Results

From statistical analysis

The results of all the demarcated experimental field and rearing- trials were same (at the latest significant difference level of P<0.01 and P<0.05 respectively by ANOVA). Bio-statistic expert used all the average data (mean with standard error of the difference between means of replicates) from the four demarcated experimental field and rearing- trials. Here, the data of first demarcated experimental field and rearing- trial were presented (in the form of Table1, Table 2 and Table 3).

Estimation of the nematode population from field trial

Initial nematode populations

The initial nematode populations [Meloidogyne incognita (Kofoid & White) Chitwood], stretching over an area of 5.6acre of mulberry plantation, were 1579±53 J₂ per 200g of soil and 820±85 J₂ per 2g of root.

Nematode populations before treatment (Day-0)

The nematode populations in the demarcated 0.16 acre, were 1930±12 J₂ per 200g of soil and  635±25 J₂ per 2g of root [before treatment (Day-0)].

Mortality test

It was observed that Cina had no toxic effects on nematodes mortality within the exposure period of 12 hours at room temperature (25±2°C). For this reason, no data were presented in the results section.

Analysis of residue

There were left no toxic residues of Cina in all the infected Cina-treated plants by thin layer chromatography plate (TLC).

Assesment of Root-knot disease

Table1 shows the effects of Cina on Meloidogyne incognita infected mulberry plants in a field trial (P<0.01 by ANOVA).

• · All infected plants treated with Cina showed increase in number and surface area of leaves and higher protein content (fresh) in leaves and root than infected untreated with Cina (control) plants.
• · In all infected Cina treated plants, the population of root-knot nematodes decreased significantly in rhizospheric soil and as well as in roots than infected untreated with Cina (control) plants.
• · The number of root galls also decreased significantly after Cina treatment.
• It is interesting that treatment effects were the most pronounced with Cina 200C.

Assesment of Rearing of silkworms

Table 2 shows the effects of Cina on the M. incognita infected mulberry plants in a rearing trial on the feeding, growth and mortality of silkworms (P<0.05 by ANOVA).

• · The average consumption of leaves by the 5^th instars, average number of feeding to cocoon formation, average number of feeding day to cocoon formation, average number of escaping – feeding during moulting and average moulting span days were lesser for Cina treated plants than for the infected untreated with Cina (control) ones.
• · The average mortality rate was nil with all Cina treated plants and 30% with infected  (control ) ones.  However, the average fresh weight of the 5^th instars larvae were higher with Cina treated plants than with infected untreated with Cina (control) one.

Assesment of Silk production and Effective Rate of Rearing

Table 3 shows the commercially increased silk production by effective treatment with Cina on the feeding of M. incognita infected mulberry leaves in a rearing trial (P<0.05 by ANOVA).

• · The average fresh silk gland weight, average fresh cocoon weight, average fresh shell weight and average shell ratio ( Shell weight / Cocoon weight X 100) were higher with Cina- treated plants than with infected untreated with Cina (control) one.

· It is interesting that the shell ratios were higher in the plants treated with Cina than with the infected untreated (control) ones.

• · It is also notable that average starting time to spinning day and average span of spinning day ( i.e. duration of span ) were fewer with the Cina treated plants than with infected untreated with Cina ( control ) ones.
• · Average effective rate of rearing (ERR%) were higher in all Cina treated groups.
• Once again the homeopathic drug Cina is confirmed as cost effective and capable  of  reducing root-knot disease as well as improving the nutritive value of the treated leaves of infected plants^7,9,11,12,14.
• · From this field trial, we once again confirm that Cina also improves the nutritive value of the treated leaves^{7,11,12,14,15} which directly influences the consumption of leaves, number of feedings and number of feeding days to cocoon formation, and indirectly affects the moulting stage in all the Cina treated groups from these trials.
• · Due to ill development of infected untreated (control) batches, larvae took more time to moult, which is demonstrated by the number escaping feeding during  moulting^{7,11,12,14,15}.
• · Higher nutritive value of treated plants contributes to higher growth of silkworm larvae, silk gland weight, cocoon weight and shell weight, which increase silk production significantly^{7,11,12,14,15} for commercial purposes.
• · The improved health of the larvae, cocoon weight, silk gland and shell weight from the Cina treated groups of  the infected  plants might have resulted in the shorter starting time to spinning , the span of spinning and the total elimination of the mortality rate^22,23,25.   Cina is too dilute to contain drug molecules^{7,11,12,15,22}. Naturally, the drug might not have affected the nematode directly^{7,11,12,14,15} and for this reason, no mortality occurs.
• · The effective rate of rearing (ERR% ) is very high in all Cina treated treatment batches, which enriches the sericulture industry in many ways,  especially for commercial purposes.
• · The mulberry leaves did not contain any toxic residues of the Cina test substances as determined by the thin layer chromatography (TLC). It is reported that Cina at ultra high dilution has physical basis in the form of charge transfer interaction and altered rate of tumbling in the specific part of the molecules of the diluents medium^14,115,22.
• · Rather, the drug Cina might have induced natural a defense response against nematode parasites in the test plants, and has conferred a defense response on the growing larvae^{4-12,14,27,28}.
• · It is surprising that all infected Cina treated plants not only are less affected by nematodes, but also have better growth than the infected untreated with Cina (control) plants^{7,11,12,14,15}.
• · From these results, one might assume that Cina acts as a fertilizer. Our explanations are that we used Cina as a foliar spray and it is already been observed from our previous experiment that the Cina treatment effects on healthy plants in order to confirm the positive effect on growth than healthy and infected untreated with Cina (control) groups^{7,11,12,14,15} .
• · The positive effects of growth may be responsible for defense resistance against pathogens^{7,11,12,14,15,27,28}.  We can say that Cina might have induced synthesis of many new proteins which have stimulated increased photosynthesis rate, stomatal activity^{7,11,12,14,15,27,28} and water retention capacity of  Cina treated plants^4,6,7.
• · The positive effects of growth on infected Cina treated plants may not only be responsible for defense resistance to the nematodes pathogen, but may also improve growth of silkworm larvae and silk gland weight, cocoon weight, shell weight and effective rate of rearing (ERR%)²⁵ which increase silk production for commercial purposes. It is proven from the result that silk production is higher in all the Cina treated groups than in the infected untreated with Cina (control) groups.

Conclusion

These results once again suggest that certain plant diseases might be effectively controlled by the potentized cost effective homeopathic medicine Cina, at an extremely low dose, and that Cina also increases silk production and effective rate of rearing commercially which directly enriches the sericulture industry as well as the agriculture sector. Cina is easily available, is non-phytotoxic, a non-pollutant and helps conserve biodiversity.

Acknowledgements

The work described here has been supported by Rtd. Prof. N.C.Sukul, Dept. of Zoology, Visva-Bharati and Joint Director, Sriniketan Sericultural Composite Unit, Sriniketan, Govt. of West Bengal and Mr. Achintya Mondal, Secretary, BIMS, BMS&BIMLS,West Bengal,India. Lastly, for help in statistical analysis we are  immensely indebted to Dr. Tapan Mondal, Asst. teacher of Secondary School. Sri Basudev Mondal, Assistant English-Teacher of Kanchannagar D.N.Das High School, Kanchannagar, Burdwan, who has revised the English of the manuscripts.

References

1     Sasser JN, Freckman DW. Crop protection: systemic and persistent effect of neem.  Pro.of 25^th Ann Meet Soc Nemat (abst), Orlando, Florida  1986; 32.

2     Datta SC. Possible use of amaranth as catch crop for root-knot nematodes intercropped   with mulberry. J Environ & Sociobiol 2005b;2(2&3):61-65.

3     Datta  SC. Possible use of amaranth as catch crop for root-knot nematodes intercropped  with okra. Phytomorphology 2006b; 56(3&4): 113-116.

4     Datta SC. Mulberry disease : a problem in sericulture. SEBA NEWSLETTER 2007; 4(1): 7-10.

5     Datta SC, Datta (Nag) R. Intercropping amaranth with mulberry for managing root-knot nematodes and improving sericulture. Sericologia 2007a; 47(3): 297-302 .

6     Datta SC, Datta (Nag) R. Increased silk production by effective treatment of naturally infected root-knot and black leaf spot diseases of mulberry with acaciasides. J Environ & Sociobiol 2007b; 4(2): 209-214.

7        Datta SC, Datta R. Potentized Artemisia nilagirica extract Cina) increases silk production and effective rate of rearing in a field trial. Hpathy Ezine, at http://www.rediffmail.com/cgiin/red.cgi?red=www%2Ehpathy%2Ecom July, 2008.

8        Paul A, Sinhababu SP, Sukul NC. Effect of Meloidogyne incognita infected mulberry plants and their treatment with nematicides of feeding silkworms, Bombyx mori L. Sericologia 1995; 35:577-581.

9    Datta SC. Plant parasitic nematodes- an agricultural problem and its solution. The Visva-Bharati Quartery 2005a; 11 (3&4): 89-100.

10   Datta SC, Sinhababu SP, Sukul NC. Improved growth of silk worms from effective treatment of mulberry diseases by Acacia auriculiformis extract. Sericologia 1997; 37(4): 707-712.

11   Datta SC, Datta (Nag) R. Liquid homeopathic medicine Cina enriches sericulture industry. J Environ & Sociobiol 2006a; 3(1): 55-60.

12   Sukul NC, Ghosh S, Sukul A, Sinhababu SP. Amelioration of root-knot disease of lady’s finger plants by potentized Cina and antonin. Homeopathy 2006; 95: 144-147.

13 Halfter G. Biodiversity conservation and protected areas in tropical countries. In Biodiver Sci and Dev Towards a New partnership (eds) Dicastri F and Younis T USA: AB Inter in associated with IUBS 1996 pp.212-223.

14 Datta SC. Effects of Cina on root-knot disease of mulberry. Homeopathy 2006a; 95(2):98:102.

15 Sukul NC, Sinhababu SP, Datta SC, Nandi B, Sukul A. Nematotoxic effect of Acacia auriculiformis and Artemisia nilagirica against root-knot nematodes. Allelopath  J 2001; 8:65-72.

16 Christie JR, Perry VG. Removing nematodes from soil. Proc Helm Soc Wash 1951; 18:106-108.

17 Sukul NC.  Soil and plant nematodes. West Bengal State Book Board Publisher 1987, pp1-271.

18 Das S, Sukul NC. Biochemical changes of some crop plants due to root-knot nematode infection. Proc Nat Symp of New Dimension in Parasitology, Allahabad, India 1986.

19 Lowry OH, Rossebrough NJ, Farr AR, Randall RJ, Protein measurement with the Folin-phenol reagent. J  Biol  Chem 1951; 193: 265-275.

20 Chatterjee A, Sukul NC. Total protein of galled roots as an index of root-knot nematode infestation of lady’s finger plants. Phytopathol 1981; 31:372-374.

21 Anonymous. The American Homeopathic Pharmacopoeia. 9^th edn, Philadelphia, USA: Boericke and Tafel 1920.

22 Sukul NC. High Dilution pharmacology and Homeopathy 1997.

23 Fenner LM. Determination of nematode mortality. Plant  Dis  Rep 1962;46:383.

24 Consden R, Gordon AH, Martin AJP.  J Biol  Chem 1944; 38:224.

25 Krishnaswamy S, Narasimhanna MN, Suryanarayana SK, kumararaj S. Manual of sericulture, vol 2 silkworm rearing. Agri Services Bull 15, FAO, Rome 1972.

26 Duncan DB. Multiple range and multiple F tests. Biometrics 1955; 11:1-42.

27 Klessig, Daniel F, Malamy J. The salicylic acid signal in plants. Plant Mol Biol 1994; 26:1439-1458.

28 Datta SC, Datta (Nag) R. Defence resistance of okra against root-knot disease by bio-nematicides. Proc Zool Soc, Kolkata, India 2006b; 59(2):

75-82.

Table Legends

Table1 Effects of Cina on Meloidogyne incognita infected mulberry plants in a field trial

Table 2  Effects of Cina on M.incognita infected mulberry plants in a rearing trial on the feeding, growth and mortality of silk worms

Table 3  Commercially increased silk production by effective treatment with Cina on the  feeding of M.incognita infected mulberry plants in a rearing trial

• · Details of Publications of Subhas Chandra Datta :

1.” Improved growth of silk worms from effective treatment of mulberry diseases by Acacia auriculiformis extract” by Subhas Chandra Datta, Santi Prasad Sinhababu & Nirmal Chandra Sukul in 1997- Sericologia 37 (4): 707-712.

2.”Acaciasides  and root-knot nematode extract suppress in tomato” by Subhas Chandra Datta, S.P.Sinhababu, N. Banerjee, K.Ghosh and N.C. Sukul in 1998- Indian J. Nematol. 28 (i): 1-5.

3.” Meloidogyne incognita extract reduces Meloidogyne incognita infection in lady’s finger plants” by Subhas Chandra Datta, R. Datta (Nag) S.P. Sinhababu & N.C.Sukul in 1998- Proceedings of the National Seminar on Environmental Biology, Edited by A.K. Aditya & P. Haldar, Visva-Bharati University, Daya Publishing House, Delhi: 205-209.

4.”Relative Attractiveness of Four Species of Vegetable Crops for Meloidogyne incognita” by Subhas Chandra Datta, Rupa Datta (Nag),Anirban Sukul, Nirmal C. Sukul and Santi Prasad Sinhababu in 2000- Environment & Ecology 18 (1): 233-235.

5.” Nematotoxic effect of Acacia auriculiformis and Artemisia nilagirica against root-knot nematodes” by N. C. Sukul, S. P. Sinhababu, Subhas Chandra Datta, B.Nandi and A.Sukul in 2001. Allelopathy Journal 8(1): 65-72.

6.”Plant Parasitic nematodes – an agricultural problem and its solutions” by Subhas Chandra Datta in2005a. Visva-Bharati Quarterly, 11 (3&4), 89-100.

7.”Possible use of amaranth as catch crop for root-knot nematodes intercropped with mulberry” by Subhas Chandra Datta  in 2005b. J. Environ. & Sociobiol., 2 (1&2): 61-65.

8.”Effects of Cina on root-knot disease of mulberry” by Subhas Chandra Datta   in 2006a. Homeopathy, 95 (2): 98-102.

9.”Possible use of amaranth as catch crop for root-knot nematodes intercropped with okra” by Subhas Chandra Datta in 2006b, Phytomorphlogy, 56 (3&4): 113-116.

10. “Liquid homeopathic medicine Cina enriches sericulture industry” by Subhas Chandra Datta  and Rupa Datta(Nag) in 2006a, J. Environ.& Sociobiol., 3 (1): 55-60.

11. “Defence resistance of okra against root-knot disease by bio-nematicides” by Subhas Chandra Datta and Rupa Datta(Nag) in 2006b, Proceedings of the Zoological Society,  59 (2) : 75-82.

12.”Mulberry disease : A problem in sericulture” by Subhas Chandra Datta in 2007. SEBA Newsletter,  Environment & Sociobiology,4 (1):7-10.

13.”Intercropping amaranth with mulberry for managing root-knot nematodes and improving sericulture”  by Subhas Chandra Datta and Rupa Datta(Nag) in 2007a. Sericologia, 47(3):1-6.

14.”Increased silk production by effective treatment of naturally infected root-knot and black leaf spot diseases of mulberry with acaciasides” by Subhas Chandra Datta  and Rupa Datta(Nag) in2007b. J Environ & Sociobiol., 4(2):209-216.

15.”Analyzing ‘Teaching Quality’ of a teacher in a school” by Subhas Chandra Datta  in 2007. Parsadbarta, West Bengal Board of Secondary Education, 34: 16-20.

16.”Potentized Artemisia nilagirica Extract (Cina) Increases Silk Production and Effective Rate of Rearing in a Field Trial” by Subhas Chandra Datta and Rupa Datta in 2008. Hpathy Ezine, at http://www.rediffmail.com/cgi-bin/red.cgi?red=www%2Ehpathy%2Ecom) July, 2008.

• · Research area and Interest:

Environmental biology, Plant protection, Plant pathology, Entomology, Sericulture, Biochemistry, Parasitology, Tissue culture, Allelopathy, Nematode control & Homeopathy.