Biochemical, microbiological and phytochemical studies on aqueous-fermented extracts from Atropa belladonna L. Part 1--biochemistry and microbiology.

Extraction methods of fresh plants into aqueous-fermented extracts according to German Homoeopathic Pharmacopoeia (HAB), regulation nos. 33 and 34 were evaluated. In the course of production, the extraction is accompanied by fermentation and the resulting preparation is stored for at least 6 months until further processing. The present work aimed at revealing the underlying biochemical reactions during manufacture and storage. In addition, the responsible microorganisms were isolated and identified. To study the robustness of the preparation method, formulation components as well as production conditions were varied. Additionally, questions were addressed at the reproducibility of the method and a comparison with an ethanolic extract was also performed. From 2006 to 2009, 110 extracts from the fresh flowering herb of Atropa belladonna var. belladonna (L.) were produced and analyzed. The results show that lactic acid bacteria (LAB) are primarily involved in the fermentation process, mainly producing lactic acid besides acetic acid and ethanol. The homofermentative Lactobacillus plantarum and the heterofermentative Lactobacillus brevis were identified as predominant lactic acid bacteria. Finally, factors for a successful fermentation are proposed.

Homeopathy treatment of Malassezia Pachydermatis in vitro

A common clinical occurrence in dogs is otitis externa caused by excessive growth of yeasts Malassezia pachydermatis, which can become chronic after wrong treatments, in which microbial resistance can occur. Homeopathic remedies can be considered a successful alternative, selecting the medicine through the similitude principle. Herein, 50 µL of a 1:1000 dilution of Malassezia pachydermatis suspension at 0.5 McFarland scale was used to seed the yeast into Sabouraud dextrose agar plates using a Drigalski spreader to proceed with colony unit counting. Before being seeded, the yeast suspensions were treated with 1% of different homeopathic treatments previously selected from a pilot study, which means Sulphur 6cH, Dolichos pruriens 6cH, and Kali carbonicum 6cH, being water, and succussed water used as controls. For comparison, a set of Sabouraud dextrose agar plates containing 1% Tween 80 was seeded in parallel. The treatments were made blind and evaluated in triplicate. Contaminated cultures were withdrawn. The number of colonies per plate was assessed, and smears were made from the cultures to classify yeast growth according to cytomorphology on ImageJ®software. The preliminary results show no significant effect of all tested medicines compared to the controls. High data variability was also observed, mainly in those cultures whose medium was prepared with Twin 80. In conclusion, at this point of the study, no evidence of the effects of the studied medicines on Malassezia pachydermatis growth in vitro could be identified. The analysis of cytomorphology is still in course.

Clinical comparison of the isolator and BacT/Alert aerobic blood culture systems.

The performance characteristics of the Isolator (Wampole Laboratories, Cranbury, N.J.) and the BacT/Alert (Organon Teknika Corporation, Durham, N.C.) aerobic blood culture systems were compared for 6,009 blood culture sets obtained from patients with suspected bloodstream infections. The BacT/Alert aerobic bottle [BTA(O2)] was continuously agitated while it was incubated in 5% CO2 at 36 degrees C; culture plates prepared from the Isolator tube [I(O2)] were incubated in 5% CO2 at 37 degrees C. From 394 blood cultures, 416 clinically significant isolates of bacteria and yeasts were recovered. The overall yields for BTA(O2) and I(O2) were not significantly different (319 versus 336; P = 0.20). I(O2) recovered significantly more staphylococcus (P < 0.05) and yeast isolates (P < 0.01). BTA(O2) recovered significantly more aerobic and facultatively anaerobic gram-negative bacilli (P < 0.05). In blood culture sets which produced growth of the same organisms in both the BTA(O2) and I(O2) systems, the BTA(O2) system detected growth sooner, but more rapid identification was possible with the I(O2) system by virtue of earlier isolation of colonies on solid media.

Comparison of BacT/Alert with Signal blood culture system.

The BacT/Alert (Organon Teknika Corp., Durham, N.C.) is an automated blood culture system. It is based on the detection of CO2 by means of a colorimetric sensor internally attached to the bottom of culture bottles. The aerobic and anaerobic media of this system were compared with one bottle of the Signal system (Oxoid Ltd., Hampshire, United Kingdom). At bedside, 20 ml of blood was drawn from each adult patient. The two BacT/Alert bottles were inoculated with 5 ml of blood each; the Signal bottle was inoculated with 10 ml. A total of 5,284 sets (2,483 patients; 2.1 cultures per patient) consisting of three bottles each were evaluated, of which 781 sets (14.8%) revealed microorganisms (n = 892); 642 of these were considered to be pathogenic. Significantly more (P < 0.0001) pathogens were isolated from the two BacT/Alert bottles together (n = 584) than from the single Signal bottle (n = 515). Escherichia coli (P = 0.007), gram-negative bacteria other than members of the family Enterobacteriaceae or Pseudomonas spp. (P = 0.006), and yeasts (P = 0.02) were isolated more often from both or either BacT/Alert bottle. Comparing the systems in terms of 388 different organisms per septic episode, the difference between BacT/Alert and Signal was significant for the total number of septicemia cases (P = 0.003). More contaminants grew in the BacT/Alert system (173 versus 116; P = 0.0001). False-positive indications were more frequent in the BacT/Alert system, 198 (3.7%) aerobic bottles and 57 (1.1%) anaerobic bottles, than in the Signal bottles, 24 (0.5%) bottles. Pathogens could be detected significantly earlier (P < 0.0001) in the BacT/Alert system than in the Signal system. The BacT/Alert instrument with two bottles allowed earlier detection as well as the isolation of more microorganisms than the manual, one-bottle Signal system.

Yeasts isolated from clinical samples of aids patients

In order to investigate yeasts in oropharyngeal secretion, urine, sputum and inguinal scales from AIDS patients, clinical samples were collected from one hundred patients interned in the Infectious and Parasitic Diseases Sector of the Hospital das Clínicas of the Universidade Federal de Pernambuco and in Hospital Universitário Osvaldo Cruz of the Universidade de Pernambuco. Yeasts were isolated from seenty-two out of one hundred and eight clinical samples. The isolated yeasts were: Candida albicans (sixty-two isolates), Candida tropicalis (four isolates), Candida glabrata (two isolates), Candida parapsilosis (two isolates), Candida krusei (one isolate) and Trichosporon pullulans (one isolate).

Aplicabilidade de Petrifilm® na enumeração de bactérias e fungos em drogas vegetais / Use of Petrifilm® for bacteria and fungi counting in herbal drugs

Técnicas alternativas aos métodos de plaqueamento em ágar para enumeração de bactérias e fungos têm sido desenvolvidas, buscando-se rapidez e simplicidade, mas sem comprometimento da sensibilidade e exatidão. O objetivo deste estudo foi investigar a eficácia e aplicabilidade de Petrifilm® AC para a enumeraçãode bactérias e de Petrifilm® YM para enumeração de bolores e leveduras em 90 amostras de drogas vegetais, comparando-se as contagens obtidas com a utilização dos sistemas Petrifilm® com àquelas obtidas com as técnicas de semeadura em profundidade em meios de cultura. Os coeficientes de correlação obtidos foram 0,9833 e 0,9231 para as contagens de bactérias e fungos, respectivamente. As contagens obtidas não mostraram diferenças significativas entre os métodos empregados. Os resultados sugerem a aplicabilidade do sistema Petrifilm® como alternativa ao método convencional de plaqueamento

The effect of homeopathic medicines on yeast growth

A reproducible method for testing the effect of homoeopathic medicines on yeast cultures is described. Potencies of Sulphur, Arnica montana, Chamomilla, Bryonia alba, Euphrasia officinalis, and Pulsatilla were tested for their effect on the rate of growth of Schizosaccharomyces pombe. No significant differences were found between the growth rate of test and control samples

Comment on the paper, "Growth of yeast cultures as in vitro model for investigationg homoeopathic remedies"

A recent paper by Steffen repeating earlier work by Jones et al. failed to confirm any effect of potencies of Pulsatilla on the growth rate of cultures of Saccharomyces cerevisiae which had previously been reported. We find that using a more specific statistical test on the results given by Steffen it can be shown that these results contain almost the same periodicity with potency that Jones et al. reported, although only corresponding to a variation of +or- l.5//about the mean value

Growth of yeast cultures as in vitro model for investigating homoeopathic medicines

1 The question of a sensitive response in the growth of yeast cell cultures to treatments with homoeopathic dilutions (potencies), has been raised in a number of recent publications. A further set of growth trials with this model performed at our Institute are reported here. 2 The actions of 8x to 16x decimal potencies of AgNO3, CuSO4, HgCl2 and NaCl on the growth of cell cultures of Schizosaccharomyces pombe were investigated. This included a set of trials with experimental poisoning of the cells by toxic levels of CuSO4 and subsequent study of the remedial action by potencies of the same substance. 3 Yeast cells were grown in batch culture in a glucose/malt extract medium. Growth was interrupted shortly before the end of the exponential growth phase and total cell concentrations were assessed by means of a Coulter Counter. The obtained data were subjected to statistical analysis. Each treatment range was tested in three separate trials. 4 The results obtained failed to indicate any response in the growth rate of Sch. pombe to the potency treatments