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This entire research had a propose and that was to examine the anti-fungal activity present in plant extract which would treat any pathogenic fungi in fish. I went to numbers of research papers and made this research review by gathering all the information I could find about my topic. These experiments were conducted by taking different plant extracts obtained from plants root and introducing it to the pathogenic fish in laboratory condition by testing it at different temperature, concentration and time exposure. The chemical agents which were used were damaging the environment of the aquaculture and were a threat for entire aquatic organism. Hence, the plant extracts, which are organic and less harmful, were tested to replaces these antimicrobial chemical agents. The extracts obtained from plant were antimicrobial in nature and were much effective and were eco-friendly.

To summarise in brief of recent development in treatment for pathogenic fungi in fishes.

To find the alternative of Malachite green an antifungal agent that has many long-lasting drawbacks on fish. The horseradish root extract Armoraciarusticana which produces AIT allyl isothiocyanates the main component if horseradish plant shows antimicrobial activity being used as an antifungal agent against Saprolegniaparasiticaby itesting iit iin idifferent itemperature, iexposure itime iand iconcentration iof iAIT i(allyl iis ithiocyanates).Members iof ithe igenus iSaprolegnia icause iSaprolegniosis, ia disease ithat iis icharacterised iby ivisible iwhite ior igrey ipatches iof ifilamentous imycelium ion ithe ibody ior ifins iof ifreshwater ifish. iIt ican icause iserious idamage ito iother ifreshwater ifishes iif ino itreatment iis idone. iHorseradish iis ia iperennial iplant iof ithe ifamily iBrassicaceae ialso iused iin icosmetics ibecause iof iits iorganic iproperties. iHorseradish iroot iis irich iin ivitamin iC iand iB1, iminerals i(iron, ipotassium, icalcium iand imagnesium), iphytoncide iand iessential ioils, isinigrin iwhich ireleases ia ivolatile iaglycone i(allyl iisothiocyanate) iidentical iwith ithe iessence iof imustard iplant iwhich ibelongs ito isame ifamily. iSinigrin iis ithe imain icomponent ipresent iin ithe ihorseradish. iMalachite igreen, ian iantifungal iagent iwas iearlier iused ito itreat ithe ioomycetes iin ithe ifishes ibut iit ihas iresidual, icarcinogenic iand iteratogenic ieffects iwhich icaused ithe ibanning iof ithe iuse iof imalachite igreen iin imost ithe icountries ilike iJapan,i USA, ietc.i Horse radish iconsistsi sinigrini whichi isi my ros in asei enzymei complex ito ithei allyliisothiocyanates iwhichi isi antimicrobiali effecti againsti Escherichiaicoli,i Listeriaimonocytogenes,iSalmonellaityphimurium,iandi Staphylococcusiaureus iasi statedi in Khomvilai i2006. iThe seitests iwerei done ibyi Khomvilai i2006 iby itakingi differenti measures iofi thei agariplugs iand itemperatures iandi the iresult iwere ishowni asi such iin imy celial igrowth.i This iwasi thei second itest idonei by iKhomvilai ion imy celiali growthi by icomparing ithei two ibyi theiri colonyi diameter iwithi onei beingi treated ibyi AIT itreatmenti (allyliisothiocyanates) iandi otheri untreatedi controli using ithei equation:i(colonyi diameter iofi thei treatedi fungus) i(colony idiameter iofi control)iX i100 i(%).iWhile itaking iini accounti ofi variancei

Table 1 Colony diameter (mm) of saprolegnia parasiticaat various incubation temperature

5 C	10 C	15 C	20 C
11.5+-0.2	28.2+-4.5	53.7+-1.3	61.5+-1.2

The different letters on the upper right side of each colony diameter indicate a significance difference between treatments (Duncan’s new multiple range test, p<0.05)

Source-Khomvilai et al 2006 of the Duncan’s new multiple range tests. The tests were compared, and the result of the latter showed better result where the time was taken that was exposed to AIT.

Table 2 Colony diameter  (mm) of saprolegnia parasitica   after allyl isohiocynate (AIT) treatment at various combination of AIT concentration and exposure time.

AIT(mg/L)	Exposure time- 30 min	Exposure time- 60 min
0	54.1+-0.4	53.3+-0.2
68	19.6+-2.5	0+-0
85	0+-0	0+-0
102	0+-0	0+-0

The different letters on the upper right side of each colony diameter indicate a significance difference between treatments (Duncan’s new multiple range test, p<0.05)

Source- khomvilaiet al 2006

Table 3 showed Zoospore germination, germinated hyphae were observed on all the hemp seeds of control but not on AIT. Hence, indicating that it was an alternative for the malachite green.

Zoospore germination(+,positive, - , negative)saprolegnia parasitica  after  allyl isohiocynate (AIT) treatments at various combination of AIT concentration and exposure time.

Table 3

AIT(mg/L)	Exposure time- 10s	Exposure time- 1 min	Exposure time- 5 min
0	+++  +++      +++  +++	+++  +++  +++  +++	+++  +++ +++  +++
34	+++  +++ +++  ---	+++   +++ +++  +++	+++  +++ +++  +++
42.5	+++  +++  +++  +++	+++  --- +--  ---	---  --- ---  ---
51	---  --- ---  ---	---  ---  ---  ---	---  ---  ---  ---

Each symbol indicates one hemp seed and a groupof three symbols shown as experimental dish with the three hemp seed.

Source- khomvilai et al 2006

The test successfully showed that the Horseradish root extract works wonder and has feweror no drawbacks compared to Malachite green. The AIT showed result faster and better with the change of temperature. Horseradish extract can be used as an antimicrobial agent due to its underlying activity of active sinigrin, AIT. The extract taken was actively antimicrobial as suggested by Mori et al in his experiment and the result showed positive effect of AIT on S. parasitica which many other researchers worked with like Isshika et al however they conducted their tests with different diameter and time periodbut with each experiment the results showed success rate of allyl isothiocyanate and using it as an antifungal agent to treat the S. parasitica.The extract taken by other scientist like Mori et al and Isshika et al conducted their test but with different colonydiameter and exposure time of ait showedpositive results.(Khomvilaiet al 2006)

Saprolegnia is a fresh water fish feeding on waste from fish or other dead animals. It produces both sexually and asexually saprolegiasis is a fungi causedin saprolegia. It can be seen in 15 degree Celsius. It could be identified as the fish skin and gills start to exhibit cotton-like growth, depigmented skin and shrunken eyes. In several cases the cotton like growth which is caused by fungus can extend into muscle tissue. Many fungicides are readily available in market but they come with the benchmark of causing environmental hazards which therefore, causes less use of these fungicides. Aqueous extract is used as it is known to show antifungal properties derived from alleopathic plants which reduces the germination of spores and mycellal growth of pathogenic fungi. As described by Ilondu et al (2009) marine seeds were used as baits to trap the fish and some other fungi from river Ethiope Delta State. Nigiria employing the method used by Ilondu et al. The culture obtained was maintained at 28 degree Celsius and to avoid bacterial contamination choloromycetin capsules were used which is antibacterial agent. They were kept in laboratory for a week or two to observe and isolate to form a new colony. They were then introduced with different concentration of the extract of vernonia amygdaline. The test were done twice, second time using the pure culture obtained from the first test. After a week the fungi which was isolated and introduced in the tank with the saprolegnia sp., the change could be noted such as skin colour turned whitish-brown another week later fluffy tufts were seen. The mycelial growth was observed that varied when introduced with different concentration of extraction. It was observed that higher the concentration, the higher is the inhibition of the fungus. The highest concentration Ilondu et al (2009) used was 50% where the fishes showed healthy conditions even though quality of water was altered proving to be an antifungal agent which helps in inhibition of fungus growth in fish.(Ilondu et al 2009)

Saprolegniaparasitica, (a disease affecting fish eggs and juvenile fish in hatcheries world wide, is caused by the pathog encioo mycete saprolegniaparasitica. This disease presents as greyish white patches of filamentous my celium on the body or fins of fish and is as sociated with tissue damage leadingi to death of the animal). Toriputitora (Hamilton) is the most important fish foodi in uplandiregion of India. But now a day due to irregular fishing the species is gettingis canty iand this speciesi is listediiniendangered species. There hence artificial breeding technology is helping in rehabilitation of this species. Pathogenic fungus saprolegniaparasiticais mainly responsible for fungal in fections in cold water fish and their eggs. The infection caused by saprolegniaispp. Is termedias “saprolegniosis”. Pathogenic fungi were isolated from the fungal infected tissues of it. Putitora collected from the experimental cemented fish tanks. Isolated fungi were in cubated in the petriiplate having the hempiseed as bait. These fungal colonies were then was hed with sterile double distilled water for i3-4 itimesi and itransferredi toi new petriiplatei with some new hemp seeds and incubatediat 14 degree Celsius for 48 hours a is mallipiece of fungus was cut out from it, rinsed with sterile distilled water and transferred to SDA (Sabouraud's dextroseiagar) plates. Again, a small piece of fungus was cut out from SDA plate, rinsed withi sterile distilled water and transferred to a new sdaiplate. This process was repeated for 4 to 5 times which finally yielded a bacteria free, pure fungal culture. Mycelial growth after treatment with plant extracts at different concentrations i.noi mycelial growth was observed in the fungal colonies in collusion treated with aqueous extracts of citrates at any of the concentrations and exposure times. Accordingly, the minimum in hibitory concentration of aqueous extract was determinedias  5% concentration for  20 min. exposure time. The research not only reveals the ant fungal effect of aqueous extract of citrates but also holds great prospect especially for the peasant local fish farmers who cannot afford the high cost of chemicals and synthetic fungicides and will also reduce the hazard caused byichemical fungicides in the environment. (Singh et al 2013)

Garlic, Galanga rizome,Betal vine, Sapindus sp., Rhinacanthus nasutus Linn. and K. galangaLinn. root extractshave proven to exist with antifungal activity against dermatophytes for veterinary and humanly treatments. They show fungicidal and fungistatic activity and with variation of concentration and exposure time they inhibit the fungus in the extracts mentioned above. Pareeyaet al used air dried leaves and roots extracts of these medical plants were extracted with the help of ethanol prepared with Soxhlet as solvent. Glucose Yeast extract was used for culturing s.pH2(1g glucose, 0.25g yeast extract, 1.5g agar and 100ml distilled water). Then it was incubated at 20 degree Celsius.Glucose yeast were used as control medium. Following the Hussein et al 2000 the extracts were diluted 20 times with sterile water and the colony formed were observed at the interval of 24, 48, 72 hours comparing it with controlled and the diameter of the colony of fungal with glucose yeast with crude extracts.When same was done with the fungicidal the fungal growth were not seen on glucose yeast agar with the interval of 48 hours. Crude extracts of betel vine leaves and K. galanga roots were observed as antimicrobial activities as the vegetative form of S. parasitica was killed at different concentration and exposure time. Extracts of betel vine leaves and K. galanga roots showed high activity in fungistatic where the other crude extracts dissolved completely in water these two were not completely suspended hence proving to show great fungistatic activity whereas R. nasutus Linn. showed slightly moderate activity and Galangarhizomes and K. galanga showed minimal fungistatic activity in different concentration. (Pareeya et al 2007)

Ini India, ilasti fewi years ihavei seeni greati advancementi iniproductivity iini fisheriesi sector.i Salmonidsi are ionei ofi the imosti commercially iimportanti fresh water ifish ispecies.i With ithei growingi India iwei have ifor gotten iabouti thei fishingi department. iBacterial iinfection iisi utmosti importanti to ius iati and idisease icaused iisimycosis i(it iis idiseasei which iisi spread iinto itissues,i bones,i andi organs).i Plantsi havei beeni known ifor imedicinali properties isincei ancient itimes. iTherei arei manyi plants iwhich iare ihighi ini supres singi growth iof ifungal iinfection. iThere iare imany ichemicals iwhich ihave irestricted iduei toi per sistence iof ienvironment iand itoxicity. iThe iaimi to istudyi wasi toi describe ithe ianti mycosisi activity iof iherb iagainst iwater imoulds iand itoi investigate itheir itoxicity. iToi prepare ithe iextract ileaves iwere icleanedi manyi times iand ileft ifor iair idrying. iThen ileaves iare iputiinig rinderi for igrinding. iThe ipowder iextracted iwasi soakedi ini stilledi water ifor i48 ihours iat ithreei differenti concentrations i(5%, i8%, iand i10%). iFinally, iextract iwas ifiltered, iandi bacteria ifreei extract iwere ipreserved iat iless ithan i4 idegree iCelsius. iiSuchi treatediagar iplus iwerei puti over iplatesi underi as eptici conditioni and iallowedi toi in cubateiat i(18-20i Celsius) ifor i48i hours. ishows iusi the iantifungal iactivityi andi holdi great iprospects iand ias ifor ithe ipeasant ilocal ifish ifarmer iwhoi cannoti afford ithe ihighi cost iof ichemicals iand isynthetic ifungicides iit ialso ihelps iini treatment iof isaprolegniasis iin ieggsi and ilarva iof ithe ifish. (Singh iet ial  i2013)iiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiI                                                                                                                                              

The iwordi phyto fungi toxic imeansi (exerting iai toxic ieffect ion ifungi). iUse iof isynthetic ipesticides ifor iquicki and imanagementi of ithe iplanti diseasei andi microbial icontaminationi ini several iagriculturali commodities. iThe ieffect ion isoil iand ibiosphere ithat icreatei healthi hazardous iin ihumani andi animalsi isi the iapplication iof ichemical ipesticides igives iresidual itoxicity. iSeverali pathogenici micro-organisms ihave idevelopedi resistanti againsti the seichemicals’ ipesticides. iPlantsi have iknowni for ithe irimedicinal iandi anti microbiali properties isincei ancienti times. iFor imany ireasons’ iattention ihasi been ishiftedi to ialternativei safe iand icheap imethodi for ithe imanagementi ofi pathogenic imicro-organism.

iTomatoesi werei collectedi and iinfectedi fruits iwere isterilizedi using i75%i ethyl ialcohol. iThei sterilizedi fruits iwere iplaced ion iPetriidish icontaining ipdaimediumi about i20 mli for ieachi plate. iThe iplates iwere iin cubatedi at i25i degree iCelsius ithe idevelopingi coloniesi werei identified ion ibasis iof imorphological icharacteristics. iFive ifishesi`

reisuspected iwith iinfectioni saprolegniasis.i Thei recognitioni ofi disease iisi done ion ipresence iofi cottoni likei whitei toi browni growth ion iskin igills,i fins. iusingi sesamei seeds iwater iculture iand ibaiting itechniques ithe igrowing ifungus iwasi isolated iandi identified. iSporei germination iof ialter iariaisalamii wasi preparedi for i7 idays ioldi culturei growing ion ipdaimedium.i Five iml iofi extractivei stocksi werei added ito i20 mli of i1% of isoili extracti agar.i Soili extractiagar iwasi use diini order itoi present ithei fungal ispores iwith iminimumi ofi added inutrient. iOf ithe itwenty iplants iextract itested iEugenia iaromatici completely iin hibitedi conical igermination. iThei remaining itestedi extracts iwere ieither iless ieffective iori exertedi simulative ieffects icompared iwithi thei control itreatment. iSimilarly, iseveral ireports idealt iwith ithei eitheri in hibitory iori simulative ieffects iofi somei plant iextracts ioni fungal ispore igermination. iAlso, iit iwas ifoundi thati thei aqueous iextracts iofi selected iplants i(except ithosei of icarroti storage iroots) ishowedi in hibitory iactioni against ithe imy celial igrowth,i produced iby iboth itestedi fungal ispeciesi wheni addedi toi the igrowth imedium.(Khallil i2001)

Based on these researches it is proved that plant extracts workwonders on pathogenic fungi in fish as compared to chemical agents. The plants extracts are much more success and useful as compared to previously used chemical agents. The new technique to treat the pathogenic fungi in fishes is by antimicrobial agent actively present in these plantsextracted. The previously existed chemicals agents were dangerous to the aquatic life and hence left long lasting effect on aquatic life. The chemical agentwas not effective as the newfound plant extracts are, it was time consuming and showed less effect on the pathogenic fish and cannot be proved effective in later stages of the pathogenic fungi on fish’s effected parts. It was either surgically removed or the fish used to die before it could be operated but because of the plant extracts used in treatment the time conservation wasreduced and the results were obtained quite fast as compared to chemicals treatment. The other untargeted aquatic organisms were not affected by plant extract. Hence it is proven to be organic and environmentfriendly low costing and much more effective than chemical agents and resolves the danger to aquaculture.

1. Khallil M, 2001, Phytofungitoxic properties in the aqueous extracts of some plant, pakistan journal of biological science, 382-394, asian network for scientific information,2001. 2. Khomvilai, C. iand iKhahiwagi, M. 2006, iFungicidal activities of horseradish extract on a fish pathogen oomycete, S. parasitica. Bull. Fac. Bio resources, Mie University. 33: 1-7. 3. Ebele M. Ilondu, Francis O. Arimoro, Adjekawen P. Sodje 2009, The use of aqueous extracts of Vernonia amygdaline in the control of saprolegniasis in Clarias gariepinus, a fresh water fish, African J. of Biotechnology Vol 8(24) pp, 7130-7132 ISSN 1684-5315, Academic Journals. 4. Michaela C, R. Dinică, L. Gitin, C. Vizireanu I,,2013, Study on herbal actions of Horseradish Armoracia rusticana, Journal of Agroalimentary Process and Techniques, 19(1), 111-115. 5. Pareeya Udomkusonsri, K. Trongvanichnam, M. L. Narumol Klangkaew and N. Kusuchari, 2007 In vitro Efficacy of the Antifungal Activity of Some Thai Medicinal-Plants on the Pathogenic Fungus, Saprolegnia parasitica H2, from Fish, Kasetsart J. (Nat. Sci.) 41 : 56 - 61 6. R. Singh, A. Kumar, N.N.Pandey, M. Gupta, P. Kumar, R.S. Haldar, S. Ali, S. Kumar and A. Pande, 2013, In vitro evaluation of antifungal activity of aqueous extract of lemon grass (Cymbopogon citrates) against pathogenic fungi Saprolegnia parasitica isolated from endangered cold-water fish Tor putitora, J. Ecophysiology. Occupy. Hlth. 1 & 2. 7. R. SINGH, A. Kumar, N.N. Pandey and M. Gupta 2013, In vitro screening of antifungal activity of aqueous extracts of some medicinal plants against pathogenic fungi saprolegnia parasitica, International Journal of Advanced Research, Volume 1, Issue 6, 116-123.