes bacteria could help overcome the threat posed by multi-drug resistant strains. The project aims to analyze and identify antimicrobial peptides isolated from entophytic bacteria and their activity against multidrug-resistant Gram-negative bacteria in South Korea. Endophytic Paenibacillus polymyxa. 4G3 isolated from the plant, Gynura procumbery exhibited considerable antimicrobial activity against Methicillin-resistant Staphylococcus aureus, and Escherichia coli. The Rapid Annotations using Subsystems Technology showed that the total size of the draft genome was 5,739,603bp, containing 5178 genes with 45.8% G+C content. Genome annotation using antiSMASH version 6.0.0 was performed, which predicted the most common types of non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS). In this study, diethyl aminoethyl cellulose (DEAEC) resin was used as the first step in purifying for unknown peptides, and then the target protein was identified using hydrophilic and hydrophobic solutions, optimal pH, and step-by-step tests for antimicrobial activity. This crude was subjected to C18 chromatography and elution with 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% methanol, respectively. Only the fraction eluted with 20% -60% methanol demonstrated good antimicrobial activity against MDR E. coli. The concentration of the active fragment was measured by the Brad-ford test, and Protein A280 - Thermo Fisher Scientific at the end by examining the SDS PAGE Resolving Gel, 10% Acrylamide and purity were confirmed. Our study showed that, based on the combined results of the analysis and purification. P polymyxa. 4G3 has a high potential exists for producing novel functions of polymyxin E and bacitracin against bacterial pathogens.

Keywords: endophytic bacteria, antimicrobial activity, antimicrobial peptide, whole genome sequencing analysis, multi -drug resistance gram negative bacteria

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Authors: Thayse Marques Passos, Brid Quilty, Mary Pryce

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The interest in developing alternative techniques to obtain safe water, free from pathogens and hazardous substances, is growing in recent times. The photodynamic inactivation of microorganisms (PDI) is a promising ecologically-friendly and multi-target approach for water disinfection. It uses visible light as an energy source combined with a photosensitiser (PS) to transfer energy/electrons to a substrate or molecular oxygen generating reactive oxygen species, which cause cidal effects towards cells. PDI has mainly been used in clinical studies and investigations on its application to disinfect water is relatively recent. The majority of studies use planktonic cells. However, in their natural environments, bacteria quite often do not occur as freely suspended cells (planktonic) but in cell aggregates that are either freely floating or attached to surfaces as biofilms. Microbes can form aggregates and biofilms as a strategy to protect them from environmental stress. As aggregates, bacteria have a better metabolic function, they communicate more efficiently, and they are more resistant to biocide compounds than their planktonic forms. Among the bacteria that are able to form aggregates are members of the genus Pseudomonas, they are a very diverse group widely distributed in the environment. Pseudomonas species can form aggregates/biofilms in water and can cause particular problems in water distribution systems. The aim of this study was to evaluate the effectiveness of photodynamic inactivation in killing a range of planktonic cells including Escherichia coli DSM 1103, Staphylococcus aureus DSM 799, Shigella sonnei DSM 5570, Salmonella enterica and Pseudomonas putida DSM 6125, and aggregating cells of Pseudomonas fluorescens DSM 50090, Pseudomonas aeruginosa PAO1. The experiments were performed in glass Petri dishes, containing the bacterial suspension and the photosensitiser, irradiated with a multi-LED (wavelengths 430nm and 660nm) for different time intervals. The responses of the cells were monitored using the pour plate technique and confocal microscopy. The study showed that bacteria belonging to Pseudomonads group tend to be more tolerant to PDI. While E. coli, S. aureus, S. sonnei and S. enterica required a dosage ranging from 39.47 J/cm2 to 59.21 J/cm2 for a 5 log reduction, Pseudomonads needed a dosage ranging from 78.94 to 118.42 J/cm2, a higher dose being required when the cells aggregated.

Keywords: bacterial aggregation, photoinactivation, Pseudomonads, water disinfection

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Authors: Christoph Hacker, Zeynep Karahaliloglu, Gunnar Seide, Emir Baki Denkbas, Thomas Gries

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A wound dressing material is designed to facilitate wound healing and minimize scarring. An ideal wound dressing material should protect the wound from any contaminations of exogeneous microorganism. In addition, the dressing material should provide a moist environment through extraction of body fluid from the wound area. Recently, wound dressing electrospun nanofibrous membranes are produced by electrospinning from a polymer solution or a polymer melt. These materials have a great potential as dressing materials for wound healing because of superior properties such as high surface-to-volume ratio, high porosity with excellent pore interconnectivity. Melt electrospinning is an attractive tissue engineering scaffold manufacturing process which eliminated the health risk posed by organic solvents used in electrospinning process and reduced the production costs. In this study, antibacterial wound dressing materials were prepared from TPU (Elastollan 1185A) by a melt-electrospinning technique. The electrospinning parameters for an efficient melt-electrospinning process of TPU were optimized. The surface of the fibers was modified with poly(ethylene glycol) (PEG) by radio-frequency glow discharge plasma deposition method and with silver nanoparticles (nAg) to improve their wettability and antimicrobial properties. TPU melt-electrospun mats were characterized using SEM, DSC, TGA and XPS. The cell viability and proliferation on modified melt-electrospun TPU mats were evaluated using a mouse fibroblast cell line (L929). Antibacterial effects of theirs against both Staphylococcus aureus strain and Escherichia coli were investigated by disk-diffusion method. TPU was successfully processed into a porous, fibrous network of beadless fibers in the micrometer range (4.896±0.94 µm) with a voltage of 50 kV, a working distance of 6 cm, a temperature of the thermocouple and hot coil of 225–230ºC, and a flow rate of 0.1 mL/h. The antibacterial test indicated that PEG-modified nAg-loaded TPU melt-electrospun structure had excellent antibacterial effects and cell study results demonstrated that nAg-loaded TPU mats had no cytotoxic effect on the fibroblast cells. In this work, the surface of a melt-electrospun TPU mats was modified via PEG monomer and then nAg. Results showed melt-electrospun TPU mats modified with PEG and nAg have a great potential for use as an antibacterial wound dressing material and thus, requires further investigation.

Keywords: melt electrospinning, nanofiber, silver nanoparticles, wound dressing

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Authors: Abdul Walusansa

Abstract:

In this paper, paper notes and coins were collected from general public in Kampala City where ready-to-eat food can be served, in order to survey for bacterial contamination. The total bacterial number and potentially pathogenic organisms loading on currency were tested. All isolated potential pathogens were also tested for antibiotic resistance against four most commonly prescribed antibiotics. 1. The bacterial counts on one hundred paper notes sample were ranging between 6～10918/cm cm-2,the median was 141/ cm-2, according to the data it was much higher than credit cards and Australian notes which were made of polymer. The bacterial counts on sixty coin samples were ranging between 2～380/cm-2, much less than paper notes. 2. Coliform (65.6%), E. coli (45.9%), S. aureus (41.7%), B. cereus (67.7%), Salmonella (19.8%) were isolated on one hundred paper notes. Coliform (22.4%), E. coli (5.2%), S. aureus (24.1%), B. cereus (34.5%), Salmonella (10.3%) were isolated from sixty coin samples. These results suggested a high rate of potential pathogens contamination of paper notes than coins. 3. Antibiotic resistances are commonly in most of the pathogens isolated on currency. Ampicillin resistance was found in 60%of Staphylococcus aureus isolated on currency, as well as 76.6% of E. coil and 40% of Salmonella. Erythromycin resistance was detected in 56.6% of S. aureus and in 80.0% of E. coli. All the pathogens isolated were sensitive to Norfloxacin, Salmonella and S. aureus also sensitive to Cefaclor. In this paper, we also studied the antimicrobial capability of metal coins, coins collected from different countries were tested for the ability to inhibit the growth of E. sakazakii, S. aureus, E. coli, L. monocytogenes and S. typhimurium. 1) E. sakazakii appeared very sensitive to metal coins, the second is S. aureus, but E. coli, L. monocytogenes and S. typhimurium are more resistant to these metal coin samples. 2) Coins made of Nickel-brass alloy and Copper-nickel alloy showed a better effect in anti-microbe than other metal coins, especially the ability to inhibited the growth of E. sakazakii and S. aureus, all the inhibition zones produced on nutrient agar are more than 20.6 mm. Aluminium-bronze alloy revealed weak anti-microbe activity to S. aureus and no effect to kill other pathogens. Coins made of stainless steel also can’t resist bacteria growth. 3) Surprisingly, one cent coins of USA which were made of 97.5% Zinc and 2.5% Cu showed a significant antimicrobial capability, the average inhibition zone of these five pathogens is 45.5 mm.

Keywords: antibiotic sensitivity, bacteria, currency, coins, parasites

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Authors: N. Hadhoum, B. Guerfi, T. M. Sider, Z. Yassa, T. Djerboua, M. Boursouti, M. Mamou, F. Z. Hadjadj Aoul, L. R. Mekacher

Abstract:

Introduction and objectives: Chlorobutanol is a raw material, mainly used as an antiseptic and antimicrobial preservative in injectable and ophthalmic preparations. The main objective of our study was the synthesis and evaluation of the antimicrobial activity of chlorobutanol hemihydrates. Material and methods: Chlorobutanol was synthesized according to the nucleophilic addition reaction of chloroform to acetone, identified by an infrared absorption using Spectrum One FTIR spectrometer, melting point, Scanning electron microscopy and colorimetric reactions. The dosage of carvedilol active substance was carried out by assaying the degradation products of chlorobutanol in a basic solution. The chlorobutanol obtained was subjected to bacteriological tests in order to study its antimicrobial activity. The antibacterial activity was evaluated against strains such as Escherichia coli (ATCC 25 922), Staphylococcus aureus (ATCC 25 923) and Pseudomonas aeroginosa (ATCC = American type culture collection). The antifungal activity was evaluated against human pathogenic fungal strains, such as Candida albicans and Aspergillus niger provided by the parasitology laboratory of the Hospital of Tizi-Ouzou, Algeria. Results and discussion: Chlorobutanol was obtained in an acceptable yield. The characterization tests of the product obtained showed a white and crystalline appearance (confirmed by scanning electron microscopy), solubilities (in water, ethanol and glycerol), and a melting temperature in accordance with the requirements of the European pharmacopoeia. The colorimetric reactions were directed towards the presence of a trihalogenated carbon and an alcohol function. The spectral identification (IR) showed the presence of characteristic chlorobutanol peaks and confirmed the structure of the latter. The microbiological study revealed an antimicrobial effect on all strains tested (Sataphylococcus aureus (MIC = 1250 µg/ml), E. coli (MIC = 1250 µg/ml), Pseudomonas aeroginosa (MIC = 1250 µg/ml), Candida albicans (MIC =2500 µg/ml), Aspergillus niger (MIC =2500 µg/ml)) with MIC values close to literature data. Conclusion: Thus, on the whole, the synthesized chlorobutanol satisfied the requirements of the European Pharmacopoeia, and possesses antibacterial and antifungal activity; nevertheless, it is necessary to insist on the purification step of the product in order to eliminate the maximum impurities.

Keywords: antimicrobial agent, bacterial and fungal strains, chlorobutanol, MIC, minimum inhibitory concentration

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