Determination of antibacterial activity of anacyclus pyrethrum extract against some of the oral bacteria: an in vitro study

Statement of Problem: There are more than 500 different bacterial species in the oral cavity which can cause tooth decay and periodontal diseases. Anacyclus Pyrethrum has been used to manage dental and periodontal diseases in traditional Iranian medicine

Purpose: The purpose of this study was to determine the antibacterial activity of Anacyclus Pyrethrum against some of the oral bacteria, such as Staphylococcus aureus, Streptococcus mutans, Streptococcus sanguis and Pseudomonas aeruginosa

Materials and Method: At first, The antibacterial effect of serial concentrations [1/10 to 1/100 mg/ml] of methanolic extract of Anacyclus Pyrethrum root were tested by using well assay method on Staphylococcus aureus, Streptococcus mutans, Streptococcus sanguis and Pseudomonas aeruginosa. In the second examination, 150-1000 mg/ml concentrations were tested and the agar dilution method, recommended by the Clinical and Laboratory Standards Institute Standards, was used. Then, the lowest concentrations of the extract which inhibited visible growth of organisms on the media plate; Minimum Inhibitory Concentration [MIC] and Minimum Bactericidal Concentration [MBC], were determined

Results: The inhibition zone was only seen in the 1.10 mg/ml concentration. The diameters were 15 and 12 mm in Staphylococcus aureus and Streptococcus sanguis agar media plate, respectively. In the second examination, the greatest inhibition zones were 27 mm for Staphylococcus aureus and 21 mm for Streptococcus sanguis in 1000 mg/ml. There was not any inhibition zone for Streptococcus mutans and Pseudomonas aeruginosa in the concentrations. The MBC was achieved as 800 mg/ml for Staphylococcus aureus and Streptococcus sanguis. Streptococcus mutans and Pseudomonas aeruginosa grew in all the concentrations

Conclusion: The antibacterial effect of Anacyclus Pyrethrum extract against Staphylococcus aureus and Streptococcus sanguis was not significant. Anacyclus Pyrethrum had no antibacterial effect against either Streptococcus mutans or Pseudomonas aeruginosa

[Effect of methanol extracts of artemisia persica on kinetic growth of S. aureus and B. subtilis bacteria]

Artemisia persica is a common species in South East Iran and A. persica is the only species in this genus and the species name persica means from Iran. Artemisia persica have been used in traditional medicine to treat various infections and inflammations. It is an effective remedy for S. aureus and B. subtilis bacterial infection. The selected methanol extracts were screened against two strain of bacteria Staphylococcus aureus and Bacillus subtilis at seven different extracts concentrations [5, 25, 100, 150, 200, 300 and 400 micro g/ml] using disc diffusion and wheel assay methods. The minimum inhibitory and bactericidal concentrations [MIC, MBC and Death kinetic] of the active extracts were tested using MH broth-dilution methods. For standard strains of S. aureus the extracts for MIC were range 100 mg/ml and MBC range were 150-200 micro g/ml. For B. subtilis standard strain the MIC extracts range were150-200 micro g/ml and MBC were 400 micro g/ml, Kinetic of death were 4 hours for B. subtilis compared to 5 hours for S. aureus. These results suggest that extracts of Artemisia persica may have the antibacterial activity against some pathogenic bacteria. It has an antibacterial action and inhibiting the growth of Staphylococcus aureus and Bacillus subtilis and it may be attributed to its content of the artemisinin, tannins, saponin, alfa- pinene and camphor. From this study it can be concluded that many extracts possess antibacterial activity Plant extracts have been used for many pharmaceuticals, alternative medicine and natural therapies

Fermentation, fractionation and purification of streptokinase by chemical reduction method

Streptokinase is used clinically as an intravenous thrombolytic agent for the treatment of acute myocardial infarction and is commonly prepared from cultures of Streptococcus equisimilis strain H46A. The objective of the present study was the production of streptokinase from strain H46A and purification by chemical reduction method. The rate of streptokinase production evaluated under the effect of changes on some fermentation factors. Moreover, due to the specific structure of streptokinase, a chemical reduction method employed for the purification of streptokinase from the fermentation broth. The H46A strain of group C streptococcus, was grown in a fermentor. The proper pH adjusted with NaOH under glucose feeding in an optimum temperature. The supernatant of the fermentation product was sterilized by filtration and concentrated by ultrafiltration. The pH of the concentrate was adjusted, cooled, and precipitated by methanol. Protein solution was reduced with dithiothreitol [DTT]. Impurities settled down by aldrithiol-2 and the biological activity of supernatant containing streptokinase was determined. In the fed -batch culture, the rate of streptokinase production increased over two times as compared with the batch culture and the impurities were effectively separated from streptokinase by reduction method. Improvements in SK production are due to a decrease in lag phase period and increase in the growth rate of logarithmic phase. The methods of purification often result in unacceptable losses of streptokinase, but the chemical reduction method give high yield of streptokinase and is easy to perform it

Crystal violet in culture media for diagnosis of neisseria species

Neisseria species are gram negative diplococci; an important characteristic of these bacteria is resistance against crystal violet. This study was done to investigate the effect of crystal violet on the growth of Neisseria, to observe the outcome of adding this substance in culture media for isolating these organisms, and finally to make a specific medium for isolation of the Nisseria species. The study was done in 3 phases: 1] Initially urethral discharge from 106 male patients with urethritis was cultured on NYC, chocolate agar and chocolate violet agar with various concentrations of violet from 1:100000 to 1:250000. We also made direct smears for gram stains. 2] Pharyngeal secretions from 230 healthy persons were cultured on chocolate agar, Muller Hinton agar, and Thayer Martin agar with different concentrations of violet between 1:50000 to 1:500000. Also direct smears were made for gram stains. 3] The standard strain of gonococcus [ATCC] were cultured on the three media. Concurrently we added various concentrations of violet from 1:50000 to 1:200000 to the above media and studied the effect of adding crystal violet on the growth of the standard strain. In first step, 69 out of 106 patients with urethritis were suspicious of gonorrhea, with positive culture of gonococcus on NYC medium from 64 patients. On chocolate agar only 54 positive cultures, [with 84% sensitivity against NCY medium], were seen together with a growth of normal flora. Chocolate agar plus violet in concentration 1:150000, showed 58 positive cultures, [with 91% sensitivity against NYC medium], with minimal growth of normal flora. In second step, 228 out of 230 healthy persons had positive culture of Neisseria, these organisms grew in different concentrations of crystal violet between 1:500000 and 1:50000. However, with minimal concentration of violet, there was a dense growth of normal flora and with gradual increase in concentration, normal flora grew sparsely. In direct exam, 228 cases of gram negative Neisseria like diplococci were observed. In third step, result of growth of the standard Neisseria gonorrhea in chocolate agar, Muller-Hinton agar and Thayer-Martin agar with and without different concentrations of crystal violet are as follow: In all media without crystal violet, the growth of the bacteria was perfect and abundant; while in media containing crystal violet, minimum colony count was observed in concentrations of 1:50000 and maximum colony count occurred at concentrations of 1:200000. To isolate pathogenic species of Neisseria, for e.g. gonococcus, we can use a specific chromogen medium like chocolate-violet agar 1:150000, or Thayer-Martin-violet agar or Muller-Hinton-violet agar with a concentration of 1:200000. Although nonpathogenic Neisseria have high resistance to crystal violet and were isolated from cultures with 1:50000 dilutions of this substance, but growth become sparse with higher concentrations. We can make chromogen media of varying strengths by adding different amounts of crystal violet in various media to get the desired results

[Evaluation of relation between Ureaplasma Urealyticum infection with abortion]

Since long time ago, abortion has been considered as one of the most important medical problems. Bacteria and infectious agents are one of its causes. Possible etiologic role of ureaplasma urealyticum in abortion has been suggested for years ago, but it has not been approved completely yet. Therefore, this study has been designed for approving of this hypothesis and prevention of spontaneous abortion. This study was a case control type of investigation done over women referred to private Gyn and Obs clinic, which had spontaneous abortion. Control group was selected among referred women who did not have any previous abortion. To investigate the presence of ureaplama urealyticum, vaginal and cervical culture was performed in both groups. Other interfering variable factors were also assessed and if there was any intervention factor in any one of two groups, sample was removed. After recording of results of study in questionnaire, central indexes were analyzed by K [2], fisher exact test. 7 persons were eliminated from this study due to intervention factors, so, study was done over 34 patients as case [27 with one abortion and 7 with recurrent abortion] and 47 patients as control group. Mean age in case group was 26.3 +/- 4.6 years and in control group was 23.4 +/- 3.6 years. Positive vaginal and cervical culture for ureaplasma urealyticum in case group was 35% [12 persons] and in control group was 6% [3 persons] [P<0.0001]. In patients with spontaneous recurrent abortion, positive culture for ureaplasma urealyticum was 71% [5 persons] and in patients with only one spontaneous abortion was 26% [7 persons] [P<0.034]. In comparison with similar studies, positive culture of ureaplama urealyticum in patients of case group were a lot more in this study. Prevalence of ureaplama urealyticum in control group was 6% which is less than similar studies. Proportion of positive culture of ureaplasma urealyticum in patients with recurrent abortion was higher than those with only one abortion. Therefore, it is recommended that pregnant women specially those who have high risk pregnancy and also in women with history of abortion, should be examined for contamination with ureaplasma urealyticum. In case of positive results, proper treatment will reduce probable recurrent abortions

[Detection and typing of human papilloma virus DNA in cervical cancer with in situ hybridization method]

Cervical cancer is one of the most frequently found cancers in women and appears to have a viral aetiology certain types of the human papillomavirus [HPV] are well established as the primary causes of cervical cancer. Clinical follow-up data, histopathologic diagnosis, in situ hybridization [ISH] and HPV DNA typing were available from 60 patients. ISH technique was performed with comercial biotinylated probes. The presence of 7 high risk HPV was evaluated in 60 cervical biopsies with squamous cell carcinoma [SCC] and Cervical Intraepithelial neoplasia [CIN] of different degrees by ISH. We analysed 60 biopsies from Iranian women. 42 of 60 [70%] carcinoma specimens were positive for HPV-DNA. HPV 31/33/51 [25%] was most frequently found, followed by HPV 16/18 [23.33%] and HPV 6/11 [21.66%] while HPV negative cases were 18[30%]. High risk HPV types appear to be most frequently associated with SCC and CIN. ISH is a sensitive test in the detection and typing of HPV DNA both in clinical and latent infections