| Abstract|| |
Background and Objective: Based on increase in individuals' inclinations toward the use of natural and traditional products in curing some diseases in a systemic and localized manner, the survey of the pathogenic factors which are transferred through such products is of a great importance from the health perspectives. Furthermore, because the emergence of infections due to nonstandard method of preparing some of these natural drugs has not been accurately determined, a poultice mixture consisting of turmeric extract and animal butter was compared in two home-based and aseptic-based preparation methods in terms of microbial load and their effects on the process of pressure ulcer healing is investigated in this study. Materials and Methods: In this study, two poultice mixtures were initially prepared in a home-based method and the other under aseptic conditions, transferred to the laboratory where the specimens were cultured under sterile conditions, in microbe-culturing plates under microbiological hood with a dilution rate of 1–10. After the specified time duration elapsed, the plates containing the culture media were evaluated in terms of bacterial and fungal growth. To prepare a poultice based on standardized methods, turmeric was sufficiently bought from the medicinal herbs market and then put in ethanol 95% v/v. Animal butter underwent separation during a 24 h period in rotary evaporation device under reduced condensation pressure, and after its water was oozed out in 70°C, the concentrate was placed in the incubator. Turmeric extract and animal butter were mixed under bacteriology hood with sterilized wheat flour in autoclave. Finally, the poultice prepared aseptically was tested on seven human specimens and the results were evaluated using pressure ulcer scale for healing (PUSH). Results: Based on the experiments, the homemade poultice mixture was found contaminated with Gram-positive microbial factors, but when the poultice mixture was prepared seven times in the laboratory, no microbial load was found (above USP standard). The mean area of the ulcer was 13.41 before the study, but in the last evaluation, value decreased to 2.21. PUSH mean score from the 1st day until the 4th week reduced from 13.42 to 5.85. It was shown that the ulcer recovery trend decreased considerably. Furthermore, regarding the tissue surveys through observing the samples at the end of intervention, it was seen that the ulcer was free of necrosis and granulated tissues were visible on the ulcer. Conclusion: The aseptic specimen was found in an standard level in terms of microbial load and practically caused effective recovery of the pressure ulcer through increasing the circulation in blood veins, reducing the secretions and edema, and is not indicative of traces of any infection; thus the consumption of this poultice was recognized as safe in improving the pressure ulcers.
Keywords: Aseptic-based, home-based, synthetic poultice, turmeric extract
|How to cite this article:|
Tazakori Z, Karimollahi M, Haghighi Z, Karimollahi M. Comparing turmeric extract plus animal butter synthetic poultice effect in two home-based and aseptic-based preparation methods in terms of microbial load. Ann Trop Med Public Health 2017;10:1686-90
|How to cite this URL:|
Tazakori Z, Karimollahi M, Haghighi Z, Karimollahi M. Comparing turmeric extract plus animal butter synthetic poultice effect in two home-based and aseptic-based preparation methods in terms of microbial load. Ann Trop Med Public Health [serial online] 2017 [cited 2019 Mar 19];10:1686-90. Available from: http://www.atmph.org/text.asp?2017/10/6/1686/222697
| Introduction|| |
For so many years, many interventions have been proposed to accelerate and improve the process of chronic ulcers and identify novel interventions for the emerging issues associated with ulcer. The oldest recorded report of the ulcer healing was discovered on the mud plates from Mesopotamia dating back to 2500 BC. During the course of history, various methods have been used in the form of the combinations of plants and animals extracts  such as the tea leaf extract by Chinese and/or honey and tree resins by ancient Egyptians to today's antiseptic solutions, all of which have been used to create the grounding for speeding up the process of tissue healing. Of course, at present, despite therapeutic effects being confirmed for synthetic drugs, individuals are increasingly losing interest to use such drugs due to their side effects  and the today's world is bearing witness to an increase in tendencies of using natural products as a result of factors such as their ready availability and inexpensiveness as well as the low level of risks and side effects. In recent years, various researches have been undertaken on natural products including the surveys on the effect of honey, olive oil, Curcuma longa L  and fish oil  as interventions for healing chronic ulcers and it can be claimed that the vast part of the country's required drugs can be supplied through the nature-based and standardized sources. One of the very important natural materials usable in healing the ulcers is the animal butter whose curing attributes have been highly underlined in Iranian medicine and Ayurveda. Another product with such features is turmeric. The use of turmeric as an analgesic leading to reduction in inflammation and treating the injuries has been pointed out in Iranian and Hindi texts., According to the daily increasing demands by individuals to use natural products for curing their diseases, inspecting and assuring the healthiness of such natural product from microbial contamination is essential. Microbe counts, the total number of the forms, the presence or absence of pathogenic organisms such as Staphylococci, Salmonella, and fungi are microbial indices of a great importance in nature-based drugs. In Ardabil region, a poultice mixture containing animal butter and turmeric is the most widely used natural medicine in curing and improving the recovery of pressure ulcers. This mixture is manually prepared at home. Due to the scarcity of the research regarding the microbial load and the effects resulting from the above-mentioned method on the recovery process of the chronic ulcers, the aim of this study is to prepare this traditional poultice mixture in laboratory in an environment free of pathogenic microbes and investigated its effects on the recovery process of the pressure ulcers.
Turmeric rhizome features in pharmaceutics
Turmeric is the native plant of southeastern Asia from ginger family. Turmeric contains protein (6.3%), fat. (5.1%), minerals (3.5%), carbohydrates (69.4%) and moisture (13.1%). One of the most important compounds found in abundance in turmeric is curcumin. This compound which is the most active element of turmeric and accounts for its yellow color constitutes 3%–4% of turmeric and its numerous pharmacological effects such as anticancer, anti-inflammation, effective as remedy against pain, accelerate healing process, and cell division boosting features have been confirmed in a number of studies. Its anti-inflammatory effect which it exert by means of prostaglandins can be compared with drugs such as phenylbutazone. A study conducted by Dua et al. reported of the antibacterial effect of a topical poultice containing turmeric extract and norfloxacin on the ulcer healing process can be compared with silver sulfadiazine 1%.
Animal butter features in pharmaceutics
Animal butter comprises of elements such as water 14.4%, fat 32.4%, protein 36%, lactose 12%, and other elements, 5.5%. Studies have shown the positive effect of animal oil on the ulcer area and different results have also been obtained but what is clear is the presence of the high amounts of Omega-3 fatty acids in animal oil exerts a direct effect on the area of ulcer inflammation which results in reduction in such a way that this fatty acid takes the place of arachidonic acid in phospholipid membrane and interrupts the production process of inflammatory prostaglandins and leukotrienes. Moreover, triglycerides extract in the animal oil act as protections for the skin and play a role in cell nutrition, immunity, and inflammatory processes and are known to play a part in treating and preventing arthritis and pressure ulcers. One of the other reasons for the positive effect of animal oil on healing is the increase in blood circulation at the periphery of the ulcer area in such a manner that, with the enrichment of the RBC membrane lipids of Omega-3 fatty acids, these globules become more flexible and reaches the surface area of the ulcer through the veins more easily. Furthermore, Vitamins A and D existing in animal oil have effects on the collagen construction and ulcer healing. It is worth mentioning that animal butter contains 4%–5% linoleic acid which plays a role in cell membrane bindings and also found to be useful effects in somatic development of the body.
| Material and Methods|| |
Preparing synthetic poultice through traditional methods
Bovine butter was placed on mild heat for 10 min and stirred continuously to prevent it from burning. In the next step, wheat flour is slowly added for about two-third of the bovine butter amount and in discontinuous intervals in order for it to become sufficiently consolidated and at the same time while adding the flour turmeric rhizome powder is added to the mixture for about one-eighth of the animal butter. After, a complete uniform mixture was obtained and allowed to cool down at an ambient temperature. The poultice is evenly applied to the ulcer area.
Preparing poultice based on laboratory methods
Turmeric rhizome was purchased from the medicinal herbs market in Ardabil which underwent an identification process by the corresponding specialist from Ardabil medical sciences university pharmacology department and milled. Turmeric rhizome powder was placed in ethanol 95% v/v in a dark environment and at room temperature for 24 hours (topical gel)., To dehydrate the animal butter, it was condensed to drying in 70°C and then placed in incubator for 2 days. Turmeric extract and animal butter were mixed with sterilized wheat flour under bacteriological hood in an autoclave. In the final stage, its microbial count was evaluated. The prepared mixture was evaluated for microbial load by the US Pharmacopeia Reference Standard (USP). The microbial load of the poultice was assessed based on a standard range and the poultices prepared based on sterile methods were put in 60 g sterilized cans after verification and then they were UV top-sealed. For complete assurance, the contamination rate was evaluated each time a new poultice was prepared. Based on American Institute of Pharmacopeia (USP) reference tables, the standard microbial load for the amount of the bacteria existing in the topical poultices should be <101 cfu/g, and for the fungus count, it should be <102 cfu/g.,
Microbial experiment explained
In this study, the synthetic poultice was prepared based on traditional methods and transferred to the laboratory. Then, under sterile conditions, it was cultured in microbe growth media with a dilution rate of 1–10. To determine the bacteria extant in the poultice, a combination of culture media, namely, MacConkey Agar, Nutrient Agar, and Mannitol Salt Agar, was used; as for determination of the molds and fungi, Sabouraud Dextrose growth agar was used. Bacteria culture media were kept at temperature of 37°C for 24 h and the fungus culturing medium was put in temperatures of 25°C and 37°C for 72 h. After the expiration of the specified time, the plates containing the culture media were investigated for bacteria and fungus growth. To differentiate the Gram-positive coccyx, catalase tests were applied and coagulase tests were used for the differentiation of the golden staphylococci (positive coagulase) from the other staphylococci.
Seven conscious patients, with Category II and Category III pressure ulcers in various parts of the body such as sacrum, trochanter, and occiput, with an age range of 50–81 years were investigated in a pilot study. The study inclusion criteria were not consuming immune system weakening drugs, absence of diabetes, AIDS, and Buerger's vascular disease whereas the exclusion criteria were the development of sensitivity to the poultice and the patient's lack of willingness to make the use of traditional poultice or stating discontent to continue participation with the study. After a written permit was obtained from the patients and the letter of agreement was filled, the patients from treatment centers associated with Ardabil's Medical Sciences University and Welfare Organization were selected based on a convenience sampling method and received dressing with the traditional poultice twice a day after their ulcers were cleansed each time. The patients' ulcers were evaluated according to PUSS scale in terms of the ulcer peripheral extensions, the amount of secretions, necrosis, granulation, and epithelialization of the tissue, and then, the patients were monitored weekly for 4 weeks and the ulcer healing results were recorded.
| Results|| |
It was found that the synthetic poultice prepared based on traditional methods was contaminated with Gram-positive microbial factors and Staphylococcus aureus accounts for the highest rate of contamination. No evidence was found indicative of Gram-negative bacteria and fungus presence.
The pathogenic agents were not higher than the standard limit announced by USP in all of the seven times the synthetic poultice was prepared. The butter was completely dehydrated after heating up to 70°C and the turmeric rhizome powder was placed in ethanol 95% (v/v) for 24 h, the Gram-positive bacteria clone count in the entire stages of preparation, and in all of the samples were in a range of 10 × 101–4.0 × 101 and no sign of Gram-negative bacteria and fungi were found.
Synthetic poultice effect on human samples
The study sample volume consisted of four male and three female patients [Table 1]. The preliminary mean area of the ulcers was 13.41 cm 2. Other quantitative variables such as the hemoglobin range, the body mass index, and the average age were 9.97, 26.68, and 63.85, respectively.
|Table 1: Gender, ulcer area, activity, nutrition and ulcer category of patients|
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The results indicate that, in the first evaluation before intervention, the mean area of ulcers was 13.41, whereas, in the last evaluation, this area reduced to 2.21. The pressure ulcer scale of healing (PUSH) from the day 1 up to the week 4 was reduced from 13.42 to 5.85. Furthermore, in terms of the tissue evaluations, the entire samples were observed visually and no sign of tissue necrosis and granulation was visible in any of the samples. Furthermore, the samples' dressing of the ulcer was found covered with the secretions in the first evaluation, whereas one case was free of any secretions and the remaining six samples had little secretions in the last evaluation. None of the study sample volume was found with infections in the ulcer area [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6].
|Figure 2: Culture medium gram-positive showed highest rate of contamination with staphylococcus aureus|
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|Figure 5: Ulcer healing after 14 days of intake traditional poultice mixture|
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|Figure 6: Ulcer healing after 28 days of intake traditional poultice mixture|
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| Conclusion|| |
Based on the result obtained, it can be inferred that none of the study sample volume had infections in the ulcer area and the obtained changes are indicative of the effective and successful recovery of the pressure ulcer by the use of synthetic poultice, thus it can be stated that the poultice considerably increases circulation in the blood veins at the periphery of the affected area, reduces the edema, secretions, and skin inflammations at the periphery of the ulcer area and generally brings about an enhancement in the recovery process. Although there is a new and ready-to-layered wound dressing for pressure ulcers' but preparation of traditional poultice is easy, convenience, free from microbial agents, with no side effects and adverse symptoms and cost-effective therefore this study has recommended it for improving diabetic foot ulcers too.
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Conflicts of interest
There are no conflicts of interest.
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Department of Nursing, Ardabil University of Medical Science, Ardabil
Source of Support: None, Conflict of Interest: None
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]