International Journal of Zoology

and Research (IJZR)

ISSN(P): 2278-8816;ISSN(E): 2278-8824
Vol. 6, Issue 2, Apr 2016, 7-10
TJPRC Pvt. Ltd.

AN ENZYMATIC STUDY IN VARIOUS FRACTIONS OF ASCARIS

SUUM FROM NATURALLY INFECTED PIGS
R. K. TRIPATHI1, AMEETA KUSHWAH2, S. M. RUDRAPPA3 & A. BANERJEE4
1,2,4
3

Veterinary Hospital, Harihar, Karnataka, India

Veterinary Officer, Veterinary Hospital, Harihar, Karnataka, India

ABSTRACT
A study was conducted to reveal the relative presence of some enzymes viz, aspartate transferase (AST),
alanine transferase (ALT), alkaline phosphatase, acid phosphatase, lactate dehydrogenase (LDH) and acetylcholinesterase in the gut, muscle, cuticle and body fluid of Ascaris suum. Amongst the tissues, the activity of acetylcholinesterase, AST and acid phosphatase was highest in gut whilst ALT and LDH was maximal in muscle. Alkaline
phosphatase was the highest in body fluid. The presence of these enzymes in the tissues was correlated with their
physiological activity and can be explored for taxonomic and diagnostic purpose.
KEYWORDS: Relative Presence of Some Enzymes, Tissues was Correlated, Physiological Activity

INTRODUCTION
Earlier studies on the biochemical nature of Ascaris suum (Tripathi, 2004) indicated the presence of some
biochemical (Tripathi et al., 2006a) and inorganic constituents (Tripathi et al., 2006b) in various fractions viz.,

Original Article

Received: Mar 15, 2016; Accepted: Mar 26, 2016; Published: Apr 07, 2016; Paper Id.: IJZRAPR20162

body fluid, muscles, gut and cuticle of this parasitic nematode. Biochemical nature of these fractions differed
markedly from each other; glucose and the lipids were maximally present in body fluid whilst the level of protein
was the highest in gut. The level of inorganic constituents also differed both in pattern as well as in magnitude
amongst these fractions. The present investigation is an extension to the above studies and deals with the relative
presence of some enzymes in these fractions providing additional information and clarity on their biochemical
nature and functions that may be explored for diagnosis as well as for taxonomic purpose.

MATERIALS AND METHODS

Mature Ascaris suum were collected from naturally infected pigs, slaughtered at local abattoirs at
Jabalpur, Madhya Pradesh. These were washed with n-saline to remove debris and excreta and were brought to
laboratory within a short span of time under cold conditions. The worms were dissected out to procure body fluid,
gut, cuticle and muscles, as indicated earlier (Tripathi et al., 2006a). The fluid from 10 ascarids was pooled and
formed one sample for enzymatic analysis. The tissues under study were homogenized separately in Potter
Elevehjem homogenizer (10% w/v) in ice-cold n-saline. These were centrifuged at low speed for 5-10 min and
respective supernatants were used for
Aspartate: 2-oxyglutarate

estimation of the activity of aspartate amino transferase (AST) (L-

transaminase

EC:

2.6.1.1),

alanine

amino

(L Glutamate: NAD oxidoreductare EC: 1.4.1.2), acid phosphatase (ACP) (Orthophospho-ric

transferase

(ALT)
monoester

phosphohydrolase EC: 3.1.3.2), alkaline phosphatase (AlP) (Orthophosphoric monoester phosphohydrolase EC:

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R. K. Tripathi, Ameeta Kushwah, S. M. Rudrappa & A. Banerjee

3.1.3.1), acetyl-cholinesterase (AchE) (Acetyl hydrolase, EC: 3.1.1.7) and lactate dehydrogenase (LDH) (L-Lactate: NAD
oxidoreductase, EC: 1.1.1.27) .
The activity of these enzymes was estimated following the instructions on the diagnostic kits, being marketed by
Reckon Diagnostic Pvt. Ltd. Baroda, India. The data were analyzed by calculating mean and standard error using
conventional formulae.

RESULTS AND DISCUSSIONS

The results of present study are presented in Table 1. It revealed presence of all the above referred enzymes in
muscles, gut, cuticle and body fluid of mature A. suum, though their activities differed differentially amongst these tissues
and body fluid. Thus, activity of both the protein metabolic enzymes namely AST and ALT involved in converting aspartic
acid and alanine to glutamic acid respectively was indicated in all tissues under study and in body fluid of this nematode.
These enzymic activities are known to be involved in protein synthetic process (Rodwell, 2000)
The activity of AST and ALT was maximally present in gut and muscles respectively and was lowest in body
fluid whilst ALT activity was the highest in muscles (Table 1). The results suggested transamination to be involved in
amino acid synthesis and relevant systems are widely distributed in this parasite. Reductive amination of keto acids was
observed in the ovaries of Ascaris lumbricoides (von Brand, 1973). Oxaloacetate, a precursor of aspartic acid, was found to
be active. Slightly different pH optima indicated involvement of both the enzymes in transamination system of ascaris.
Highest activity of AST in gut correlates well with highest level of protein in this tissue (Tripathi et al., 2006a). The
activity of the other protein metabolic enzyme ALT, was also moderately present in gut and was maximally present in
muscles. Muscles, too, contained a good activity of AST that appears to be actively involved in protein metabolic
processes.
The results of the present investigation also reflected activity of organophosphatases in the tissues of this parasite
(Table 1). Thus, activity of acid phosphatase was abundantly present in gut whilst activity in muscles was more than that in
cuticle (Table 1). However, activity of alkaline phosphatase was the maximum in body fluid and lowest in cuticle. The
enzymic activity in gut was slightly higher to that of muscle (Table 1). Many studies dealing with the presence of
phosphomonoesterases in parasites have been published (von Brand, 1973). The distribution of acid and alkaline
phosphatases has been studied frequently by means of histochemical methods allowing exact localization of the enzymes
and thereby, providing possible clues to their biological significance.
Phosphatases have been found rather frequently associated with structures of absorptive or excretory nature. Their
localization in gut of this parasite may be correlated with physiological and biochemical activities of these organs. Notably,
the activity of alkaline phosphatase was highest in body fluid (Table 1), suggesting enzymic participation in catalyzing
hydrolytic cleavage of pyrophosphates. In ascaris, occurrence of pyrophosphatases was reported in coelomic fluid and in
body wall. The enzyme had been partially purified and its properties were studied (Islam et al., 2003). The activity has also
been indicated in Ascaris lumbricoides, Acaridia galli and Moniezia expansa (Holguin and Monteolive, 1965).

CONCLUSIONS
In A. suum, presence of an anaerobic carbohydrate catabolic enzyme, lactate dehydrogenase (LDH) was also
indicated in muscles. The activity was beyond detectable limits in gut and cuticle. Body fluid, too, exhibited a low

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An Enzymatic study in Various Fractions of Ascaris suum from Naturally Infected Pigs

enzymic activity (Table 1). The enzymic activity has earlier been indicated in A. lumbricoides and Ascaridia galli (SoonHyung, 1967). In adult helminthes, various pathways of carbohydrate catabolism are essentially anaerobic and
carbohydrates are more or less broken down quantitatively to lactic acid (Barrett, 2004). The presence of LDH activity in
muscles is justified since muscles are well known to be involved in anaerobic carbohydrate catabolic process. The presence
of acetyl-cholinesterase (Ach E) activity is also indicated maximally in gut and minimally in muscles and was moderately
low in cuticle and body fluid (Table 1). The enzymic activity in nematodes was reported to play a functional role in nerve
activity of parasites (von Brand, 1973). Both specific and non-specific cholinesterases were reported in various organs of
many helminthes. Acetylcholine itself, or any acetylcholine like substance, has been reported in nematodes.
Pharmacological analysis of ascaris also indicated the presence of acetylcholine, a substrate for acetylcholinesterase
activity (del Castillo et al., 1963).
Table 1: Relative Activity of Some Enzymes in Various Fractions of Mature Ascaris suum
Enzymes
Aspartate amino transferase (IU/L)
Alanine amino transferase (IU/L)
Acid phosphatase (IU/L)
Alkaline phosphatase (IU/L)
Lactate Dehydrogenase (IU/L)
Acetyl choline esterase (U/L)

Body Fluid
62.247a
4.327
65.280a
3.872
00.00
00.00
96.560a
5.151
6.573a
0.334
406.386a
16.510

Muscle
507.640b
26.370
468.640b
16.152
6.706a
0.480
61.520b
2.293
26.113b
1.260
328.446a
16.562

Gut
1091.50c
43.198
173.326c
12.631
35.920b
3.283
74.800c
1.613
0.00
0.00
1015.460b
33.967

Cuticle
142.160a
11.161
113.846ac
8.641
2.533c
0.160
80.460a
1.902
0.00
0.00
382.280a
11.522

Values are S.E. of 10 observations.

REFERENCES

1.

Barrett, J. (1994). Biochemistry of helminthes. In: Chowdhary, N. and Tada, I. (ed.) Helminthology. Narosa
Publishinh House, New Delhi. pp 211-223.

2.

Del Castillo, J., de Mello, W. C. and Morales, T. (1963) Arch. Int. Physiol. Biochem. 71:741-757.

3.

Holguin, V. L. and Monteoliva, M (1965) Rev. Iber. Paraitol. 25: 101-118.

4.

Islam, M. K., Miyoshi, T., Kasuga-Aoki, H., Isobe, T., Arakawa, T., Matsumaoto, Y. and Tsuji, N. (2003)Inorganic pyrophosphatase in the round worm Ascaris and its role in the development molting process of the
larval stage parasite. European J. Biochem. 270: 2814.

5.

Rodwell, W. V. (2000)- Catabolism of proteins and amino acid nitrogen. In: Murry, R., Granner, D. K., Mayes, P.
A. and Rodwell, V. W. (ed.) Harpers Biochemistry. Appleton and Lange Stanford, Connecticut. pp 313-322.

6.

Soon-Hyung, L. (1967) - Studies on lactic dehydrogenase activity in parasitic helminthes. Korean J. Parasitol. 8:
70-80.

7.

Tripathi, R. K., Kushwah Ameeta, Sharma, R. K. and Chaudhary,R. K. (2006a) - Inorganic constituents in various
fractions of Ascaris suum of pigs (Sus scrofa domestica). Indian Vet. Med. Jour.

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R. K. Tripathi, Ameeta Kushwah, S. M. Rudrappa & A. Banerjee

8.

Tripathi, R. K., Kushwah Ameeta, Sharma, R. K. and Chaudhary,R. K. (2006b)- Biochemical studies in various
fractions of Ascaris suum. Indian Vet. Med. Jour.

9.

Von Brand, T. (1973) - Biochemistry of parasites. Academic press, New York.

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