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A New Species of the Genus Metacystis (Ciliate, Prostomatida) from a Wastewater Treatment Plant of a Health Resort Arregui, Arregui, L1, PerezPerez-Uz, Uz, B1, Zornoza, Zornoza, A 2 and Serrano, S 1 1Departamento

de Microbiología III. Facultad de Biología. Universidad Complutense, Complutense, Madrid, Spain. 2EGEVASA. EDAR QuartQuart-Benager. Benager. EPSAR. Valencia , Spain.

INTRODUCTION Ciliates, the most significant group of protists within the ecosystem of biological reactors of sewage treatment plants, can be divided into crawling and sessile ciliates that are associated with flocs and, swimming ciliates that move freely in the mixed liquor. Species of Prostomatids, usually free-swimming ciliates, are scarce in activated sludge WWTP, they do not usually reach high abundances and have never been reported as the dominant population in stable conditions. The aim of this work is to describe a new species of a sessile prostomatid found associated with flocs in the biological reactor.

MATERIAL AND METHODS Metacystis sp. was found in the biological reactor of a waste water treatment plant of a health resort located in Cofrentes, Valencia (Spain). 50 μl aliquots containing living cells were placed on a coversliped slide and viewed by phase contrast microscopy using either an Olympus BH-5 or a Zeiss Axiostar microscope. Specimens were stained with Flutax-2 (as described in Arregui et al. 2003) and DAPI (final concentration of 10 μg/ml), and were observed under a Zeiss Axioplan 2 epifluorescence microscope with a CCD Spot Camera. Physico-chemical parameters were measured according to standard methods (APHA 1989). Scale bar means 20 μm.

RESULTS and DISCUSSION Species Diagnosis

Different stages of the life cycle

67.6--51.6 μm

Metacystis galiana n. sp.

Caudal cilium

“In vivo” 18.8-13.6 μm

Bipartition process

9Elongate cell shape 9Cytostome at the anterior apical end of a retractile proboscide 9Sac-like lorica tapered toward the aperture with a smooth neck and 11--16 annular ridges 9Somatic ciliature formed by longitudinal kineties 9Circumoral ciliature constituted by perioral girdles 9One single posterior contractile vacuole 9One large vacuole protruding at the cell terminal end 9Rounded secreted particles often observed inside the lorica

Swimming form

Stained Cells FLUTAX

DAPI Ck Ok

OC

CIRCUMORAL INFRACILIATURE 9One circumoral kinety (dikinetids) 9Two perioral girdles (fiveand two densely packed kinetosomes)

9One spherical macronucleus

LSk

SC

Transversal bipartition inside the lorica

TSg

9Single micronucleus

920-30 somatic kineties, each constituted by eight kinetosomes 9Contractile vacuole pore 9 PHYSICAL-CHEMICAL CONDITIONS IN THE BIOLOGICAL REACTOR OF WWTP High conductivities due high values of sulfates, carbonates, magnesium, calcium and sodium ions and unusually high water temperature (rounding 30ºC) S pecie s

GENUS Metacystis

D escriptio n

S hap e

S ize (μm )

C yto stom e

N um ber of som atic kinetie s

M etacystis annulata

S tokes, 1895

elo ngate

55

_

_

N um be r of tra nsv erse ciliary ring s _

_

_

M etacystis borro ri

A la dro-L ube l and M artinez M urillo P ena rd, 1922

ovo idcylind ric a l

10-35 x 10-18

centra l

22-30

5-7

2 perio ra l 1 circ um ora l

3-7

M etacystis daphnicola

C irc um ora l ciliature

Ma (μm )

Mi

_

T erm ina l V acuo le

C auda l cilium

pre se nt

abse nt

abse nt

Lorica S hap e

cylind rica l, w ith a nterior neck cylind rica l

H abitat S ize (μm )

N um b er of gird le s

140

pre se n t

brack ish

18-61 x 11-26

abse nt

m arine

elo ngate

75-17 0

_

_

24

_

_

_

pre se nt

abse nt

_

freshw ate r

K ahl, 1928

cylind ric a l, elo ngate

40-12 0 x 10

centra l

30

18-21

1 perio ra l

5,5

1

pre se nt

abse nt

tub ular, curve d anterio rly _

_

M etacystis elongata

_

_

m arine, brack ish

M eatcystis exig ua M etac ys tis ga lia n a

P ena rd, 1922 n.sp.

sacca te

13-20

ace ntric

_

5-6

2 perio ra l

_

_

abse nt

va rio us

sacca te

_

abse nt

freshw ate r

elo ngate to ovo id

44-60.5 x 15-16

apic a l

20-30

8

sing le, ovo id

sing le, sp haeric al

pre se nt

am pho ra-like

11-16

bio log ica l reactor of W W TP

M eacystis hyperh alina M etacystis lagenula M etacystis m egapterae

M etacystis recurva M etacystis striata M etacystis tesselata M etacystis truncata

D ietz, 1964

elo ngate

92

4

2 (5+ 2) perioral 1 circ um ora l (dik inetid s) _

_

_

pre se nt

pre se nt in the sw im m ing sta ge abse nt

P ena rd,1 922

va se -like

30

ace ntric

pre se nt

_

2 perio ra l

sm all

4

abse nt

va rio us

K ahl, 1930

elo ngate

80-12 0

_

_

_

_

_

_

abse nt

abse nt

centra l

63-66

20-24

20-26

9

2 perio ra l 1 circ um ora l 1 perio ra l

10

centra l

ab unda nt perioral cilia

714 μm

P ena rd, 1922 S tokes, 1893

elo ngate

45-68

cylind ric a l

K ahl, 1926

cylind ric a l

50-11 0 x 2 5-39 50

C ohn, 18 66

ovo idcylind ric a l

10-91 x 1 1-30

_

_

15

centra l

10-18

1

pre se nt

1

like M . annulata V ase-lik e cylind rica l, sem ia nnular, ro und ed poste rio rly va se -like

66.4 x 2 0

_

_

m arine,

50-62

abse nt

freshw ate r

_

_

freshw ate r

freshw ate r, m arine brack ish, m arine freshw ate r

1

pre se nt

abse nt

tub ular

100 x 48

abse nt

1

pre se nt

1

va se -like

84 x 25

abse nt

1

pre se nt

abse nt

cylind rica l, annular

77-14 0 x 2 4.531.5

15-26

brack ish, m arine

COMPARATIVE DATA


A new species of the genus Metacystis