Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study / Avaliação antimicrobiana e toxicológica de Origanum vulgare: um estudo in vivo

Abstract Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Resumo Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Evaluation of triclosan toxic effects on the methanogenic activity

BACKGROUND: Triclosan (TCS) is an antimicrobial agent widely used in health care and consumer products. This compound is present in sludge of wastewater treatment plants (WWTPs), and because of its bactericidal characteristics, it can inhibit the methanogenic activity in anaerobic digestion (AD) technology. The aim of this study was to evaluate the toxic effects of TCS on the methanogenic activity. RESULTS: Batch anaerobic reactors were used with TCS concentrations of 7.8, 15.7, 23.5, and 31.4 mg/L. These assays consisted in three successive feedings (I, II, and III), wherein the sludge was exposed to each TCS concentration and volatile fatty acid (VFA) substrate. For evaluation of the residual sludge activity during feeding III, only VFA was used. The results showed that the increase in TCS concentrations correlated with the reduction in methane (CH4) production. In this case, the minimum values were achieved for TCS concentration of 31.4 mg/L with CH4 levels between 101.9 and 245.3 during feedings I, II, and III. Regarding the effect of TCS on VFA consumption, an inhibitory effect was detected for TCS concentrations of 23.5 and 31.4 mg/L, with concentrations of acetic, butyric, and propionic acids at the end of the assay (37 d) between 153.6 and 206.8, 62.5 and 60.1, and 93.4 and 110 mg/L, respectively. Regarding the removal of TCS during AD, these values were above 47%. Conclusion: TCS is an inhibitor of methanogenic activity with a decrease between 63 and 70% during the different feedings. The CH4 production was not recovered during feeding III, with inhibition percentages of 21­72%.

Acute and chronic toxicity and antimicrobial activity of the extract of Stryphnodendron adstringens (Mart) Coville / Toxicidade aguda e crônica e atividade antimicrobiana do extrato de Stryphnodendron adstringens (Mart) Coville

This study evaluated the antimicrobial activity and acute or chronic toxicity of the extract of Stryphnodendron adstringens. The stem bark dry extract was obtained by static maceration with ethanol. Quantification of tannins was performed by the Folin-Denis method, which indicated a total tannin content of 32.7%. The antimicrobial activity of the dry extract of S. adstringens was evaluated by agar-based disk diffusion assay with Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) in the concentration of 200, 400 and 600µL/mL. The results indicated that 600µL/mL inhibited microbial growth, i.e. had antimicrobial activity against these species. Acute and chronic toxic effects of S. adstringens was evaluated in Wistar rats treated with 200, 400, 600 and 800mg/kg of extract, administrated by gavage. Liver degeneration was observed in the group of rats receiving 800mg/kg in chronic exposure, what may indicate some degree of toxicity at this concentration. However, no systemic toxicity was observed at lower doses. Considering the broad use of S. adstringens as a phytotherapeutic agent for various human and animal diseases and the livertoxicity observed at high concentrations, attention should be paid to the possible adverse effect of using the extract from this plant at high concentration.(AU)

Avaliou-se neste estudo a atividade antimicrobiana e a toxicidades aguda e crônica do extrato da entrecasca de Stryphnodendron adstringens. A partir do extrato seco, obtido através da maceração estática da casca do caule em etanol de cereais, foi realizada a quantificação de taninos totais (32,7%) pelo método de Folin-Denis. A atividade antimicrobiana do extrato seco extraído de cascas do caule de S. adstringens foi avaliada pela técnica de disco-difusão para os micro-organismos Escherichia coli (ATCC 25922) e Staphylococcus aureus (ATCC 25923) nas concentrações de 200, 400 e 600µL/mL. Os testes de concentração inibitória mínima revelaram que 600µL/mL inibiu o crescimento dos dois micro-organismos testados; o mesmo resultado foi observado para atividade bactericida na concentração de 600µL/mL sobre essas espécies. Efeitos tóxicos sistêmicos agudos e crônicos do extrato seco de S. adstringens administrados por gavagem foram estudados em ratos Wistar, utilizando as doses de 400, 600 e 800mg/kg. Foi observada degeneração hepática no grupo de animais que receberam 800mg/kg tanto no estudo da toxicidade aguda quanto crônica, que pode indicar algum grau de toxicidade de S. adstringens nessa concentração. Considerando o amplo uso de S. adstringens como fitoterápico para humanos e animais, atenção deve ser dispensada para ingestão em altas doses mediante os efeitos tóxicos observados neste estudo.(AU)

Identification and antimicrobial resistance of members from the Enterobacteriaceae family isolated from canaries (Serinus canaria) / Identificação e resistência antimicrobiana de membros da família Enterobacteriaceae isolados de canários (Serinus canaria)

The Enterobacteriaceae family contains potentially zoonotic bacteria, and their presence in canaries is often reported, though the current status of these in bird flocks is unknown. Therefore, this study aimed to identify the most common genera of enterobacteria from canaries (Serinus canaria) and their antimicrobial resistance profiles. From February to June of 2013, a total of 387 cloacal swab samples from eight domiciliary breeding locations of Fortaleza city, Brazil, were collected and 58 necropsies were performed in canaries, which belonged to the Laboratory of Ornithological Studies. The samples were submitted to microbiological procedure using buffered peptone water and MacConkey agar. Colonies were selected according to their morphological characteristics on selective agar and submitted for biochemical identification and antimicrobial susceptibility. A total of 61 isolates were obtained, of which 42 were from cloacal swabs and 19 from necropsies. The most isolated bacteria was Escherichia coli with twenty five strains, followed by fourteen Klebsiellaspp., twelve Enterobacterspp., seven Pantoea agglomerans, two Serratiaspp. and one Proteus mirabilis. The antimicrobial to which the strains presented most resistance was sulfonamides with 55.7%, followed by ampicillin with 54.1% and tetracycline with 39.3%. The total of multidrug-resistant bacteria (MDR) was 34 (55.7%). In conclusion, canaries harbor members of the Enterobacteriaceae family and common strains present a high antimicrobial resistance rate, with a high frequency of MDR bacteria.

A família Enterobacteriaceae possui bactérias com potencial zoonótico e a presença destas bactérias em canários é relatada na literatura, porém a realidade dos plantéis de criadores de canários é desconhecida. Portanto, este trabalho teve como objetivo isolar enterobactérias de canários belga (Serinus canarius) com o intuito de conhecer os gêneros mais comuns nestas aves e suas respectivas resistências a antimicrobianos. De fevereiro a junho de 2013 foram coletadas 387 amostras de swabs cloacais de canários de oito propriedades da cidade de Fortaleza, Brasil e de 58 necropsias de aves do acervo próprio do Laboratório de Estudos Ornitológicos. As amostras foram submetidas a isolamento microbiológico utilizando-se água peptonada e ágar MacConkey. As colônias foram selecionadas de acordo com suas características morfológicas nas placas, submetidas à tipificação bioquímica para identificação e ao teste de sensibilidade a antimicrobianos. Foram isoladas 61 cepas, sendo 42 de suabes cloacais e 19 de necropsias. A bactéria mais isolada foi Escherichia coli com vinte e cinco cepas, seguida por catorze Klebsiella spp., doze Enterobacter spp., sete Pantoea agglomerans, duas Serratiaspp. e uma cepa de Proteus mirabilis. As cepas apresentaram maior resistência a sulfonamidas com 55,7%, seguidas por ampicilina com 54,1% e tetraciclina com 39,3%. Além disso, o total de cepas resistentes a múltiplas drogas (RMD) foi 34 (55,7%). Portanto, conclui-se que os canários albergam enterobactérias e que as cepas apresentam alto índice de resistência a antimicrobianos, com alta frequência de cepas RMD.

Time- and concentration-dependent cytotoxicity of antibiotics used in endodontic therapy

OBJECTIVE: New drugs have to be assessed in endodontic therapy due to the presence of microorganisms resistant to therapeutic procedures. Thus, this study evaluated the time- and concentration-dependent cytotoxicity of different antibiotics used in endodontic therapy. MATERIAL AND METHODS: Human gingival fibroblasts were treated and divided into the following experimental groups: Group I - control; Group II - ciprofoxacin hydrochloride; Group III - clyndamicin hydrochloride; and Group IV - metronidazole. Each drug was used at concentrations of 5, 50, 150, and 300 mg/L for 24, 48, 72, and 96 h. Cytotoxicity was evaluated by the MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and spectrophotometric reading of ELISA plates. The results were analyzed by BioEstat 4.0 software using Kruskal-Wallis and Dunn's tests at a signifcance level of 5 percent. Cell viability was assessed for the different concentrations and times. RESULTS: All drugs presented dose-dependent cytotoxicity. Concentrations of 5 and 50 mgjL produced viable fibroblasts at all experimental times in all groups. CONCLUSIONS: Cell viability at 24 h was greater than in the other experimental times. Comparison between the same concentrations of antibiotics at different times showed that metronidazole presented the highest cell viability at 72 and 96 h compared to the other antibiotics, whereas clyndamicin hydrochloride showed higher cell viability at 72 h than ciprofoxacin hydrochloride.

Recovery of Salmonella using a combination of selective enrichment media and antimicrobial resistance of isolates in meat in Thailand.

From November 2004 to March 2005, 50 samples (chicken, pork and beef) of registered meat and non-registered meat were purchased from supermarkets and retail markets located in Bangkok, Thailand. Each sample was evaluated for Salmonella spp by a conventional method using combination of selective enrichment media (RV+MSRV) and compared with selective enrichment medium alone (DIASALM). Our study revealed the performance of RV+MSRV for the detection of Salmonella spp was significantly better than those of DIASALM alone since the recovery of Salmonella spp in both groups of meat was high using RV+MSRV, particularly in the registered meat. In addition, the recovery of serovars in registered meat was significantly higher than those in non-registered meat. Antimicrobial resistance of 62 Salmonella isolates in both groups of meat was determined for 10 antimicrobial drugs using the disk diffusion method. The results show that 100% of isolates from both groups were susceptible to amoxicillin/clavulanic acid, ciprofloxacin, cefotaxime and norfloxacin and 50-60% of isolates from both groups were resistant to tetracycline, streptomycin and ampicillin. Sixty percent of Salmonella isolates from meat showed multiresistance antimicrobial patterns.

Isolation, phage typing and antibiogram of Salmonella from man and animals in northeastern India.

BACKGROUND AND OBJECTIVE: Salmonella is an important zoonotic pathogen and its prevalence in the animals acts as a continuous threat to man. The present study was carried out to report the isolation along with the serotypes, phage types and antibiogram pattern of Salmonella among man, livestock and poultry in the northeastern India. METHODS: A total of 654 samples from diarrhoeic livestock and humans were processed for the isolation of Salmonella. All the isolates were subjected to antibiogram studies against 15 antimicrobials. Representative isolates of S. Typhimurium and S. Enteritidis were phage typed. RESULTS: Ninety five isolates of Salmonella enterica belonging to 5 serotypes- S. Typhimurium, S. Enteritidis, S. Gallinarum, S. Paratyphi B and S. Bareilly were obtained with an overall prevalence rate of 14.40 per cent. S. Typhimurium isolates were distributed among four phages- DT003, DT004, DT096 and DT193 and all the S. Enteritidis isolates belonged to a single phage type, PT13a/7. Interspecies sharing of the phages was observed. Norfloxacin, enrofloxacin, gentamycin and ciprofloxacin were most effective, whereas, doxycycline, ampicillin, amoxycillin and tetracycline were relatively less effective. INTERPRETATION AND CONCLUSION: Our findings showed that three of the five serovars as well as some of the phage types of these serovars were shared by animals and humans indicating the zoonotic potential of the organism. Thus, it is imperative that salmonellosis control measures adopted for humans should give adequate importance to its control in the animals particularly their products.

Effect of metronidazole on spermatogenesis and FSH, LH and testosterone levels of pre-pubertal rats.

Metronidazole, a 5-nitroimidazole drug has been reported to decrease testicular weight, testicular and epididymal spermatid counts and causes abnormal sperm morphology with degeneration of seminiferous tubules with 6 weeks treatment of metronidazole (400 mg/kg, day). In contrast to DNA flow cytometry (FCM), the histological and gravimetric parameters do not allow a rapid, sensitive, objective and multiparameteric evaluation of reproductive toxicants on spermatogenesis. Moreover, the exact mechanisms for such an effect are not entirely clear. The present study was therefore undertaken to assess the effects of intraperitoneal (i.p.) administration of metronidazole 400 mg/kg daily for 30 days on testicular germ cell changes assessed by DNA (FCM) and hormone levels of testosterone, FSH and LH in pre-pubertal rats. A significant reduction in the haploid cell population in metronidazole treated groups as compared to saline treated controls was observed. The mean serum FSH, LH and testosterone value were also lowered in treated animals. Thus, the spermatotoxic effects of metronidazole were probably mediated by decrease in the circulating hormones responsible for spermatogenesis.

Toxicology of Lapachol in rats: embryolethality

Lapachol is a naphtoquinone with therapeutic potential against enterovirus, Chagas disease and is also used as an antimalarial and antiinflamatory agent. In order to study teratogenic potential of Lapachol, pregnant Wistar rats were treated with 0.5 ml of distilled water (control group); 0.5 ml of hydroalcoholic solution (vehicle group) and 10 mg of Lapachol in 0.5 ml of hydroalcoholic solution (treated group) by oral gavage from the 8th to the 12th day of pregnancy. The following variables were observed: maternal body weight on days 1, 6, l5 and 21 and food intake on days 2, 6, 15 and 21 of pregnancy. The number of live and dead fetuses and the sites of resorptions were counted. The ovaries were weighed and the corpora lutea were counted. Data were analyzed by ANOVA-one way, Dunnett test and the chi square test. Significance level test alpha = 0.05. Results have shown that mothers were unaffected but there were a 99.2 percent of fetus mortality, indicative of a strong abortifacient effect of Lapachol in rats

Esparfloxacino e as fluorquinolonas: uma revisäo de sua classificaçäo, atividade antibacteriana, mecanismo de açäo, características farmacocinéticas, tolerabilidade e perspectivas farmacêuticas / Sparfloxacin and the fluoroquinolones: a review of classification, antibacterial activity, mechanism of action, pharmacokinetic properties, tolerability and pharmaceutical perspectives

Foi realizada uma revisäo na literatura sobre as quinolonas, classe antibacteriana que possui amplo espectro de açäo, enfocando, principalmente, o esparfloxacino, fluorquinolona de terceira geraçäo que possui potente atividade contra bactérias Gram-positivas, como Streptococcus pneumoniae e Staphylococcus aureus inclusive cepas metilina resistentes (MRSA), bactérias Gram-negativas, anaeróbios, Legionella spp, Mycoplasma spp, Chlamydia spp e Mycobacterium spp.

Elucidacion de los mecanismos de fototoxidad inducida por farmacos / Mechanisms of phototoxicity induced by pharmaceutical products

First of all some general concepts are given on phototoxic activity of pharmaceutical products which full fill the structural characteristics required to decompose by light and to cause biological damage, either themselves, their photoproducts or the products of their metabolism. These considerations are important due to the fact that this field of research is fairly new. Next, a review is given on recent research carried out in this laboratory on the photochemistry and phototoxicity of fibric acid and their derivatives and finally a refc5r cview is made as well on the photochemistry and phototoxicity of antibacterial quinolones. Mechanisms are postulated for the photochemical decomposition of the substances investigated and possible mechanism for the in vitro activity at cellular level are also presented

In vitro short-term and long-term cytotoxicity of fluoroquinolones on murine cell lines.

Short-term and long-term cytotoxicity of four fluoroquinolones (ciprofloxacin, rufloxacin, ofloxacin, lomefloxacin) on 7 established murine cell lines (WEHI-3B, L1210, EL4, P388D1, 32DC13, L929, SR-4987) by a microtiter MTT assay have been studied. In short-term cytotoxic test (24 hr), cell lines with a high proliferating cell rate (as leukemias) showed a greater sensitivity to quinolones than other cell lines. In long-term cytotoxic test (7 days) no different sensitivity was observed among the cell lines. In short-term test ciprofloxacin and rufloxacin were more toxic than lomefloxacin and ofloxacin whereas in the long-term test the activity of the four quinolones was similar. Ratio between IC50 on cell lines and MIC50 against gram negative bacteria evidenced remarkable differences when short-term or long-term cytotoxic tests were considered. The results confirm toxic activity of quinolones on mammalian cells evidencing that the sensitivity to quinolones, in short-term cytotoxic test, correlates with the doubling time of cell population. The results further suggest that long-term cytotoxic tests measure better the antiproliferating activity of quinolones providing a more powerful assay to investigate their in vitro toxicity.