Rezumatul tezei de doctorat - USAMV Cluj-Napoca

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The experimental model used, included the collecting of 5 water samples from each fishpond, from the 4 lake sides and from centre, and 50 cm medium depth. From the 5 samples we made a single water sample, which was subsequent analyzed in laboratory conditions. For the identification of different species from Saprolegniaceae family, we collected and analyzed water samples, carp species and spawns affected by disease, from Transylvania monitored fishponds. The water samples were collected in 2 litres recipients, from the 5 areas of each fishpond, then the 5 samples were mixed in a 15 litres tank, collecting for the analyze a single blended sample from each fishpond, in a 2 litres recipient. The transport was made in the first 12-24 hours from the collection and the analysis were performed in the next day. I.3.2. The initiation of Saprolegnia culture in laboratory conditions To ensure that the possible differences, which could appear between the fungal species from Saprolegniaceae family present in the water, could be strict attributed to the differences given by water nature and the species which populates it, we performed as it follows: from each water sample, in a quantity of 2 litres, we made up 3 samples of 100 ml, in plastic sterile recipients. In each recipient there were introduced 10-15 spawns from the same carp female. Because the literature offers contradictory data respecting Saprolegnia’s development temperature in different world areals, we suggested the testing of the fungus growing and development mode, at 3 levels of temperature: 10°C, 15°C and 22°C. Water samples, containing 10-15 spawns for artificial infestation, were introduced in a incubator, following the temperatures mentioned, between 3-7 days. In this period we made daily observations on each sample, to find out the starting moment of fungus infestation upon spawns and the intensity of its development at that temperature. There was constituted a number of 207 samples for each collecting month, 3 samples for each fishpond, which were monitored at the 3 temperature levels, and making an 48
estimation of the spawns infestation degree, with marks from 0 to 4, at 48, 96 and 144 hours. The data interpretation was made by estimating the medium score of all the samples from the same thermal gradient, the differences being percentual expressed. The marks used have the following significations: 0 – uninfested 1 – very weak infested 2 – weak infested 3 – medium infested 4 –strong infested I.3.3. Culture media used The scientific literature underlines the fact that, generally, in the fungi cultures development could be used either solid media, or liquid ones. Liquid media allow the development of fungal mycelium in a large quantity, that could be used in techniques for DNA extraction (Dieguez-Uribeondo and al., 2007; Fernandez-Benitez and al., 2008). In this research we utilized comparatively 2 solid media: Potato Dextrose Agar (PDA) and Sabouraud 2% Glucose Agar (SGA), and 2 liquid media: Potato Dextrose Broth (PDB) and Sabouraud Dextrose Broth (SDB). The spawns were sterile collected in Petri dishes and washed with distilled water containing 100 mg L -1 Penicillin G potassium salt. Then, were inoculated in two solid media: Potato Dextrose Agar (PDA) and Sabouraud 2% Glucose Agar (SGA), and for preventing the bacterial growing, we added the same antibiotic, in the recommended concentration. The colonies were preserved then on PDA and SGA media, in the incubator for 3 days, at the mentioned temperatures (10, 15 and 22°C). In this period we observed Saprolegnia colonies growing speed and their diameter (adapted after Fernandez-Benitez and al., 2008). The original contribution consisted in the fact that we used two comparative solid culture media and three temperature gradients for each medium. 49
Page 1 and 2: UNIVERSITATEA DE ŞTIINłE AGRICOLE
Page 3 and 4: II.6. REZULTATELE RESTRICłIEI ENZI
Page 5 and 6: diverse specii piscicole. De aici s
Page 7 and 8: Fig.1. LocaŃiile în care s-au efe
Page 9 and 10: A fost constituit un număr de 207
Page 11 and 12: Sabouraud Dextrose Broth (SDB-bulio
Page 13 and 14: Echipament, materiale şi reactivi
Page 15 and 16: Primerii folosiŃi de către noi î
Page 17 and 18: CAPITOLUL II REZULTATELE CERCETĂRI
Page 19 and 20: Mediile diametrului coloniilor de S
Page 21 and 22: laborator specifice metodologiei de
Page 23 and 24: Deoarece am utilizat în experiment
Page 25 and 26: fragmente: primul la aproximativ 45
Page 27 and 28: II.7. REZULTATE PRIVIND STUDIUL ÎN
Page 29 and 30: mai mare genetică, indivizii despr
Page 31 and 32: MenŃionăm faptul că în toate ba
Page 33 and 34: CAPITOLUL III CONCLUZII ŞI RECOMAN
Page 35 and 36: isexualis (761 pb, o identitate de
Page 37 and 38: BIBLIOGRAFIE SELECTIVĂ 1. Colao Ma
Page 39 and 40: 19. Oroian, T.E., 2006, SelecŃia a
Page 41 and 42: UNIVERSITY OF AGRICULTURAL SCIENCES
Page 43 and 44: SPECIES FROM SAPROLEGNIACEAE FAMILY
Page 45 and 46: The specific literature presents up
Page 47: Fig.1. The locations where the rese
Page 51 and 52: pellets were filtred, washed with d
Page 53 and 54: The DNA rapid extraction using PBS
Page 55 and 56: We used a comparatively mixture of
Page 57 and 58: II.1. THE RESULTS OF SAPROLEGNIA CU
Page 59 and 60: Reading time 24 48 72 n Culture med
Page 61 and 62: asexual one. We can precisely concl
Page 63 and 64: 1 2 3 4 5 6 7 8 9 700pb- Fig.3. Ele
Page 65 and 66: pairs length. The sequencing result
Page 67 and 68: II.7. RESULTS REGARDING THE GENETIC
Page 69 and 70: higher genetic distance, the indivi
Page 71 and 72: We mention the fact that in al the
Page 73 and 74: extraction have values between 1,10
Page 75 and 76: 13. In researches on the aquatic fu
Page 77 and 78: 40. Frisvad, J.C., Bridge, P.D., Ar
Page 79: 61. *** http://www.sigmaaldrich.com

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