4. Result - INFLIBNETshodhganga.inflibnet.ac.in/bitstream/10603/9641/8/08...The posterior region of...
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Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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4. Result
4.1 Qualitative data of microzooplankton from Gulf of Mannar coastal and
brackish waters, India.
Great varieties of planktonic and benthic forms were present in the Gulf of
Mannar, region in East coast of India. Nearly 116 species of planktonic and benthic
forms were identified based on viable and different staining techniques.
1. Uronema marinum (Dujardin, 1841) (Plate-1,1)
Elongate ovoid structure, the body is 45-50µm long, slightly flattened, anterior
region not ciliated, inconspicuous peristome with ciliated right edge cytostome on the
ventral side close to left border in the anterior half, with a small tongue-like membrane
also present, cytopharynx indistincted, macronucleus spherical, contractile vacuole
terminal, and mostly in marine water.
2. Urostyla sp. (Ehrenberg, 1831) (Plate-1,2)
Ellipsoid structure, 65-70µm, ends round flatten ventral surface with four to ten
rows of small cirri, two marginal rows, three or more frontals, five to twelve anals,
macronucleus a single body or in many parts and marine water species.
3. Coleps sp. (Ehrenberg, 1831) (Plate-1, 3)
Cytostome at the anterior end, body barrel-shaped, 100-105µm size with
pellicular plates and uniform cilia were present. Anterior mouth some time not visible ,
posterior spines not observe. The body width size is 38-40µm. and mostly in brackish
water basins.
4. Eschaneustyla sp. (Stokes, 1891) (Plates-1, 4)
Elliptical or ovate shaped. Narrow peristome, and one–third of the body length
is extended. Size of the body is 100-110µm. Frontals were present. About ten to
twenty membranes observe in anterior margin, ventrals small and numerous in three
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oblique rows observed, no anals, marginal uninterrupted. Contractile vacuole a long
canal near left border notclearly visible. Mostly in brackish water.
5. Keronopsis sp. (Ehrenberg, 1831) (Plate-1, 5)
Reni form structures, five or six oblique rows of ventral cirri were observed.
140-145µm in size. In some species have two ventral rows of cirri reaching frontal
region. Caudals are variable in size. Macronucleus usually observed. Mostly in marine
water.
6. Tintinnopsis beroidea (Hada,1932 ) (Plate-1, 6)
Characterised by the presence of a long bullet shaped lorica, which has pointed
aborally. Lorica elongated and agglomeration has moderately present. In aboral part,
the pointed terminal sometime clearly no visible, without agglomeration. The
measurements were made at a magnification of x100-150. Lorica length is 75-80µm
and oral diameter 25-30µm.
7. Leprotintinnus nordquisti (Brandt, 1906) (Plate-1, 7)
Lorica tubular. Opening at the aboral end. Collar absent. Lorica elongated,
Cylindrical and posterior end inflated, with conical base. Often openings on both ends.
Lorical wall with „bulli‟ form fiber and grains on its surface. This is widely distributed
in Gulf of Mannar region, India.
8. T. campanula (Ehrenberg,1831) (Plate-1, 8)
Lorica is cylindrical campanulate. Aboral region convex or horn conical shaped.
Oral rim present in adoral region. Rose Bengal and lugol‟s iodine adopted. Sometime
„cell-membranelles were raised out in adoral part‟. Lorica length is 185- 200µm. Width
is 70-80µm. One or two blade like oral polykinetids, form a full circle around the
anterior end of the cell.This part called as External Polykinetid Zone. (EPZ).
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9. T. kofoidi (Hada, 1932) (Plate-1, 9)
Lorica with aboral horn structure. Often lorica cylindrical with small horn,
opening at the end region. Present both neritic and oceanic waters. It forms many times
saccular nature. Lorica wall is often covered by fine grains, soil and other matter.
10. T. aperta (Brandt, 1906) (Plate-1, 10) (Plate-4, 8)
The agglomeration of sand particles on lorica is not intense. Lorica‟scylindrical
end is an indistinct or sometime distinct bowl. Oral region without a flare. An aboral
part of lorica is pointed. Length of lorica is 160-165µm.Oral diameter 64-68µm. One or
two macronuclei were present. The size of bowl in adoral region is 30-35µm.
11. T. turgida (Brandt, 1906) (Plate-1, 11 & Plate 2, 11)
Lorica is cylindrical or subcylindrical shaped in anteriorly, expanding in
aborally and oral diameter is 13µm. Aboral end broadly rounded, sometime with a
„faint indication of a pointed‟. Length is 80-110µm. Often found in coastal waters. It
differs from T. gracilis in distended structure and rounded aboral region.
12. Favella ehrenbergii (Lachmann, 1856) (Plate-1, 12,15)
Lorica very elongated, more or less cylindrical shaped. Bowl rounded aborally
and highly variable in length. Mostly „Hyaline lorica‟. Pedicel was prominently
present.Lorica has no spiral turns. Two or three macronucleiwere observe clearly. One
external polykinetid present. Length ofthe lorica about 150-210µm and width 85-
110µm. Macronuclei size 5-12µm. The macronuclei were oval-shaped and variable in
position in their cytoplasm. Aboral region has conical shaped, not elongated in
cylindrically. Lorica‟soral rim has denticulate structure. Diameter of oral rim is 30-
35µm.
13. Philasterides sp. (Kahl, 1932) (Plate-1, 13) (Plate-3, 19) (Plate-5, 9)
Oral groove elongate triangular structure, adoral ciliature is tetrahymenal shape,
and three membranelles are replaced by three ciliary structure. The body size is 50-
60µm. Sometime bodyis cylindrical shape and peristome about one-third to two-fifths
of the body length. Broader „cytostome‟ with a series of longer cilia, uniform ciliation,
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a caudal cilium, and clearly identified oval macronucleus with a micronucleus
structure. Mostly in brackish water.
14. T.dadayi (Kofoid & Campbell, 1929) (Plate-1, 14 & Plate-3, 10)
Moderately erected lorica, which is campanulate anteriorly. Aboral part of
lorica is subpherical nature. Agglomeration evenly distributed. Rose Bengal staining
procedures were adopted. Lorica length is 70-180µm. Diameter of lorica is 40-47µm.
Lorica‟s sand particles and algae matters were observed by Sodium Thiosulphate
staining procedures.
15. Tiarina fusus (Ehrenberg, 1831) (Plate-1, 16 & Plate-3, 4)
Cytostome at the anterior side with „uniform ciliation‟. The body length is 120-
125µm. More or less similar to Coleps species structure. But clearly indicated in the
posterior end is tapering to a pointed in structure. This is also in marine waterspecies.
16. Spirostomum minus (Ehrenberg, 1831) (Plate-1, 17)
Body elongated, large, highly contractile nature and uniform body ciliation.
AZM observed in long. Mostly in yellowish to brown in colour. Cilia are short.
Peristome closely lined with short membranellae. The size of the body length is 200-
210µm. Sometime observed in 300µm long, while culture raised above in the culture
medium. Macronucleus is chain form structure. Often in marine water nature.
17. Euplotes balticus (Kahl, 1932) (Plate-2, 1)
Kahl first described this species as E. vannus while later, other scientists were
considered it to be an E.crassus. It is a medium-sized, 60-72µm long ovoid marine
species. The peristome is long and narrow extending down to about 50 AZM
membranelles.There is no fronto-ventral, 5 transverse and 4 or 5 caudal cirri. The
ventral argyrome consists of a few large irregular polygons. The dorsal argyrome is of
the single-vannus type with 6 dorsolateral kineties carrying about 10 dorsal cilia in the
central rows. The macronucleus is C-shaped with a club-like extension on the posterior
arm. The micronucleus is situated in a depression of the left anterior edge of the
macronucleus. The contractile vacuole has several satellite vacuoles were observed by
lugol‟s iodine stain.
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18. Favella taraikaensis (Hada,1932) (Plate-2,2)
Lorica is medium size about 70-90µm in width and highly variable in length
(145-150µm). Bowl sometime slightly expanded below the middle, rounded aborally
and joined by wings like structure with a short, „blunt or turned‟ pedicel.
19. Strombidium turcicum (Lachmann, 1856) (Plate-2, 3)
Body ciliation is greatly reduced or absent, adoral membranelles are well
developed and extend beyond the anterior region of the body. Ovoid to spherical
shaped nucleus. Adoral zone is very conspicuous sickle shaped, frontal membranellae
and adoral membranellae extend down up to cytopharynx, the first section of
membranelles surrounding an apical process, with no body bristles or cirri. Oval or
band-form micronucleus also observed. Always presence in marine coastal water only.
20. T. patula (Kofoid & Campbell, 1929) (Plate-2, 4)
This species is characterized by an agglomerated bowl, which is enlarged
anteriorly into a flare similar to that of a „flower vase‟. The agglomeration is moderate
on the bowl but dense on the rim of the flare. Length is 60- 70µm and oral diameter
has 40-45µm.
21. Dichilumsp. (Ehrenberg, 1831) (Plate-2, 5)
Ovoid to ellipsoidal in shaped. Medium and large size body. Uniform and
dense ciliation rows were observed. In this species, cytopharynx have conical shaped
with „undulating membrane‟ and contractile vacuoles were observe. About 40-50µm in
size. Similar to Monochilum sp., but membrane on both edges of the cytostome differ.
Often presence in brackish habitat.
22. F.ehrenbergii strain -1, (Plate-2, 6)
Lorica very elongated, often in cylindrical shaped. Bowl rounded aborally and
„highly variable‟ in length. Hyaline lorica was observed. Pedicel was prominently
present. Lorica has no spiral turns. This species observe only in Rameswaram areas and
also Kurusadai island of Gulf of Mannar, region.
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23. T. patula strain-1, (Plate-2, 7)
An agglomerated bowl which is enlarged anterior into a flare similar to flower
vase. The agglomeration is moderate on the bowl but not dense on the rim of the flare
and „rim has distinctly‟ observe. Length is 60- 70µm and oral diameter has 30-40µm.
Present only in Tutocorin coastal water.
24. T. patula strain-2, (Plate-2, 8)
This species is characterized by highly agglomerated bowl, which is „more
enlarged in the middle part of the lorica‟. Sometime agglomeration is moderate on the
bowl but dense on the rim of the flare. Length is 140- 150µm and oral diameter has40-
50µm. Observed in Tutocorin areas.
25. T. entzil (Kofoid & Campbell, 1929) (Plate-2, 9)
This species is characterized by „very short perfect cylindrical structure‟. Lorica
is distincted. The total length of the lorica is 30-40µm and width has 25-30µm. No
distinct oral rim was observed. Collected in the Mugathuvaram region of Rameswaram
areas.
26. T. brandti (Kofoid & Campbell, 1929) (Plate-2, 10)
This species is similar to Leprotintinnus nordquisti, but the length of the lorica
is very short with dense agglomeration (sand particles were observed on the lorica).
Recorded in the Mugathuvaram region of Rameswaram areas.
27. T. turgida (Strain 1) (Plate-2, 12)
The lorica is cylindrical shaped with adoral part of the oral cavity „just
extended‟. The posterior region of the lorica has flower bowl arrangement. The
diameter of bowl is 40 to 50µm. Such variety of this species never observed in the
Kofoid‟s classification. This variety of sample were observed only in the region of
Mandapam camp and Kelakkarai areas of Gulf of Mannar. Staining procedures
adapted by lugol‟s iodine and 5% Rose Bengal.
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28. T. minuta (Kofoid & Campbell, 1929) (Plate,2 13)
Lorica is short tubular and oral diameter of the lorica is also very short. Oral rim
have no collar structure, cylindrical bowl, aboral end is hemispherical wall with sparse
agglomerations. Length is 25-30µm. It is presence in the region of Rameswaram areas.
29. Strombidium pelagicum (Taniguchi, 1997) (Plate,2 14)
Body subspherical, anterior part is slight cylindrical, posterior more subconical
and fixed with one side flattened structure. Cell surface often covered by small rods
visulised by Protargol stain only. The body has conical shaped. The body size is 195-
200µm. Oral cavity is funnel-shaped. Five somatic kineties occasionally observed. S.
neptuni has more similar to this species.
30. Trachelocerca sp.(Ehrenberg, 1831) (Plate 2, 15) (Plate-5, 20)
Elongated structure, vermiform or flask shaped, more or less extensible, and
drawn-out anterior region.The body without any ring-furrow which marked on „head‟
similar to Lacrymaria sp.. When contract, the pellicular is not spiral shaped and no
neck structure appeared like Chaenea sp.Often presence in marine habitat.
31. Uronema sp. (Kudo, 1986) (Plate 2, 16) (Plate 4, 17)
Cell typically elongate structure, cytostome in anterior region of the cell, three
adoral polykinetid are longitudinally arranged and one distinct oral kinety, bearing long
cilia, runs from the anterior end of the cell. It describes a slight curve around the
cytostome, somatic kineties, extend to the end of the cell, one to several caudal cilia
and one macronucleus, ovoid to spherical shaped. The length of the body is 45-47µm
and width has 23-25µm. Lugol‟s fixation staining highly adopted. The key feature of
this species one to several „caudal cilia‟ with distinct mucocysts are present.
32. Poroecus sp. (Kofoid & Campbell, 1929) (Plate 2, 17)
This species more similar to Codonella variety. The lorica width is 40-50µm
and length has 175-180µm. The aboral part of the lorica is developed a small prominent
projection. This is a key feature for identification of Poroecus sp. Often occurrence in
Rameswaram coastal areas and Kelakkarai coast of the Gulf of Mannar, region.
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33. Euplotes crassus (Curds, 1975) (Plate 2, 18)
Dorsolateral kineties are 10 in numbers, with 26-30 dorsal cilia in central rows.
E. crassus is a large,100-130µm long, elongate oval marine species. The dorsal surface
is strongly sculptured by 8 longitudinal ridges. The peristome is long and narrow while
the AZM consists of about 50 membranelles and extends down the body length. The
argyrome is simple single type and there are „10 dorsolateral kineties‟ with the central
one bearing about 26 dorsal cilia. There are 10 frontoventral, 5 transverse and 5 or 6
caudal cirri. The macronucleus is C-shaped with the posterior arm bearing a foot-like
extension. The compact micronucleus is situated anteriorly near the left edge of the
macronucleus. Often occurred in sedimented areas of Mandapam and Mandapam
Camp.
34. Codonellopsis sp. (Taniguchi, 1997) (Plate 2, 19)
Lorica consisting of long hyaline collar, bowl thickly agglomerated, oval to
spherical , with aboral horn. The lorica width is 50-60µm and length has 120-125µm. It
is distributed in Arthukkarai brackish water areas of Gulf of Mannar. This species
similar to C. schabi.
35. Ascampbelliella sp. (Taniguchi, 1997) (Plate 2,20)
This is more similar to Favella variety. The adoral lorica has double layer
structured. The lorica length is 35-40µm. Present in Mandapam camp areas in the Gulf
of Mannar region. The aboral part of hyaline lorica small prominent pedicel was
observed.
36. C. frigida (Taniguchi, 1997) (Plate 2,21)
The length of the lorica is 62-70µm and width has 25-30µm. Collar thin,
transparent and length slightly changed in the posterior end. Collar with five to six rows
of elliptical or circular fenestrae. Species identified by collar with two or three collar
belts. Bowl globular, or sub globular, with large and irregular grains attached. This
species is widely distributed in the Tuticorin areas of Gulf of Mannar region.
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37. Euplotes latus (Curds, 1975) (Plate3, 1)
There are six to seven dorsolateral kineties were present. Kahl,(1932) has
described as E. patella. The only difference between the E.patella have single
eurystomus type of dorsal argyrome, whereas E.latus have double-patella type.
Kineties, dorsal cilia and macronuclei are very different shapes. The size of body has
70µm long. Mostly found in marine water. The main characteristic feature is „broad and
rounded posteriorly but narrows anteriorly‟. The peristome is large. The AZM is
broadly curved and has composed of 25-30 membranelles. Few authors were observed
as 9 frontoventral cirri are situated very closely together. The macronucleus is an open
angular C-shapedstructure with a micronucleus situated close to the left anterior border.
Present only in the region of Rameswaram areas (Sedimented Areas).
38. E. mutabilis (Curds, 1975) (Plate 3, 2)
There are 11 dorsolateral kineties, with 11-15 dorsal cilia in central kineties.
Dorsal argyrome is single type. Highly kinetosomal networks were observed.
Maximum size is 95µm long and oval shaped marine species. The peristome is quite
large. The right margin extends unevenly down to the body like a spike projection. And
also 10 or 11 frontoventral, 5 transverse and anal cirri five were observe. The
micronucleus is situated in anterior region.
39. Favella taraikaensis (Taniguchi, 1997) (Plate 3, 3)
Lorica is bell or cup shaped. Aboral end with pointed angle. Lorica with
networks structure. It is a common species in inshore water of Rameswaram areas,
notpresent in Tuticorin region. The size of body width is 60-65µm and length has
150-155µm. The key features of the this species „pedicel sometime bended‟ in the
aboral region. Lorica morphology evenly cylindrical shaped from anterior to posterior
region.
40. E. eurystomus (Curds, 1975) (Plate 3, 5)
There are nine frontoventral cirri, sigmoidal AZM with 60-65 membranelles
present. Body is 100-110µm long, 50-60µm width, ovoid and brackish water
hypotrichs. The buccal aperature is triangular and there is a single anterior peristomial
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pouch. The Adoral Zone of Membranelles collar is high and prominent structure. The
dorsal argyrome is typical of the double-eurystomus type with 10 dorsolateral
kineties.Although two strains have variable number (8-12) of kineties. The
macronucleus is typically 3 in shaped and sometime a definite „concave notch‟ which
contains the micronucleus. This species widely distributed in the Thonithurai and
Rameswaram regions of Gulf of Mannar.
41. Ileonema dispar (Kudo, 1986) (Plate 3, 6)
Body flattened, flask-shaped, somewhat similar to Trachelophyllum sp. but,
there is a remarkable flagellum-like projection extending from anterior end.
Cytopharynx have trichocysts present. Often present in brakish areas. The key features
of this species are highly contractile nature, an anterior flagellum half body length, with
basal portion spirally furrowed, and cytostomehave flagellum. Cytopharynx spindle-
form with trichites, two contractile vacuoles with cytopyge posterior, ovoid
macronucleus, and 120µm long. Present in Mandapam camp.
42. E. mutabilis (Strain 1) (Plate 3, 7)
There are 6-7 dorsolateral kineties. Dorsal argyrome is single type. Highly
kinetosomal networks were observed. Maximum size is 60µm long, oval and marine
species. The peristome is large. The right margin extends unevenly down the body to
terminate in a spike-like projection. And also 6-7 frontoventral, 5 transverse and 5anal
cirri were observed. The micronucleus is situated in anterior region. This data was not
observed in earlier report. Very prominentfrontoventral kineties were observed.
Generally, „short shaped‟ not match with any other species of benthic ciliates of
hypotrichs.
43. Trachelophyllum sp. (Kudo, 1986) (Plate 3, 8) (Plate 5, 7)
Elongate and flattened body structure. Flexible, ribbon-like anterior end, neck-
like structure, tips truncate, cytopharynx narrow and ciliary rows evenly distributed.
Two macronuclei were clearly observed. Contractile vacuole is present in terminal
region. Mostly in coastal water species, not in brackish water areas. Maximum size of
body length is 200µm.
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44. Favella meunieri (Striider-Kypke, 2003) (Plate 3, 9)
Aboral region has conical shaped and no wings. In oral rim, „no spiral turns‟
were observe. Pedicel is well developed. It is distribute only in the region of
Rameswaram coastal areas.
45. Strombidium stylifer (Striider-Kypke, 2003) (Plate 3,11)
It is aloricate ciliate groups. The body width is 20µm and length has 65-70µm.
Body truncate to conical shaped. Posterior end pointed anterior end forms dome shaped
and no peristomial collar. Adoral polykinetid zone (APZ) distinctly separated from
ventral polykinetid zone (VPZ). One oval to spherical shapemacronucleus was
observed. Commonly distribute in coastal areas in the Gulf of Mannar region.
46. Pleuronema sp. (Striider-Kypke, 2003) (Plate 3, 12) (Plate 5, 8)
Celltypically elongate-ovoid shape. The cytostome is an equatorial or
subequatorial structure. Three adoral polykinetids were longitudinally arranged. The
length is 30µm to 33µm and width has 15-20µm size. The key features of the species is
„sail structure membrane‟ was observed in the lateral side the of the organisms. One
hyper-telic par-oral kinetyand long cilia run from the anterior end.It describes a distinct
curve, around the cytostome. One to several caudal cilia were present. One
macronucleus, was observe. Mostly, observe in the Rameswaram waters.
47. Parauronema sp. (Striider-Kypke, 2003) (Plate 3, 13)
Generally, oral cavity typically shallow type nature. Cytostome is anterior
region. Three adoral ploykinetids on the left side of the oral region present
longitudinally arranged and appear as field of kinetosomes, not with rows. The size of
the body is 30-32µm length. Only one long cilia, runs from the anterior end. 30 to 40%
cell length is a slight curve around the cytostome.
The key feature is typical „zigzag arrangement of kinetosomes‟, scutica
posterior to the par-oral kinety and visible only in protargol stainig. Mostly in brackish
water.
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48. Amphorellopsis quadrangular (Kofoid & Campbell, 1929) (Plate 3, 14)
The body size of the lorica is 20-25µm in length and width has 75-80µm. One
of the smallest clean loricate species, which is acute posteriorly. The aboral part is
sometime a simple „groove structure‟ was appeared. This species is distributed in the
region of Mandapam camp in the Rameswaram areas.
49. S. capitatum (Striider-Kypke, 2003) (Plate 3, 15)
Cell is squat shaped. Conspicuous peristomial collar was present. The length is
34-40µm and width is 25-30µm size. The girdle kinety is subequatorial structure.It
divides conical bottom from hemispherical top, oral cavity wide and deep structured.
APZ and VPZ clearly separated. One macronucleus was observed. Sometime nucleus
has elongated. It is commonly distributed in inshore water of Gulf of Mannar. India.
50. Heterosigma inlandica (Jomas, 1997) (Plate 3, 16) (Plate 5, 12)
This is marine heterotrophic flagellates. Zoomastigophora groups. The cell
length is 10-18µm. The flagella are approximately equal to cell length, only one was
observed. Few authors recorded as chloroplasts may be presented in the cytoplasm as
colour of greenish brown, but this study not observed such type of colours. Cell more
or less compressed with a branched and ventral groove. Present in Arthukkarai water
areas of Gulf of Mannar, India.
51. S. elegans (Taniguchi,1997) (Plate 3, 17)
The key features of this species, 12 adoral polykinities zones were observed.
Cell almost heart- shaped. Oral cavity has complex structure. Both zones were
separated. Ventral kinety is spirals clockwise direction. Multiple macronuclei were
observed. Nuclei scattered throughout the cytoplasm. Lugol‟s iodine with sodium
thiosulphate wasadopted.
52. Amphileptus sp. (Kudo, 1986) (Plates 3, 18)
Cytostome is slit like structure, located in the adoral region. Body is always
flask- shaped. It is laterally compressed and ciliation is uniform. The maximum size is
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100-105µm. The key feature of this species many contractile vacuocles were observed.
Two or more macronuclei also observed. Present in the Tuticorin region coastal
waters.
53. S. macronucleatum (Taniguchi, 1997) (Plate 3, 20)
This species present only in the coastal waters. The size of the body is 80-
85µm. About 12-15 adoral polykinities were observed. Identification based on viable
technique only possible. The key feature is „zigzag movement and jumping from one to
another by rapid displacement‟. Prominent one macronucleus was observed. Mostly
distributed in the inshore region of Rameswaram in summer season only.
54. Euplotes vannus (Kahl, 1932) (Plate 4, 1)
Key feature of this species is about 20-22 dorsal cilia in central kineties and 60-
70 membranelles in AZM. It is a medium–sized marine species. The length of body
75-80µm. It has an overall oval configuration but is slightly curved towards the right.
The peristome is narrow but large sized and the AZM which extend and contains over
60 membranelles. The dorsal argyrome is single type with dorsolateral kineties. There
are 10 strong frontoventral, 5 transverse and 4 fine but „rigid caudal‟ cirri. The
macronucleus is an open C-shaped with a twisted foot-like extension to the posterior
arm. The micronucleus is compact and lies close to and sometime overlapping the
macronucleus. It is very common in brackish areas species. Mostly occurred in
aquaculture tank as an annoying parasite.
55. Codonellopsis ostenfeldi (Kofoid & Campbell, 1929) (Plate 4, 2)
Collar thin and transparent, length slightly less than width. The posterior end of
the collar has five to six rows of elliptical or circular fenestrae or layers. The length of
the body is 150-160µm and width has 70-80µm. The anterior end of the collar with
two or three collar belts. Bowl globular, or subglobular, with large and irregular
grains attached. This species is widely distributed in the coast of Gulf of Mannar,
particularly in summer seasons.
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56. Epiplocyloides reticulata (Taniguchi, 1997) (Plate 4, 3)
Hyaline lorica present. Oral rim is completely absent. The bowl is irregular
shaped. Pedicel is sharp and pointed. The diameter of the lorica is 40-43µm and length
is 60-65µm. Often distributed in coastal waters of Tuticorin areas. Sometime, sand
particles or algae matters were attached on the lorical part naturally.
57. C.schabi (Taniguchi, 1997) (Plate 4, 4)
Characterized by a densely agglomerate posterior bowl a number of moderately
sand particles are attached. The anterior part of the lorica, which is clean and window-
like, has provided with about 4-8 of fenestrae. The anterior most of the lorica also has 6
or 8 annuli. Length is 110-122µm, and oral diameter has 40-50µm.
58. Eutintinnus tubulosus (Kofoid & Campbell, 1929) (Plate 4, 5)
This is characterized bynon-agglomerate variety. The presence of a tubular and
clean lorica in which both ends are open. A slight flare is present at the anterior end of
the lorica. Length is 160-165µm and oral diameter has 40-45µm. It is distributed in
Rameswaram waters.
59. Undella columbiana (Taniguchi, 1997) (Plate 4, 6)
This species otherwise called as Proplectella columbiana. It is simple cup
shaped hyaline lorica. The length is 30-35µm and width has 25-30µm. The oral rim is
distinct. Rose Bengal „staining mixtures‟ are highly adopted.
60. F. azorica (Taniguchi, 1997) (Plate 4, 7)
The lorica length is 80-90µm and width has 40-44µm. This species more similar
to F. philippinensis but size is differ. It is characterized by the presence of a fairly
long, clean and campunulated lorica. The posterior end of the lorica a „short pedicel‟
like structure called the aboral horn, which is provided with two small ears or wing-like
structures, one on either side of the aboral horn.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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61. T. corniger (Taniguchi, 1997) (Plate 4, 9)
The size of the lorica is larger than other Tintinnopsis groups. The length is 180-
190µm and width has 25-40µm. Many spiral turns were observe on the lorica.
Sometime lorica has bented. Pedicel a small projection comes out from the aboral part.
62. Strombidium sulcatum (Taniguchi, 1997) (Plate 4, 10)
This is aloricated variety. Mostly present in coastal waters. The key feature is
11-13 adoral polykineties membranelles present, and ventral polykineties 13-15 were
observe. Macronucleus as small fragments. More or less similar to S. conicum.
63. Leprotintinnus nordquisti (Brandt, 1906, Zong, 1989) (Plate 4, 11)
Lorica elongated, tubular or cylindrical, „posterior end inflated,‟ with conical
base. Lorica opens on both ends. Lorica wall with bulliform fiber and grains on its
surface. This species is widely distributed in Gulf of Mannar, coastal waters. Always
many sand particles attached on the lorical surface. This species slightly differ from the
plate 1-7.
64. Favella taraikaensisstrain-2 (Plate 4, 12)
Lorica is medium size. The body is 50-55µm in width and highly variable in
length of size 170-175µm, bowl sometime slightly expanded below the middle,
rounded aborally and joined by wings to a short, „blunt or turned‟ pedicel.Many times
the middle of the lorica has „invaded‟ structure appear. Very commonly distributed in
Rameswaram areas.
65. Turborotalita humilis (Taniguchi, 1997) (Plate 4, 13)
This species related to foraminifera of protozoa. Always the shell is spiral,
chamber angular, and oval or spherical shaped. Umbilicus present. Sutural line curvate
or radial on the dorsal face. It is a good indicator of warm water species. Mostly present
in the Tuticorin waters.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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66. E.minuta (Curds, 1975) (Plate 4, 14)
This species characterized by about 13 dorsal cilia in central kineties and 30-40
membranelles in AZM. It is a small size benthic sedimented ciliates. Size is 50µm
long, width has 25µm. Oval shaped. The dorsal argyrome is single type with 9
dorsolateral kineties. The macronucleus was observed under the Rose Bengal staining
techniques with viable culture. The micronucleus is situated anteriorly on the left edge
of the macronucleus.
67. U. hadai (Taniguchi, 1997) (Plate4, 15)
Kofoid and Campbell, 1929 redescribed as U. hemispherica. The lorica length
is 60-63µm and diameter has 40-45µm. Bowl is cup shaped. Oral rim has two layers
appearance. The aboral part of the lorica not pinpointed with terminal end. Curved
structure. No agglomeration observed.
68. F. azorica (strain 1) (Plate 4, 16)
The lorica length is 100-104µm and width has 80-82µm. The characterized by
the presence of a fairly long, and campunulated lorica. The posterior end of the lorica is
„short pedicel‟ like structure. Taniguchi, (1997) has described as F. azorica species.
The oral part of the lorica is „without any differences‟ for identification.
69. T. dadayi strain 1 (Plate 4, 18)
It is campanulate anterior region. Aboral part of lorica is small bowl shaped.
Agglomeration evenly distributed. Rose Bengal staining properties are adopted. Lorica
length is 70-82µm. Diameter of lorica is 30-37µm. Few agglomerations are observe.
Recorded in the coastal water of Kurusadai Island of Rameswaram areas in the Gulf of
Mannar, India.
70. Litonotus faciola (Curds, 1975) (Plate 4, 19)
Flask-like structure, elongate, flattened, anterior region is neck like view and
cilia present only right side. Contractile vacuoles were clearly observed in the
cytoplasm. The key feature of this species is „bent toward the dorsal side‟ and
recognized in the culture medium fast moving ciliates.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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71. Favella ehrenbergii (strain 2) (Plate 5, 1)
Lorica very elongateand cylindrical shape. Bowl roundaborally and highly
variable in length. Hyaline lorica. One external polykinetid present.Length ofthe lorica
about 190-220µm and width has 85-110µm. Macronuclei size is 5-12µm. The
macronuclei are oval-shaped and variable in position. Aboral region has conical
shaped. Lorica‟s oral rim denticulate structure. Oral rim diameter is 30-35µm with
6-7 layers formed. Aboral part 50-55µm range. One pedicel with spiral turns
waspresent in posterior part. Predominantly present inthe Pamban areas of
Rameswaram in the all seasons.
72. T. brevicollis (Taniguchi, 1997) (Plate 5, 2)
This species more similar to T. fimbriata. The length of lorica is 58-65µm and
width has 40-45µm. Macronucleus is elongate structure. The aboral part small flat
surface base. The oral rim has distinct with bristles. More or less small pot shaped.
Rose Bengal adopted.
73. Neogloboquadrina conglomerate (Taniguchi, 1997) (Plate 5, 3)
Shell tower spiral or twisted structure. Chamber globular, oval or bacilliform,
Aperture opens at the umbilicus or extends from the umbilicus to the spiral face,
secondary sutural aperture, areal aperture or accessory infra-laminal aperture present
and bulliform substance. The major feature is „aperture opens at the umbilicus‟ or
extends from the spiral face, sometime with a sutural secondary aperture. Recorded in
Tuticorin coastal waters.
74. T. butschlii (Taniguchi,1997) (Plate 5, 4)
The aboral part is bowl or cylindrical shaped. Oral diameter is 120-122µm.
Dense agglomeration present on the lorica. This variety observed only on summer
season of coastal waters. The adoral is highly extended. Rose Bengal adopted.
75. T. lohmanni (Taniguchi, 1997) (Plate 5, 5)
This species otherwise called as T. turbo. The width of the lorica is 35-37µm
and length has 105-108µm. The adoral part has many spiral turns presence. About 5-6
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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spiral belts were observe under the lugol‟s iodine staining. It is commonly distributed in
the Rameswaram water.
76. Laboea strobila (Striider-Kypke, 2003) (Plate 5, 6)
Cell conical shape. Screw-like appearance, girdle kinety as helix of
approximately „5 whorls‟, and multiple macronuclei were observed. While culture, this
species the swimming pattern is zigzag movement with sharp turns. Lugol‟s iodine
adopted. AZM is observing in theadoral part.
77. Globorotalia sp. (Zhong, 1989) (Plate 5, 10)
Shell spiral, either side convex, or convex only at the umbilicus, and the spiral
face is flat or sub concave structure. Umbilicus present. Periphery with or without a
single keel. Chambers are oval shape. Sutural line curvate structure. Such variety have
been found in the Rameswaram coastal waters.
78. G. crassaformis (Taniguchi, 1997) (Plate 5, 11)
The shell is spiral structured. The key feature is elongate shell with „less spiral
turns‟. Chambers were clearly identified with ×100. Sometime, Crystal violet stain
adopted. It is commonly distributed in the coastal of Rameswaram areas.
79. T. cornigerstrain 1 (Plate 5, 13)
The size of the lorica is very small,when compare with other Tintinnopsis
groups. The length is 80-90µm and width has 20-24µm. Many spiral turns are observe
on the lorica. Sometime lorica has bented. Pedical a small projection with curved
structure come out from the aboral part.Lugol‟s iodine adopted. Rarely observe in
the Tuticorin coastal water.
80. Canthariella pyramidata (Taniguchi, 1997) (Plate 5, 14)
This species re-described by Kofoid & Campbell, 1929 as Canthariella brevis.
Lorica length is 20-22µm and width has 12-15µm. Some of the projections come out
from anterior side, like flower vase. Rarely observe in the Tuticorin sample.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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81. T. nana (Striider-Kypke,2003) (Plate 5, 15)
Lorica is small with cylindrical shape. Aboral end is round to slightly point. The
key feature of this species is „degree of agglomeration varied‟. The lorica‟s length is
14-17µm.
82. T. gracilis (Kofoid & Campbell, 1929) (Plate 5, 16)
Lorica slender, no oral flare, subconical shape and posterior part is „blunt point‟.
The wall is thick not spiraled. This species is rarely distributed in the coastal water
basin of Rameswaram.
83. Myrionecta rubra (Striider-Kypke, 2003) (Plate 5, 17)
This was collected from Rameswaram coastal areas with QPS fixation. The cell
body is aloricated planktonic forms. The length of body is 96-98µm and width has 40-
45µm. Cytostome rudimentary and oral tentacles are observe. Somatic kineties also
wereobserved. In viable method, special jumping movement then stay in one position
for certain minutes were seen. Morphological characteristics seen by lugol‟s iodine
staining method.
84. G. inflata (Taniguchi, 1997) (Plate 5, 18)
The test is very low spiral structure. The equatorial periphery is slightly
lobulated and the periphery has rounded in sometime. The spherical chambers are
arranged in „many whorls‟, whichis consists of five chambers. The wall is perforated
and rough structure. The sutures are radially depressed and the aperature is inter-
margional and extra-umbilical. Often found in Tuticorin coastal waters.
85. Amphorides amphora (Taniguchi, 1997) (Plate 5, 19)
This is one of the smallest clean loricate species, found in Rameswaram coastal
and brackish waters. The aboral part of the lorica is furrow or groove like structure
always occured. Length of the lorica is 30-32µm and oral diameter has 15-19µm.
Crystal violet is adopted.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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86. Favella franciscana (Striider-Kypke, 2003) (Plate 6, 1)
Characteristics feature of this species, oral rim of the lorica is „denticulated‟.
The bowl with nuchal constriction present. One side of the lorica is plain structure
another side of the lorica irregularly arranged. Pedicel seen under the high power
magnification. Lugol‟s iodine adopted. Rarely presented in Kelakkarai coastal region,
particular in summer season only.
87. Amphorellopsis acuta (Kofoid & Campbell, 1929) (Plate 6, 2)
It is medium sized lorica with hyaline nature. The lorica‟s length 80-83µm and
width has 30-35µm. Bouin‟s fluid adopted. Sometime lorica‟s body observed or stained
in the anterior region only. The oral lip has extended like cup shaped. Aboral part is
groove structure. Mostly present in Kurusadai Island.
88. T. angulate (Kofoid & Campbell, 1929) (Plate 6, 3)
Lorica more or less cylindrical shape. Middle part of the lorica have wing like
structure appear, and end with pointed. The oral rim is more thickness. Length is 70-
72µm, and width has 50-55µm. The key feature of this species present only in summer
season of the coastal region, not in brackish water basins.
89. Amphilonche sp. (Zhong, 1989) (Plate6, 4)
It is group of Radiolaria species. Spicules 20 in number, equal or unequal in
length and shaped with two to four major spicules. „Central capsule elongate oval or
spindle-shaped‟. Cytoplasm always transparent. This species moderately distributed in
the area of Tuticorin coastal waters.
90. T. lobiancoi (Kofoid & Campbell, 1929) (Plate 6, 5)
It is one of the largest lorical structure. The length of lorica is 190-220µm and
width has 35-50µm. Lorica consisting of a bowl, without an aboral horn, oral region
with a flare and wall composed of a fine primary structure and also dense agglomerated
materials presence. Lorica cylindrical gradually into a „stout aboral horn‟. Rarely
distributed in Rameswaram coastal waters.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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91. Epiplocyloides sp.(Taniguchi, 1997) (Plate 6, 6)
Characteristics feature of this species is „double layered structure of lorica‟ with
dense irregular agglomeration or sometime hyaline structure presence. More or less
bowl shaped lorica. The aboral of part of the lorica has prominent clean pedicel
presence. Macronucleus clearly identified with the help of QPS staining techniques.
The oral diameter of the lorica is 40-44µm. This species is recorded in the Arthukkarai
brackish water of Rameswaram areas.
92. Undella clevei (Taniguchi, 1997) (Plate 6, 7)
Lorica length is 72-80µm and width has 20-25µm. Mostly in hyaline nature.
The key feature is lorica have more transparent structure, inside the matters were
identified without staining procedures. Sometime, bowl has large size. Adoral part of
the lorica, parallel distributed. This species is widely distributed in the Tuticorin, and
Sewage rich areas industrialized like Tuticorin TPS areas.
93. Leprotintinnus nordquisti strain 1 (Plate 6, 8)
Lorica elongate, tubular or cylindrical shape. Middle part of the lorica is
swelling structure appeared. Not recorded in the Kofoid & Campbell, (1929)
classification. The present observation, not based on culture techniques. The mid
swelling,not caused by ingestion of food particles. The reason is mainly environmental
parameter changes such as, may be „temperature infliction‟. The lorica „posterior end is
an inflated,‟ with conical base. Lorica opens on both ends. Lorical wall with bulliform
fiber and grains on its surface. This species is widely distributed in Gulf of Mannar,
coastal waters. Always many sand particles attached on the lorical surface.
94. T. beroidea (Taniguchi, 1997) (Hada, 1932) & (Godhantaraman, 2003)
(Plate 6, 9)
The key feature of this species is lorica have one extention, with bullet shaped
structure and aboral part „always pointed‟. Dense agglomeration present. Aboral part is
always opening. The length of the lorica is 70-72µm and width has 20-26µm. Mostly
seen in the Mandapam camp coastal region of Rameswaram in summer season only.
This species slightly differ from plate 1-6.
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95. Homalozoon sp. (Kudo, 1986) (Plates 6, 10)
This species have slit- like cytostome at the anterior end, on non-ciliated
ridge, and uniform body ciliation. Vermiform, flattened, one side with short cilia with
longitudinal rows, and the opposite side without cilia, but marked by a conspicuous
longitudinal ridges. Widely distributed in sewage areas of TPS, Tuticorin.
96. Gallitellia vivans (Taniguchi, 1997) (Plate 6, 11)
This species come under the foraminifera groups. The test may be many
chambers. When the shell is about 150µm long, it is sinisterly coiled and two
rudimentary lappets have formed at the sides of the foot. Later a symmetrical structure
develops along with a „bilobed velum‟. Rarely present in Rameswaram sample.
97. Euplotes crassus strain 1 (Plate 6, 12)
Dorso-lateral kinetiesare 8-11 in numbers, with 17-20 dorsal cilia in central
rows. E. crassus is a large (110-130µm long) elongate oval marine species. The dorsal
surface is strongly sculptured by 8 longitudinal ridges. The peristome is long and
narrow with AZM membranelles and extends to the body length. The argyrome is
single type and there are 10 dorsolateral kineties with 26 dorsal cilia. There are 8-10
frontoventral, 5 transverse and 5 or 8 caudal cirri. The macronucleus is C-shaped
with the posterior arm bearing a foot-like extension. The compact micronucleus is
situated anteriorly near the left edge of the macronucleus. Often occurred in
sedimented areas of Mandapam and Mandapam camp. Not recorded in the Curd, (1975)
Hypotrichs classification and his monograph literatures.
98. Litonotus fasciola (Kudo, 1986) (Plates 6, 13)
Elongate structure, hyaline with flattened neck and both of which are
moderately contractile, posterior end is „bluntly rounded‟. Neck is stout structure, bent
toward the dorsal side, cytostome a long slit, contractile vacuole posterior, and two
spherical macronuclei between micronucleus. In the culture medium, it moves very
rapidly by „straight direct‟. Length is 100-110µm long. Very rarely observe in
Rameswaram waters.
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99. Amoeba radiosa (Kudo, 1986) (Plates 6, 14)
It is small size Amoeba forms. Outline is globular or oval shape. It characterized
by 3-8 radiating slender pseudopodia which is varying in length and degree of rigidity.
When pseudopods are withdrawn, the organism may be similar to A. proteus. Usually
the diameter of the body is 30µm. Mostly in brackish water of Arthukkarai of
Rameswaram.While culture, „Usually inactive condition‟ are observe. This is key
character of the species.
100. 2nd
nauplius of Lucifer sp. (Zhong, 1989) (Plate 6, 15)
Non segmented, with 3 setae distally, second one with spinule dorsally, each
with 1 seta on both sides, and mid-interior side with 2 spinules. Sometime „spinule of
second one is disappeared‟. Always lugol‟s iodine adopted. More abundant in
Sethukkarai, brackish water of Rameswaram areas. Total length of body is 120µm to
150µm.
101. 3rd
nauplius of Calanus sp. (Zhong, 1989) (Plate 6, 16)
The body is oval, with three pairs of appendages and an ocellus similar to that
of Cirripedia. The key characters of this stage are the „larva lives on the yolk‟ and
movement is less compare to other stages. The body length is 190-200µm. Calanus sp.
abundant in Kuraikulam brackish areas. Different concentration of crystal violet
staining procedures adopted. This species more useful forms, for aquaculture
hatchery.
102. Early nauplius of Evadne sp. (Zhong, 1989) (Plate 6, 17)
This is an early stage of „instars‟ of Cladoceran variety. Evadne sp. is marine
strain. After, parthenogenesis of each molt (Ecdysis), the animal increases in size and
enters into another instar. Evadne sp.growth is also highly influenced by environmental
factors, such as temperature.Crystal violet stain easy adopted. Mostly present in
Kuraikulam brackish water basins.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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103. Early nauplius of Cypridina sp. (Zhong, 1989) (Plate 6, 18)
This is an Ostracoda (marine) variety. It is a bioluminescent organisms.
Carapace is an elliptical in shapeand non-transparent. The key feature is early stage
have „caudal portion of the carapace is narrow‟, distally tapered, slightly bend dorsally.
Sometime the same parts occurred in adult organism also. Lugol‟s iodine easy adopted.
Often present in Arthukkarai sample. In the summer season, the organism reach above
the 200µm.
104. 3rd
nauplius of Verruca sp. (Newell, 1963) (Plate 6, 19)
This is barnacles variety. Mostly occured in offshore waters. It is triangular
inshape. The labrum has a rounded tip. The caudal region is longer than other species.
Third stage of nauplius is approximately 200µm long. Often present in Kelakkarai
sample.
105. 3rd
nauplius of Calanus finmarchicus (Zhong, 1989) (Plate 7, 1)
The species differ from the adult, like smaller body, fewer body segments and
pereiopods were observed. The characteristic feature of this species, the body length is
„above the 170µm, antennule 10‟, antenna 9, exopod of mandible 6, thoracic somites 5,
abdominal somites 2 and pereiopods 4. Lugol‟s iodine adopted.Abundantin
Kuraikulam brackish areas.
106. 3rd
nauplius of Penaeus sp. (Zhong, 1989) (Plate 7, 2)
An ocellus is present at the anterior median region of the body. This stage is
characterized by the presence of a pair of dorsally curved caudal setae at the posterior
end of the body and 3 paris of appendages. The first pair is uniramous, the other 2 pairs
are biramous. The last pair, however is shorter than the other appendages. The setae are
„non-plumous‟. Length is 190-220µm.
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107. 3rd
nauplius of Euphausia sp. (Zhong, 1989) (Plate 7, 3)
Euphausiids, known as krill are small shrimp like crustaceans (commercial
prawn). This nauplius are oval form with two or three pairs of appendages. After
ecdysis two pairs of appendages are developed. The key feature is „mandibular leg
disappeared‟. The metanauplius enters the first stage of calyptosis. This species widely
distributed in the Kuraikkulam areas of brackish water basin. The posterior part of the
appendages always closed condition. The lugol‟s iodine easily adopted.
108. 1st larva of Galathea sp. (Newell, 1963) (Plate 7, 4)
The species belong to Anomuran groups. The „spiny tip to the antennal scale‟ is
unique among Galatheid larvae. Anterior region consists of all the appendages. The
very earlier or first larval stage is the smallest one of the Galathea sp. After two
ecdysis, the larvae antennal scale is quite board. More abundant in Sethukkarai
brackish water.
109. 3rd
nauplius of Pseudeuphausia sp. (Zhong, 1989) (Plate 7, 5)
The larvae of this stage resemble the protozoea of Decapoda. Most nauplii are
oval in form with three pairs of appendages in the anterior region. The key feature of
this species, „budding processes of the mandible‟, the first and second maxilla and the
first pereiopoda appear in succession. The diameter of the body is maximum 90-
100µm. Abundant in Kuraikkulam sample.
110. 1st nauplius of Pseudeuphausia sp. (Zhong, 1989) (Plate 7, 6)
The appendages of the anterior region are not emerging out from the body. It is
characterized by the presence of „triangular oval body and no appendages‟. The size of
the larva is 80-90µm. Larvae hatched into metanauplius or pseudo-metanauplius. After
one period of ecdysis, the metanauplius enter the first stage of calyptopsis. Sometime,
crystal violet adopted.
111. Glochidia of Meretrix sp. (Zhong, 1989) (Plate 7, 7)
The Glochidium is a parasitic larval form that develops from the fertilized egg
of mussels. Glochidia have an average size of between 100-200µm. They have two
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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shellhalves, that can be closed with a closing muscle.On each mantle lobe sense organs
arise, and having a cluster of bristles. The key characteristics of the Glochidia is a
triangular shell, relatively thicksize, with a sharply defined, broadly expanded posterior
slope, with yellow oryellowish green with green raysand in the anterior region
biflagellate projection come out. It is widely distributed in the brackish water basin of
Mugathuvaram of Rameswaram particularly in „spring‟ season.
112. Veliger of Crepidula sp. (Zhong, 1989) (Plate 7, 8)
The veliger larva is developed from trochophora. The telotroch extends to form
two semicircular vela with long cilia as swimming organs. The rotate like wheels were
observed. At the same time, foot, eyes and antennae appeared. It is followed by the
torsion of the visceral mass. The prototroch develops into a characteristic swimming
organ, the velum which is bilobed circlet. Sometime, the shell has more torsion and
turns to the right close to the mouth. It is widely observe in the coastal water basin of
Rameswaram. Lugol‟s iodine adopted.
113. Veliger of Crassostrea madrasensis (Santhanam, 1993) (Plate 7, 9)
Straight hinge stage was observed. The hinge size is 70-72µm with a
semitransparent velum clearly appeared. The velum has not shrinkaged, even after
staining. The key feature is larvae are vigorous swimming and „circular movements‟.
The umbo is sometime oval shaped. Rose Bengal and Crystal Violet often adopted.
Present in coastal waters of Tuticorin.
114. 3rd
setiger of Pseudopolydora kempi (Santhanam, 1993) (Plate 7, 10)
Total length of the body is 170-175µm and width has 95-105µm. It has 3 pairs
of setae. Sometime, the anterior region of the body has triangular shaped and black
chromatophores developed on the mid-dorsal line of the 1st segment. Prototroch and
telotroch clearly appeared under the Lugol‟s iodine staining procedures. Recorded from
Kurusadai Island, Rameswaram sample.
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115. Brachionus rubens (Ehrenberg, 1831) (Plate 7, 11)
The characteristic features of this species occipital spines have arranged in
„saw-tooth‟ (peculiar asymmetrical shaped) structure. Lorica is stippled forms. Clearly
observe the anterior dorsal margin with six spines. Posterior spines are absent. The
length of lorica 130-140µm in size and maximum width of lorica is 90-94µm. Present
in Koraikkulam brackish water basins in all the seasons.
116. Embryo of Clupea sp. (Newell, 1963) (Plate 7, 12)
The key characteristic features of this species are „oil globules‟ are present in
the center region, of the embryo. The center diameter of oil lobes size is 32-35µm. It is
one the commercial fish species in the East coast region. The outer covering membrane
is very thickness. Gulaldehyde solution and formaldehyde adopted. This species
moderately distributed in the areas of Kurusadai Island, Rameswaram.
4.2 Quantitative measurement of microzooplankton from Gulf of Mannar
coastal and brackish water, India.
Microzooplanktons are quantified by standard stocker‟s formula and Shannon-
Wiener diversity index. Plankton samples are enumerated by Sedgwick procedures.The
total varieties of microzooplanktonic forms present in a litre of water samples could be
calculated using the following standard diversity index formula.
v
N = n x –––––
V
Where, N is total number of microzooplankters per litre of filtered sample, n is
average number of plankters in 100µl of plankton sample, v is vol. of concentrate
sample and V has total water filtered. The following tables explain the details of
diversity index of microzooplankton from the Gulf of Mannar, region.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
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Table -1: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
Groups
Common name Class Order Family Species
Details
Total
org./m
L
1. Protozoa
Radiolarians
Foraminiferans
Strombidiids
Sarcodina
Polyhymenophora
Acantharia
Amoebida
Foraminifera
Oligotrichida
Acanthometridae
Amoebidae
Globorotallidae
Guembelitridae
Globigernidae
Strombidiidae
Halteriidae
Amphilonche sp.
Amoeba radiosa
Globorotalia crassaformis
G. inflata
Neogloboquadrina conglomerate
Globorotalia sp.
Gallitellia vivans
Turborotalita humilis
Laboea strobila
Strombidium stylifer
S. capitatum
S. elegans
S. turcicum
S. pelagicum
S.macronucleatum
S. sulcatum
Myrionecta rubra
2
4
4
2
3
1
3
1
1
1
1
1
2
8
3
1
2
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
84
Table -2: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
groups
Common name Class Order Family Species
details
Total
org./mL
Tintinnids Tintimnida
Tintinnididae
Codonellidae
Leprotintinnus nordquisti
L. nordquisti (S-1)
Canthariella pyramidata
Amphorellopsis quadraugula
A. acuta
Amphorides amphora
Poroecus sp.
T. brevicollis
T. beroidea
T. butschlii
T. dadayi
T. campanula
T. gracilis
T. kofoidi
T. nana
T. aperta
T. angulate
T. lohmanni
T. lobiancoi
2
2
1
2
2
2
2
5
5
2
6
3
2
4
3
3
1
1
2
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85
Table -3: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
Groups
Common name Class Order Family Species
details
Total
org./
mL
Codonellopsidae
T. patula
T. patula (S-1)
T. patula (S-2)
T. entzil
T. corniger
T. corniger (S-1)
T. brandti
T. turgida
T. turgida (S-1)
T. minuta
Codonellopsis sp.
C. schabi
C. ostenfeldi
C. frigida
2
2
3
1
1
2
3
4
2
1
6
2
4
2
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86
Table -4: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
Groups
Common name Class Order Family Species
details
Total
org./
mL
Ascampbelliellidae
Ptychocylididae
Epiplocylididae
Ascampbelliella sp.
Favella azorica
F. azorica (S-1)
F.ehrenbergii
F.ehrenbergii (S-1)
F. ehrenbergii(S-2)
F. taraikaensis
F. taraikaensis(S-1)
F. taraikaensis(S-2)
F. meunieri
F. franciscana
Epiplocyloides reticulata
Epiplocyloides sp.
1
1
1
5
4
3
5
4
3
2
1
1
1
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Table – 5: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
Groups
Common name Class Order Family Species
details
Total
org./
mL
Suticociliates
Holotricha
Hymenostomatida
Gymnostomatida
Tetrahymenidae
Cohnilembidae
Amphileptidae
Salpingellinae
Undellidae
Eutintinnus tubulosus
Undella columbiana
U. hadai
U. clevei
Dichilum sp.
Uronema marinum
Philasterides sp.
Uronema sp.
Pleuronema sp.
Parauronema sp.
Tiarina fusus
Amphileptus sp.
Litonotus fasciola
2
2
2
1
1
6
5
4
3
2
2
1
1
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88
Table – 6: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
Groups
Common name Class Order Family Species
details
Total
org./
mL
Hypotrichs
Spirotricha
Heterotrichida
Hypotrichida
Colepidae
Spathidiidae
Holophryidae
Spirostomatidae
Oxytrichidae
Euplotidae
Coleps sp.
Trachelocerca sp.
Homalozoon sp.
Trachelophyllum sp.
Ileonema dispar
Spirostomum minus
Eschaneustyla sp.
Urostyla sp.
Keronopsis sp.
Euplotes balticus
E. crassus
E. crassus (S-1)
E. latus
E. mutabilis
E. mutabilis (S-1)
E. eurystomus
1
1
1
1
1
1
1
5
4
3
2
1
1
1
1
1
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89
Table – 7: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
Groups
Common name Class Order Family Species
details
Total
org./
mL
2.
Microcrustaceans
Heterotrophic flagellates
Nauplius
Crytophyceae
Crustacea
Cryptomonadales
Branchiopoda
Decapoda
Myodocopa
Copepoda
Eucarida
Raphidophyceae
Podonidae
Penaeidae
Galatheidae
Cypridinidae
Calanidae
Euphausiidae
E. vannus
E.minuta
Heterosigma inlandica
Early nauplius of Evadne sp.
3rd nauplius of Penaeus sp.
2nd nauplius of Lucifer sp.
1st larva of Galathea sp.
Early nauplius of Cypridina sp.
3rd nauplius of Calanus sp.
3rd nauplius of Calanus finmarchicus
3rd nauplius of Euphausia sp.
3rd nauplius of Pseudeuphausia sp.
3
1
1
1
1
1
1
1
1
4
2
2
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90
Table – 8: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas
during period of 2006-08.
S.No Diversity
Groups
Common name Class Order Family Species
details
Total
org./
mL
3.
4.
5.
Larval forms
Rotifers
Pelagic fish egg
Larval of Polychaetes
Larval of Lamellibranches
Larval of Gastropods
Rotatoria
Embryo of fish
Polychaeta
Bivalvia
Gastropoda
Monogononta
Osteichthyes
Verrucomorpha
Spionida
Veneroida
Pterioida
Mesogastropoda
Ploima
Clupeiformes
Verrucidae
Spionidae
Veneridae
Pteriidae
Cerithidae
Brachionidae
Clupeidae
1st nauplius of Psedeuphausia sp.
3rd nauplius of Verruca sp.
3rd setiger larva of Pseudopolydorakempi
Glochidia of Meretrix sp.
Veliger of Crossostrea madrasensis
Veliger of Crepidula sp.
Brachionus rubens
Embryo of Clupea sp
2
1
1
1
1
3
2
1
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Table-9: The details of protozoan ciliate diversity and abundance from
Mugadhuvaram coastal and brackish water, Tirupulani, Ramanathapuram
district, Gulf of Mannar.
S.NO. SAMPLE
NUMBER
CILIATE
GROUPS
FAMILY SPECIES
DETAILS
NUMBERS
/mL
TOTAL SPECIES
PER
LITER
1. 01 1.Tintinnids Codonellidae Tintinnopsis beroidea 05 59 11.8 (12)
T.dadayi 10
T.minuta 30
T.brandti 04
Ptychocylididae Favella ehrenbergii 10
F. azorica 12 12 2.4 (2)
2. 02 1. Tintinnids Codonellidae T.beroidea 25 67 13.4 (13)
T.dadayi 20
T.turgida 04
Ptychocylididae Favella ehrenbergii 02
Codonellopsidae Codonellopsis schabi 08
Salpingellinae Eutintinnus tubulosus 08
3. 03 1.Stombidiids Stombidiidae Strombidium sp. 02 02 0.4
2.Tintinnids Codonellidae Tintinnopsis beroidea 35 60 12.0 (12)
T.dadayi 25
4. 04. 1.Tintinnids Codonellidae Tintinnopsis patula 28 60 12.0 (12)
T.beroidea 06
T.minuta 04
T.dadayi 20
T.aperta 02
5. 05 1.Cyclotrichs Halteriidae Myrionecta sp. 03 03 0.6
2.Tintinnids Codonellidae Tintinnopsis nana 08 30 6.0 (6)
T.dadayi 04
T.brevicollis 18
6. 06 1.Tintinnids Ptychocylididae Favella meunieri 07 80 16.0 (16)
Codonellidae Tintinnopsis nana 28
T.dadayi 30
T.brevicollis 15
7. 07 1.Tintinnids Codonellidae Tintinnopsis corniger 13 28 5.6 (6)
T.dadayi 15
8. 08 1.Tintinnids Codonellidae Tintinnopsis patula 28 76 15.2 (15)
T.dadayi 22
T.beroidea 14
T.turgida 12
9. 09 1.Scuticociliates Cohnilembidae
Philasterides sp.
(Philaster groups)
05 05 1.0 (1)
2.Tintinnids Codonellidae Tintinnopsis minuta 24 36 7.2 (7)
T.dadayi 12
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92
Table-10: The details of interpretation data of protozoan ciliates in
Magadhuvaram brackish water samples
Table -11: Themean value and standard deviation of protozoan ciliate diversity
and abundance from Mugadhuvaram coastal and brackish water, Tirupulani,
Ramanathapuram district, Gulf of Mannar
S.NO CILIATES
GROUPS
TOTAL
NUMBERS
MEAN
VALUE/
SAMPLE
CUPIC
METER
(M3)
SD-
DISPERSION
VALUE
1. Tintinnids 496 55.11 11.022/M3
±28.70 2. Stombidiids 7 0.7 156/M3
3. Cyclotrichs 12 1.3 267/M3
4. Scuticociliates 3 0.3 66/M3
S.NO GROUPS TOTAL NUMBERS IN
MUGADHUVARAM
AREAS
PERCENTILE
VALUE
A. Tintinnopsis
1. Tintinnopsis aperta 198 89.60%
2. T. dadayi 157
3. T.beridea 25
4. T.brevicollis 20
5. T.nana 02
6. T.campanula 12
7. T.corniger 15
8. T.patula 19 (448)
B. Favella
1. Favella ehrenbergi 12 3.80 %
2. F.meunieri 7(19)
C. Eutintinnus
1. Eutintinnus tubulosus 08 (8) 1.60%
D. Codonellopsis
1. Codonellopsis schabi 08 (8) 1.60%
E. Epiplocylidids
1. Epiplocyloides sp. 13(13) 2.60%
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
93
Graph-1: The details of percentail value of protozoan ciliate diversity
Graph -2: The details of ciliates diversity per liter value from Mugadhuvaram
coastal and brackish water, Tirupulani, Ramanathapuram district, Gulf of
Mannar
0
2000
4000
6000
8000
10000
12000
Cubic value of diversity indexTotal numbers
Tintinnopsis
Favella
Eutintinnus
Codonellopsis
Epiplocylidids
0.00% 20.00% 40.00% 60.00% 80.00% 100.00%
Percentile values of diversity index
Percentile values of diversity index
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
94
Table -12: Average of different season’s microzooplanktonic forms recorded at
different stations of the Gulf of Mannar area in 2006-07 and 2007-08.
S1. Rameswaram; S2. Pamban; S3. Kurusadai island; S4. Thonithurai; S5. Mandapam S6.
Mandapam camp; S7. Koraikulam(BW); S8. Sethukkarai(BW); S9.Arthukkarai(BW);
S10.Tirupulani mugathuvaram; S11. Tuticorin; S12. Tuticorin TPS.
S.NO
MICROZOOPLANKTON
S1
S2
S3
S4
S5
S6
S7
S8
S9
S10
S11
S12
Protozoans
1. Radiolarians 06 10 01 06 02 12 - - - 01 03 -
2. Foraminiferans 11 08 06 06 06 08 - - - 02 17 -
3. Strombidiids 07 03 02 02 01 02 - - - 08 04 03
4. Halterids 03 - 03 01 01 03 - - - 03 06 -
5. Tintinnids (Agglomerated) 26 29 29 23 13 17 11 10 08 34 26 17
6. Tintinnids (Non-
agglomerated)
17 23 20 24 27 19 04 09 09 22 27 13
7. Suticociliates 01 - 01 01 - 01 - 02 02 01 01 -
8. Heterotrophic flagellates 03 02 03 08 04 06 10 07 03 06 08 03
9. Cyclotrichs - - - 01 - - - - 01 - - -
10. Hypotrichs - - - - - - 34 28 35 - - 02
Microcrustaceans
11. Nauplii of Penaeus sp. - 08 - 06 06 13 17 08 02 05 05 -
12. Nauplii of Calanus sp. 16 07 26 10 10 18 38 28 23 10 26 02
13. Nauplii of Pseudeuphausia
sp.
- 01 - 01 - - 08 - - 01 01 -
14. Nauplii of Lucifer sp. - - - - - - 05 - - 01 - -
15. Nauplii of Verruca sp. 07 04 04 04 10 04 16 16 26 14 03 -
16. Nauplii of Evadne sp. - - 02 03 03 01 03 - 02 - 02 -
17. Nauplii of Cypridina sp.
Rotatorians
18. Rotifers - - - - - - 22 08 06 - - -
Larval forms
19. Polychaete larvae 04 - 02 02 - 03 - - - - 02 02
20. Gastropod larvae - - 04 06 02 03 01 - - 04 01 -
21. Lamellibranch larvae 07 - 08 07 08 04 02 - - - - -
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
95
Table-13: Analysis of variation between microzooplankton quantity and different
stations in the Gulf of Mannar areas (One-way method).
Tintinnids were abundant, in high temperature. Lower temperature, salinity and
other nutrient trace elements like magnesium, nitrite, and nitrate were essential for
abundant growth of Hypotrich (Benthic ciliates in brackish water). Lamelli branch
larvae were present only in coastal waters. Nauplii of Calanus sp. were abundant in
stations-7,8 and 9. Rotifers were present only in the brackish water. From this analysis,
the density of protozoan ciliates ranged from 1400-23200m3. In One way analysis of
variation (ANOVA) shows that there is a significant difference between stations. At
.05% level of F -valve is 2.04(df-11,36) 1.967 insignificant whereas .01 level of F valve
as highly significant.
4.3 Physico–chemical properties of coastal and brackish water basins from
Gulf of Mannar (Rameswaram to Tuticorin).
Physico-chemical properties of the marine environment have played a provital
role in determining the different type of ecosystem behaviour. Seasonal variations of
different parameters were analyzed (2006-08).
Source of variation
df
SS
MS
F
Between stations
Within Samples
11
36
3390
25271
1130.00
574.34
1.967
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96
Table 14: Average of different seasons physico-chemical parameters recorded at
different stations of the Gulf of Mannar area in 2006-2007.
Table 15: Average of different seasons physico-chemical parameters recorded at
different stations of the Gulf of Mannar area in 2007-2008
S1. Rameswaram; S2. Pamban; S3. Kurusadai island; S4. Thonithurai;
S5. Mandapam S6. Mandapam camp; S7. Koraikulam(BW);
S8. Sethukkarai(BW); S9.Arthukkarai(BW);S10.Tirupulani mugathuvaram;
S11. Tuticorin; S12. Tuticorin TPS.
Parameter S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12
Surface
water
temperature
28.4 33.2 31.4 30.1 35.1 31.0 26.5 32.0 32.0 28.4 28.2 26.2
PH 7.4 7.3 7.3 7.0 7.0 8.2 8.2 8.3 7.1 7.8 7.3 7.4
Salinity 32.2 28.0 36.0 32.2 31.0 28.0 27.0 25.5 32.1 33.0 32.4 33.4
DO 4.4 4.0 3.8 4.0 3.8 4.0 3.1 2.8 2.6 4.6 2.6 2.6
Ca mg/l
580 585 690 420 425 290 330 335 600 580 440 410
Mg mg/l
1500 840 830 840 810 1280 800 750 755 1200 1225 1600
PO4 µl/l
0.8 0.8 1.1 1.1 1.0 1.7 0.7 2.4 2.8 1.5 1.0 4.25
SiO4 µl/l
72.0 74.3 73.0 69.0 69.1 60.2 61.0 76.8 72.0 63.5 65.8 85.3
NO2 µl/l
0.07 0.01 0.04 0.04 0.02 0.02 0.03 0.04 0.04 0.05 0.04 3.10
NO3 µl/l
0.3 0.2 0.4 0.3 0.3 0.5 2.80 3.65 1.4 0.2 0.2 3.50
Parameter S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12
Surface
water
temperature
26.0 32.3 31.2 29.1 28.2 28.6 27.2 31.4 30.2 31.2 26.8 27.2
PH 7.2 7.4 7.5 7.8 7.8 7.5 8.0 8.2 7.5 7.4 7.4 8.2
Salinity 33.6 32.2 34.2 32.0 33.2 33.0 22.3 22.2 23.3 32.1 29.0 34.1
DO 4.1 4.4 4.2 4.6 4.3 4.6 3.0 3.8 3.2 4.3 4.2 2.8
Ca mg/l
370 640 600 490 400 680 270 280 270 400 480 410
Mg mg/l
1200 1210 1180 1296 1217 1201 806 810 810 1220 1240 1610
PO4 µl/l
1.0 0.9 0.9 6.1 3.1 1.1 2.6 2.4 2.6 1.1 1.3 4.78
SiO4 µl/l
63.3 69.2 68.3 68.4 68.0 63.4 75.2 73.2 74.3 73.3 72.2 70.2
NO2 µl/l
0.04 0.04 0.02 0.02 0.04 0.04 0.02 0.02 0.01 0.04 0.01 4.05
NO3 µl/l
0.2 0.3 0.2 0.2 0.4 0.4 3.15 3.10 3.04 0.3 0.4 3.28
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
97
Graph-3: The negative correlation co-efficient between microzooplankton
quantification and parameter of temperature at different stations of
Gulf of Mannar.
Graph-4: The negative correlation co-efficient between microzooplankton
quantification and parameter of salinity at different stations of Gulf of Mannar.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
98
Graph-5: The negative correlation co-efficient between microzooplankton
quantification and parameter of Dissolved Oxygen (DO) at different stations of
Gulf of Mannar.
Graph-6: The negative correlation co-efficient between microzooplankton
quantification and parameter of silicate at different stations of Gulf of Mannar.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
99
Graph-7: The positive correlation co-efficient between microzooplankton
quantification and parameter of pH at different stations of Gulf of Mannar.
Graph-8: The positive correlation co-efficient between microzooplankton
quantification and parameter of phosphate at different stations of
Gulf of Mannar.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
100
Graph-9: The positive correlation co-efficient between microzooplankton
quantification and parameter of nitrate at different stations of
Gulf of Mannar.
Physico-chemical parameters were recorded during the study were shown in
tables 14 &15. The present results showed that the plankton diversity is highly changed
with the changes in the environmental conditions and comparable with the previous
reports on the tropical nature of the Gulf of Mannar Biosphere Reserve database.
Surface water temperature ranged between from 26.2o C to 33.2
o C in 2006-07 and
26.0o C to 32.2
o C in 2007-08. Salinity varied between 28.0 and 36.0
% among various
stations of coastal waters. In brackish waters salinity ranged between 22.2 and 32.1%.
.In general, pH recorded at all the stations of Gulf of Mannar waters are neutral to
alkaline nature (7.1-8.3). High salinity was recorded at station-3 and station-12 in
2006-07 and station-3 in 2007-08. High level of dissolved oxygen (DO) was recorded
at station-2 and station-6 in 2007-08 whereas station-01 and station-10 in 2006-07 (4.4-
4.6) respectively. Station-12 in 2007-08 and stations-8, 9, 11 and 12 in 2006-07 was
slightly lowered in DO level. Phosphate content varied from 0.8 to 6.1 mg/L. Nitrate
and nitrite slightly varied in all stations of both years. Positive correlation co-efficient
occured between microzooplankton and pH in 2006-07, phosphate in 2006-07, and
nitrate in 2007-08 (Graphs: 8, 9, 10). Negative correlation co-efficient were present
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
101
between microzooplankton and temperature in 2006-07, salinity in 2006-07 & 2007-08,
dissolved oxygen in 2006-07 and silicate in 2006-07 & 2007-08 (Graphs: 4,5,6,7).
4.4 Analysis of feeding activity of Seabass (Lates calcarifer).
Lates calcarifer, commonly known as giant sea perch or Asian seabass, is an
economically food fish in the tropical and subtropical regions in the Asia-Pacific.
Seabass is a euryhaline nature and catadromous species. It spends most of its growing
period in freshwater bodies such as rivers which are connected to the sea. Early stages
of larval forms consumed a variety of food items, preferably to take primary consumer
products such as rotifers and other microzooplanktonic forms. When compared with
other ingestion, preferably to take only cultured species of Euplotes sp. rather than
rotifers (Brachionus plicatilis). Each stages were verified by micro-dissection
techniques (Gut analysis). All aquaculture forms preferably considered rotifers for early
stage food, but loss of more hatchlings. The following photograph gives the fluorescent
picture of hatchling stages of seabass for micro-dissection of gut analysis.
Fig.7. Hatchling stages of Lates calcarifer
Many first feeding activities of larval marine fish are too small to take rotifers.
From this experimental analysis, the the larvae of some species of finfish like seabass
would not feed on rotifers. The following picture gives the detail of culture of seabass
hatchlings.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
102
Fig.8. Different vessels for culture of hatchlings
Fig.9. Matured forms of Lates calcarifer
In the present experiment, it was found that Brachionus plicatilis were „leastly‟
ingested by the Lates calcarifer sp. on 3rd
to 5th
day of hatchling. In the early stages (3-
5 days) the larvae may not be in a position to ingest the large sized rotifers
(Thirunavukkarasu et al., 2009).
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
103
Table -16: Details of feeding activities of seabass (Lates calcarifer) recorded on
RGCA, Sirkali, Tamil nadu, India.
S.
N.
Microzooplankton
communities
36
hrs
42
hrs
48
hrs
72
hrs
4th
day 5th
day
6th
day
7th
day
1. Brachionus
plicatilis (Control)
7 7 6 5 5 4 4 0
2. Euplotes sp. 12 10 6 5 2(49%) 3 0 0
3. Uronema sp. 30 28 27 15 12 8 0 0
4. Lionotus fasciola 25 24 24 18 12 0 0 0
5. Strombdium sp. 30 27 27 25 24 (50%) 12 10 8
4.5. Culture and Biochemical characterization of selected strains of Protozoan
ciliates.
Recently, the nutritional requirements of marine finfish larvae have received
considerable attention. Marine microzooplankton are a potential source of live feed for
marine fish larviculture. The technology to mass culture the marine microplanktonic
forms at a low-cost technology is still being developed. Certain selected strains are
being promoted as a food-organism in some marine fish hatcheries. Marine benthic
forms are natural feed, which can act as alternatives or supplements to rotifer
(Brachionus plicatilis) and artemia nauplii, doubtful nutritional suitability. From this
result, Euplotes vannus fed preferably baker‟s yeast and Isochrysis galbana where,
collected from CIBA (Central Institute for Brackish water Aquacultute). Chennai.
Euplotes crassus fed Chlorella salina and nutrient broth of E.coli.Uronoma sp. and
Parauronema strains fed only Staphylococcus aureus and E.coli. Another groups, such
as Strombidium based culture fed Dunaliella salina (Green micro algae) and nutrient
broth of E.coli. Successfully, cultured strains were only used for nutritional analysis.
The following the pictures give the details of culture of different strains for nutrient
analysis.
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
104
Culture of different microzooplankters from the Gulf of Mannar samples for
nutritional analysis
Fig.10. Euplotes sp.
Fig. 11. Homalozoon sp.
Fig. 12. Uronema sp.
Fig. 13. Heterosigma sp.
Fig. 14. Amoeba radiosa Fig. 15. Lionotus fasciola
Diversity and biotechnological applications of microzooplankton from Gulf of Mannar, India
105
Table-17: Details of nutritional profile of selected strains of protozoan ciliates
S.No. Species Total
Carbohydrates
Mg/mL
Total
(cons.)Protein
Mg/mL
(Wet weight)
Total
Lipid
Mg/mL
1. Euplotes vannus 0.112 mg/mL 7.24 mg/mL 4.1 mg/mL
2. E. crassus 0.71 mg/mL 7.231mg/mL 6.2 mg/mL
3. Uronema sp.(Non-
pathogenic free living
ciliate)
0.623 mg/mL 8.11 mg/mL 4.2 mg/mL
4. Parauronema sp. 1.10mg/mL 10.8 mg/mL 0.8mg/mL
5. Strombidium sp. 0.70mg/mL 15.3mg/mL
(43.71%)
4.9mg/mL
Therefore, from the present analysis, the biochemical constituents are varied
depends on „many factors‟ including food items of the prey.
Fig.16. Culture of Strombidium sp., Uronema sp. and Euplotes sp.
Proteins are the major component that may serve as the main metabolic reserve.
The microzooplankton do not appear to have an extensive storage lipid and
carbohydrate and this might be due to availability of phytoplankton food. From this
experiments the Strombidium sp. rich in protein (43.71%) than rotifers (Brachionus
plicatilis).
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Fig.17. Culture containers of Moneuplotes crassus
Fig. 18. Pure culture of Moneuplotes crassus (3 days starved condition)
Euplotes stains were collected from Rocky shore areas of Rameswaram,
Pamban and Thirupulani for this purpose. Pure culture, based on prey size of the
particles was done. Often Chlorella salina used for successful culture. The following
picture shows the pure culture of Moneuplotes crassus.
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Fig. 19. Structure of Moneuplotes crassus (40-42µm length).
4.6. Compound isolation, structure prediction and tested against fish pathogens
The extract was subjected to the flash chromatography and 26 fractions of
30mL each were collected. Preparatory and analytical thin layer chromatography were
analyzed (Fig.10,11 & 12) with different solvents, by addition of 1% dimethyl
sulfoxide (DMSO). All fractions were purified by High Pressure Liquid
Chromatography (HPLC), then compound was verified by Nuclear Magnetic
Resonance Spectroscopy (NMR) by Protons counting and C13 counting were analyzed
(Fig.25-,29). After many trials, single, pure compound was isolated and identified with
silica gel powder. The following pictures give the details of pure compound isolation
techniques.
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Fig. 20. TLC (Ultra violet observation)-Pure compound isolation
Fig. 21. Single peak identification by Hydrogen Counting techniques (NMR )
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Fig.22. Single compound expansion structure Hydrogen Counting techniques (NMR)
Fig. 23. Details of the single compound carbon counting techniques (C13)
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This compound (1,2 dimethoxyl benzone unit) exhibits the typical benzene
properties of Liquid Alkaloids. The following structure predicted by Chemdrawn
Software Package (Fig. 30).
Fig.24. 1,2 di-methoxyl benzone unit
Another third group of precusors such as OCH3 and CO also were presented.
Compound should be related to papaverine, mescaline, often used for sedation purpose
(Induce drowsiness). The compound could be used for anti-microbial activity of fish
pathogen of Lates calcarifer.
Isolation of pathogens from infected gills of Lates calcarifer (Scuticociliatosis)
Fig. 25. Infected skin lesion
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Uronema marinum are ciliate parasites that infect the skin of many variety of
Lates calcarifer marine fish. Uronema marinum lose the colours and appeared as pale
in nature. Clinical signs of scuticociliatosis are „thickening of the skin‟s mucus(Fig.
32).
Fig. 26. Culture of pathogenic parasite Uronema marinum Strain -1 (Scuticociliates )
0.1:10mL measurement scale (100µl) used
Fig. 27. Culture of pathogenic parasites (Scuticociliatosis disease)
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External signs include skin lesions or pigmentation changes, but the parasite
frequently invade the body muscle and the internal organs. It can be accompanied by
„erratic swimming‟ or lethargy. Diagnosis is based on the finding of ciliates in
„scrapings of different organs‟. The typical morphology of Scuticociliatida can be
easily observed in fresh preparations at light microscope, in stained smears or
histological sections.
Fig. 28. Treatment of scuticociliatosis (in-vitro)
The following pictures give the details of„No‟ zones formed against bacterial
„resistant‟ species.
Fig. 29. No effects on bacterial resistant species
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Fig. 30. MIC on Proteus strain
(Nosocomial collection)
Fig.31. MIC on Escherichia coli
(High resistant strain)
From the analysis, DMSO and 10 µl Tetracyclin were used dissolved serial
dilution 150-170 µl concentration killed 50% of Uronmea marinum of seabass Strain-1.
(Scuticociliatosis). Approximately 200µl pure dose in the sterile water of 1 ml to killed
50% parasites. About 1.3 1.5 ×105
pathogenic parasites concentrated in the medium.
Parasites were measured by haemocytometer glass. Culture of parasites maintained in
25-26‟C at room temperature. The drug not responds against bacteria (Gram negative,
Gram positive and resistant strains from UTI) and fungi.
4.7 Single dose acute toxicity studies
Pharmacological animal experiments were performed according to OECD guide
lines 1997 and after the approval from the Institutional Animal Ethics Committee
(IAEC) of the Directorate of Medical Education, Chennai (Letter Ref. No.
2254/ME1/2009 dated 07.05.2009), experiments were conducted in accordance with
the standard guild lines. Healthy male and female (Wister strain) albino rats with body
weight 150-250g were obtained from the animal house of Periyar College of
Pharmaceutical Sciences, Tiruchirappalli, TN. Animals were feed with standard chaw
diet and water adlibitum. They were housed in polypropylene cage maintained under
the standard conditions. (12h light/12h dark cycles 22±3‟C, 35, 60% humidity). Albino
rats of either sex were starred overnight and divided into five groups. Each containing
six animals. After seven days of acclimatization period, animals were randomly
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selected for the in-vivo determination of acute toxicity studies. During the course of the
experiment the animal‟s behavior was normal. Animals were orally feed with an
increasing dose of 75, 100, 125 mg/kg (P.O) body weight of compound of Moneuplotes
crassus (LA-6), with the help of an oral catheter, but adequate supply of water was
given to the rats before the experiments. After oral administration, the rats were
observed the symptoms of toxicity, gross behavioral changes and mortality up to 14
days. The following pictures give the details of acute toxicity behavior pattern 1,Oral
admistration of newly prepared drug 2, Lacrimation 3 & 4, Sedation.
Fig. 32. Behavioural pattern of Lacrimation
Fig. 33. Sedation condition
The test compound should be administered to albino rats to identify doses
causing “adverse effect and doses causing major toxicity”. The acute toxicity behavior
parameter test on sample test is Positive (One hour extend). The lacrimations from the
eyes are more excessive and conjunctival vasculature is hyperemic nature and high
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sedation was observed. (Fig. 39. & 40). Compounds with low toxicity, the maximum
feasible dose should be administered.
4.8 Studies on micro-technical procedures for kidney tissues (histopathology)
of albino rats.
4.8.1 Microscopic lesions
In addition, the toxicity studies should be designed to assess the dose-response
relationships and pharmacokinetics. Clinical histopathology should be monitored at an
early time and termination period. The rats were treated with the Liquid Alkaloid of
Euplotes sp. that contained 50% doses.
Fig.34. 14th day (acute toxicity)
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Fig. 35. Left lobe of liver swelling
Fig. 36. Visceral parts of adverse affect
Fig. 37. Kidney swelling measured by
vernier measurement
Fig. 38. Liver lobe transfer Fig. 39. Tissues and organ transfer to PBS
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The kidneys of the rat from the animal group had focal areas of glomerular and
tubular necrosis with periglomerular mononuclear cellular infiltration. There were
renal casts and some calcified mass collecting tubules were observed. The following
pictures give the details of histopathogical tissue of kidneys of albino rats.
Fig.40. Section of glomerular part (Adverse condition)
Photomicrograph of a kidney from rat not treated with dose containing 50%
liquid alkaloid of Euplotes sp., H & E stain, (×100). The renal tissue cells were even
distributed in all the areas of the kidney part.
Fig.41. Section of oral tissues
(Cortex- adverse condition)
Fig. 42. Section of middle part
(Medulla- adverse condition)
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Photomicrograph of a kidney from rat treated with dose containing 50% liquid
alkaloid of Euplotes sp., H & E stain, (×100). The renal tissue cells were uneven
distributed in all the areas of the kidney part, by adverse effect of drug (LA-6).
4.9 Measurement of acute ingestion toxic activity.
The LD50 is the dose that kills half (50%) of the animals tested of 14 days
observation. The measurement value of LD50 is 950mg –1000mg/ weight (200mg.) was
observed. The following the table revealed that LD50 data of different extracts from
liquid Alkoloid of Euplotes sp.
Table -18:Data LD50 values of albino rats
S.No Substance LD50 Oral rat (mg/kg)
1. Methonolic extract 600mg/kg.
2. Ethyl acetate extract with chloroform 710mg/kg.
3. Pure extract of LA-06 (test sample) 950-1000mg/kg.
4.10 Anti-inflammatory activity of 1,2 dimethoxy benzene unit (LA-06)
After seven days of acclimatization period, animals were again randomLy
selected for the in-vivo determination of anti-inflammatory activity.
Standardindomethacin 10mg/kg. was used. Test extract at a dose of 100mg/kg was
used. Paw volume of the rat was measured. The following pictures give the details of
paw injection (Fig.50), after injection (Fig.51), paw edema and oral administration of
Indomethacin (Fig. 52 & 53).
Fig.43. Paw injection Fig.44. After injection
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Fig.45. Paw edema Fig.46. Oral administration Indomethacin
Table. 19: Anti-inflammatory activity of test sample against carrageen an induced
paw edema in albino rat.
There was significant and dose dependent anti-inflammatory activity of
compound in the acute carrageenan induce rat paw edema model was conducted. Orally
administered dose of 100 mg/kg (P.O.) of absolute ethanolic based liquid alkaloid (LA-
6) compound of M. crassus produced 82.12% inhibition respectively after 3hrs, as
compared to Indomethacin (standard 10mg/kg (P.O.), which showed 83.36% inhibition
after 3hrs (P<0.0001). Results of anti-inflammatory activities are showed in (Table-
19). Carrageenan induced rat paw edema study was conducted. The remaining
petroleum ether, methanol and chloroform based compound of M. crassus produced
72.00% inhibition respectively after 3 hrs. The following table gives the result obtained
from the anti-inflammatory activity in Albino rats. (Anandakumar & Thajuddin, 2011).
S. No. Treatment
Paw thickness Percentage
inhibition At 0
hr.(mm)
At 3
hr. (mm)
1. Control 0.567±0.08 1.967±0.29 -
2. Test sample
(100mg/kg./P.O) 0.617±0.15 0.876±0.08 82.12
3. Indomethacin
(100mg/kg./P.O) 0.65±0.10 0.883±0.15 83.36
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4.11. Sequencing of Moneuplotes crassusfrom Hypotrichs family.
Pure culture of Moneuplotes crassus cells were rinsed three times with sterile
marine water after being starved overnight and the pelleted by centrifugation. Standard
methods were used for DNA extraction from Moneuplotes crassus (Dawson et
al.,1998).
Additionally, ciliates were removed from the coral associated specimens, after
processing of microscopic analysis fixed bybouin‟s solution and stored in the dark
room at 4oC. The conserved eukaryotic primers were used for this study. PCR primers
and 18S rRNA-targeted oligonucleotide probes used.
18S-6-CIL-V AAYCTGGTTGATCCTGCCAG.
18S-1511-CIL-R GATCCWTCTGCAGGTTCACCTAC.
The following picture gives the amplified 18S rRNA gene. Generated the
primers were checked against the GenBank database by a standard nucleotide-
nucleotide BLAST search.
Fig. 47: Amplified 18S rRNA gene
The nucleotide sequence data have been submitted to the GenBank nucleotide
sequence database got accession number for this species. (HQ256516).
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Moneuplotes crassus strain NTNA17 18S ribosomal RNA gene, partial sequence
GenBank: HQ256516.1
FASTAGraphics
LOCUS HQ256516 816 bp DNA linear INV 01-MAR-2011
DEFINITION Moneuplotes crassus strain NTNA17 18S ribosomal RNA gene, partial
sequence.
ACCESSION HQ256516
VERSION HQ256516.1 GI:324036116
KEYWORDS .
SOURCE Moneuplotes crassus
ORGANISM Moneuplotes crassus
Eukaryota; Alveolata; Ciliophora; Intramacronucleata; Spirotrichea;
Hypotrichia; Euplotida; Euplotidae; Moneuplotes.
REFERENCE 1 (bases 1 to 816)
AUTHORS Thajuddin,N. and Anandakumar,N. TITLE Molecular characterization of marine hypotrichous ciliates from
Gulf of Mannar, India
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 816)
AUTHORS Thajuddin,N. and Anandakumar,N.
TITLE Direct Submission
JOURNAL Submitted (13-SEP-2010) Microbiology, Bharathidasan University,
Palkalaiperur, Tirchirappalli, Tamil Nadu 620024, India
FEATURES Location/Qualifiers
source 1..816
/organism="Moneuplotes crassus"
/mol_type="genomic DNA"
/strain="NTNA17"
/db_xref="taxon:5936"
/collection_date="06-Jan-2008"
/collected_by="N. Ananadakumar"
/note="PCR_primers=fwd_name: 18S-6-CIL-V, rev_name:
18S-1511-CIL-R"
rRNA<1..>816
/product="18S ribosomal RNA"
ORIGIN 1 gggatcgcct gtctcaagat aagccatgca tgtctaagta taaaggttac atacaatgaa 61 actgcgaatg gctcattcaa acagttatag tttatttgga tttacacatt agttaaatgg
121 ataaccgtag taattctagg gctaatacat gcgttacggg ggacttcacg gaaccccagt
181 atttattaga ttcaaaccaa tattccgaag gtctacttga gatgattcat gataactgat
241 cgaattgctg gtctaccggc aataagtcat tcatgtttct gcttcccatc agcttgatgg
301 tagtgtattg gacaaccatg gcattcacgg gctatcgggg gattagggtt cgattccgga
361 gagggagcct gagaaacggc taccacttct acggaaggca gcaggcgcga aaattatcca
421 atcctgattc agggaggtag tgaaacaaat aatgaactag gatttatcct ggggtcacaa
481 tgggcttgat ttgcaaactt tatttagcga ggaacaattg gagggcaagt ctggtgccag
541 cagccgcggt aattccagct ccaatagtgt atattaatgt tcctgcagtt attcgatgct
601 cgtagttgga tttctggagg ctgagatcgg agggtagcca aggttaccgc tgaactcttc
661 cttcatccac ctgttaacgt tgtccgggat tcgtttctcg gcttcgggct cacgcatata
721 cttttaccct ttttcaattt attgttttga gtaaattata gtgtttcagc aggcgtgcgc
781 ggaatactta gcatggaata atcgaattgg accgtc
//
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4.12. Phylogenetic position of Moneuplotes crassus from the Order: Euplotida
This evolutionary tree was inferred by using the neighbor-joining method. The
optimal tree with the sum of branch length (= 15.00098249) is shown. The tree is
drawn to the scale, with branch lengths in the same units as those of the evolutionary
distances used to infer the phylogenetic tree. All positions containing gaps and
missing data were eliminated. There were a total of 470 positions in the final dataset.
This evolutionary analysis was constructed by using MEGA, version-5 software
package.
The nucleotide sequences in this work are available from the GenBank/EMBL
databases under the following accession numbers: Euplotes vannus, AJ305243; E.
minuta, AY361908; E.minuta, AY361904; E.vannus, AJ305241;E. crassus, AJ305240;
E. crassus, AJ305255; Sterkiella histriomuscorum, AB437309; Moneuplotes crassus,
AY361894; M.crassus, AY361899; Strombidium sulcatum, FJ377546; M. crassus,
HQ256516; Oxytricha granulifera, AM412769; O. elegans, AM412767 and Halteria
grandinella, AY007442 were used as the out group species. Hence, the subclass
Hypotrichia, as well as the order Euplotida, is supported as a paraphyletic clade.