Antimicrobial activity of two medicinally important plants (Juniperus excelsa and Ephedra intermedia) of Gharshinan Rodh Malazai area, District Pishin, Balochistan-Pakistan

Antimicrobial activity of two medicinally important plants ( Juniperus excelsa and Ephedra intermedia ) of


Introduction
Utilization of medicinal plant species as medication is one of the routinely used practices in subcontinent. Plants have consistently been a cure for various infections because of quality of numerous phytochemical compounds. Medications that are acquired from home grown sources have a wonderful impact in forestalling and restoring deadly sicknesses. Individuals in most created nations utilize conventional medication in medical services [1]. Locally, different plants are utilized as drugs at the preliminary levels in various areas of the world. Mostly of individuals are subject to natural plants for medical care [2]. Authenticated by WHO, that Gentamycin like biological drug further not useful for prevention of any microbes. There are multiple names of drugs which are effective once but do not show its activity against bacterial strains [3]. Some medication by plants has emblems that causes a various abnormal feature as like as side effects in human beings. These abnormal characteristics of plants compelled the ethno botanist for seeking drugs which are affordable, biologically degradable and with least side effects [4]. Traditionally ethno medicinal plants have a vital role in the treatment of developed nations. Plants which belong to herbs have been utilized for the treatment of human diseases due to these is medicinally important [5]. Cassia alata (L.) is an sin qua non medicinal plant indigenous to Asian states, particularly South Asian States. The research parts of the included plant were experimented to show ethno botanical properties such as antimicrobial and painkiller. The treatment of ring worm took place with the help of leaves of aforementioned plant. It is traditionally high beneficial against dermal diseases in human beings and cattle [6]. Over population may cause the needy components of life such as food, cloth and hygiene. The homeopathic uses of plants have a great concern over highly effective and least side effects in biota as compared to allopathic medicine [7] concluded that the use of ethno botanical plants by observing their medical application through update research. Long range of medicinally important plant parts are utilized as raw components for medicines.
However, numerous plants have been observed for antibacterial activity but still some local plants of some areas remain under exploration which is unexplored for the diverse antimicrobial potential that the local community is carelessly utilizing [8].
The growing scene of microbes resistance to the affordable antibiotics has felt the need to examine for coming and new antimicrobial agents. Gharshinan Rodh Malazai Pishin is the study area which is located located in the northwest of Baluchistan, in the east of the province near the border with Afghanistan. In summer season temperature can reach 40 °C (104 °F). In the winter, temperatures can down to freezing point. The Latitude is 30°34′54″ N and Longitude is 66°59′38″ E. Pishin elevation above sea level is 1536 m = 5039 ft. This area has extremely low rainfall and evaporation is very high on the one hand and on the other hand with the increasing demand for urban, agricultural and industrial, faces a severe water stress [9]. Psammogeton comprises of 6 species, majority is found in sandy hills of deserts in various parts of Turkmenistan, Pakistan, Afghanistan and Iran (Nasrabadi et al., 2009). P. biternatum Edgew normally known as "izbotk" in Kalat and "sparki" in different locales. It is a yearly species that goes from 15 to 22cm long. It has little blossoms that are white or pink in shading. It blossoms from March to April. It is a yearly plant that is situated in a few locales of District Kalat. Psammogeton biternetum is privately utilized for the fix of intestinal sickness, challenging hack, typhoid and chest infections by individuals of district Kalat [10]. P. biternatum Edgew seeds are extremely useful for post pregnancy contaminations (after conveyance) if ground and blended in water and taken between suppers [11].
Researchers reported about those plants which increases the antimicrobial activity against the resistant strains of bacteria. Researchers also studied the antimicrobial agents evolution of Solanum incanum, and Hydnora johanis against Enterococcuss feacalis by utilizing disc diffusion method [12]. Ephedra sinica is the principle plant beginning of the Chinese home grown medication Mahuang, Ephedra Herb. 85 examples of E. sinica, gathered across eastern China, Mongolia, and Buryatia (Russia), were concentrated anatomically and artificially to clarify the neighborhood varieties and the connection between ecological elements and the varieties. The outcomes showed that examples filled in more dry conditions would in general have a more twisted epidermis, more circular tubers, and more sub epidermal fiber packages anatomically, and contained more all-out ephedrine alkaloids. These examples likewise had a high pseudoephedrine content. These achievements suggest that Ephedra herb with great quality ought to be gathered from parched fields, and the substance quality can be assessed by noticing the anatomical attributes [13]. A quantitative survey was conducted in 32 stands of juniper (Juniperus excelsa) in four adjacent districts of Baluchistan [Baluchistan] Pakistan. No other tree species was recorded in the study area. Data are tabulated on the herb, shrub and grass composition of the stands -only 11 species were found apart from juniper. Mean density of juniper was 105/ha and basal area 18.4 m 2 /ha. Density and basal area were significantly correlated. Density of female trees was higher than that of male trees. Healthy trees produced only 21% of heavy seeds. Trees were slow growing (18 years/cm radial growth rate), and many were unhealthy and over-mature; there was no seedling regeneration. Details are given of 5 morphological tree forms found in the area. Leaf and soil analysis were also performed. Sandy clay loam with a calcareous nature was the dominant soil. No correlation was observed between tree density, basal area and soil characteristics. It is concluded that these forests are rapidly degrading due to human disturbance [14]. In pakistan 80% individuals are living in provincial regions. They are reliant upon therapeutic plants. Artemisia indica is utilized for asthma, uncontrollable and anxious impact however Medicago falcata contains enormous quantities of nutrients and proteins [15]. Wild Garlic (Allium ursinum) tissues (leaf, stem and root) were meant something negative for microorganisms which are microbes and growths [16].

Result
Leaves and fruits of J. excelsa were commonly moderate in amounts of organic contents like alkaloids tannins, flavonoids and saponins. The result of the antibacterial activity of the J.excelsa contents by the disc diffusion method is given in the (Table 1 &  Fig. 1). The microorganisms tested were Escherichia coli, Thiobacillus ferrooxidans (TBF), Leptospirillum ferrooxidans (LSF) Thiobacillus Thiooxidans and Staphylococcus aureus. All bacterial species tested were found sensitive to the essential oils contents of J. excelsa berries, leaves, and mixture with the inhibition zones ranging from, 12.6 mm to 23.3 mm, 11.8mm to 22.6mm and 12.7mm to 23.1 mm respectively. The higher value was found for tested Gram-negative bacteria as compared to tested Gram positive bacteria. As per above discussion, it is concluded that the highest inhibition zone reported for Escherchia coli which are 22.6mm, 23.1mm and 23.3mm for leaves, mixture (leaves + Berries) and berries respectively. As well as the lowest inhibition zone accounted for Staphylococcus aureus which are 11.8mm, 12.6mm and 12.7mm for leaves, berries and mixture. The average inhibitory zones of leaves, berries and mixture are 17.5mm, 18.2mm and 18.56mm respectively against tested five species of bacteria. The three parts of Juniperus excelsa has slightly difference against the microbes, especially bacteria. On the basis of average, the leaves have the lowest (17.5mm) inhibitory contents against bacteria, but the berries have the highest (18.56mm).    (LSF) Thiobacillus Thiooxidans and Escherichia coli. All bacterial species tested are sensitive to the essential oils of J. excelsa leaves, berries and mixture with the inhibition zones ranging from 11.9 mm to 22.7 mm, 12.7 mm to 23.4 mm and 12.8mm to 24.1 mm respectively. The lower values were found for the tested Gram-positive bacteria as compared to the tested Gram-negative bacteria. Omani authors reported that the essential oil of J. excelasa berries does not contain any antimicrobial activity against Staphylococcus aureus and Escherichia coli which is contradictory to the present study [21]. In another study the authors reported that J. excelsa essential showed less antimicrobial activity against the tested microorganisms [22]. It has been reported that the essential oils are usually believed to be active when they show minimum inhibitory concentration (MICs) of 128 mg/ml and/or below [23]. The essential oils isolated from leaves and twigs of J. excelsa growing wild in Lebanon showed antimicrobial activity against Staphylococcus aureus but Escherichia coli was resistant [24]. Therefore, the presented results of antimicrobial activity of J. excelsa essential oils similarities and dissimilarities are found in literature that could probably be due to the diverse percentage of chemical constituents present in essential oils. All bacterial species tried are touchy to the fundamental oils substance of J. excelsa berries, leaves, and blend with the hindrance zones going from, 12.6 mm to 23.3 mm, 11.8 mm to 22.6 mm and 12.7mm to 23.1 mm respectively. The higher worth was found for tested Gram-negative microbes when contrasted with tested Gram positive microscopic organisms. According to above conversation it is clear that the most elevated hindrance zone detailed for Escherchia coli which are 22 (11) and the most noteworthy for S. aureus (19). The normal of roots substance have the higher antibacterial movement when contrasted with shoots which are 18.25mm and 15mm. The roots and shoots of E. intermedia have the greatest antimicrobial (Antibacterial) movement against the S. aureus of microorganisms. Notwithstanding, the roots and shoots of E. intermedia have recorded the base action according to tried to B. subtilis. The normal of roots and shoots substance against the tried microscopic organisms: Staphylococcus aureus has the most noteworthy hindrance zone however the Bacillus subtilis has the least one when contrasted with different types of microorganisms.

Conclusion
The aforementioned research reviewed that the selected plants (Juniperus excelsa and Ephedra intermadai) have highly impacts against bacterial strains. Consequently, the outcomes display the chance of utilizing these concentrates for the illness of destructive diseases. The research further presumes that the berries, leaves and their mixture of Juniperus excelsa is better dynamic for microbial exercises. As well as, Ephedra intermadia has also a high effect against bacterial strains. Thus, it is very well may be reasoned that the two plants have restorative properties against wide scope of bacterial and contagious strains and can be utilized as better substitute to costly medications accessible in market.