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STUDY OF AQUATIC PROTOZOAN IN NEHER-E-AMBARY IN AURANGABAD VICINITY
Shaikh Afreen S. M, Shaikh J. D., *Prashant Bangale and Mohammed Ehtesham
Department of Zoology, Dr. Rafiq Zakaria Campus Maulana Azad College
Aurangabad -431001 (M.S.)
*Rashtramata Indira Gandhi College, Jalna.
(Corresponding Author: Shaikh J.D.)
ABSTRACT
The present research work covers systematic study of morphology and prevalence of free living protozoan from
fresh water bodies of Neher-e-Ambary in Aurangabad region of Maharashtra (India). During this study total
Number of 15 species have been recorded i.e. 10 species of ciliates, 5 species flagellates and some actinopods,
rhizopods have been reported. Most of the ciliates and flagellate species which are obtained directly from water
samples or from various type of culture Viz; boiled egg infusion, yeast infusion, Phaseala radiatus, Lens culinaris
(lentil) infusion, rice infusion, wheat infusion, Hay infusion,etc. we’re not different. The hematoxyline stain was
allowed for the detailed identification of species. The same method was also used for those species, which could not
be determined by direct observation. Majority of species (about 60%) are identified by means of hematoxyline
stain. The cell volume and the population of most of ciliates have been found to be increase as per the
inoculation of respective nutrient in the specific sample. Thus the study was found helpful for
identification/presence of the advantageous protozoan of Neher-e-Ambari by employing the specific nutrient
enriched media.
KEY WORDS: Free Living Protozoan (FLP), Neher-E-Ambary (Aurangabad), Prevalence.
INTRODUCTION
The protozoa are single celled (Dobell), but it not appropriate to think these organisms are as simple as they are
observed. Many protozoa resemble structural complexity to that of multi-cellular organisms. Being complex internal
structure and carry out complex metabolic activities the protozoologist considered this organism as a part of modern
sanitary science. Protozoa are considered to be a subkingdom of the kingdom Protista. Protozoa are important entity
of every possible ecosystem (Sheikh and Shaikh, 2012). The majority of protozoa live freely in various bodies of
water they are adaptive to various habitats. Finlay (1988) and even in the circumpolar region at extremely high
altitude certain protozoa are found in fairly large numbers in the water. As protozoa are one of the important entity
like other organism and it plays unique and specific role in ecosystem and being smaller size, rapid growth and high
metabolic rate they are an important link between food web.
When appearing in low abundance protozoa has short generation time that allow many species to respond
immediately to changing environmental condition, thus protozoa are sensitive and highly valuable bio-indicator
organism in water quality analysis (Gerald Pfister, 1999). It would be difficult to overestimate their importance in the
food chain of soil organism, ocean organism and freshwater organism. According to their food habit protozoa are
classified as bacteria eating protozoans, and non-bacteria eating protozoans. By Virtue of their grazing activities, they
provide particulate organic matter to higher trophic levels and dissolves nutrients to bacteria. Protozoa plays an
important role in the life of mankind. The protozoans are more and more intensively and extensively studied. Single
cell protozoans compete to greater extent with the multicellular invertebrate.as protozoans represent all types of
nutritional habit namely holozoic, halophytic, saprophytic or saprozoic parasitic mixotrophic etc. Protozoans are
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heterotrophic they obtained energy and nutrient by uptake of complex organic molecules either soluble or particulate
matter. Protozoa often show discrimination in food selection whether this discrimination depends upon the shape size
of the prey or also upon its chemical quality. Protozoan during grazing in the freshwater consumes bacteria and algae.
The giant protozoa are filter feeders they ingest large size other ciliate. The freshwater protozoon occurs in
specialized biological environment, including fungal colonies bacterial biform and in chalk medium. In abundance of
bacterial population oxygen demand is decreased and water body becomes more oxygenated. The grazing of protozoa
directly effects on population of aquatic bacteria. (Martin W. Hahn). The study help here to know more about the
selective scavenging of protozoa that directly and indirectly influences on the bacterial growth.
The bacterial filament in the size range of 4 to 10 u m would not be grazed by the majority of bacteriovorous nanoflagellates but predated by the larger flagellate. The protestian grazing can impact the size distribution population of
bacterial species. In the absence of the (predators the bacteria grew in edible size range of predators. Experimental
field studies have demonstrated that natural bacterial communities could respond within 1 to 2 days to enhanced
protestian grazing pressure.
Neher-e-Ambary (water carriage system)
Malik Amber introduced the system of water supply for the public utility during 1612 AD. This well organized system
of water supply is known as Nehar-e-Ambary Malik amber discovered subterranean water table for mountainous
elevated valley in north of Aurangabad city. Appearance of natural water stream underneath the river-bed has got
number of man-holes from different region. The old water supply system is the reminiscent of medieval period.
Aurangabad and its nearby area not only receives pure mineral subterranean drinking water but some channels
supply water to the lake of city like salim Ali lake, Amkhas,Town hall, Himayat bag lake, Harsool lake.
Availability of adequate supply of water directly affect to the biodiversity of area. There were 12 channels which were
sufficient to supply ample water to town some of them are still functioning properly. Like Neher-e-palsi Neher-e-koila,
Neher-e- dergah shaali. Neher-e-panchakki ‘Neher-e-causar, Neher-e-lalmahal, Neher-e-kiradpura, begampura,
Garkheda, Neher-e-chhavani, etc.
Freshwater habitat with defined factors enhances the chances for specific type of protozoa, but the freshwater habitat
of Neher -e- Ambari in which the survival factors changes from open area to underground surface increases the
chances of abundance of species. That’s why the study area has been chosen for study of protozoans. The important
factor that affect the survival as well as species number of protozoa is food and oxygen the abundant areas for the
survival and growth of protozoa. The fresh water resource available with moderate amount of bacteria and dissolved
oxygen plays important role for the presence of specific type of protozoa. Since protozoa are generally observed as
microorganism the study of freshwater of Neher-e- Ambari provides biological evidences for identification of useful
protozoa.
MATERIALS AND METHODS
The protozoa are tend to be abundant in habitat where productivity is high the subterranean water system with lotic
water system has the potential importance. The hypoheric zone provides nutrient to the stream organism. The
subterranean habitat along with the lotic water i.e. the dimension of revering ecosystem. The major shift occurs in the
kind of organism present due to the availability of food. The 50% of organic energy is for the macro inverbrate and
fishes through an algal and bacterial grazer like rotifer and protozoa. The hydraulic water system Neher-e-Ambari
and its channel like Neher-e- panchakki and Himayat bag are choosen as a study area for the month of June, July,
August and September. The water sample for this study are collected in simple sterile plastic bottle and brought to the
laboratory and maintained for direct and indirect observation.
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Direct Live Observation:
The water sample stored in bottle kept in open place under low intensity of light. For two days and observed the
specimen on simple compound microscope. In this method for immobilization of live organism, a special technique
applied. First the water droplet were taken with the help of ordinary syringe on slide and covered with the Vaseline
petroleum jelly, later covered the droplet with the coverslip. This technique is found to be effective for the observation
of different parts of protozoa without loss of integrity of the organism. The visibility of cell decreases with increase in
the pressure. The staining technique of using Indian ink, 3% Acetocaramine, 3% metylene blue tarphan blue and direct
grinded vegetable color like Beetroot, Acetocaramine are found more effective for the identification of ciliates like
vorticella.
Indirect Observation:
Before applying the permanent stain it is necessary to relax the organism, 3% solution of copper acetate is used and the
specimen is subjected for permanent staining by using the different range of protozoan stain such as Hematoxyline
stained. For the protozoa especially iron hematoxyline deflaid hematoxyline tungestophosphoric hematoxyline eosin
are found effective.
Methodology
 Selection of location:
 Collection of water sample :∙ simple plastic bottle .jars
 Immobilization of specimen for observation for direct and indirect observation.
 Preperation of permanent slide.
 Analysis of water sample for the study of physico chemical parameter
 Isolation of specific protozoa species.
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Culturing protozoa
Laboratory equipment for protozoa culture: sterilized, 150ml petridish with cover micropipette of various
sizes, compound microscope stereomicroscope. Simple 2ml pipette etc., are chosen.
Maintenance of light is necessary for the cultivation of protozoa. The precaution has been done by
providinging cover on culture as per need.
Prevention of contamination: For preventing contamination in culture the spring water used. For avoiding
fungal growth few crystals of naphthol added to 5ml stock solution of sucrose. These solutions are found to
be effective in all the culture medium.
Care and feeding: According to the feeding habit the egg infusion, milk infusion yeast infusions are found
more effective for bacteria grazing protozoa. Lentil infusion (Phaseolus radiatus and Lens culinaris), boiled
rice infusion solid rice infusion are found more effective.in most of the species.
Observation and re-observation has been done time to time before culturing and after culturing the protozoa.
RESULTS AND DISCUSSION
The short diurnal study (June-Sept) month is found positive for the study of freshwater protozoa of Neher-e-Ambari. The
Results shown in Table 1, 2 and 3 and Figure 1 and 2. While during the instant study, the sampling ratio in each month has been
kept constant. During this period of rainy season the estimation of Physico-chemical parameter of this freshwater habitat found
to be fluctuating .The chlorinity of water increased during the month of August.
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Table 1. Percentage Prevalence of protozoa and Physicochemical Parameters
Duration
No. of Sample
Percentage of Prevalence
PH
Temperature
Chlorine
June
5
60%
6.6
27.0
7.5
July
5
80%
6.5
27.0
7.5
August
5
100%
7
28.0
10.6
September
5
80%
6.5
26.0
8.8
Dissolved Oxygen
2.0
2.5
2.5
2.6
Table 2 Morphology of Free Living Protozoa (Cilliate
Almost all the freshwater invertebrate phyla are representative of hypoheric zone. One of the contributing factors for
this study is quantity and availability of food. There are detrital inputs from surface water and even from the
sediment, and interstitial spaces. Thus the organism density strongly related to the amount of nutrients present in the
medium. The same principal kept in practice while applying different culture methods through proportionate artificial
spring water. The spring water plays an important role as growth medium for protozoa. The availability of food and
optimum limit of Physico-chemical parameter directly affect to the density of ciliate Protozoa. Besides identification
of protozoa, further step has been taken for the culturing of the protozoa by different simple methods such as.
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Boiled egg infusion: Hypotrichis.
Yeast infusion: Tetrahymina Vorticella
Phaseolus radiatus infusion: Vorticella euplotes
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Lens culinaris infusion: T. vorex, T. pyriforms, Stokesia vernalis
Milk infusion: Loxophylum species
Hay infusion: Vorticella stentor
Wheat infusion: Vorticella,Oxytricha
Solid rice infusion: Tetrahymina urostyla
Boiled rice infusion: Urostyla trichogaster
From live observation of these infusions it is concluded that, these different infusion found effective for the growth of
protozoa as the cell volume and population of most of the species such as T. pyriformis, T. vorex, and Paramecium
oxytricha, Litonatus, loxophylum, Euglena, Urostyla found to increase after inoculation of nutrient medium. In
artificial spring water the pH were maintained at 6.5 are found most useful for the growth of Tetrahymina, and
Urostyla trichogaster. The nutrients through Phaseala radiatus, Lens culinaris infusion are best sources for providing
carbohydrate, and protein for the growth of protozoa.
Figure 1. Shown the Ciliates
A
B
C
D
E
F
A ) V. campanula, B) urostyla trichogaster,C)T. vorex, D) Stokesia vernalis E)Cytotricha convixa, F) Euplotes
erystmus
A
B
A) Amphicilia thaiophaga
A
B
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B)Litonatus species
C
D
E
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F
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CILLIATES (STAINED)A) T. vorex, B)Cytotricha convixa C)Stokesi vernalis ,T. rostratum
D)Litonatus specie, F) Amphicilia thiophaga,F)Urostyla trichogaster
A
B
A) V. microstoma B) V.campanula,
C
D
C) loxophylum sincium
D) Didinum nasutum,
Figure 2. Shown the Flagellate
A
B
C
A ) Notosolenus apocamptus and ,Euglina pyriformis
Ocellatum pocheti
E.) E. deses
D
B) Paranema
E
C) Paranema trichophorum
D)
Table 2. Morphology of free living protozoa (ciliates)
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Table 3. Morphology of Free Living Protozoa (Flagellate)
CUNCLUSSION
The surface run-offs and the wastages along the course of water in monsoon season Provides nutrient for most of the
organism (protozoa).The high species number of organism is one of the indications of suitability of habitat for the
organisms and to keep the good water condition. The short durational study in monsoon season is found significant to
increase the protozoan population by different culture methods. From the estimated account of Physico-chemical
parameter it is difficult to understand about the abundance of protozoa but it helps to understood about the optimum
limits of nutrient for most of the useful protozoan species.
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