PROJECT WORK ON

Prevalence of Argulosis in Indian Major Carps in selected fish

farms of Nagaon district of Assam, India.

Name of the Student: Sunayan Gogoi

Registration No: 17-F-14
Programme of Study: B.F.Sc.
Course Code: PW 422
Selected Area of Work: Fish Pathology
Advisor: Dr. Pabitra Kumar Saharia, Assistant Professor

COLLEGE OF FISHERIES

ASSAM AGRICULTURAL UNIVERSITY

RAHA, NAGAON, ASSAM, 782103

COLLEGE OF FISHERIES, ASSAM AGRICULTURAL UNIVERSITY, RAHA,
NAGOAN, ASSAM

STUDENTS'S RESEARCH PROJECT PROPOSAL ON PREVALENCE OF

ARGULOSIS IN INDIAN MAJOR CARPS IN SELECTED FISH FARMS OF
NAGAON, ASSAM

• NAME OF THE DEPARTMENT: AQUATIC ANIMAL AND HEALTH

MANAGEMENT
• NAME OF STUDENT: SUNAYAN GOGOI (17-F-14), B.F.Sc, 4th year
• NAME OF THE MAJOR ADVISOR: DR. PABITRA KUMAR SAHARIA,
ASSISTANT PROFESSOR, AQUACULTURE
• DATE OF THE PROJECT: MARCH 2021
• COMMENCEMENT: MARCH 2021
• EXPECTED COMPLETION: MAY 2022
1. Title

Prevalence of Argulosis in Indian Major Carps in selected fish farms of Nagaon district of
Assam, India.

2. Introduction

One of the major problems of fish culture is the parasitic infestation and disease. Fish parasites
cause morbidity and mortalities of fishes in culture operations. They attack fishes and destroy
them or make wounds or disease on their flesh, thus making them inedible (Woo, 1999). Hence,
in order to control fish diseases caused by the parasites, it is essential to study their taxonomy
to identify them, their infestations, as well as their effect on hosts. Argulus is one of the
problems in fish culture practices in Assam. Fish farmers raise several complains to protect
their crop from Argulus both in nurseries and culture ponds. Argulus sp. belong to Sub-class:
Branchiura (meaning - Gill tails) of crustaceans. It is also called fish lice or common fish louse.
It causes the disease called argulosis in most of fishes. Argulus, the crustacean feeds by first
inserting a pre-oral sting which injects digestive enzymes into the body. They then suck out the
liquidated body fluids with their proboscis-like mouth. The various spines, suckers and hooks
that lice use for attachment may also cause additional tissue damage (Shimura and Inoue,
1984). In India, loss due to argulosis in freshwater aquaculture was estimated about US$ 615
(Rs.39,268.24) per hectare per year (Sahoo et al., 2012) which accounts nearly 30% to the net
profit per hectare per year.

Except some fragmentary reports, detailed information of argulosis from Assam is not
available. Hence, the research work is proposed to accomplish the following objectives.

3. Objectives

i. To identify the parasites by phenotypic and genotypic methods.

ii. To find the prevalence and frequency Index of Argulus sp. in Indian Major Carps
(IMCs) of the selected fish farms of Nagaon district.
iii. To study the histopathological alterations of infested fish species with argulosis.
iv. To assess the economic implications due to argulosis.

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4. Review of literature

Das and Das (1997) reported Argulus foliaceus, Argulus bengalensis and Argulus siamensis
infesting Indian major carps L. rohita, C. mrigla and C. catla.

Padmavathi and Prasad (1998) studied on the control measures of argulosis using Nuvan and
Ekalux.

Ahmed (2004) worked for development of environment friendly measures for the treatment of
argulosis in carp brood ponds.

Taylor et al. (2005) reported that the parasites cause a reactive hyperplasia of the epithelium
and increased mucus production.

Kalita et al. (2009) reported the epidemiology of Argulus sp. infections in fish culture ponds in
Nagaon district. The infestation potential of Argulus foliaceus in cultured carp species varied
from 68.36% in Labeo rohita to 23.45% in Catla catla.

Kalita et al. (2010) reported frequent infection and occurrence of Argulus foliaceus in
freshwater ornamental goldfish, Carassius auratus in Assam. Argulus foliaceus was identified
by the size of their cephalothorax, with distinct thoracic areas and posterior lobes that do not
reach the base of the abdomen, and distinct lobes are separated by a clear cleft. Infected fishes
were lethargic, ceases feeding and loose condition. Attachment sites were moved with
haemorrhagic spots and was spread all over the body surface of the fish.

Al-Darwesh et al. (2014) reported focal deposition of melanocytes in the epidermal layer,
skeletal muscles, with edema and hyalinization affecting of skeletal bundles and degenerative
with necrotic changes in the epithelial cell and hyperplasia of fin filaments by histopathological
examination of Argulus japonicas in goldfish (Carassius auratus).

Kalita et al. (2019) reported the occurrence of Argulus throughout the year in Assam. The
incidences of EUS is highest followed by Argulus, Myxobolus and fin-rot.

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5. Materials and Methods

5.1.Species and Area of Study:

The study will be carried out on Indian major carps namely Mrigal (Cirrhinus mrigala),
Catla (Catla catla) and Rohu (Labeo rohita) for a period of 15 months from March 2021
to May 2022. The diseased fish samples will be collected from Nagaon District of Assam.
5.2.Facilities available:
The facilities available in Department of Aquatic Animal and Health Management, CFSc,
AAU, Raha will be fully utilised during the study.
5.3.Collection, fixation, preservation and study of Argulus sp.:
Samplings will be carried out from the farms at monthly intervals between March 2021 to
May 2022 from Nagaon district. Samples will be kept alive in plastic water container.
Samples consisted of moribund fishes with clinical signs will be transported to Fish Disease
Diagnostic unit of COF, AAU, Raha. Samples with clinical signs of ulcer, haemorrhages,
red spot with attachment of Argulus sp. will be packed in insulated boxes with gel ice pack
during transportation from the sampling site to the laboratory. The strategies used for
collection and preservation of the samples for parasitic examination will be followed as
described by Soota, 1980. The fishes with clinical symptoms will be used within 3-4 hrs.
for analysis and detection. Prior to collect the affected fish samples, its behaviour and
clinical signs will be recorded.
5.4.Methodology for objective I:
The length and body weight of the fishes along with date and site of collections will be
recorded. The gills and body surface will be checked thoroughly. Microscopic
examinations will be done from the smears taken from gills and body surface &
photomicrographs of ectoparasites will be taken using microscope. Phenotypic
characterizations of parasites will be studied as defined and described by Soulsby (1982).

Genotypic identification: Genotypic identification of selected parasitic isolates will be

done by 18S rDNA sequencing for further confirmation of the parasite. The genomic DNA
of parasitic isolates will be extracted by using genomic DNA isolation kit (Macherey-
Nagel, Germany) as per the manufacturer’s protocol. The 18S rDNA gene will be amplified
through PCR reaction. The PCR products will be analysed on 1.0% agarose (HiMedia,
India) gels containing 0.5 μg/ml ethidium bromide in 1X Tris-acetate- EDTA (TAE) buffer.

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5.5.Methodology for objective II:
Parasitic Prevalence Study: The parasitic prevalence will be calculated with the aid of
Parasitic Frequency Index (PFI, %), the formulae suggested and proposed by Margolis et
al. (1982).
No.of hosts infected
Prevalence (%) = ×100
No.of hosts examined
Srivastava (1980) suggested that the frequency index could be further classified into rare
(0.1-9.9%), occasional (10-29.9%), common (30-69.9%) and abundant (70-100%).

5.6.Methodology for objective III:

Sections will be taken from the affected skin and gills of diseased fish and will be fixed in
10% formol saline for twenty-four hours. Sections will be washed in tap water and will be
passed in serial dilutions of alcohol for dehydration. Specimens will be cleared in xylene
and will be embedded in paraffin. Paraffin wax tissue blocks will be prepared for sectioning
at 5-7microns thickness by sledge microtome. The obtained tissue sections will be collected
on glass slides, deparafinized, stained by hematoxylin and eosin and then will be examined
by using light microscope (Roberts, 2001).

5.7. Methodology for objective IV:

Standard questionnaire will be prepared for generating maximum data on loses encountered
during the period of investigation. Data will be collected on various aspects, such as size
of the ponds, stocking density, mortality during infection, loss in weight and total loss in
income through personal interview. Mortality will be recorded according to the category of
the pond size. The overall average stocking density, average mortality (%), average number
of infected fish, average loss (kg ha−1 yr−1) and average loss in income (Rs ha−1 yr−1) will
be calculated according to the equation by (Atallah & El-Banna, 2005) for each category
of ponds in the district.

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6. Expected Outcomes

• The predominant Argulus parasite is expected to be evaluated for the selected sites.
• Expected prevalence and frequency index of Argulus sp. will be estimated for different
months and seasons.
• Highly susceptible species for Argulus infectivity is expected to be evaluated among the
cultured species.
• Economical loss due to the Argulus infestation is expected to be assessed indicating the
impact on the income of the farmers, which is usually ignored by the farmers.

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7. Reference:
Ahmed ATA (2004). Development of environment friendly measures for the treatment of
argulosis in carp brood pond. Bangladesh Journal of Fisheries 27: 3-9.
Al-Darwesh, A.A., Al-Shabbani, M.A.A. and Faris, B.H., 2014. Diagnostic and
pathological study of Argulus japonicas in goldfish (Carassius Auratus). Glob. J.
BioSci. Biotechnol, 3(4), pp.384-387.
Alom, M.Z., Yasmin, M.S. and Rahman, M.A., 2019. Status, occurrence, intensity and
impact of Argulosis in different brood stock ponds. MOJ Eco Environ Sci, 4(5),
pp.225-229.
Atallah, S., & El-Banna, S. (2005). Effect of fish diseases on economic and productive
efficiency of fish farms under Egyptian conditions. In 4th Int. sci. conf. (pp. 104).
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Das MK and Das RK (1997). Fish and prawn Diseases in India. Diagnosis and control.
Inland Fisheries society of India, Barrackpore, West Bengal, India, pp. 139.
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Aquaculture diversification: Current status and prospects in North East Region.
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Kalita, B., Ali, A., Islam, S., Hussain, I.A. and Pokhrel, H., 2019. Incidence of fish diseases
in Assam.
Kalita, B., Tamuli, K.K., Baishya, S. and Dutta, O.K., 2010. Occurrence of parasites in
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