A
PROJECT REPORT ON
EFFECT OF DRETERDENT ON BIOCHEMICAL
PARAMETERS OF THE FRESHWATER FISH, LABEO
ROHITA
SUBMITTED TO
SAVITRI BAI PHULE PUNE UNIVERSITY
PUNE – 411007
BY
KHAN ZOYA JAVED
MSC-II ZOOLOGY
In partial fulfillment of the requirement for
Master of Science Degree (Zoology)
Under guidance of
Ms. SHABISTA SHAIKH
Assistant professor
(Department of Zoology)
AKI'S Poona College of Arts Science and Commerce, Camp, Pune-41100
[2023– 2024]
This is to certify that project work entitled “Effect Of Detergent On
Biochemical Parameters Of The Fresh Water Fish, Labeo Rohita” has been
successfully carried out by Ms. Zoya Javed Khan during academic year 2023
– 2024 in partial fulfillment of the course of study for the Master degree in
Zoology as per syllabus prescribed by Savitribai Phule Pune University,
Ms. Shabista Shaikh Dr. Mujeeb A.J. Shaikh
Project Guide Head, Department of Zoology
Date :-
Seat No. :- 501005 Dr. Iqbal N. Shaikh
Principal
Examiner :-
1. _____________
2. _____________ 2
ACKNOWLEDGMENT
3
ACKNOWLEDGMENT
I am extremely grateful to Dr. Iqbal N. Shaikh, Principal, AKI’S Poona College of
Arts, Science & Commerce, Camp, Pune for providing necessary research facilities with
his gracing and blessings.
I would be falling in my duty if I do not extend my sincere thanks to Dr. Mujeeb
Shaikh, Head Department of Zoology, AKI’S Poona of Arts, Science & Commerce,
Camp, Pune for his valuable suggestions, comments and altitudinal support.
I express my deep sense of gratitude to my project guide Ms. Shabista Shaikh, for her
immaculate guidance, immense help and encouragement throughout the duration of this
project.
I am highly obliged to Dr. Kalim Shaikh, Mr. Wasim Shaikh, Dr. Shoeb Ahmed
Mrs. Anjum M. Pathan, Ms. Sameen Gazikhan & Ms. Raniya Shaikh, Department
of Zoology, AKI’S Poona of Arts, Science & Commerce, Camp, Pune for their insightful
comments and encouragement.
I would like to express my deep sense of gratitude to my Family for their infinite
patience, kind blessing and regular moral support without which the successful ending of
the present work was not possible.
Place : Zoya Javed Khan
Date : MSC- II
4
INDEX
Sr. No. Particular Page No.
1. INTRODUCTION 9-10
2. REVIEW OF LITERATURE 12
3. MATERIAL AND METHOD 14
4. RESULT 16
5. DISCUSSION 20
6. CONCLUSION 22
7. REFERENCES 24
5
INTRODUCTION
6
Introduction
Freshwater fish are the most abundant and cost-effective source of protein for human
consumption. The quality and quantity of protein generally determines the nutritive value of
fish. These values gets decline due to environmental contaminants present in aquatic habitats.
Various primary pollutants contaminate our air, water, and land, comprising matter, metals,
agricultural chemicals, organic compounds, photochemicals, solid waste, and detergents. Major
freshwater bodies of the world are becoming polluted due to the addition of various pollutants.
These pollutants include domestic and industrial discharges such as heavy metals, organic and
inorganic chemical substances, insecticides and pesticides, heat effluents, radioactive substances
and household waste containing human waste. A detergent is a multifaceted structure of the
chemical substances, produced by the surfactant such as Linear Alkylbenzene Sulfonate (LAS),
and alcohol derivatives like Alcohol Sulfates (AS), Alcohol Ether Sulfates (AES) and
Alcohol Ethoxylates (AE). There are also a large number of different brands of detergents
powders, flakes and soaps used for washing a wide range of objects. Detergents increase
alkalinity and produce foam on the surface of the water, stimulate excessive growth of
submerged and hovering aquatic weeds, surface weeds reduce the exchange of gases between
the air and water and penetrate sunlight to the bottom of the water body. This results in a lack of
dissolved oxygen for the aquatic organisms.
A detergent does more than just alter the chemical makeup of water bodies and the metabolic
cycle of biota; it also hastens the degradation and mortality of aquatic organisms. Detergents are
widely used without the knowledge of its effect on receiving water bodies (Mukherjee and
Pankajakshi, 1995). Detergents cause damage to buccal linings and gill epithelium. This can
lead to poor food utilization and interfere with oxygen uptake. It can also affect the liver of
aquatic organisms indirectly through absorption of certain tissue, as liver acts as detoxicant of
any toxic substances enters the body (Yatim, 1990). Liver can also alters its structure and is
significant in the evaluation of fish health (Ogundiran et al., 2010) These could potentially
disrupt the enzymatic activities and normal functions of different organs, consequently affecting
food digestion and the overall physiology of fish. It was mentioned further that the first liver
damage found was congestion, i.e. the increase of the blood volume in the blood capillaries. The
failure of oxygen intake by the fish and liver damage result in the growth retardation (Himawan,
1998). The surfactants are one of the major contents of detergents, this surfactants are
extensively employed in detergents, soaps, fabric softeners, paints, shaving creams, leather and
textile products, pesticides, defoliants, antiseptics, disinfectants, as well as food additives, often
in substantial quantities. This information can also be extrapolated for application in diagnosing
and treating toxic reactions in humans.
Water pollution refers to the contamination of water sources such as lakes, rivers, oceans,
aquifers, and groundwater. Water pollution has emerged as a significant global concern. It's
regrettable that rivers are increasingly serving as natural dumping grounds for various
7
community and industrial wastes. Information on acute toxicity can aid in determining how a
substance acts toxically and may offer insights into the doses linked with organ-specific toxicity
and lethality, which are valuable for establishing dosage levels in repeated dose investigations.
It has become essential to provide farmers and crofters and those involved in activities such as
agriculture contractors and companies involved in spreading organic measures to land, with
practical guidance on how to prevent pollution.The aim of the study is to Effect of detergent on
biochemical parameters of the fresh water fish, Labeo Rohita.
Classification /Type of deterge:
Detergents are categorized based on the electrical charge of their surfactants. There are four
primary classes of detergents: Anionic detergents, Cationic detergents, Non-ionic detergents,
and Amphoteric detergents. s including dishwashing liquids, laundry detergents, and carwash
detergents. They are often referred to as neutral detergents. Cationic detergents, with a positive
charge, have relatively low detergency and are primarily used for germicides, fabric softeners,
and specialized emulsifiers. Non-ionic detergents do not ionize or carry a charge when dissolved
in water, hence the absence of ionic constituents. Amphoteric detergents have a net zero charge
due to equal numbers of positive and negative charge chemical groups. They are commonly
found in shampoos, skin cleansers, and carpet shampoos. Furthermore, there are various types of
detergents such as laundry detergents, dishwashing liquids, and biological detergents, each
serving specific cleaning purposes.
Occurrence of detergent in freshwater:
All detergents destroy the external mucus layers that protect the fish from bacteria and
parasites; plus they can cause severe damage to the gills (Ikele et al., 2011). Most fish will die
when detergent concentrations approach 15 parts per million (Alaa Eldin et al., 2012). Detergent
concentrations as low as 5 parts per million (ppm) can be detrimental to fish eggs, leading to
their destruction. Surfactant detergents are known to diminish the reproductive capacity of
aquatic organisms and reduce the surface tension of water. Additionally, organic chemicals like
pesticides and phenols are readily absorbed by fishes, posing significant risks to their health and
ecosystem balance. Phosphates found in detergents can contribute to the occurrence of
freshwater algal blooms. These blooms release toxins and consume oxygen in waterways during
decomposition, which can lead to oxygen depletion harmful to aquatic life. The risk of acute
aquatic toxicity is relatively low from the direct release of secondary or tertiary sewage effluents
containing laundry detergent residues. However, the primary entry point of detergent pollutants
into water is typically through sewage systems, ultimately reaching surface water bodies.
8
Labeo Rohita.
Labeo rohita, commonly known as Rohu fish, is a type of carp primarily inhabiting rivers and
ponds. Rohu is characterized as a column feeder, meaning it feeds within the water column. Due
to its feeding behavior, it is frequently cultivated alongside two other carp species: Cirrhina
mrigala, which is a bottom feeder, and Catla catla, which is a surface feeder. This combination
of species allows for efficient utilization of different ecological niches within the aquaculture
environment. The adult Labeo fish is about 0.5 to 2 metres in length and weigh around 4 kg
(maximum up to 45 kg).
Scientific classification:
Kingdom Animalia
Phylum Chordata
Subphylum Vertebrata
Class Osteichthyes
Subclass Actinopterygii
Family Cyprinidae
Genus Labeo
Species rohita
Natural distribution and habitat:
Rohu, scientifically known as Labeo rohita, is a freshwater fish commonly found in rivers and
ponds. It has a wide geographical distribution across temperate and tropical regions, including
countries like Vietnam, Pakistan, Nepal, India, Myanmar, and Bangladesh. Labeo rohita
primarily breeds in bund-type tanks and rivers, showing a preference for open water habitats
rather than confined environments
Environmental tolerance ranges:
Rohu grows rapidly and reaches a total length of 35-45cm and a weight of 700-800g per year
under normal culture conditions. Rohu is an eurythermic species, meaning it does not tolerate
temperatures lower than 14°c.
9
Figure: Labeo rohita fry
Figure: Adult Labeo rohita
10
Figure: physiological effects
11
REVIEW OF LITERATURE
12
REVIEW OF LITERATURE
Fish serve as excellent biosensors of aquatic contaminants, acting as bio-indicator species that
exhibit high sensitivity to changes in their aquatic environment. Despite this, an examination of
relevant literature indicates that research on the effects of detergents on fish remains sparse,
primarily focusing on acute toxicity assessments.
Though, a number of workers have studied the effects of pollutants on physiology and
biochemistry of fishes (Rani et al., 1998). The effect of detergent on the growth rate of
C.gariepinus h (syaku, B. and *Solomon, J. R. 2006).
The biochemical components like protein, carbohydrate and lipid were estimated quantitatively
in the tissue of gills, liver, kidney and muscle of control and detergent treated fishes. Role of
Synthetic Detergents in Damaging the Tissues of Labeo rohita, Kalaiselvan K, Govindasamy C
(2022)
Studies on the effects of commercial detergents on freshwater fishes are meager except the
works of Abel (1974), Bhaskaran et al.(1991), Rani et al.,(1998), Bakirel et al.(2005), Saxena et
al. (2005), Mukherjee and Pandey (2005), Manoj et al. (2007), Topale et al.(2013), Sarma et al.
(2015) and Kalita et al. (2016). Aravind kumar, 1995)
The present study focusing on effect of detergent on biochemical parameters of the fresh water
fish, Labeo Rohita
13
MATERIAL AND METHOD
14
MATERIAL AND METHOD
Rohu grows rapidly and reaches a total length of 35-45cm and a weight of 700-800g per year
under normal culture conditions. Rohu is a eurythermic species, meaning it does not tolerate
temperatures lower than 14°c.
Fish Samples:
Healthy Rohu fishes were transported in a container to the laboratory. The fishes were
acclimatized for 14 days and their water was changed every two days to remove fecal, and
unconsumed feeds. The fish were fed twice daily.
Preparation of test media:
An appropriate concentration range of 10-50mg was used to determine the median lethal
concentration with a minimum of 10 fish for each concentration and mortality was recorded
every 24 hours for 96 hours. The median lethal concentration was found at 27mg for 96 hours
using Probit Analysis according to Finney 1971. Four groups of fish were exposed to a
concentration of 2.7 mg (1/10 th of 96 h LC50 value) of the detergent 'Tide' for 24 hours, 48
hours, 72 hours, and 96 hours respectively.
Exposure of test organism:
5 fishes were exposed each to detergents (Tide) concentrations in plastic containers. Feeding
was discontinued 24 h before the start of the experiment. Control fishes were in de-chiorinated
tap water.
Probit Analysis Method (Finney 1971):
Probit analysis is a method of analyzing the relationship between a stimulus (dose) and the
quantal (all or nothing) response. (Finney, 1971), determining the LC50 and LD50 values. Groups
of fishes were exposed to 2.7mg (1/10thof 96hours LC50 value) concentration of the detergent,
Tide‟ for 24, 48, 72, and 96 hours respectively. Another group was maintained as control. For
each experimental study tissue samples were collected from the fish for the analysis of
biochemical parameters like protein and carbohydrate.
15
RESULT
16
RESULT
The gill tissue of fishes exposed to 2.70 mg of detergent for 24, 48, 72, and 96 hours was found
to contain 1.70 mg/g, 1.63 mg/g, 1.54 mg/g, and 1.20 mg/g of protein, respectively. In
comparison, the control group of fishes maintained without exposure to detergent had a mean
protein content of 1.78 mg/g. Liver tissue was found to contain 1.85 mg/g, 1.73 mg/g, 1.68
mg/g, and 1.55 mg/g of protein after exposure to a concentration of 2.70 mg of detergent for 24,
48, 72, and 96 hours, respectively.
The carbohydrate content in the gill tissues of fishes exposed to short-term exposure periods of
24, 48, 72, and 96 hours were measured at 12.88 mg/g, 11.50 mg/g, 11.00 mg/g, and 10.70
mg/g, respectively.
In liver tissue, the carbohydrate content was found to be 18.00 mg/g, 17.70 mg/g, 17.21 mg/g,
and 16.55 mg/g after short-term exposure periods of 24, 48, 72, and 96 hours, respectively. In
the control group, the mean carbohydrate value was 18.50 mg/g. Additionally, the kidney tissue
of control fishes recorded a carbohydrate content of 17.33 mg/g. The fishes exposed to short-
term periods were found to have carbohydrate contents of 16.50 mg/g, 16.00 mg/g, 14.23 mg/g,
and 12.56 mg/g, respectively.
The amount of carbohydrate in the muscle tissue of fishes exposed to 2.70 mg of detergent after
24, 48, 72, and 96 hours exposure periods were recorded as 14.34 mg/g, 12.72 mg/g, 11.50
mg/g, and 10.20 mg/g, respectively. In comparison, the control fishes were found to contain
15.77 mg/g of carbohydrates in their muscles.
17
DISCUSSION
18
DISCUSSION
Proteins play crucial roles in various cellular functions, serving as essential biomolecules within
organisms. They represent one of the most vital and comprehensive groups of biological
materials, contributing to the nitrogen constituents of the body and carrying out diverse
biological functions. Environmental stress triggers compensatory metabolic adjustments in an
animal's organs by altering both the quality and quantity of proteins present, thereby facilitating
adaptation to challenging environmental conditions.
The gradual decrease of protein from various days of exposure may be due the influence of
exogenous factors like toxic environment, Sapna Srivastavaet al. (2004) reported that the
decreased level of protein, globulin and serum enzyme activity in vaccinated Tilapia nilotica
exposed to sub lethal concentration detergent. The decreased trend of the protein content in most
of the tissues may be due to metabolic utilization of the ketoacids to gluconeogenesis pathway
for the synthesis of glucose (Veeriah, 2013).
Carbohydrates, which provide the primary source of metabolites for energy in a typical
individual, are oxidized to fulfill energy needs. They can also be converted into glycogen for
storage or directed within the metabolic pathway to provide carbon chains for amino acids, or
they may be transformed into fat for storage.
The low values of carbohydrates recorded in the present study could be due to the fact that
glycogen, in many marine animals, does not contribute much to the reserves in the body
(Jayasreeet al., 1994) A significant decrease in carbohydrate content suggests a higher
utilization of carbohydrates to manage heightened metabolism during stressful conditions.
Despite the liver's continuous and rapid release of glucose through glycogenolysis to meet the
increased energy demands for muscular activity, a decline in the overall carbohydrate content in
fishes exposed to pesticide treatment is likely.
19
Conclusion
20
Conclusion
The current study aimed to assess the impact of Tide detergent on biochemical parameters,
including protein and carbohydrate levels, in the freshwater fish Labeo rohita. A consistent
decline was observed in the protein and carbohydrate contents of the gill, liver, kidney, and muscle
tissues of Labeo rohita, following short-term exposure to Tide detergent, compared to the control
group. These effects were observed across all exposure durations. Fish exposed to various
concentrations of the detergent exhibited different behavioral changes, such as reduced swimming
activity and bleeding through the gills. Hemorrhaging was also observed at the base of the body
appendages (fins) and along the belly. Loss of nervous control was observed in fishes along the
lateral side of the body. Additionally, the body appeared slimy due to mucus secretion from the
epithelium of the gills. Even at low concentrations, fishes experienced mortality due to bleeding
through the gills. This observation holds particular significance due to the fact that fishes serve as
a staple diet for many people and are of considerable commercial value.
21
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