Acta hydrochim. rt hydrobiol. 12 (1984) 0, 609-613

OMKAR, R. >IURTI and G. 8. SHURLA

Research Laboratory, Department of Zoology, Vniveraity of Gorakhpur

Aldrin Induced Changes in the Activities of Three Phosphstases of Mucrobruchium Znmarrei (H. Mane EDWARDS)

S ' w n m z r y : Xucrobrachiun~~ Zu,mzn.ei is exposed to 0.008, 0.017 and 0.021 mg/l aldrin (0.4. 0.8 and 1.0 of the LCjO,Stih) for 24 ... 96 h and aft.er exposure the activities oftheacid, alkaline and glurose-6-phosphatascs are determined. With concentration and t,ime of exposure the acid phos- pliatase shows an increasing activity up to 170 o:'o compared with the control, whichisinterpreted as a part of prenecrotic changes in the cells. With concentration and time of exposure the alkaline phosphatasc shows a decreasing activity down to 20 0,'11 of the control, which is indicative of considerable disturbances of the t.otal metabolism. The glucose-ti-phosphatasesiio~~aan inereasins act,ivit.y with the a.ldrin concentration up to 150 9;) during 72 h ; then the activity drops t o 60 ... 80 "',,. The initial rise in t.hc glucose conccntrat.ion of blood eai isd by t.hat is t.he ctlear consequence of a stress reaction and thc change of metabolism resulting from it.

Introduction

The increasing contamination of the aquatic environment by the indiscriminate arid widespread use of pesticides is a serious problem for environmental biologists. Organochlorine pesticides are more hazardoils since t,hey are not only highly toxic h i i t ,

are also residual in nature. The deleterious effects of aldrin on several criistaceans have been studied (NEBRKER and GACFIN; K1sLE:H: SANDERS: SHCKLA and OXIRAR). But these studies are confined to the determination of its niedian lethal conc,entr:i- tions. Studies on the effects of uldrin on the biochetnical aspect.s of c~ristaceans a re meagre. Sonie reports on tkese aspects in a fish Iletero~3iei'atI's fossilis are availa1)lP ( VEIIMA, TOKK and DALELA; SRIVASTAVA and SJXGIJ). While studying the :Iciite t.oxicit>- of aldrin to a n ecoiioiiiically important frcshwater prawn having a nutritive value, the authors felt, t.lie need to study the effects of this cheniical on the hio- c.hemical aspects in various tissues of N . lrmctrrcii. Therefore, an attempt was niacle to investigate the effects of a lethal and two sublethal exposiirrs to aldrin 011 acid, alkaline and glucose-6-phosphatase activities in t,he hepatopancreas of Jf. / (ouurre; which has R significant role in the physiology of t.he organisms, for iintlerstandinp the nature of changes causing t,he death of organisnis due to aldrin stress.

Material and Methods

111. lccnwrrri were collected froiii the Raiiigarh lake of (;orakhpur city and iiiinie- cliately brought to the laboratory. They were kept in large reservoirs and accliniatized for three days at room temperature (20.05 1.5 "C). Aldriri solritions of 0.008, 0.017 42'

610 Scta hydrochiin. et hydrobiol. It? (1984) (i

arid 0.021 riig/l (0.4, 0.8 ant1 1.0 of 96 h IAcl3,,) were prepared in 10 1 dechlorinatetl t ap water in pl:tst.ic containers on the day of exposure and t,wenty healt,hy specimens were transferred carefully to each container. Controls were also set sirnult aneoiisly. cloni- pressed air was supplied continuously to the prawns during a(,Cliiriat,izat,ion and ex- perinients. The speciniens were taken froin esperiniental and corit.ro1 sets, antl hepa- topancwas was dissected out . A weighed quantity of tissue was homogenizetl in 0.9 S KaCl solution for acid and alkaline phosphatase and in 0.25 M sucrose solution for glucose-6-phosphRtase, in glass homogenizers. These hoiuogenates were centrifuged a t 5000 rpni for 20 niin. The supernataiits were collected arid used as the enzyme source. 1'-nitrophenyl phosphate of pH 5.0 and 9.3 for acid antl alkaline pliosphat.ase, respect.ively, was used as siihstrate, while sodirirn salt of glricose-6-phosphate (pH =.

6.5) was the siihstrate for glir~ose-6-~~hospIlntase. The activities of acid and alkaline phosphatase were nieasirred hj- the niethods of HERGMEYER. The Iiiethod of SWANSUN was used for t,he determination of the activity of glucose-6-phosphatase. Each experi- ment was replicated six t,iriies and the data were statistically analysed for significant difference (CAMPBELL).

Results

The data in Tab. 1 show an elevation in the acid phosphatase activity after aldrin treatment. The increase in the acid phosphatase activity showed a time- and dose- dependent relationship. The increase in the acid phosphatase activity was less signi-

Table 1 . ('hanges in the Activities of Phosphataxes in the Hepatopancreas of :2ldrin Rxposed M . Lamnrrei Tabrllr 1 . VerLndcrungen der ;\ktivitat vonI'hospllntnsen in der Hrpatopankreas von M. Zainarrei Linter der Eiii\virkung von Xldriri

Kxposurc ('oncen- period trations I1 m g i l __ - . . .. . -. .. .-

('ontrol

0.021

OMKAR, et al . : .\ldrin Induced Changes of I’hosphatasrs 61 1

continued Table 1

Exposure Coneen- Phosphatase activity period trations Acid Alkaline Glucose-& h mg/l Phosphntase Phosphetase Phosphat m e

0.008 10.63 :50.6O* 3.52 k0.48* 5.75 f0.51 ( 1 34) (61) (121)

(1.50) (43) (142)

(158) (35) (149)

72 0.017 11.91 :k0.71*** 2.47 50.72** 6.54 *0.44**

0.021 12.54 C0.74*** 2.02 *0.58** 7.08 f0.45**

96

0.008

0.017

0.021

11.27 k0.69** 2.87 ?O.61** 3.75+0.4i

12.78 f0.72*** 184.50.66*** 3.23 50.48*

13.49 f0.76*** 1.15 ? 0.71 *** 2.98 S0.47*

(142) (50) (79)

(161) (32) (68)

(170) (20) (03)

b o l e : (a) =The yalues (Mean f SEM, of six replicate$) of acid and alkaline phospliatase are expressed in y nioles of 1)-

( t i ) =The values (JlrJan f SEM, of six replicates) of glucose-a-pliciPphatasc are rxprrssrd in y iiiolesofinorgmic plior-

(c) =Figirru in brackets indicate 1111: par-cent ncf ivity. ((1) = *, ** h * * * indicate yalues significant at P ~ 0 . 0 5 , P ~0.01 nntl Y -.0.001, rwpwtively.

~iitrophenol lilieratt!d/g wet weight. of tissue.

phale lilirrated/g wet weight of tissue.

ficnnt (P-=O.05) in two higher concentrations after 24 11 of esposiire and in t.he lowest. concent,ration (0.008 nig/l) after 48 and 72 h of exposure. Elevation in the activityof enzyme was significant (P<O.Ol) in two higher concentrabions after 48 h and in the lowest one after 96 h while in others the increase in t,he enzyme activity was highly significant. (P- 0.001).

The alkaline phoxphat.ase activity was inhibited in relation to exposure time and dose of aldriri (Tab. 1 ) . The inhibition in alkaline phosphat;ase was less significant (P-= 0.05) in an acute concent,r;ttion after 24 11, two higher concent.rat,ions after 48 11 and in the lowest concentration aft.er 72 and 96 t i , significant (P-c 0.01) a.nd highly significant (1’-=0.001) in two higher concentrat,ions after 72 h and 96 h of cxpowre, respectively.

The activity of glucose-6-phosphatase was enhanced up to 72 h of exposiire, there- after a decreasing trend in the enzyme activity was recorded (Tab. 1) . The elev n t . ion in glucose-Ei-phospliat,alle was significant (P-= 0.05 and P-= 0.01) only in two higher concentrations after 48 and 72 h of exposiire after which the inhibition in enzyne activity was less significant (I.’-= 0.05) in two higher concentrations.

Organochlorine insecticides are known t,o affect the activities of various enzynies in the fish (VEnnra; TONE and DALELA; SASTRY and SHARMA 1978, 1979; SHAKMA, CHATUKVEDI and ISASTRY ; THOMAS and MURTHY), but there is a lack of such studies in freshwater prawns. In the present investigation the elevation in the acidphosphatase

612 Acta hydrochim. et hydrobiol. 12 (1984) 6

activity was recorded. A similar increase in t.he acid phosphatase activity in the liver of a teleost Chunna punctatus after endrin exposure (SASTRY and SHARMA 1978, 1979 a, b) and in nine freshwater teleosts exposed to thiodan (SHAFFI) was reported, too; SHAFFI suggested that the increase in the acid phosphatase activity might be attributed to the rupture of cellular and lysosoinal membranes. The activity of alkaline phosphatase was inhibited after aldrin exposure to M . Znwarrei, which is similar to the findings of other workers (SASTRY and SHQRMA 1978, 1979b; THOMAS and MURTHY : SIIAFFI; UPADHY,AY and S H ~ K L A ) . The inhibition in t'he alkaline phosphatase activity may he ascribed t'o the fall in pH due to the rupture of the inenihranes ( SHAFFI).

Although the physiological role of acid and alkaline pliosphatases are not clearly uncterstood yet, some possible role has been assigned to them by the workers. Acid phosphatase is a lysosoinal enzyme (DE DUVE et al.: ARUNA et al.) which helps inthe aiitolysis of cell after it.s death. The increase in the acid phosphatase activity in the injured cells occiirs as a part of prenecrotic changes (NOVIKOFF). Alkaline phosphatqse is a brush border enzyipe, splits varioiis phosphate esters a t an alkaline pH and me- diates nieinbrane transport (GOLDFISHER, EWER arid XOVIIIOFF). Alkaline phospha- t a . G : e also takes part in transphosphorylation reactions, protein synthesis (PILO, ~ ~ W A X I and SrrA~r) thereby involved in t.he s\-nt.hesis of certain enzymes (SUMNER) and secretary activit;v ( IDRAHIM, HIGAZI and D E X I ~ N ) . Thus the inhibition of alkaline phosphatase indicates a general distiirb:rnce of various physiological ;irocesses of the orgnnisni.

(.;luc.ose-B-pliosI,Iiatase catalyses the hrealidown of glucose-B-pliospl.iate to glucose and phosphoric acid. The hF-drolysis of plucose-6-phosphate is a kej- step I n gluconeo- genesis and in the conversion of liver glycogen to blood glucose (HOCHACIIAKA). Thus the elevation in the glncose-6-phosphatnse actirit7- accelerates thc conversion of liver glycogen to blood glucose, while the inhibition in enzyme act,ivitj- adversely affect the process. O N I L ~ et. al. (1983, iinpiihlished findings) recorded a decrease in t'he hepatic glycogen content with a convoniitnnt increase in the Mood gliicose leveliip to 7 2 11 of aldrin exposure to M . Imnrirre i , after 7% 11 a decrease in the glucose level was noted, which support the present findings.

Thus i t is concliided that the exposure of aldrin to Jf. lanzccrrei creates a widespread disturbance in the general physiological process which i11tiniatelj- valises the death of the organisni.

A c kiiowledgeirieiit

>luthors (OYKAR orid RAX JIuRTI) are grateful to L-niversity Grants Commission, SelT Delhi for financial assistanoe.

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;Mnnrcskripteingang: 19. 9. 1983.

Anaehrift der Verfasser:

Dr. OSIKAR, RAM MURTI and G. S. SIIUKLA, Pollution Relevant Research Laboratory, Department of Zoology, University of Gorakhpur, IND - 273001 Gorakhpur.