Marisia - Maros Megyei Múzeum Évkönyve 32-34. (2014)

Zoology

A novel halophilic archaeon of Haloferax genus isolatedfrom Telega (Prahova county, Romania) rock salt crystals

nitrite were performed as previously described [6]. Also, the test for starch, casein, Tween80 and gelatin hydrolysis followed previously described methods [6].

Sensitivity to bile salt sodium deoxycholate and antibiotics (chloramphenicol, novobiocin, bacitracin, erythromycin, penicillin, ampicilin and neomycin) was tested according to previously described protocols [6].

The membrane lipids profile was investigated performed using thin layer chromatography [11]. Haloarchaeal cells from 100 ml cultures were used to obtain genomic DNA following the method ofTamaoka [18].

The PCR technique was used to amplify the l6S-rRNA genes, using archaeal specific forward and reverse primers, 5-TCCGGTTGATCCTGCCG (position 8—24 in E. coli) and 5-GGAGGTGATCCAGCCG (position 1540—1525), respectively. The amplified DNA fragment was sequenced using BigDye Terminator Cycle Sequencing Kit (Pharmacia Biotech) and ABI Prism DNA genetic analyzer (Applied Biosystems) [6].

Results and discussions

The RF1 strain was isolated from rock salt crystal from Telega area, Prahova county, Romania, and provided by courtesy of Dr. Sergiu Fendrihan. The rods cells of investigated strain lyses in distilled water and culture medium devoided of sodium chloride, and showed a Gram-negative staining.

Table 1. Characteristics of Haloferax genus members

Properties/ 12345678 9 10 11 12

Strains_____________________________________________________________________________________

alexandrinus + + + + - + + 1.7-5.2_______43_______Saltern A

chudinovii - - + nd + + - - 1.2-4.6 2.5-3.0 Silvinite N

_____________________________________________________________________________deposit_______

denitrificans + + + + 1.5-4.5 1.5-4.5_____Saltern A

elongans - -+/- + + + + + 1.7-5.1 1.7-5.1 Microbial N

___________________________________________________________mats________

gibonsii________+ + + + - + + + 1,5-5.2 1.5-5.2 Saltern A

larsenii + + + + + + + - 1.0-4.8 1.0-4.8_____Saltern A

lucentense + - + + - + -- 1.8-5.1_______43_______Saltern_____A

mucosum - -+-- + + 1.7-5.1 2.6-3.4 Microbial N

______________________________________________________________________________mats________

mediterranei_____+ + + + + + + 13-4.7_______23_______Saltern_____A

prahovense_____+ - + + + + -- 23-5.2_______33______Salt lake N

sufurifontis + + + + - + + 1.0-5.2 2.1-2.6 Zodletone N

____________________________________________________________________________spring________

volcanii______ + + + + ---- 1,0-4.5_____1.7-2.5 Dead Sea N

RF1__________ + + 1 + I - I - I - 1 - 1 2.0-5.2 2.0-4.0 Rock salt N

1 = Formation of sulphide from sodium thiosulphate; 2 = Nitrate reduction; 3 = oxidase; 4 = indole from tryptone; 5 = starch hydrolysis; 6 = Tween80 hydrolysis; 7 = Gelatin liquefactions; 8 = casein hydrolysis; 9 = range of NaCl for growth (M); 10 = optimum NaCl for growth (M); 11 = isolation site; 12 = origin of isolation site; “+” = positive;

= negative; = variable results; nd = no data available; N = natural environment; A = man-made environment

Colonies grown on JCM 168 medium are circular, smooth, having elevated surface with right margins. The colour of the colonies was red-orange. The major differences between RF1 and other members of Haloferax genus are detailed in Table 1. RF1 strain was able to grow in a

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