Variation in the Muscle Tissue Protein of Some Fish Parasitic Isopods
Parasitic attack commercially important fishes are a matter of great concern. Parasites were collected from the Nagapattinam coastal environment, Southeast coast of India. The concentration of protein in the muscles shows variation among the isopods parasites. The muscle proteins from 5 species of isopods parasites was studied, among the species maximum protein content of 18.26% was observed in Nerocila loveni and the minimum protein content of 3.44% was noticed in Catoessa boscii. After the Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), their clear bands were detected in the gel that represents proteins from five species of isopods. The present study reported for the first time that muscles proteins of isopods were isolated and considered for protein quantification and identification.
Received: June 13, 2012;
Accepted: August 31, 2012;
Published: November 28, 2012
Knowledge of fish diseases and parasites is essential for successful aquaculture,
particularly in a country like India with long and highly productive coastal
waters. However, the tropical coastal and brackish waters which likely contain
many unknown parasites and pathogens, are poorly studied. Parasites play a pivotal
role in the biology of fishes and can affect their behavior, health and distribution
(Rohde, 1993). Isopods are one of the most diverse orders
of crustaceans. Marine isopods are poorly studied animals in many regions. The
parasite may be detrimental to the host simply by being there. As a consequent
the normal functioning of the host structures can easily be disturbed, especially
fins, operculum, mouth and lateral line canal. Pressure atrophy can often be
seen in the tissues surrounding the larger isopod parasites (Kabata,
1985). Isopods absorb their nourishment directly from the hosts body
and they depend upon their hosts for feeding (Pillai, 1958).
However, these isopods also possess positive qualities. A nutrient such as protein
is needed to fulfil its energy requirement rather than of growth maintenance,
is an expensive component in a diet. The reported estimate of protein requirement
must be carefully examined because it is dependent on quality (essential amino
acid profile) of dietary protein, age or physiological state of crustaceans
(DAbramo and Sheen, 1994). Nerocila is a
large genus of Cymothoidae including at least 65 species living usually attached
on the skin or on the fins of fishes (Bowman, 1978).
The Nerocila, were collected from the body surface, on the head, on the
pectoral fin or from the caudal peduncle of hosts. The Catoessa boscii
and Cymothoa eremita are buccal parasites. So, the present study was
carried out to evaluate and update the protein value of fish parasite in different
species of isopods such as Nerocila phaeopleura, Nerocila loveni, Nerocila
sundaica, C. boscii and C. eremita and also to extent which SDS-polyacrylamide
gel electrophoresis (SDS-PAGE) used to discern interspecies differences in five
species of isopods parasites. Data is provided to characterize these species
on the basis of species-marker bands in SDS-PAGE patterns.
MATERIALS AND METHODS
Collection of animal: The parasites were collected from Nagapattinam coastal environments, Southeast coast of India were examined during the years 2011. Species were collected and transported to the plastic cover and washed with fresh water. They were separated into different species. After homogenizing with pestle and mortar, the samples were transferred to centrifuge tubes. The homogenate was centrifuged for 20 min at 4000 xg and the supernatant was collected. Samples were maintained on cool condition at all times to avoid protein denaturation.
Estimation of protein: The amount of protein was measured by spectrometry
according to the method of Biuret (Raymont et al.,
1964). Biuret reagent was used as a colour reactant and concentration was
calculated in response to absorbance at 540 nm in spectrophotometer (Spectro
Determination of molecular weight by using SDS PAGE: SDS-PAGE was used
to find out the molecular weight active fractions of the sample. SDS-PAGE was
performed in separating gels, according to the method described by Laemmli
Estimation of protein: Variation in protein substance of the muscles of isopods parasites were shown in Fig. 1. Among the muscles protein from five species of isopods parasites, maximum protein content of 18.26% was recorded in N. loveni and the minimum content of 3.44% was noted in C. boscii parasite.
SDS-PAGE: The muscle tissue samples were used to estimate the Molecular
Weight (MW) of proteins present in it by SDS-PAGE. Different samples were used
to determine the molecular weight of proteins in the muscle tissue. The muscle
tissue containing proteins were detected. Protein marker with molecular weight
in the range of 20 to 50 kDa was used. The bands were detected in the gel that
|| Comparison of protein content in the isopod parasites
||SDS-PAGE gel showing variation in molecular weight of muscle
protein in different isopod parasites. Lane 1: N. loveni, Lane 2:
N. sundaica, Lane 3: C. eremita, Lane 4: C. boscii,
Lane 5: N. phaeopleura, Lane 6: Marker
|| Molecular weight of muscle protein in isopod parasites
|-: Not detected bands
SDS-PAGE was shown to have protein patterns of parasites samples (Fig.
2). Seven bands with molecular weight of 54, 47, 39, 32, 23, 20 and 19 KDa
were found in N. loveni. N. sundaica also seven bands were detected with
molecular weight of 53, 45, 39, 32, 22, 20 and 19 KDa. C. eremita had
five bands with molecular weight of 50, 41, 33, 22 and 20 kDa. C. boscii
had only one bands were detected with molecular weight of 18 kDa. N. phaeopleura
showed six bands with MW of 56, 50, 34, 27, 23 and 19 kDa. Only few bands
were detected and they were basically similar for all species except C. boscii
Information about these five isopod species is important because each infects
and reduces the productivity of commercially important species of fishes. Proteins
were, in proportion, the main body component and also the most stable. Parasites
were found to have protein, maximum content of protein in the muscles of parasites
N. loveni (18.26%) and C. eremita (15.24%) followed by N.
phaeopleura (11.51%), N. sundaica (6.83%) and the lowest content
of (3.44%) was reported in C. boscii. Among the five parasites body surface
parasites of Nerocila genus, N. loveni show maximum value in both
estimation of protein and molecular weight. In the buccal parasites C. eremita
shows Maximum value in protein estimation and molecular weight. N. phaeopleura,
N. loveni and N. sundaica had maximum bands were detected in compared
with C. boscii and C. eremita. Most of the researchers have concentrated
on the quantification, identification and isolation of protein from edible crustaceans
like crabs, shrimps and lobsters. Isolation of protein has been earlier reported
in the hemolymph of crustaceans in the brachyuran crabs (Rameshkumar
et al., 2009). The present study was observed percentage protein
in the muscles of parasites. The protein content of P. pelagicus and
P. sanguinolentus was 0.47-15.91% and 12.81-13.6% (Radhakrishnan
and Natarajan, 1979). Reported on the protein value in blue crab was 17.17%
(Anonymous, 1999). In general, the average body proteins
of the crab were directly related to the levels of protein in the diet (Khan,
1992). Similar pattern of protein changes was reported by Alava
and Lim (1983) in P. monodan and in M. malcolmsonii by Soundarapandian
and Ananthan (2008). The results of the present study indicate that the
concentration of protein in the muscles showed wide inter specific variation
among the parasites. In the present investigation samples showed that protein
level was subjected to SDS-PAGE to estimate the molecular weight. After electrophoresis
band were detected in the gel which represented proteins of molecular weight.
Further, the identification of protein pattern of the host fishes could give
sufficient knowledge about the feeding, energy requirement and developmental
needs of isopods which might pave way for devising a strategy to reduce the
infection. To date, although progress has been achieved in this respect, knowledge
on the protein requirements of isopod parasites was still too insufficient to
provide reference for effective diet levels. In addition experiments presented
in the study represent a preliminary analysis to compare the information obtained
by electrophoretic methods, namely SDS-PAGE. This method allowed to separate
proteins and to determine their molecular weights.
The results of the present study indicated that muscle tissue protein content
shows wide inter specific variation among the fish parasitic isopods. No study
has been previously dealt with a trial to estimate the protein content in isopods.
They were considered to be least important but the present study has initiated
a new baseline in parasitological. The study means more as the parasites acquire
energy from the host in the form of proteins.
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