Mesquite ( Prosopis juliflora ) plant prevalence in different districts of Balochistan & antibacterial activity of its pods

This study was conducted to evaluate in vivo multiple uses of mesquite plant, identification of selected areas in Balochistan and anti-bacterial activity of mesquite pods (alkaloid-enriched extracts). For this purpose a survey was conducted in areas where mesquite plant prevails. Pods of mesquite were collected from different trees standing at Yaro Pishin Roadside Plantation and were brought to the physiology laboratory of Centre for Advanced Studies in Vaccinology & Biotechnology (CASVAB) University of Balochistan Quetta. Pods were dried and extract was obtained which was subjected to check antibacterial activity through two different methods. The inhibition zones of bacteria were measured in mm. It was concluded that mesquite, which was once viewed as weed, its pods have antibacterial components. Furthermore, this study indicated that mesquite is a useful plant if properly managed.


Introduction
Mesquite (Prosopis juliflora) is woody legumes found in arid and semiarid areas of Northern and Southern America, Mexico, Northern Africa and Eastern part of Asia.Mesquite have more than 40 species, originated from South America [1].Although the geographical dispersion of mesquite plant is stable, its density have amplified in the recent past which have been attributed to anthropogenic activities or dissemination of mesquite seed by the livestock [2].Actually mesquite (genus Prosopis) is a thorny wild shrub occurring in arid and semiarid zones of the world.Mesquite species is multipurpose evergreen and at cold areas deciduous shrub / tree with straight and thorny arm producing fruit called pods.In shallow soil mesquite grows as a small shrub whereas in deep soil it gets height as tall as 50ft making rounded canopy with availability of adequate moisture.Thus mesquite plants have different size and shape, subject to growing conditions and water availability.If seedlings of mesquite plants are not damaged by weather or animals, trees may get 40-50 feet height, with a large canopy.If the seedling is injured, the plant grows into an extensive multi-trunked shrub.Mesquite is one of the most economically and ecologically important shrubs or tree, which are found in arid and semi-arid zones of North and South America, North Africa and East Asia.More than 40 species of mesquite are originated from South America [1].Mesquite dispersal has been attributed to anthropogenic activities such as suppression of natural fires or dispersion of mesquite seed by the herding and journey of domestic livestock [2].Mesquite are also referred to as Algarroba.The genus Prosopis has 44 species which that have been known [3].The genus Prosopis is highly adapted to arid lands of Northern, Southern America, Central America and the Caribbean [4].It is adapted to a wide range of differences in temperature i.e. 12 to 50 o C .It can also tolerate drought condition i.e. regions where rainfall is less than 500 mm per year.Mesquite pods are regarded as highly sugar content pods and it is said that pods are included in human diet and cattle fodder for thousands of years.About 100 G/ Kg of dried material, with 40 G/Kg of rough protein on a dried material base (DM), pods have been used in semi-arid and arid areas of the world for centuries by human and cattle in diet.Mesquite pods contains toxins and polyphenolics, limiting uses of the pods as fodder for animals.Algaroba is noticed in cattle in many parts of the World.Medical symptoms, includes mandibular sensations, lethal saliva, problems in assimilation and twisted head, with many more symptoms such as cranial nerves function interruption , are common in small ruminants.Extended combination of algarroba in cattle foods is much lethal.Cattle are given diets having less than 40 g of dry algaroba beans/100 g DM, to avoid intoxication particularly when the nourishing duration surpasses two months.Mesquite perhaps originated in Peru or Mexico, now it is found as wild weed in semi-arid and arid zones of North America, South America, Central America, Mexico and Southern parts of United States of America.Mesquite has been brought into tropical regions such as North-eastern Brazil, Africa, Southeast Asia and Southern parts of Asia.Basically mesquite was transported in barren arid zones of the world as a valuable feed, source of energy and shade shrub.However, mesquite with the passage of time evolved into an unwanted invader weed species, especially in regions where water is scarce.Algarroba, honey mesquite, mesquite.

Mode of distribution pattern
In several parts of Pakistan, Prosopis juliflora occurs and grows in following ecological conditions.Altitude: -0 -1500m Mean annual temp: 14-34 o c Mean annual rainfall: 50 -1200 mm.

Seed ecology and seedling development
Mesquite propagates only through seed.Mesquite seed occurs in long pods (legumes) around 6-10 inches long and comprise 8-24 seeds per pod.Most of the pods which falls to the ground are eaten by ruminants or destroyed by insects.Within Soil seed remain viable for many years i.e. why much suitable for dry areas of the province.Mesquite pods do not burst out at maturity.Foraging animals that eat pods distribute mesquite seed through fecal after eating.Seeds when taken out of pods germinated in Pishin within 24 hours after processed in Soda.Germination is boosted if seed are processed by passage through animal digestive system [2].The author passaged it through digestive tract of donkey and germination was marvellous.

Growth of mesquite
Mesquite can raise up to 25 feet in height with main support stems as much as 2 feet in diameter mesquite which depend on the site and climate.Sapling mature as single to few-stemmed plants.Flowering begins shortly after leaf development.Leaf development starts shortly after March.In flowering period, mesquite trees have thousands of flowers but only few of these produce pods.Mostly pods are matured at August -September period.

Root characteristics and water use patterns
Mesquite is deep-rooted, water-using "phreatophyte" plant that resist drought [24].In drought affected areas of Balochistan mesquite depends on shallow lateral roots system which is extended as much as 30 ft from the plant .The author calculated roots up to 10 feet at district Pishin.Mesquite plant is extremely hard, scare water using plant that avoid drought .It can get water from the water table through its long taproot system.Taproots of the mesquite tree may reach a depth of 200 feet down into the soil as recorded.Furthermore, mesquite can also use water from surface part of the ground, liable to availability of water.Mesquite plant can easily and quickly shift itself from using one water source to the other.For several people, particularly farmers, mesquite plant is a pest as the plant competes with rangeland grasses for moisture.As mesquite trees have a long taproot which are used to find sufficient moisture for growth purpose.This feature allows mesquite plant to survive through severe drought.There are recorded instances of taproots of the mesquite tree reaching a depth of almost 200 feet down into the soil.The roots of the mesquite can redevelop into a plant even if the tree is completely cut down above the ground that make mesquite plant one tough tree to get rid of completely .Farmers believe that the mesquite removes water from the land that could be used for livestock and farming, thus getting title of notorious plant.

Determining the prevalence of mesquite plants within the selected areas
Study comprises of field data Acquisition and data assessment.Both North and South part of Balochistan were selected.Samples of mesquite pods were also collected from the field area for determining out antibacterial activity.

Evaluation of in vivo multiple uses of mesquite plant through field survey
Survey is an important method regarding collection of information on a particular object.Vegetation Survey is the assessment of occurrence of a particular (under study) plant in an area.A survey was carried out to assess the multiple uses of mesquite plant in vivo.

Obtaining extracts
The powdered plant was first extracted by filtration with hexane (5 L).The hexanic extract (HE, 6.50 g) was obtained after passing the concentration in a rotary evaporator (40 °C).Then, the plant material was extracted with ethanol (5 L) by filtration, and this solution was concentrated under reduced pressure to yield the crude ethanoic extract (EE, 435 g).Ethanoic extract (EE, 285 g) solvent was made in aqueous 1.6 M acetic acid (AcOH, 250 mL), and the subsequent solution was filtered completely to obtain the acid aqueous solution (AAS).Then, neutralization of acid aqueous solution was done with NaOH 2.0 M up to pH 9.0, converting it into the basic aqueous solution (BAS), which was later on extracted with CHCl3, and the resultant organic layer was washed out with brine, desiccated with Na2SO4 and concentrated under low pressure to produce the basic chloroformic extract (BCE, 0.72 g).

Antibacterial activity was carried out by two different methods
Bacteria Culture were carried out in nutrient broth (Oxoid) for 24 hours at temperature 37 °C.The basic chloroformic extract (BCE) was assayed against the Grampositive bacteria in plates having 5 well.The primary microorganisms' inocula were adjusted to the turbidity of 0.5 McFarland, and the last concentration was 0.9 × 10 6 cells/well.Samples were assayed in triplicate.The extract being diluted in a normal solution prepared in water-dimethyl sulfoxide (80:20 v/v) and examined at the final concentrations of , 50, 25, 6.3, 6.3, and 0.8 μg/mL respectively.Samples were assayed in triplicate.

The Plate agar diffusion (Inhibition zone) method
In this method Nutrient Agar (NA) was used.First of all 500 micro liter of a uniform bacterial cell suspension of M. luteus, S. aureus, S. mutans and other were spread over on sterile Petri-dishes which was solidified at room temperature for 24 hours.The wells made after drying the plates were filled with extract suspension.The plates were later on incubated at 37°C for 24 hours and the inhibition zones were measured [28].Three replicates were made for the treatment.

Mesquite prevalence Mesquite prevails both in North and
South of Balochistan Among North parts of Balochistan mesquite plants are scattered all over Loralai district, good patches of mesquite plants are found inside Loralai and its adjacent areas.Mesquite is spreading with the passage of time .The core zone is District Duki.In district Musakhel mesquite plants are scattered all over the District.Mesquite plants are found on large scale in district Barkhan with water channels which flow from Musakhel, mesquite plants reach Naharkot.Disrict Harnai which is located at 30* 6 N latitude and 67* 56" longitude and at elevation around 2952 above sea level also had good patches of Mesquite plants.In Pishin, Quetta, Killa Abdullah and Killa Safullah naturally grown mesquite plants are rarely found instead mesquite are planted on roadside through plantation.Among Southern districts, mesquite plants are found in Bibi Nani, Badhar, Sigugar and Gokurt areas of Bolan around water channels and seasonal streams.In whole of Jaffer Abad district mesquite are found scattered.In Jhal Magsi Gajan, Gandava and Panjuk are main areas of mesquite distribution.Mesquite plants are distributed all over the district, mainly inside Sibi Thal, Quat, Terri and in Mall.Mesquite plants grow highly in Lasbella where it is considered as weed plant.
Mesquite plant is one of the most common tree of the legume family found both in North and South parts of Balochistan.This plant is ideally adaptable for arid zones of Balochistan as it requires less water and highly drought resistant.Mesquite grows in the Northern and Southern parts of Balochistan.Mesquite is found in areas having a low annual rainfall.The advantages of the mesquite plant is more than the title of notorious weed plant given by ranchers.The public had pluralistic views about mesquite plant.Which is elaborated in figures 1, 2,3,4,5 and 6.

Outcome of the survey
The bean pods of the mesquite is edible.Mesquite pods provides good feed for cattle and is an excellent fuel wood.It should, therefore, be worthwhile to improve its stocking by artificial sowings and encouragement of natural regeneration through establishment of seed reserves at potential sites.The pods of mesquite palatable, and digestible for the animals' especially small ruminants.The pods of mesquite are rich in nutrient and fulfils the body requirement of animals.Since the availability of forage has become reduced pods of mesquite have become major source animal diet.Mesquite is a good fuel wood and local population meets their energy requirement from the collection of mesquite branches etc. Mesquite burns very slowly .It is also used in brick and charcoal making industry.
Mesquite is hard and it could be used in making furniture and tool handle.The wood is used for decoration items.Mesquite tree play important role in the environment, and provide habitat for many wildlife including animals and migratory birds.It is interesting to be noted that mesquite trees provide shade and wildlife habitat where other trees will not grow.
Mesquite is very good for sand dune stabilization in desert .In fact mesquite is tree of dry region.It could be used in desert areas for reclamation of sand dunes and generating a valuable source of income to the poor of the desert area.Exactly mesquite (Kabol) is a very resilient tree against drought thus is very suitable for dry areas.
Mesquite's flowers offer a nectar source for honey bees to produce mesquite honey, which has a characteristic flavour.Figure 7 showing mature mesquite tree with full of flowers on Pishin Quetta highway.Mesquite (Kabol) plays an important role in supporting the livelihood of the rural poor, tenants, farmers and artisans, due to all these it is called the tree of 'the poor'.In many rural areas, mesquite plant is usually the single major source of fuel, and fodder for dry season and generate income for many poor families.The large size branches and trunks of mesquite tree yield good quality timber.Mesquite meal is obtained from the pods of the tree.Mesquite flour is sweet in taste, low in carbohydrate and fat concentration.It is rich in protein and have about 20 per cent protein as compared to the seeds which have up to 40 per cent protein).Mesquite has good concentration of calcium, manganese, potassium, iron, zinc and high in fiber and lysine.Root of mesquite plant at Pishin, Balochistan was calculated roots up to 10 feet at District Pishin by the authors which shows its resilience against drought as showed in figure 8.   Results of the present study for screening antibacterial activity showed that the extract of mesquite (kabol) pods was effective against all tested bacteria.Which has also been reported in some previous works on the same [33-37].The antibacterial substances not only protects mesquite against microorganisms but could also help pharmaceutical industries.Survey indicated that mesquite was found in almost all selected districts of Balochistan either as wild or planted by Balochistan Forest and Wild Department for afforestation and sand dunes stabilization as was found in Gwadar district.Which has also been reported in previous studies [38].The public had pluralistic views about mesquite plant which negates the previous title of being declared noxious weeds [39].It was observed that 98 per cent land of Balochistan is barren land and dry as Balochistan is water scarce province.Balochistan has arid to hyper arid climate with very low precipitation levels.If an aerial campaign of mesquite sowing is started in these barren areas in collaboration with the Balochistan Forest and Wildlife Department, the meagre forest cover could be enhanced and pressure on reserve tree cutting would be reduced greatly.We can improve our range lands to a great extent in a short period of 5 to 10 years and improve the socioeconomic conditions of the local poor on one side and enhance the poor ratio of trees on the other by planting this multipurpose tree in drought affected

Conclusions
The present study pointed out that mesquite prevails in Balochistan and its pods have antibacterial components.Besides this mesquite could be an alternative source of food for grazing animals if planted on large scale with proper management.Acknowledgement This research paper could not have been written without the wisdom, keen intellect, and unwavering commitment of Prof Dr. Shakeel Babar, and Dr. Zia Uddin, members and colleagues from CASVAB.They have drawn upon their scholarship and extensive practical experience not only to produce this research paper but also to provide valuable advice and support to my efforts on my research work.Prof Dr. Muhammad Masod Tariq Kiyani in particular, has guided during whole research work, I am honored and grateful for his cooperation.I also owe a considerable debt of gratitude to my Forest and Wildlife Department, Balochistan officers for their extensive contributions and advice to the progress of this research work.This research work has benefited from these partnerships and input from dozens of farmers /interviewers from every walk of life.

Figure
Figure 1.Showing agriculture damage Figure 2. Showing respondent views about mesquite plant useful or weed

, 13].Still mesquite has been considered as weed in many parts of the world [3, 14, 15]. Mesquite origin and dispersion in Pakistan
[22]uite is one of the major exotic intrusive species in Pakistan[18].In Gwadar, a district of Balochistan, Mesquite is propagated, for the stabilization of sand dunes movement[22].Now Mesquite pods are a major source of forage for cattle in many parts of Balochistan.Mesquite grows in the arid and semi-arid areas of Balochistan.Districts of Pishin, Harnai Bolan, Loralai, Sibi, Jaffarabad, Jhalmagsi, Barkan, Musa Khail, and Barkan, are main districts where mesquite dispersal was observed.

Table 1 . Province wise forests cover statistics
Table2) so as to mainly satisfy the objectives of the survey.The survey was carried out in the selected areas mostly which had mesquite plant distribution.The data was collected both from North and South parts of Balochistan.The main aim was to know the general community judgment about mesquite plant.Survey was done on the basis of interviews.