A New Species of the Genus Metacystis (Ciliate, Prostomatida) from a Wastewater Treatment Plant of a Health Resort

Arregui, L1, Perez-Uz, B1, Zornoza, A 2 and Serrano, S 1 Arregui, Perez- Uz, Zornoza,
1Departamento

de Microbiologa III. Facultad de Biologa. Universidad Complutense, Madrid, Spain. 2EGEVASA. EDAR Complutense, Quart-Benager. EPSAR. Valencia , Spain. Quart- Benager.

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 l 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 g/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 m.

RESULTS and DISCUSSION Species Diagnosis

67.6--51.6 m

Metacystis galiana n. sp.

Different stages of the life cycle

In vivo
18.8-13.6 m

Caudal cilium

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

Bipartition process

Swimming form

Stained Cells FLUTAX

Ck

DAPI
Ok

OC

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

LSk

One spherical macronucleus

SC
TSg

Transversal bipartition inside the lorica

Single micronucleus

20-30 somatic kineties, each constituted by eight kinetosomes Contractile vacuole pore 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 30C)
S pecie s D escriptio n S hap e S ize (m ) C yto stom e N um ber of som atic kinetie s N um be r of tra nsv erse ciliary ring s _ C irc um ora l ciliature Ma (m ) Mi T erm ina l V acuo le C auda l cilium Lorica S hap e S ize (m ) N um b er of gird le s pre se n t abse nt brack ish H abitat

M etacystis annulata M etacystis borro ri

S tokes, 1895

elo ngate

55

pre se nt

abse nt

GENUS Metacystis

M etacystis daphnicola M etacystis elongata M eatcystis exig ua M etac ys tis ga lia n a

A la dro-L ube l and M artinez M urillo P ena rd, 1922 K ahl, 1928

ovo idcylind ric a l

10-35 x 10-18 75-17 0

centra l

22-30

5-7

2 perio ra l 1 circ um ora l

3-7

abse nt

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

140

18-61 x 11-26

m arine

elo ngate

24

pre se nt

abse nt

cylind ric a l, elo ngate sacca te elo ngate to ovo id

40-12 0 x 10 13-20 44-60.5 x 15-16 92 30 80-12 0

centra l

30

18-21

1 perio ra l

5,5

pre se nt

abse nt

tub ular, curve d anterio rly _

freshw ate r

m arine, brack ish freshw ate r bio log ica l reactor of W W TP m arine, freshw ate r freshw ate r

COMPARATIVE DATA

P ena rd, 1922 n.sp.

ace ntric apic a l

_ 20-30

5-6 8

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

_ sing le, ovo id

_ sing le, sp haeric al _ 4 _

abse nt pre se nt

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

sacca te am pho ra-like

_ 66.4 x 2 0

abse nt 11-16

M eacystis hyperh alina M etacystis lagenula M etacystis m egapterae

D ietz, 1964 P ena rd,1 922 K ahl, 1930

elo ngate va se -like elo ngate

_ ace ntric _

_ pre se nt _

4 _ _

_ sm all _

pre se nt abse nt abse nt

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

_ 50-62 _

_ abse nt _

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

P ena rd, 1922 S tokes, 1893 K ahl, 1926 C ohn, 18 66

elo ngate cylind ric a l cylind ric a l ovo idcylind ric a l

45-68 50-11 0 x 2 5-39 50 10-91 x 1 1-30 centra l centra l centra l 63-66 20-26

15 20-24 9 10-18 2 perio ra l 1 circ um ora l 1 perio ra l ab unda nt perioral cilia 10

1 1 1 714 m 1

pre se nt pre se nt pre se nt pre se nt

1 abse nt 1 abse nt

100 x 48 84 x 25 77-14 0 x 2 4.531.5

abse nt abse nt 15-26

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