Soil moisture status under traditional agroforestry ...



Haemagglutination as a rapid tool to differentiate Saraca asoca bark from the adulterant

Polyalthia longifolia

C BEENA* AND V V RADHAKRISHNAN

All India Coordinated Project on Medicinal , Aromatic Plants and Betalvine, College of Horticulture, Kerala Agricultural University, P.O. Vellanikkara, Thrissur -680656, Kerala, India.

E mail: beenac2@

ABSTRACT

Saraca asoca(Roxb.) Wilde, the asoka tree is one of the red listed plants of the Western Ghats. . The bark of asoka tree is the source of the ayurvedic medicine “asokarishtam” used in the treatment of gynecological disorders. The rising demand has led to its widespread adulteration. It is widely adulterated with the bark of Polyalthia longifolia an ornamental tree. This paper presents a quick and easy method to determine the adulteration in asoka bark. Haemagglutination method using the phosphate buffered saline ( PBS) extract of the stem barks and o positive human erythrocytes was proved to serve as an effective ,quick, easy and cheap tool in differentiating the raw bark of asoka from its major adulterant Polyalthia longifolia .This can be recommended as a tool for the floor level checking of the market samples for ensuring the quality .

Key words: Adulteration, haemagglutination, Saraca asoca, Polyalthia longifolia, PBS ( phosphate buffered saline) .

Saraca asoca (Roxb.) de Wilde commonly known as Asoka (Figure 1) is a sacred tree of India, famous for its use in the treatment of gynaecological disorders. Asoka belongs to the family Caesalpiniaceae. It is one of the red listed plants of the Western Ghats. Asoka is especially relied upon as an astringent to treat excessive uterine bleeding from various causes including hormone disorders, fibroids and for regulating the menstrual cycle. It was estimated that the domestic demand of the bark of Saraca asoca was more than 15,000 tonnes for the year 2007-08. This high annual demand of the bark needs to be obtained from this medicinal tree which is now in an endangered stage. As there is a wide gap between demand and availability, it is clear that some other plant material is collected and utilized instead of Saraca asoca. There are reports that the bark of asoka is widely adulterated with the bark of Polyalthia longifolia (Sonn.) (Figure2) which is known as Bangali ashok. belonging to the family Annonaceae. Polyalthia is having different medicinal properties and uses and it cannot be used as a substitute to asoka. Active ingredients that contribute to the medicinal property of asoka are phenols and tannins where as that of polyalthia are alkaloids. Substituting asoka with polyalthia may not be effective in treating gynaecological disorders or it may lead to some serious health hazards whose symptoms will develop only later.

Adulteration of herbal products has clinically relevant effects. Health problems related to herbal drugs are observed too often due to the contaminants rather than the declared ingredients. As these adulteration cause serious health hazards later ,it is important to have a floor level checking for the market samples for avoiding the adulterants. Under this circumstances we have taken up this study to find out an easy ,quick and reliable method for the identification of common major adulterant of the important ayurvedic herbal drug asoka bark and the results of the study are presented here.

MATERIALS AND METHODS

Stem barks of Saraca asoca and Polyalthia longifolia were collected from College of Horticulture, Kerala Agricultural University, Thrissur, Kerala, India.and authenticated by the botanists. The samples were shade dried. 1 g sample of each was put in 10 ml Phospahte buffered saline ( PBS, pH 7.4) overnight ( 10%). This extract was used for the HA ( haemagglutination) assay using standard methodology10. Double fold serial dilutions of 50 ul extract in 50 ul Phosphate buffered saline( PBS) was prepared in micro titer plates (ELISA plates)and mixed with 50ul of 2% PBS washed human erythrocytes of O positive blood group taken from human volunteer. Microtiter(ELISA) plates were incubated at room temperature for about 2 hours and the HA titer for each sample was recorded. Haemagglutination titer (HA titer) is the maximum dilution of the sample giving a visible agglutination. Agglutination is the clumping together of blood cells due to the network like linkage between the Red Blood Cells (RBCs) and the specifically reacting molecules present in the samples. As the RBCs are coloured there is no need of any other colouring agents. It was noted that all the S.asoca samples (4 different tree samples taken) gave positive haemagglutination with an HA titer ranging from 8 to 36 where as no agglutination was given by any of the four different Polyalthia longifolia bark samples ( HA =0) tried .(Figure-3). This revealed that the genuine S.asoca bark can be easily differentiated from the adulterant Polyalthia. using this technique.

Haemagglutination assay (Figure 3)

RESULTS AND DISCUSSION

In most of the cases of drug adulteration, the adulterant will have similar morphology as that of the genuine samples. It is very difficult to distinguish them physically. If the drug in question is spurious or adulterated, or is from an entirely different biological source it may still contain similar confusing compounds. Hence chemical fingerprints also will be confusing. Fingerprinting experiments by TLC conducted showed that there were a lot of similarities between asoka and polyalthia rather than differences. Remashree et al has reported that the comparative anatomical study can be taken up for the differentiation between the original and spurious bark samples of asoka. Very recently S.Khatoon et al has reported that HPTLC profile studies using the methanol extract of bark samples can be depended. All these techniques require costly equipments , chemicals and cumbersome procedures. But the present study revealed that HA assay using O + human RBCs is a good technique,practically very simple, cheap and less cumbersome. It can be used as a quick reliable and effective tool for the authentication and quality assessment of S. asoca and this method can be recommended for the floor level checking of market adulterant of the important herbal raw drug Saraca asoca. The work was carried out during 2009- 2010.

Haemagglutination technique has never before tried adulterant identification in herbal drugs. Usually chromatographic techniques are reported for standardization and to control the quality of both the raw material and the finished products. We tried a different biological technique that can be used for differentiating asoka from polyalthia. The presence of an entity- a haemagglutinin- was found in the stem barks of saraca asoca which causes agglutination of RBCs whereas it was found to be absent in polyalthia . Detailed studies are required to find out the specific molecule causing haemagglutination in asoka samples.

Acknowledgement

Authors thank the financial support from ICAR.

REFERENCES

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Assessment of seasonal soil moisture under traditional agroforestry systems in

Garhwal Himalaya, India

ARVIND BIJALWAN

Faculty of Technical Forestry, Indian Institute of Forest Management (IIFM), Bhopal, M.P., India.

Email: arvind276@

ABSTRACT

The seasonal soil moisture content was assessed under three agroforestry systems viz. Agrisilviculture (AS), Agri-hortisilviculture (AHS) and Agri-horticulture (AH) systems in northern and southern aspects of Garhwal Himalaya, India. The soil moisture under these agroforestry systems was higher compared to sole agriculture (treeless or control) system. The soil moisture ranged from 8.66 per cent (AS) in summer to 35.96 per cent (AH) in monsoon season (0-15 cm depth) and 11.39 per cent (AS) in summer to 31.71 per cent (AS) in monsoon (16-30 cm depth). The soil moisture status in sole agriculture system reported significantly different in all agroforestry systems under 0-15 and 16-30 cm depths. The influence of northern aspect obtains more moisture than the southern.

Key words: Moisture content, Agroforestry, Sole cropping, Agrisilviculture, Agrihortisilviculture, Agrihorticulture

Agroforestry systems are considered more sustainable and favourable to improve the soil properties. Presence of trees in combination with annual crops is believed to offer systematic plant cover to protect the soil from erosion as well as enhancement of moisture status of soil. In traditional agroforestry systems, trees are used to improve the soil fertility, maintain the hydrological balance and conserve the soil, moreover the tree-crop combination used the soil water more efficiently than the sole cropping. The presence of multipurpose trees as an essential component of traditional settled agriculture on terraced slopes, and indicated the importance of trees in rehabilitation, improvement of degraded wastelands and mitigating drought (Dhadwal et al 1986). In situ retention of rainfall on the land itself by agronomic measures in the rhizosphere for better plant growth is one of the essential factors which can be achieved through agroforestry practices and by suitable agronomic measures.

The productivity in agroforestry systems is higher as compared to sole cropping systems, because higher yield of crop has been observed in forest influenced soil than in ordinary soil (Chaturvedi, 1981; Sanghal, 1983; Verinumbe, 1987). Agroforestry systems based on traditional kn owledge with water management as an integral component are more effective for rehabilitation of degraded community lands than afforestation with plantation crops (Maikhuri et al. 1997). Keeping in view the appraisal and assessment of soil moisture status under agroforestry systems, the present study was carried out in the traditional agroforestry systems of Garhwal Himalayan region of India.

MATERIALS AND METHODS

The study was carried out in six different villages of Tehri Garhwal district of Garhwal Himalayan region of India, ranging between the elevation of 1000m to 2000m asl during 2004 to 2006. The selection of these villages (sites) varied in elevation, aspects and biodiversity. The selected sites stretched between sub-tropical to temperate zones. The study area receives 1240 mm annual rainfall with the mean monthly maximum temperature varies from 11.6 0C in January to 26.0 0C in June, whereas the mean monthly minimum temperature ranges from 2.3 0C in January to 16.8 0C in July (Fig. 1).

The soil analysis was performed in the soil samples taken from agroforestry systems and sole agriculture system (controll) to compare the insitu moisture status of the soil. The soil samples were randomly collected from 0-15 and 16-30 cm depths during winter, summer and monsoon seasons, using soil auger. The soil samples were collected thrice in a season with one month interval. The soil samples were collected from different agroforestry systems and sole agricultural fields and immediately weighed using mobile digital weighing balance to obtain the fresh weight of the soil. Later the soil samples were brought to the soil science laboratory of G. B. Pant University of Agriculture and Technology, Hill Campus, Ranichauri, Tehri Garhwal, Uttarakhand, India and kept in the oven at 105 0C for 24 hours till constant weight was achieved and weighed. Further the soil moisture was calculated using gravimetric method.

RESULTS AND DISCUSSION

The soil moisture percentage under agroforestry systems was observed to be higher as compared to sole agriculture system, which is thought to be beneficial for the growth and development of agriculture crops. The soil moisture ranged in different existing agroforestry systems varied from 8.66 per cent (AS) in summer to 35.96 per cent (AH) in monsoon season at 0-15 cm of depth and 11.39 per cent (AS) in summer to 31.71 per cent (AS) in monsoon (16-30 cm depth) on different study sites (Table 1).

It was observed that the soil moisture % in winter varied from 22.38 to 29.87 %, 19.30 to 27.95 %, 14.69 to 20.97 % in 0-15and 20.38 to 28.15 %, 18.95 to 30.12 %, 19.15 to 25.00 % in 16-30 cm depth for AS, AHS, AH systems respectively. In summer season, the soil moisture % ranged from 8.66 % (AS) to 17.87 % (AH) under 0-15 cm depth and 11.39 % (AS) to 20.33 % (AH) under 16-30 cm depth in different study sites. It was recorded that the soil moisture % varied from 8.66 to14.19 %, 10.88 to 16.53 %, 11.47 to 17.87 % in 0-15 cm depth and 11.39 to 17.80 %, 14.58 to 19.83 %, 14.70 to 20.33 % in 16-30 cm depth for AS, AHS, AH systems respectively in summer season (Table 1). In monsoon season the soil moisture % ranged from 25.01 to 35.96 % both in AH system (0-15 cm depth) and 22.71 % (AH) to 31.71 % (AS) under 16-30 cm of depth on different study sites. It was found that the soil moisture % varied from 28.89 to 35.62 %, 28.78 to 31.28 %, 25.01 to 35.96 % in 0-15 cm depth and 28.48 to 31.71 %, 27.57 to 30.30 %, 22.71 to 31.03 in 16-30 cm depth for AS, AHS, AH systems respectively in monsoon season (Table 1).

Table 1: Seasonal soil moisture content (%) under traditional Agroforestry systems

|AF system/ Site |Winter Season |

| |AS |AHS |AH |

| |0-15 |16-30 |0-15 |16-30 |0-15 |16-30 |

|N1 |29.34 |24.28 |27.95 |30.12 |19.94 |24.88 |

|S1 |25.13 |21.99 |19.30 |18.95 |15.81 |23.35 |

|N2 |22.46 |20.38 |25.41 |28.29 |15.04 |19.40 |

|S2 |22.38 |21.38 |21.43 |23.05 |14.69 |19.15 |

|N3 |29.87 |28.15 |25.93 |28.27 |20.97 |25.00 |

|S3 |27.43 |23.36 |21.15 |20.40 |17.83 |23.31 |

|Mean (N+S) |26.1 |23.26 |23.53 |24.85 |17.38 |22.52 |

|Mean (N) |27.22 |24.27 |26.43 |28.89 |18.65 |23.09 |

|Mean (S) |24.98 |22.24 |20.63 |20.80 |16.11 |21.94 |

|Control |18.23 |22.42 |15.12 |18.37 |14.63 |18.6 |

| AF system/ Site |Summer Season |

| |AS |AHS |AH |

| |0-15 |16-30 |0-15 |16-30 |0-15 |16-30 |

|N1 |12.10 |16.25 |11.21 |15.92 |10.52 |16.43 |

|S1 |10.02 |12.26 |10.88 |14.58 |11.47 |14.70 |

|N2 |14.19 |16.51 |12.04 |19.83 |15.84 |19.85 |

|S2 |13.95 |16.00 |16.53 |17.80 |16.86 |18.49 |

|N3 |8.66 |11.39 |14.61 |14.73 |17.87 |20.33 |

|S3 |13.85 |17.80 |14.61 |14.73 |16.87 |18.30 |

|Mean (N+S) |12.13 |15.04 |13.31 |16.27 |14.91 |18.02 |

|Mean (N) |11.65 |14.72 |12.62 |16.83 |14.74 |18.87 |

|Mean (S) |12.61 |15.35 |14.01 |15.70 |15.07 |17.16 |

|Control |9.55 |13.73 |8.42 |9.12 |9.06 |10.35 |

|AF system/ Site |Monsoon Season |

| |AS |AHS |AH |

| |0-15 |16-30 |0-15 |16-30 |0-15 |16-30 |

|N1 |31.39 |29.02 |33.54 |29.59 |32.34 |22.71 |

|S1 |28.89 |28.48 |30.53 |29.09 |35.96 |27.23 |

|N2 |34.44 |30.46 |30.33 |30.30 |33.58 |29.42 |

|S2 |35.62 |31.71 |30.88 |29.27 |30.41 |31.03 |

|N3 |31.04 |29.43 |31.28 |28.39 |25.63 |23.07 |

|S3 |33.91 |30.75 |28.78 |27.57 |25.01 |25.11 |

|Mean (N+S) |32.55 |29.98 |30.89 |29.04 |30.49 |26.43 |

|Mean (N) |32.29 |29.64 |31.72 |29.43 |30.52 |25.07 |

|Mean (S) |32.81 |30.31 |30.06 |28.64 |30.46 |27.79 |

|Control |35.58 |24.19 |33.54 |29.73 |30.83 |29.36 |

AS = Agrisilviculture system, AHS = Agrihortisilviculture system, AH= Agrihorticulture system

N (Northern aspect) = N1, N2, N3

S (Southern aspect) = S1, S2, S

The soil moisture status in sole agriculture system (control or without trees) is significantly different in all agroforestry systems on different sites under 0-15 and 16-30 cm depths. The statistical analysis (Table 2) shows that there is a significant difference (p ................
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