Applied Science and Engineering Journal for Advanced Research

Introduction

Since traditional medicine has always been the most budget-friendly and easily accessible treatment type, it has been the most commonly preferred primary health care system, especially among poor communities.

Throughout the history, traditional plants were used by the local people for therapeutic purposes. As we can understand from the writings, the therapeutic use of medicinal plants, which was firstly used by the Chinese who use the plants as the natural herbal medicines, dates back to ancient times, circa 4000 - 5000 B.C.

(1) However, the Rig-Veda, which is known to be written between 1600 - 3500 B.C., states that plants were firstly used for medicinal purposes in India(2). The ancient physicians focusing on ancient medical treatments in India as a local medicine system studied the characteristics and remedial usage of medicinal plants in detail and recorded the empirical data related to their studies.

Throughout the world, medicinal plants are an integral component of endemic medical systems. The ethno botany stands out as a rich reference guide in terms of the research and development activities with respect to the natural drugs.

The significant historical use is meant by “Traditional” use of phytomedicines (herbal medicines), and this definitely applies to many products that are accessible as “traditional herbal medicines”.(3) A great number of the people in many developing countries mostly depend on traditional practitioners and their treatment techniques using plants therapeutically to deal with their health care needs(8).

Modern medicine can be applied along with such traditional treatment procedures; however, use of plants for medicinal purposes has not fallen from grace thanks to cultural and historical reasons(4). Natural products have been instruments in treating and preventing human diseases all around the world(7).

Photo 2: pinus nigra

Photo 1: Clevenger apparatus oil One Kg from pinus nigra leaves gave oil about 1.65ml as (Photo :2). One Kg from Abies equi-trojani gave oil about 2ml as (Photo :3), One Kg from Pinus sylvestriys gave oil about1.0ml as (Photo:4).

Photo 4: Pinus sylvestris

Photo 7: Cedrus libani

Microbial Material

Strains of fungi and bacteria obtained from the biology Laboratory in Kastamonu University:

Srains Gram-positive bacteria: Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis DSMZ 20044, Enterococcus faecium, Enterococcus faecalis ATCC 29212, Bacillus subtilisA DSMZ 1971 (Table 3.2).

Strains Gram-negative bacteria: Salmonella typhimurium, Salmonella kentucky, Salmonella infantis, Salmonella enterlidis SL 1344, coli ATCC 25922, Enterobacter aerogenes ATCC 13048, Pseudomonas aeruginosa, Pseudomonas fluorescens DSMZ 50071, Klebsiella pneumonia ATCC 7544 (Table 3.3).

And fungi: Candida albicans DSMZ 1386.

Prepare Microorganisms

A bacteria culture (which has been adjusted to 0.5 McFarland standards (1.0 x 108 CFU/ml) deliberated using the Turbidometer (Oxoid, UK)). Took swab of the bacterial and fungal suspensions by sterile swab then mixed ( saline solution ) in the test tube (Photo :8). After that, wrote the bacteria and fungi names on the tubes then mixed by the shaker befor used.(9)

Photo 8: Prepare microbs in test tube

The MS examine conditions utilized incorporated an exchange line temperature of 250˚C, an interface temperature of 250˚C, a particle source temperature of 200˚C. Recognizable proof of the constituents depended on examination of the maintenance times and on PC coordinating against Wiley Data library. At the point when conceivable reference mixes were cochromatographed to affirm GC maintenance times.(13)

Results

Table 2: MIC results µ/ml.

“-’’= No affact

Table 3: MBC results µ/ml.

“-’’= No affact

nigra have affected against Gram-positive bacteria: Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis DSMZ 20044, E.durans E. faecium, E. facalis and Bacillus subtilis ADSMZ 1971, L. monocytogenes and L.innoeua where were 8 µ/ml, 4µ/ml, 1µ/ml, 6 µ/ml, 4 µ/ml, 2 µ/ml, 4 µ/ml and 1 respectively. On the other hand have no affect against C. albicans. And Gram-native bacteria Salmonella typhimurium, Salmonella kentucky, Salmonella infantis, S. enterlidis, E.coli, E. aerogenes, P.aeruginosa, P. fluorescens and K. pneumonia where were 1µ/ml, 1µ/ml, 3µ/ml, 1µ/ml, 1µ/ml, 2µ/ml, 7µ/ml, 2µ/ml and 1µ/ml. As shown (Graphic 1).

Photo 11: MBC of P. nigra

Photo 13: MBC of P. sylvestris

libani have affected against Gram-positive bacteria: S. aureus, S. epidermidis, E.durans E. faecium, B. subtilis L.monocytogenes and L.innoeua where were 9µ/ml, 10 µ/ml, 3 µ/ml, 1µ/ml, 3µ/ml, 6µ/ml and 2 µ/ml respectively.

But have no affect against E. facalis and fungi of C. albicans. And Gram-native bacteria Salmonella typhimurium, Salmonella kentucky, Salmonella infantis, S. enterlidis, E.coli, E. aerogenes, P.aeruginosa, P. fluorescens and K. pneumonia where were 2µ/ml, 1µ/ml, 5µ/ml, 2µ/ml, 2µ/ml, 2µ/ml, 5µ/ml, 2µ/ml and 2µ/ml. As shown (Graphic 3).

Photo 15. C. libani.

Photo 17: MBC of P. orientalis

equi-trojani have affected against Gram-positive bacteria: S. aureus, S. epidermidis, E.durans, B. subtilis, L.monocytogenes and L.innoeua where were 10 µ/ml, 8µ/ml, 4 µ/ml, 8 µ/ml, 2 µ/ml, 8 µ/ml and 2µ/ml respectively. But have no affect against E. facium . And Gram-native bacteria Salmonella typhimurium, Salmonella kentucky, Salmonella infantis, S. enterlidis, E.coli, E. aerogenes, P.aeruginosa, P. fluorescens and K. pneumonia where were 1 µ/ml, 4µ/ml, 4µ/ml, 4µ/ml, 5µ/ml, 3µ/ml, 4µ/ml, 2µ/ml and 3µ/ml. On the other hand have no affect against C. albicans. As shown in (Graphic 5).

Conclusion and Discussion

Czerwińska, E., & Szparaga, A. (2015) tested the antimicrobial activity of the leaves, flowers and young sprouts of Pinus sylvestris flowers and leaves essential oil against Gram-positiveand 12 different microorganisms of fungi. They have found a disc-diffusion method inhibition zone of Staphylococcus aureus ( 12.08 mm), Lavandula vera (10.30 mm), Listeria monocytogenes(12.08 mm) and Escherichia coli (ø 8.22 mm) [14].

In our study we determined of MIC value for Staphylococcus aureus 9µg/mL, Listeria monocytogenes 6 µg/mL and 1 µg/mL of E. coli, but no affect againsy fungi. This difference is possibly related to they used 5 g of driedplant poured over 100 ml of cold water and left for 24 h at 20°C, then filtered.(19)

Hakkı Alma et al. (1999) tested the antimicrobial activity of many parts of various trees grown in the Kahramanmaraş region of Turkey of Pinus brutia, Cedrus libani, Pinus nigra and other Chloroform, CH3)2CO and methanol concentrates of leaves in opposite to 15 microorganisms.

Picea orientalis aganst Staphylococcus aureus, Staphylococcus aureus (MRSA), Escherichia coli, Klebsiella pneumonia and Pseudomonas aeruginosa werw 7mm, 7mm, 7mm 8mm and 7mm, respectively, but have no affect against Candida albicans with all extract [16]. In our study we determined of MIC value for Pinus brutia, Cedrus libani, Pinus nigra Abies equi-trojani and Picea orientalis were (1-9 µg/mL), (1-10 µg/mL), (1-8 µg/mL), (1-10 µg/mL) and (1-8 µg/mL) respectively. Except Candida albicans have no affect.(17)

References

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2. Ghimire, B., Lee, C., Yang, J., & Heo, K. (2015). Comparative leaf anatomy of some species of Abies and Picea (Pinaceae). Acta Botanica Brasilica, 29(3), 346-353.
3. Tumen, I., Hafizoglu, H., Kilic, A., Dönmez, I. E., Sivrikaya, H., & Reunanen, M. (2010). Yields and constituents of essential oil from cones of Pinaceae natively grown in Turkey. Molecules, 15, 5797-5806.
4. Šarac, Z., Matejić, J. S., Stojanović-Radić, Z. Z., Veselinović, J. B., Džamić, A. M., Bojović, S., & Marin, P. D. (2014). Biological activity of Pinus nigra terpenes—Evaluation of FtsZ inhibition by selected compounds as contribution to their antimicrobial activity. Computers in Biology and Medicine, 54, 72-78.
5. Maciąg, A., Milaković, D., Christensen, H. H., Antolović, V., & Kalemba, D. (2007). Essential oil composition and plant-insect relations in Scots pine (Pinus sylvestris ).
6. Saab, (2011). Phytochemical analysis, antiproliferative against k562 humam chronic myelogenus leukemia, antiviral and hypoglycaemic activities of cedrus species and medicinal plants native from Libanon. (Doctoral Dissertation, Università Degli Studi di Ferrara).

16. Šarac, Z., Matejić, J. S., Stojanović-Radić, Z. Z., Veselinović, J. B., Džamić, A. M., Bojović, S., & Marin, P. D. (2014). Biological activity of Pinus nigra terpenes—Evaluation of FtsZ inhibition by selected compounds as contribution to their antimicrobial Computers in biology and medicine, 54, 72-78.