<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article  PUBLIC "-//NLM//DTD Journal Publishing DTD v3.0 20080202//EN" "http://dtd.nlm.nih.gov/publishing/3.0/journalpublishing3.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" dtd-version="3.0" xml:lang="en" article-type="research article"><front><journal-meta><journal-id journal-id-type="publisher-id">ABB</journal-id><journal-title-group><journal-title>Advances in Bioscience and Biotechnology</journal-title></journal-title-group><issn pub-type="epub">2156-8456</issn><publisher><publisher-name>Scientific Research Publishing</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.4236/abb.2015.61003</article-id><article-id pub-id-type="publisher-id">ABB-53408</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Biomedical&amp;Life Sciences</subject></subj-group></article-categories><title-group><article-title>
 
 
  Effect of Homoeopathic Drugs to Control Growth and Production of &lt;i&gt;A. flavus&lt;/i&gt;
 
</article-title></title-group><contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>.</surname><given-names>N. P. Singh</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Sunita</surname><given-names>Kumari</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>M.</surname><given-names>M. Prasad</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib></contrib-group><aff id="aff1"><addr-line>Post Graduate Department of Biotechnology, T. M. Bhagalpur University, Bhagalpur, India</addr-line></aff><author-notes><corresp id="cor1">* E-mail:<email>drhnpspgbiotech@gmail.com(.NPS)</email>;</corresp></author-notes><pub-date pub-type="epub"><day>12</day><month>01</month><year>2015</year></pub-date><volume>06</volume><issue>01</issue><fpage>18</fpage><lpage>21</lpage><history><date date-type="received"><day>27</day>	<month>December</month>	<year>2014</year></date><date date-type="rev-recd"><day>accepted</day>	<month>16</month>	<year>January</year>	</date><date date-type="accepted"><day>21</day>	<month>January</month>	<year>2015</year></date></history><permissions><copyright-statement>&#169; Copyright  2014 by authors and Scientific Research Publishing Inc. </copyright-statement><copyright-year>2014</copyright-year><license><license-p>This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/</license-p></license></permissions><abstract><p>
 
 
  Five common homoeopathic drugs viz., Belladonna, Bryonia, Colchicum, Colocynth and Lathyrus sat were selected and tested against growth and aflatoxin production of 
  Aspergillus flavus. The result indicates that all five drugs suppressed the growth of 
  A. flavus. The lower concentration of all the tested drugs induced maximum growth of fungi and maximum production of aflatoxin. However, the growth as well as aflatoxin production potentiality was considerably decreased when the concentration of the drug was increased. Out of all the five drugs, Bryonia was comparatively less effective with respect to inhibition in aflatoxin production. But Belladonna was found to be most effective drug on growth and aflatoxin production.
 
</p></abstract><kwd-group><kwd>Effect Homoeopathic</kwd><kwd> Drugs</kwd><kwd> Growth</kwd><kwd> Production</kwd><kwd> Aflatoxin</kwd></kwd-group></article-meta></front><body><sec id="s1"><title>1. Introduction</title><p>A wide range of fungi are known to produce mycotoxin on a number of substrates. Conusumption of such contaminated substrates causes different symptoms in human beings and animals. Mycotoxicosis, a toxic syndrome resulting after intake of mycotoxins by men and animals, is well known since long, yet its economic value remained neglected until aflatoxin was discovered. Aflatoxins are most harmful mycotoxin produced by toxigenic stains of Aspergillus flavus and A. parasiticus. Studies carried out during last two decades have established aflatoxins as a strong carcinogen, mutagen and teratogen on animals and human beings [<xref ref-type="bibr" rid="scirp.53408-ref1">1</xref>] [<xref ref-type="bibr" rid="scirp.53408-ref2">2</xref>] . An elaborated work has been done on the prevention of aflatoxin by fungicides, chemicals, phenols, plants/leaf extracts and antibiotics. However only few reports are available on use of homoeopathic drugs as preventing agents for aflatoxin production. So the present investigation is carried out to evaluate the efficacy of some homoeopathic drugs like Belladonna, Bryonia, Colchicum, Colocynth and Lathyrus sat on aflatoxin production by toxigenic isolates of A. flavus.</p></sec><sec id="s2"><title>2. Materials and Methods</title><p>Screening of Aflatoxin procluction was studied by the method of [<xref ref-type="bibr" rid="scirp.53408-ref3">3</xref>] . Subsequently quantitative estimation of aflatoxin was done by the method of [<xref ref-type="bibr" rid="scirp.53408-ref4">4</xref>] .</p><p>The SMKY (Sucrose Magnesium sulphate, Potassium nitrate and Yeast extract) medium was used to test the efficacy of homoeopathic drugs on growth and aflatoxin production potentiality of A. flavus. The mother tincture of five homoeopathic drugs viz., Belladonna, Bryonia, Colchicum, Colocynth and Lathyrus sat were separately added to make the concentration 50, 100, 150 and 200λ/25 ml medium [<xref ref-type="bibr" rid="scirp.53408-ref3">3</xref>] in 250 ml conical flasks. The flasks were inoculated at 28˚C &#177; 1˚C for 10 days. At the end of incubation period, the medium was filtered using Whatman’s filter paper No. 42. The dry weight of the mycelia was determined on monopan balance after washing and drying the same at 60˚C for 48 hours. The filtrate was extracted with chloroform and the extract was evaporated to dryness in a waterbath. Quantitative estimation of aflatoxin was done on TLC plates using toluene:isoamylalcohol:methanol (90:32:2 v/v) solvent systems. The chromatoplates were observed under long UV light at 360 nm. The spot of aflatoxin B&#173;<sub>1</sub> was scrapped and quantitative estimation was done by spectrophotometer [<xref ref-type="bibr" rid="scirp.53408-ref5">5</xref>] .</p></sec><sec id="s3"><title>3. Results</title><p>A perusal of <xref ref-type="table" rid="table1">Table 1</xref> shows that growth as well as aflatoxin production potentiality of A. flavus were considerably influenced by different concentrations of five homoeopathic drugs viz., Belladonna induced maximum growth inhibitions (75.84%) and aflatoxin production (56.42%) at the concentration of 200l/25 ml, however its lowest concentration 50l showed only 26.44 and 27.85 percent inhibition respectively over control. Similarly lower concentration (50l/25 ml) of colocynth induced 17.75% and 22.14% inhibition in growth and aflatoxin production which gradually increased with increase in the concentration of drugs. The maximum reduction in growth (63.76%) and aflatoxin production (51.42%) was noticed at 200l/25 ml concentration.</p><table-wrap id="table1" ><label><xref ref-type="table" rid="table1">Table 1</xref></label><caption><title> Effect of Homoeopathic drugs on growth and aflatoxin production by A. flavus</title></caption><table><tbody><thead><tr><th align="center" valign="middle"  rowspan="2"  >Name of the drug</th><th align="center" valign="middle"  rowspan="2"  >Concentration (λ/25 ml)</th><th align="center" valign="middle" >Mycelial</th><th align="center" valign="middle" >Growth</th><th align="center" valign="middle" >Aflatoxin</th><th align="center" valign="middle" >Production</th></tr></thead><tr><td align="center" valign="middle" >Amount (mg)</td><td align="center" valign="middle" >% Inhibition over control</td><td align="center" valign="middle" >Amount in ppm</td><td align="center" valign="middle" >% Inhibition over control</td></tr><tr><td align="center" valign="middle" >Control</td><td align="center" valign="middle" >0</td><td align="center" valign="middle" >828</td><td align="center" valign="middle" ></td><td align="center" valign="middle" >140</td><td align="center" valign="middle" ></td></tr><tr><td align="center" valign="middle" >Belladonna</td><td align="center" valign="middle" >50</td><td align="center" valign="middle" >609</td><td align="center" valign="middle" >26.44</td><td align="center" valign="middle" >101</td><td align="center" valign="middle" >27.85</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >100</td><td align="center" valign="middle" >496</td><td align="center" valign="middle" >40.09</td><td align="center" valign="middle" >93</td><td align="center" valign="middle" >33.57</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >150</td><td align="center" valign="middle" >400</td><td align="center" valign="middle" >51.69</td><td align="center" valign="middle" >85</td><td align="center" valign="middle" >39.28</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >200</td><td align="center" valign="middle" >200</td><td align="center" valign="middle" >75.84</td><td align="center" valign="middle" >81</td><td align="center" valign="middle" >56.42</td></tr><tr><td align="center" valign="middle" >Bryonia</td><td align="center" valign="middle" >50</td><td align="center" valign="middle" >800</td><td align="center" valign="middle" >3.38</td><td align="center" valign="middle" >130</td><td align="center" valign="middle" >07.14</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >100</td><td align="center" valign="middle" >725</td><td align="center" valign="middle" >12.43</td><td align="center" valign="middle" >108</td><td align="center" valign="middle" >22.85</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >150</td><td align="center" valign="middle" >662</td><td align="center" valign="middle" >20.04</td><td align="center" valign="middle" >95</td><td align="center" valign="middle" >32.14</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >200</td><td align="center" valign="middle" >535</td><td align="center" valign="middle" >35.38</td><td align="center" valign="middle" >80</td><td align="center" valign="middle" >45.85</td></tr><tr><td align="center" valign="middle" >Colchicum</td><td align="center" valign="middle" >50</td><td align="center" valign="middle" >690</td><td align="center" valign="middle" >16.66</td><td align="center" valign="middle" >109</td><td align="center" valign="middle" >22.142</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >100</td><td align="center" valign="middle" >500</td><td align="center" valign="middle" >39.61</td><td align="center" valign="middle" >103</td><td align="center" valign="middle" >26.42</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >150</td><td align="center" valign="middle" >446</td><td align="center" valign="middle" >46.13</td><td align="center" valign="middle" >89</td><td align="center" valign="middle" >36.42</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >200</td><td align="center" valign="middle" >335</td><td align="center" valign="middle" >59.54</td><td align="center" valign="middle" >71</td><td align="center" valign="middle" >49.29</td></tr><tr><td align="center" valign="middle" >Colocynth</td><td align="center" valign="middle" >50</td><td align="center" valign="middle" >681</td><td align="center" valign="middle" >17.75</td><td align="center" valign="middle" >109</td><td align="center" valign="middle" >22.14</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >100</td><td align="center" valign="middle" >535</td><td align="center" valign="middle" >35.38</td><td align="center" valign="middle" >101</td><td align="center" valign="middle" >27.85</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >150</td><td align="center" valign="middle" >338</td><td align="center" valign="middle" >59.17</td><td align="center" valign="middle" >87</td><td align="center" valign="middle" >37.85</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >200</td><td align="center" valign="middle" >300</td><td align="center" valign="middle" >63.76</td><td align="center" valign="middle" >68</td><td align="center" valign="middle" >51.42</td></tr><tr><td align="center" valign="middle" >Lathyrus sat</td><td align="center" valign="middle" >50</td><td align="center" valign="middle" >750</td><td align="center" valign="middle" >9.42</td><td align="center" valign="middle" >115</td><td align="center" valign="middle" >17.85</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >100</td><td align="center" valign="middle" >689</td><td align="center" valign="middle" >16.78</td><td align="center" valign="middle" >105</td><td align="center" valign="middle" >25.00</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >150</td><td align="center" valign="middle" >549</td><td align="center" valign="middle" >33.69</td><td align="center" valign="middle" >90</td><td align="center" valign="middle" >35.71</td></tr><tr><td align="center" valign="middle" ></td><td align="center" valign="middle" >200</td><td align="center" valign="middle" >468</td><td align="center" valign="middle" >43.47</td><td align="center" valign="middle" >76</td><td align="center" valign="middle" >45.71</td></tr></tbody></table></table-wrap><p>Colchicum exhibited 16.66 and 39.61 percent inhibition in growth when the concentration was 50l and 100l/25 ml respectively and the percent inhibition with respect to aflatoxin production was also lowered (22.14 and 26.42 percent). The maximum loss i.e. 59.54 and 49.25 percent in growth and aflatoxin production was recorded at the concentration of 200l/25 ml. In case of Bryonia lower concentration did not induce any marked inhibitory effect on growth of A. flavus. The maximum inhibition in growth and aflatoxin production was noted at 200l/25 ml concentration where it was 35.38% and 42.85% respectively. Subsequently Lathyrus sat exhibited similar trend with respect to inhibition in growth and aflatoxin production. It was noticed maximum i.e. 43.47 and 45.71 percent at 200l/25 ml followed by 150 and 100l/25 ml of medium. Minimum growth and aflatoxin production was recorded at 50l concentration where it was 9.42 and 17.85 percent respectively in Lathyrus sat.</p></sec><sec id="s4"><title>4. Discussion</title><p>A comparative review of results shows that addition of all the five homoeopathic drugs suppressed the growth of A. flavus. The lower concentration induced maximum growth of fungi and subsequently maximum production of aflatoxin. The growth as well as aflatoxin production potentiality was considerably reduced when the concentration of the drug was increased. <xref ref-type="table" rid="table1">Table 1</xref> also indicates that out of all the drugs, Bryonia was comparatively less effective with respect to inhibition in aflatoxin production. From the above observations efficacy of all the test homoeopathic drugs may be graded in descending order as Belladonna &gt; Colocynth &gt; Colchicum &gt; Lathyrus sat &gt; Bryonia.</p><p>In the light of present findings it may be stated that production of aflatoxin is directly related to the growth of A. flavus and lesser growth of fungi induced less synthesis aflatoxin in media [<xref ref-type="bibr" rid="scirp.53408-ref6">6</xref>] [<xref ref-type="bibr" rid="scirp.53408-ref7">7</xref>] . Some workers like [<xref ref-type="bibr" rid="scirp.53408-ref8">8</xref>] - [<xref ref-type="bibr" rid="scirp.53408-ref10">10</xref>] also used chemicals and fungicides against fungal growth and aflatoxin production and was also found effective in inhibiting the growth of fungi and aflatoxin production. [<xref ref-type="bibr" rid="scirp.53408-ref11">11</xref>] - [<xref ref-type="bibr" rid="scirp.53408-ref13">13</xref>] also worked on homoeopathic drugs against growth of fungi and aflatoxin production and concluded their inhibitory role in certain intermediate biosynthetic steps. Similar observation was also noted by [<xref ref-type="bibr" rid="scirp.53408-ref14">14</xref>] [<xref ref-type="bibr" rid="scirp.53408-ref15">15</xref>] working with different food, feed and medicinal plants. [<xref ref-type="bibr" rid="scirp.53408-ref16">16</xref>] also stated that level of aflatoxin production depends on the genetic makeup of the host and the mould involved.</p></sec><sec id="s5"><title>5. Conclusion</title><p>Homeopathic drugs are cheaper and have no side effects so effort was made to study the effect of Belladonna, Bryonia, Colchicum, Colocynth and Lathyrus sat on growth and aflatoxin production potentiality of A. flavus. It was observed that 200λ/25 ml concentration of Belladonna was most effective and showed 75.84% inhibition in the mycelial growth. This was followed by Colocynth, Colchicum, Bryonia and Lathyrus sat respectively. Similar results were obtained by [<xref ref-type="bibr" rid="scirp.53408-ref17">17</xref>] - [<xref ref-type="bibr" rid="scirp.53408-ref19">19</xref>] .</p></sec></body><back><ref-list><title>References</title><ref id="scirp.53408-ref1"><label>1</label><mixed-citation publication-type="other" xlink:type="simple">Scott, P.M. and Kennedy, B.P.C. (1973) Analysis and Survey of Ground Black White and Capsicum Peppers for Aflatoxins. 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