4. result - inflibnetshodhganga.inflibnet.ac.in/bitstream/10603/9641/8/08...the posterior region of...

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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54


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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.


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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.


68

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.


69


70

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


71

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.


72

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.


73


74

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.


75

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.


76

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.


77

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.


78

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.


79


80

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


81

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.


82

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.


83

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


84



S.No Diversity

groups


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


85



S.No Diversity

Groups


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


86



S.No Diversity

Groups


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


87

Table – 5: Total average of species composition and abundance of microzooplankton from Gulf of Mannar coastal and brackish water areas


S.No Diversity

Groups


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


88



S.No Diversity

Groups


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


89



S.No Diversity

Groups


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


90



S.No Diversity

Groups


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


91

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


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%


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


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 - - - - -


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


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


97

Graph-3: The negative correlation co-efficient between microzooplankton

quantification and parameter of temperature at different stations of

Gulf of Mannar.


quantification and parameter of salinity at different stations of Gulf of Mannar.


98


quantification and parameter of Dissolved Oxygen (DO) at different stations of

Gulf of Mannar.


quantification and parameter of silicate at different stations of Gulf of Mannar.


99

Graph-7: The positive correlation co-efficient between microzooplankton

quantification and parameter of pH at different stations of Gulf of Mannar.


quantification and parameter of phosphate at different stations of

Gulf of Mannar.


100


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


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.


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).


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.


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


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).


106

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.


107

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.


108

Fig. 20. TLC (Ultra violet observation)-Pure compound isolation

Fig. 21. Single peak identification by Hydrogen Counting techniques (NMR )


109

Fig.22. Single compound expansion structure Hydrogen Counting techniques (NMR)

Fig. 23. Details of the single compound carbon counting techniques (C13)


110

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


111

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)


112

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


113

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


114

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


115

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

//

http://www.ncbi.nlm.nih.gov/nuccore/324036116?report=fasta

http://www.ncbi.nlm.nih.gov/nuccore/324036116?report=fasta

http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=5936

http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=5936

http://www.ncbi.nlm.nih.gov/nuccore/324036116?from=1&to=816


122

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.


123

Fig. 48. Phylogenetic tree relationship between the 18S rRNA gene ciliate sequence obtained from a sample of the corals of Gulf of

Mannar, India inferred by neighbor-joining method.

4. result - inflibnetshodhganga.inflibnet.ac.in/bitstream/10603/9641/8/08...the posterior region of...

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