Comparative analysis of the phytochemical and antibacterial activity of the root extracts of Euphorbia heterophylla and Vitellaria paradoxa

Background: Over time, herbal plants and their various components have been major sources of therapeutic medicine for man. A comparative study was carried out to determine the phytochemical components and antibacterial activities of the different crude extracts of Euphorbia heterophylla and Vitellaria paradoxa roots on four enteric bacteria; Salmonella typhi, Shigella flexneri, Escherichia coli and Proteus vulgaris. Methodology: Root samples of E. heterophylla and V. paradoxa were collected, washed, air dried and processed to fine powder in the microbiology laboratory of Federal University of Technology, Minna, Nigeria. Crude extract of the root samples was done by the cold maceration technique using four solvents (chloroform, methanol, petroleum ether and water). Phytochemical analysis of the extracts was done using previously described technique, and in vitro antibacterial activities of different concentrations of the extracts (50-200 mg/ml) and a standard antibiotic (ciprofloxacin) were tested on four enteric bacteria (S. typhi, S. flexneri, E. coli, P. vulgaris) by the agar diffusion test. In vivo antibacterial activities of the two plants were also tested by daily oral administration of 2000 mg/kg bodyweight (for 7 days) of each extract on inbred mice infected through intraperitoneal inoculation of an infective dose of each of the four enteric bacteria. Data were computed as mean ± standard error and analysed by the Statistical Analysis System (SAS) version 9.4. Associations between variables were determined using analysis of variance (ANOVA), with p < 0.05 considered as significant value. Results: Phytochemical analysis of the crude extracts of both plants revealed the presence of cardiac glycosides, saponins, alkaloids, flavonoids, and tannins but V. paradoxa contain more carbohydrates and starch, and less phlobatannins, compared to E. heterophylla. In vitro assay showed dose-dependent antibacterial activity of the methanol, aqueous and chloroform (but not petroleum ether) extracts of the two plant roots. The in vitro antibacterial activities of the different extracts of V. paradoxica extracts were significantly higher (higher mean diameters of inhibition zones) than those of E. heterophylla (p<0.05), and methanol extracts gave the highest antibacterial effects. However, the root extract of E. heterophylla gave a higher antibacterial activity with the in vivo assay on inbred mice than V. paradoxa, and methanol extracts of the two plant extracts gave the highest in vivo activity, followed by aqueous extract and least activity was obtained with the chloroform extract. Conclusion: Crude extracts of E. heterophylla and V. paradoxa roots produce antibacterial activity against enteric Gram-negative bacteria pathogens involved in diarrhoea illnesses. Further researches should be directed towards isolation and characterization of the active compounds in the crude extracts.

Clinical effect of Metformin on inflammation and intestinal flora imbalance in elderly patients with type 2 diabetes mellitus / 中华老年医学杂志

Objective To investigate the clinical effect of Metformin on inflammation,intestinal flora imbalance,and metabolic impairment in elderly patients with type 2 diabetes mellitus(T2DM) Methods A total of 74 elderly patients with T2DM admitted to our hospital from October 2014 to October 2016 were included in this study and assigned into an intervention group(n=37)and a control group(n=37)according to the sequence of admission.The intervention group was treated with 0.5 g Metformin once a day for 2 weeks,while the control group was given a placebo with the same frequency as in the intervention group.Fresh pre-and post-treatment stool samples were collected in sealed sterile anaerobic growth bags for the detection of changes in patient intestinal flora.Serum inflammatory factor levels,blood glucose levels,and blood lipid levels were measured.Results Before treatment,the numbers of beneficial intestinal flora(bacillus bifidus,bacillus acidi lactici,and bacteroides)in the intervention group were lower than in the control group(all P＜0.05);The levels of serum inflammatory factors(CRP,IL-6,and TNF-α)in the intervention group were higher than in the control group(all P ＜0.05);The levels of parameters for glucose and lipid metabolism(HbA1c,FPG,TC,TG,and LDL-C)in the intervention group were higher than in the control group,while the level of HDL-C was lower than in the control group (all P ＜ 0.05).After treatment,the number of beneficial intestinal flora significantly increased in the intervention group(all P＜0.05)but was not different from that in the control group(all P＞0.05);The levels of serum inflammatory factors had significant reductions in the intervention group compared with pre-treatment levels(all P＜0.05)but demonstrated no difference from those in the control group(all P＞0.05);Parameters for glucose and lipid metabolism in the intervention group showed improvement(all P ＜0.05)after treatment but no difference was observed between these two groups(all P＞0.05).Conclusions Metformin can effectively control blood glucose and blood lipid levels in elderly patients with T2DM,improve the chronic inflammatory state,increase the number of beneficial intestinal bacteria,and regulate flora imbalance.

Ecotypes and virulence factors of Salmonella spp. detected in shrimp farms in State of Ceara, Brazil / Ecotipos e fatores de virulência de Salmonella spp. detectadas em fazendas de carcinicultura do Ceará, Brasil

Our objective was to group in ecotypes 12 serovars of Salmonella isolated from shrimp farming environments in the State of Ceara (Northeast Brazil). Grouping was done based on genotypic virulence factors. Two groups based on the similarity of the Box-PCR were identified: a group consisting of three strains (01 S. ser. Madelia serovar and 02 S. ser. enterica subs. houtenae) and another group consisting of nine isolates (02 S. ser. Saintpaul serovars; 03 S. ser. Infantis; 02 S. ser. Panama; 01 S. enterica subs. enterica; and 01 S. enterica subs. houtenae). Distribution pattern of the serovars was not influenced by the origin matrices (water and sediment). Plasmid virulence genes pefA and invA were detected, unrelated to the serovar and environmental origin of the isolates. The presence of virulence genes in the isolates underlines the potential to trigger salmonellosis events via shrimp consumption. Biomonitoring of these sources of contamination should be encouraged as a protective measure, minimizing health risks and economic losses for the industry.

Nosso objetivo foi agrupar em ecotipos 12 sorovares de Salmonella isolados em ambientes de carcinicultura no Estado do Ceará. O agrupamento foi feito a partir da pesquisa de fatores genotípicos de virulência. Constatou-se a formação de dois grupos baseados na similaridade do Box-PCR: um grupo com três estirpes (01 sorovar S. ser. Madelia e 02 sorovares S. enterica subs. houtenae) e outro constituído por nove isolados (02 sorovares S. ser. Saintpaul, 03 sorovares S. ser. Infantis, 02 sorovares S. ser. Panama, 01 sorovar S. enterica subs. enterica e 01 sorovar S. enterica subs. houtenae). O padrão de distribuição dos sorovares não sofreu influência das matrizes de origem (água e sedimento). Os genes de virulência plasmidial pefA e invA foram detectados independente do sorovar e da origem ambiental dos isolados. A presença desses genes de virulência nos isolados de carcinicultura evidencia o potencial para desencadear eventos de salmonelose relacionados ao consumo de camarão. O biomonitoramento dessas fontes de contaminação deve ser incentivado como medida protetiva, minimizando os riscos do ponto de vista sanitário e das perdas econômicas para o setor da carcinicultura.

Antimicrobial effect of Malaysian vegetables against enteric bacteria

Objective: To investigate the antibacterial activities of green vegetables (pennywort, mint, garlic, parsley and celery) against four common enteric bacteria [Salmonella enterica (ATCC 25957) (S. enterica), Shigella flexneri (ATCC 12022) (S. flexneri), Escherichia coli (ATCC 43889) (E. coli) and Enterobacter cloacae (ATCC 13047) (E. cloacae)] as an alternative medicine for controlling food borne diarrhea disease and the synergistic effect of green vegetables against those bacteria. Methods: Five common vegetables (pennywort, mint, garlic, parsley and celery) were purchased and extracted. The antimicrobial activities of these extracts were tested against four common enteric bacteria (S. enterica, S. flexneri, E. coli and E. cloacae). Ten different concentrations of the extracts (from 640 to 1.25 mg/mL) were prepared and used for the study. The minimal inhibitory concentration (MIC) was determined by the broth dilution method. The antimicrobial activities were assessed by using both well diffusion and disc diffusion methods. Results: Garlic extract showed excellent inhibitory effects on all enteric bacteria. Other plants (parsley, celery, mint and pennywort) were not effective against enteric bacteria. The MIC of garlic against S. flexneri and E. cloacae was 40 mg/mL. The MIC of S. enterica and E. coli were 20 and 10 mg/mL, respectively. The performance of the well diffusion method was better than that of the disc diffusion method with clear and sharp inhibition zones of tested bacteria against plant extracts. Conclusions: Garlic had excellent antimicrobial effects against enteric bacteria and was recommended to be given to patients with gastroenteritis. The other vegetables (pennywort, mint, parsley and celery) showed no inhibitory effects on enteric bacteria but still can be used for its richness in vitamins and fibers. The performance of the well diffusion method was better than that of the disc diffusion method in detecting the antibacterial effects of green vegetables.

Incidence and transferability of antibiotic resistance in the enteric bacteria isolated from hospital wastewater

This study reports the occurrence of antibiotic resistance and production of β-lactamases including extended spectrum beta-lactamases (ESβL) in enteric bacteria isolated from hospital wastewater. Among sixty-nine isolates, tested for antibiotic sensitivity, 73.9% strains were resistant to ampicillin followed by nalidixic acid (72.5%), penicillin (63.8%), co-trimoxazole (55.1%), norfloxacin (53.6%), methicillin (52.7%), cefuroxime (39.1%), cefotaxime (23.2%) and cefixime (20.3%). Resistance to streptomycin, chloramphenicol, nitrofurantoin, tetracycline, and doxycycline was recorded in less than 13% of the strains. The minimum inhibitory concentration (MIC) showed a high level of resistance (800-1600 µg/mL) to one or more antibiotics. Sixty three (91%) isolates produced β-lactamases as determined by rapid iodometric test. Multiple antibiotic resistances were noted in both among ESβL and non-ESβL producers. The β-lactamases hydrolyzed multiple substrates including penicillin (78.8% isolates), ampicillin (62.3%), cefodroxil (52.2%), cefotoxime (21.7%) and cefuroxime (18.8%). Fifteen isolates producing ESβLs were found multidrug resistant. Four ESβL producing isolates could transfer their R-plasmid to the recipient strain E. coli K-12 with conjugation frequency ranging from 7.0 x 10-3 to 8.8 x 10-4. The findings indicated that ESβL producing enteric bacteria are common in the waste water. Such isolates may disseminate the multiple antibiotic resistance traits among bacterial community through genetic exchange mechanisms and thus requires immediate attention.

Distribution of enteric bacteria in the sediments of Parangipettai and Cuddalore coast of India.

Distribution of major groups of enteric bacteria viz., Escherichia coli, Shigella spp., Salmonella spp., Vibrio parahaemolyticus and Vibrio cholerae was studied in the sediment samples collected from different depths of 5, 15, 25 and 35 m in Parangipettai and Cuddalore coast during March, 2009 to February, 2010. Among the bacterial population in Parangipettai, V. parahaemolyticus was found to be maximum with an occurrence of 55% followed by Shigella spp. (15%), V. cholerae (12%), Salmonella spp. (12%) and E. coli (6%). In Cuddalore, as an oddity, Vibrio cholerae emerged as dominant species with an incidence of 60% of the total. V. parahaemolyticus came next in the order with an incidence of 14%, followed by E.coli (11%), Shigella spp. (8%) and Salmonella spp. (7%). Similarly, monsoon and premonsoon seasons registered higher bacterial populations in both the regions. When the depth wise results were viewed, 5 and 15 m depths showed maximum (V. parahaemolyticus 3.7x 10 CFU g-1 in Parangipettai; V. cholera - 8.6x10 CFU g-1 in Cuddalore) bacterial population compared to subsequent depths. Cluster and MDS also showed distinct spatial and seasonal variations of bacterial populations in both the regions. Biota-Environment (BIO - ENV) method revealed the combinations of temperature, salinity and TOC as the best match influencing bacterial population.

Distribution of potentially pathogenic enteric bacteria in coastal sea waters along the Southern Kerala coast, India.

This study evaluated the relationship between the traditional indicators of faecal pollution, total coliforms (TC), faecal coliforms (FC) and Faecal streptococci (FS), and the presence of few potentially pathogenic enteric bacteria, Vibrio cholerae (VC), Vibrio parahaemolyticus (VP), Shigella spp. (SH) and Salmonella spp. (SL) in coastal sea water. The distributional statuses of these bacteria were also studied along the Southern Kerala coast. Cluster analyses were done to identify similar groups of indicator as well as enteric pathogenic bacteria. Kochi was found to be highly polluted with enteric pathogens and indicator bacteria (TC of 4700, VC of 820, FC of 920 and FS of 410 CFU ml-1). Percentage incidence of VC (97.42%) was comparatively higher than the traditional indicator bacteria (TC 95.04%, FC of 63.64% and FS of 47.64%). VC found to be rather stable and showed significant relationship with all the traditional indicator bacteria (R² > 0.370), suggests that both quantitatively and qualitatively the abundance of Vibrio cholerae can determine faecal pollution, could be used as a faecal pollution indicator bacterium, especially in the marine environment where traditional indicator bacteria failed to survive. It would be advisable to always perform the detection of SH and VP beside the traditional indicators as no significant relationship (R²<0.076, p>0.05) exists among them.

Bactérias entéricas presentes em amostras de água e camarão marinho litopenaeus vannamei oriundos de fazendas de cultivo no estado do Ceará, Brasil / Enteric bacteria in water samples and marine shrimp litopenaeus vannamei cultured in the state of Ceará, Brazil

Foram realizados a quantificação de coliformes totais (CT) e termotolerantes (CTT), isolamento e identificação de coliformes, e pesquisa de Salmonella em 28 amostras de água e 28 de camarão da espécie Litopenaeus vannamei, oriundas de duas fazendas de cultivo localizadas no Estado do Ceará, Brasil. Nenhuma amostra de água apresentou índice de CTT acima do limite de 2.500/100 mL preconizado pela legislação para águas salobras destinadas ao cultivo de organismos para fins de consumo. O Número Mais Provável (NMP/g) de CTT das amostras de camarão variou de <3 a 2,9 x 104. A maior frequência de isolamento de coliformes nas amostras de água e camarão foi a da espécie Escherichia coli. Em apenas três (5,35%), das 56 amostras analisadas, foi detectada a presença de Salmonella sorovar Newport e S. Saintpaul. Apesar do baixo índice de CTT e da baixa incidência de salmonela, a presença dessas bactérias entéricas em ambientes de cultivo de peneídeos é preocupante, uma vez que podem provocar infecções em humanos.

Samples of water (n = 28) and Litopenaeus vannamei (n = 28) from two shrimp farms in Ceará state, Brazil were evaluated for total coliforms (TC), total thermotolerant coliforms (TTC), coliform species diversity and Salmonella. No water sample presented TTC levels above the maximum level (2,500 MPN/100 mL) established by regulation for brackish water aquaculture producing seafood for human consumption. The most probable number (MPN) of TTC in shrimp samples ranged from <3 to 2.9 x 104 CFU/g. The coliform species most frequently isolated from water and shrimp was Escherichia coli. Only three (5.35%) of the 56 samples tested were positive for Salmonella (Newport and Saintpaul serovars). In spite of the low TTC levels observed, the presence of potentially pathogenic enteric bacteria in shrimp culture is a disquieting finding.

Resistance to tetracycline and β-lactams and distribution of resistance markers in enteric microorganisms and pseudomonads isolated from the oral cavity

This study evaluated the occurrence of enteric bacteria and pseudomonads resistant to tetracycline and β-lactams in the oral cavity of patients exhibiting gingivitis (n=89), periodontitis (n=79), periodontally healthy (n=50) and wearing complete dentures (n=41). Microbial identification and presence of resistance markers associated with the production of β-lactamases and tetracycline resistance were performed by using biochemical tests and PCR. Susceptibility tests were carried out in 201 isolates of enteric cocci and rods. Resistance to ampicillin, amoxicillin/clavulanic acid, imipenem, meropenem and tetracycline was detected in 57.4 percent, 34.6 percent, 2.4 percent, 1.9 percent and 36.5 percent of the isolates, respectively. β-lactamase production was observed in 41.2 percent of tested microorganisms, while the most commonly found β-lactamase genetic determinant was gene blaTEM. Tetracycline resistance was disseminated and a wide scope of tet genes were detected in all studied microbial genus.