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Assessment of Antimicrobial Activity of Punica Granatum against Antibiotic-Resistant Clostridium Perfringens Type (D)
Abstract
The search for new antibiotics and alternative products to solve the increasing number of bacterial resistance to customary antibiotics has become an urgent need. To investigate the effectiveness of the extracts prepared from different parts of Syrian Punica granatum Linn (family Punicaceae), against Clostridium perfringens type (D), which is resistant against many antibiotics, 684 samples were isolated from intestines and livers of death goats by using blood agar, and a selective agar for growing of Clostridium perfringens(SPS agar). The isolated bacteria were typed by using ELISA apparatus. Many parts of Punica granatum was extracted with water, absolute alcohol, then ether by using soxhlet apparatus and rotary evaporator. The Antibiotic susceptibility testing of many antibiotics was conducted by using disc diffusion method in anaerobic atmosphere and break points method. The alcoholic extracts prepared from many parts of punica granatum (pericarp, leaves, flowers, seeds) showed different antibacterial effect against Clostridium perfringens type(D),whereas the studied antibiotics had not shown any antibacterial effect, except Clindamycin which showed partial effectiveness.
Despite the abundance of promising studies, developments, and improvements about the elimination of microbial toxins from food matrices, they are still considered as one of the major food safety problems due to the lack of their complete avoidance even today. Every year, many crops and foodstuffs have to be discarded due to unconstrained contamination and/or production of microbial toxins. Furthermore, the difficulty for the detection of toxin presence and determination of its level in foods may lead to acute or chronic health problems in many individuals. On the other hand, phenolic compounds might be considered as microbial toxin detoxification agents because of their inhibition effect on the toxin synthesis of microorganisms or exhibiting protective effects against varying damaging mechanisms caused by toxins. In this study, the effect of phenolic compounds on the synthesis of bacterial toxins and mycotoxins is comprehensively reviewed. The potential curing effect of phenolic compounds against toxin-induced damages has also been discussed. Consequently, phenolic compounds are indicated as promising, and considerable natural preservatives against toxin damages and their detoxification potentials are pronounced.
Punica granatum L, cv. Hicaznar and three different genotypes, which are named genotype 19-121, 17-67and 19-66 which have been obtained at Batı Akdeniz Agricultural Research Institute (BATEM) in Antalya, have been studied for determination of their antibacterial and antifungal activities. The antibacterial activity of peel extracts was tested against three bacteria strains, which were named Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922) and Salmonella Enteritidis (ATCC 13076). The antifungal activity was tested against two fungal strains, which were named Aspergillus parasiticus NRRL 2999 and Aspergillus parasiticus NRRL 465. All extracts possessed remarkable antibacterial and antifungal activities against all tested bacterial and fungal strains. Among the extracts genotype 19-66 peel extracts had lowest antimicrobial activity, although its peel had highest total phenolic content. Also its acidity was determined to be remarkably lowest as compared other Punica granatum L peels. This result may explain that antibacterial effect of phenolic compounds could be increasing with presence of organic acids.
Clostridium perfringens is a gram-positive, anaerobic bacterium that causes a wide range of diseases in humans and animals. Th e purposes of this study were to determine, using the enzyme-linked immunosorbent assay (ELISA), the clostridial toxins in intestinal samples of lambs with suspected enterotoxemia; to conduct molecular typing of C. perfringens isolates; and to investigate the presence of the genes of 4 major toxins (α, β, ε, and ι) in the isolates by polymerase chain reaction (PCR). According to the ELISA results, clostridial toxins were determined in 32 intestinal samples. Of the samples, 13 (40.62%) were C. perfringens type A, 1 (3.125%) was C. perfringens type B, 9 (28.125%) were C. perfringens type C, and 9 (28.125%) were C. perfringens type D. C. perfringens was isolated from 13 (8.66%) out of 150 intestinal samples. A total of 13 C. perfringens isolates from lambs were genotyped by PCR. Of the isolates, 10 (76.92%) were type A, 2 (%15.38) were type D, and 1 (7.69%) was type C. Type B and type E were not identifi ed. Th e results showed that C. perfringens type A was the most common type by both ELISA and PCR in lambs with enterotoxemia.
Nowadays, uncontrolled and frequent use of antibiotics may cause emergence of microbial resistance among pathogenic agents. Therefore, the use of new synthetic and natural antimicrobial compounds is inevitable. One source of natural compounds in this respect comes from plants. The purpose of this study was to examine the antibacterial effects of peel extracts from sour and sweet pomegranate. Methanolic extracts of sour and sweet pomegranate peels and aqueous solutions of tetracycline and chloramphenicol were prepared. Antibiogram tests using disk diffusion technique and serial dilution method were performed against ten pathogenic bacteria isolated from animals, and relative minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were also determined for the above compounds. The greatest zone of inhibition induced by the action of pomegranate peel extracts was obtained for Staphylococcus aureus (about 25 mm) and the smallest zone of inhibition was obtained for Pasteurella multocida (about 9 mm). In addition, the lowest MIC and MBC values of pomegranate peel extract were obtained for Staphylococcus aureus (7.8 and 62.5 mg/ml, respectively). Results of serial dilution tests indicate that bactericidal effect of sour pomegranate peel extract was more than that for sweet pomegranate peel extract; and sweet pomegranate peel extract exerts a bacteriostatic action against bacteria. The antibacterial effect was greater against Gram-positive bacteria compared to that for the Gram-negative bacteria. Effects of these extracts were considerably lower than those for tetracycline and chloramphenicol. In conclusion, methanolic extracts of pomegranate peels exhibit relatively good bacteriostatic and bactericidal effects.
Automatic face annotation (or tagging) facilitates improved retrieval and organization of personal photos in online social networks. In this paper, we present a new collaborative face recognition (FR) method that aims to improve face annotation accuracy. The proposed method makes efficient use of multiple FR engines and databases that are distributed over an online social network. The performance of our collaborative face recognition method was successfully evaluated using the standard MPEG-7 VCE-3 data set and a set of real-world personal photos from the Web. The efficacy of the proposed method is demonstrated in terms of comparative annotation performance against non-collaborative approaches utilizing a single FR engine and a single database only.
Inflammation can cause various physical dysfunctions. Punica granatum Linne (pomegranate), a high phenolic content fruit, is widely used as an antipyretic analgesic in Chinese culture. Pomegranate has shown potential nitric oxide (NO) inhibition in LPS-induced RAW 264.7 macrophage cells. Moreover, pomegranate (100 mg/kg) significantly decreased carrageenan-induced mice paw edema for 1, 3, 4, and 5 h. Therefore, column chromatography combined with in vitro bioassay-guided fractionation was used to isolate the active anti-inflammatory components from the pomegranate. Punicalagin (1), punicalin (2), strictinin A (3), and granatin B (4) were obtained with yields of 0.093%, 0.015%, 0.003%, and 0.013%, respectively. All these hydrolysable tannins inhibited NO production and iNOS expression in RAW 264.7 cells. Among them, 4 showed the strongest iNOS and COX-2 inhibitory effects, and exhibited these effects in the inhibition of paw swelling and the PGE2 level in carrageenan-induced mice. Taken together, we suggest that 4 could be used as a standard marker for the anti-inflammatory effect of pomegranate.
The in vitro antimicrobial and antioxidant activities of the crude methanol extracts of nine Hypericum species (H. barbatum Jacq., H. hirsutum L., H. linarioides Bosse, H. maculatum Crantz, H. olympicum L., H. perforatum L., H. richeri Vill., H. rumeliacum Boiss. and H. tetrapterum Fries) growing spontaneously in the Balkans were screened using a disk diffusion assay against a panel of standardized bacteria and fungi and a phosphomolybdenum method, respectively. The most active extract against all microbial strains was that of Hypericum hirsutum (collected at Vranjska Banja, South Serbia), which showed remarkable antimicrobial activity at a dose of 5 μg per disk and a diameter of growth inhibition zone reaching values more than two times larger than that of standard antibiotics. The antimicrobial activity of H. barbatum, H. hirsutum, H. olympicum, H. richeri and H. tetrapterum has not hitherto been published. All the extracts possess a very broad spectrum of strong antimicrobial activity. All the investigated species, except H. perforatum, have been tested for the first time in an antioxidant capacity assay. The results are correlated and discussed with respect to the until now known constituents of the investigated species.
Improper cooking of meat and poultry products may cause food borne gastroenteritis by Clostridium perfringens due to its enterotoxin. In this study we isolated Clostridium perfringens from cooked meat, pork and poultry samples from different areas of Coimbatore District, Tamilnadu, India. The presumptive, confirmed and completed characterization showed that isolated organism was Clostridium perfringens and it produces various toxins during sporulation. The toxin detected by simple immunodiffusion and various temperatures treatment showed its hyperthermophlicity. The antibiogram has indicated resistance of the organism to conventional antibiotics hence in silico study is significant. Comparative modeling of the δ-enterotoxin, most important toxin in gastroenteritis, was carried out with its protein sequence present in NCBI as no crystallographic structure of the same is available. PSI-BLAST analysis indicated the δ-toxin showed perfect homology (31% identity and 51 % similarity) with Staphylococcal β-hemolysin. This has confirmed by MSA and phylogentic analysis. The ORF has 1300 coding frames and predicted secondary structure consist of mainly randomcoil. The model refinement and validation is done by various empirical force fields and RMSD value, 1.43 A°, showed that the model is good. Most residues of the toxin are located in allotted regions of Ramchandran plot. The modeled structure could give novel ideas for molecular docking, development of new agonist and potential target for studies of drug- receptor interaction.
Arils from six pomegranate (Punica granatum L.) varieties grown in the Mediterranean region of Turkey were tested for their antimicrobial properties by the agar diffusion and minimum inhibitory concentration (MIC) methods against seven bacteria: (Bacillus megaterium DSM 32, Pseudomonas aeruginosa DSM 9027, Staphylococcus aureus Cowan 1, Corynebacterium xerosis UC 9165, Escherichia coli DM, Enterococcus faecalis A10, Micrococcus luteus LA 2971), and threefungi (Kluvyeromyces marxianus A230, Rhodotorula rubra MC12, Candida albicans ATCC 1023). It has been observed that the pomegranate aril extracts had antimicrobial effect on all microorganisms, giving inhibition zones ranging in size from 13 to 26 mm. The MIC values for active pomegranate extracts ranged between 30 and >90 microg/mL. The results obtained appeared to confirm the antimicrobial potential of the Punica granatum varieties.
The present study investigated the effects of Punica granatum aqueous extract (PgAq) on streptozotocin (STZ) induced diabetic rats by measuring fasting blood glucose, lipid profiles (atherogenic index), lipid peroxidation (LPO) and activities of both non-enzymatic and enzymatic antioxidants. Diabetes was induced by single intraperitoneal injection of STZ (60 mg/kg) to albino Wistar rats. The increase in blood glucose level, total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), very low density lipoprotein (VLDL), LPO level with decrease in high density lipoprotein cholesterol (HDL-C), reduced glutathione (GSH) content and antioxidant enzymes namely, glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) were the salient features observed in diabetic rats. On the other hand, oral administration of PgAq at doses of 250 mg/kg and 500 mg/kg for 21 days resulted in a significant reduction in fasting blood glucose, TC, TG, LDL-C, VLDL-C and tissue LPO levels coupled with elevation of HDL-C, GSH content and antioxidant enzymes in comparison with diabetic control group. The results suggest that PG could be used, as a dietary supplement, in the treatment of chronic diseases characterized by atherogenous lipoprotein profile, aggravated antioxidant status and impaired glucose metabolism and also in their prevention.
There have been considerable changes to the format of the recommendations since the previous version (version 7). The majority of the footnotes to the tables have been removed and the notations added to the end column; it is hoped that this change will avoid confusion in interpretation. Antibiotics have been separated into groups, e.g. beta-lactams, aminoglycosides, etc. Recommendations for urinary tract infections (UTIs) have been removed for most agents except for those that are administered solely for the treatment of uncomplicated UTIs or where there are limited recommendations for specific organisms, e.g. trimethoprim. For agents that previously had dual recommendations, systemic recommendations remain and the intermediate category can be used for interpretation for UTIs because intermediate susceptibility infers that the infection may respond as the agent is concentrated at the site of infection. This change will also avoid errors in interpretation when an organism is isolated from multiple sites, e.g. blood and urine. The changes that have been made to version 7 are as follows: MIC and zone diameter breakpoints (BPs) for trimethoprim, fosfomycin and nitrofurantoin for UTIs (Table 7); MIC and zone diameter breakpoints (BPs) for doripenem (Tables 7-9); colistin MIC BPs for Pseudomonas spp. (Table 9), co-trimoxazole MIC BPs for Stenotrophomonas maltophilia (Table 10); staphylococci MIC and zone diameter BPs for clarithromycin, clindamycin, erythromycin, quinupristin/dalfopristin, trimethoprim UTI, nitrofurantoin UTI and rifampicin (Table 11); Streptococcus pneumoniae MIC and zone diameter BPs for azithromycin, clarithromycin, erythromycin, co-trimoxazole, linezolid, rifampicin and telithromycin (Table 12); addition of streptomycin recommendations for enterococci (Table 13); enterococcal MIC and zone diameter BPs for quinupristin/dalfopristin, nitrofurantoin UTI and trimethoprim UTI (Table 13); beta-haemolytic streptococci MIC and zone diameter BPs for azithromycin, clarithromycin, erythromycin and telithromycin (Table 15); clarithromycin and erythromycin MIC and zone diameter BPs for Moraxella catarrhalis (Table 16); azithromycin MIC BPs for Neisseria gonorrhoeae (Table 17); chloramphenicol and rifampicin MIC BPs for Neisseria meningitidis (Table 18); azithromycin MIC BPs for Haemophilus influenzae (Table 19); MIC BPs for metronidazole for Bacteroides fragilis, Bacteroides thetaiotaomicron and Clostridium perfringens (Tables 23-25, respectively); susceptibility testing of Listeria spp. (Appendix 3); the acceptable range for ATCC 25923 to a 10 microg tobramycin disc (Table 26).