Cr (VI) induced changes in the activity of few ion dependent A TPases in three vital organs of mudskipper Periophthalmus dipes (Pisces: Gobidae)

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1 Indian Journal of Marine Sienes Vol. 28, Marh 1999, pp r (VI) indued hanges in the ativity of few ion dependent A TPases in three vital organs of mudskipper Periophthalmus dipes (Pises: Gobidae) Jayesh Thaker, Jignasa hhaya, Renu Mittal, Sheeba Nuzhae & Rahul Kundu 2 Department of Biosienes, Saurashtra niversity, Rajkot 36 5, Gujarat, India Reeived 7 Otober 1997, revised 21 September 1998 xperiments were arried out to assess the dose and duration dependent influene of r() to few ion dependent A TPases in liver, brain and musle of a oastal euryhaline teleost Periophthalmus dipes. Fishes were exposed to three sublethal onentrations (5, 1 and 15 mg/i) of r(vi), prepared by using K2rO. with normal seawater, for three exposure duration (2, 4 and ). In the present report, effets of r(vi) toxiity on total, (Na+,K+)-, (a ++)-, (Mg ++)-, (a++,hoj -)- and (Mg++,HOJ-)- in liver, brain and musle were evaluated. Though results indiated a general dose and duration-dependent inhibitory trend, exposure duration was predominant over dose in the inhibition of the enzyme ativity. hromium ompounds are onsidered highly ar'inogeni and an produe potential hazards at a very low onentration I. The priniple soure of hromium in the aquati environment is from disharge of industries using large amount of hromate and dihromates. Toxiity of potassium hromate on some parameters on some fresh water fishes were earlier reported 2 Heavy metals depress ion transport in several important osmoregulatory systems of teleosts leading to strutural damage to various organs like gills, brain and kidney3. Lakshmi et al. 4 reported the effet of merury toxiity on six ion dependent in the gill 's tissue of B. dentatus. Therefore, in the present ommuniation an attempt has been made to evaluate the dose and duration dependent effets of r(vi) as K 2 r 4 on the A TPases in liver, brain and musle of a oastal amphibious fish the mudskipper, Periophthalmus dipes Bleeker (Pises: Gobidae). Materials and Methods A total of 275 numbers of a oastal teleost, ommonly known as mudskipper, Periophthalmus dipes, were seleted for the present investigation. Other details regarding fish are same as reported earlier5. Analytial grade of K2r 4 (. Merk) was.dissolved in normal seawater to a desired stok solution. Different onentrations of this toxiant were prepared with seawater (salinity ppt). Other details regarding toxiant are as reported earliers. On the basis of L5 value (47.84 mg/l) three different sublethal onentrations were prepared (5, 1 and 15 mg/l i.e 96 1 Department of Biohemistry, Allahabad niversity, Allahabad , India. 2For orrespondene M, 192 M and 288 M respetively) with seawater having salinity5 of ppt. At least 1 fishes were put in eah onentration and exposed for three different durations (2, 4 and ). Separate ontrol groups were also maintained along with eah treatment. The toxi media were renewed every 12 h. Animals were sarified after sheduled exposure and seleted tissues were quikly disseted out and plaed 111 hilled surose - DTAimidazole buffer (PH 7.1) to remove exess blood and adhering tissues. Piees of liver and myotomal musle were taken from approximately same portion of the tissue from all killed speimens, these samples were also washed and blotted. The brain was removed from the skull, washed and blotted. ah tissue sample from all the killed animals was ombined together and 2 mg of eah tissue was taken for enzyme extration. The tissue preparation, enzyme estimation and other assays were arried out by the methods desribed earlier The ativities of (Na+-K+)-, (a++)-, (Mg++)-, (a++, H 3 -)- and (Mg++, HOJ-)- were estimated 4. All experiments were repeated at least thrie and obtained data were subjeted to various statistial analyses 8. Results and Disussion Results of the present investigations showed the behavioural manifestations upon exposure to the toxiant. As soon as the fishes were introdued to the toxi media, they started jumping and tried to ome out of the media. After this unsussful attempt fishes showd errati swimming for sometime. Then the fishes settled down. It

2 46 INDIAN 1. MAR SI VOL. 28, MARH 1999 was observed that fishes beame sluggish as the time elapsed. Generally, in maximum time duration (6 d) speimens beame absolutely sluggish with slight hange in their body olour (bluish gray to yellowish gray). In liver, the speifi ativity of most of the enzymes showed dose and duration dependent inhibition. The ativity of total and (Na+, K+)- are effeted mostly. In these ases a gradual inhibition was observed. The ativity of (Mg++, H 3 -)- showed initial stimulation in lower dose (Smg/l). In all other ases less inhibition was observed. However, the ativity of all enzymes studied showed maximum inhibition in fishes exposed to highest dose for longest duration (Table I). Varied hanges in the ativity of different enzymes in the whole brain were evident from the present study. Most of the enzymes showed inhibition. However, in few ases slight stimulation in the enzyme ativity, after an initial fall in lower onentration, was also evident (Table 2). Marked hanges in the ativity of different enzymes in the myotomal musle tissue was evident. The ativity of total and (Na;, K;)- A TPase fell sharply in fish exposed to S mg/i and 1 mg/i onentrations, but stimulated in the highest dose. However, inhibition of varied magnitude in the ativity of other s was observed (Table 3). In the present study a general inhibitory trend was observed in all the ion dependent A TPases, in all the tissues studied of r(vi) intoxiated fishes. However, slight irregularities were observed in some ases on exposure to r(vi). (Na ' K;)- is one of the major enzymes responsible for movement of Na+ and K+ ions aross membrane and in the proper funtioning of the Na+, K+ pump. It is apparent that inhibition of (Na+, K+)- by r(vi) may ause alteration in the ion transport proesses in the ell membrane and impair metabolite uptake and transfer in the liver tissue 9 Marty & Houston l O reported signifiant deviations in plasma sodium and hloride levels as well as ti ssue sodium and water ontent on exposure to sublethal onentrations of heavy metals. Strik et al. II observed a derease in the Na+ onentrations of the blood of rainbow trout intoxiated with lethal doses of hromium. This drasti inhibition of (Na+, K+)-A TPase may disrupt the ative Na; reabsorption whih explains for the Na+ loses observed in the ells 9. Brain is always in a state of onstant eletrial ativity and most of the effetor ativity taking plae is initiated from the brain. hromate ions I are also known to inhibit eletrial potential aross the membrane. The resulting impairment of neurotransmitter release and transport ould be responsible for the paralyti symptoms observed in this study. The drasti inhibition in the ativity of musular (Na+, K+)- in the present investigation may have resulted from the binding of these metals with the enzyme protein whih usually denatures them l 2 The inhibited (Na+, K+)- ativity may ause a blokade of Na +, K+ oupled pump. Sine this pump is ; ti -a..". N ;; 1::'... e : u > 9 ';; ;;; ;;; X '5....>< ::l ;;; ;;: -5 '- "2 ::l v; > ;,; ;:: OJ P.. N OJ ::l >< -;;; > '-,.:::: v -;- : :::!. ;;: ti u t;: u I : I- N u.j... -,,- 1-1-«i ",,,- l- «:I:" t I «, :I::. 1 I- 1) «::

3 '(.. n/yms Tota l. \ TPas Na- K' ATP as a Mg' a HO,' Mg HO,' Table 2 - Mean speifi ativity ( 11 mol Pi mg ' Protein h ') o f six e nzymes in th e brain tissue of the mudskippers exposed to varying onentrations ofr(vi) for 2,4 and respetively Table 3- nzymes Total Na- K ' ATP ase a Mg a HO,' Mg HO,' " l> \ ontro l days I 'v'a lu s rprsnlmean standard deviation o f tripliate data] 5 mg! I Mean speifi ativity (11 mol Pi mg " Prote in h " ) of six enzymes in the musle tissue of the mudskippers exposed to varying onentrations ofr(vi) for 2, 4 and respetively ontrol I : R mg I I [Values represent mean standard deviation of tripliate data] 5 mg II \ mg / l \ \ mg I I mg / l i > r;; ;:.::I a I:l!'-- ('). z o (") ttl o (") ::t ttl I) Z I:<. -So -...J

48 INDIAN J. MAR. SI VOL. 28, MARH 1999 responsible for the ation potential the ondutivity of musle may be signifiantly redued making the whole musular tissue nonfuntional6.

4 48 INDIAN J. MAR. SI VOL. 28, MARH 1999 responsible for the ation potential the ondutivity of musle may be signifiantly redued making the whole musular tissue nonfuntional6. Inhibition in the ativity of (a...)- and (Mg++)- in liver may be due to the aumulation of large amount of r (VI) in the liver ells whih ombine with the enzyme protein thus altering its ativity. Similar inhibitions in the ativity of the (a O-+)- and (Mg++)- in the liver of fish exposed to Hgl 2 have been reported13. In the present investigation, inhibition of these two vital enzymes learly indiates a blokade in the transport of these ions. Mg++ together with a++ is essential for neurotransmittory fu ntion espeially reflex ation. Therefore, the redution of Mg++ might ause delay in transmission at the neural juntion 7. Results of the present investigation showed duration dependent inhibition in the ativity of (a++)-a TPase and (Mg++)-A TPase in the musular tissue of intoxiated fishes. This inhibition probably redued the uptake and transport of a++ and Mg++ whih are essential for the ontratile property of the musle. It is also possiblel 4 that this imposed stress impairs funtions of saroplasmi retiulum and the transport and storage of a... Inhibition in the ativity of the anion dependent A TPases learly revealed the disrupted transport of anions through the membrane whih eventually might have aused other funtional disorders in the liver. However, the slight stimulation in the enzyme ativities observed in some ases ould be due to the initial effort made by the animals to adapt to the imposed stress. The role of biarbonate ions in the entral nervous system (NS) is not very lear at this point. However, they are believed to be assoiated with axoplasmi flow and normal maintenane of nervous tissue. Heavy metals are reported to ause irregularities in their release and uptake of neurotransmitters in the brain whih may be due to the inhibition of H 3 - s l5. The observed neurobehavioural disturbanes in the mudskippers may also be due to this impaired neurotransmission. However, sudden enhanem e t in the ativity of (a++, HO) -)- A TPase in r(vi) exposed fishes was probably due to nerosis in the membl \ne where the entry of anions was not ontrolled by the ative transport l6. The drasti inhibition of (Na" K' )- reated a blokade in the transport of Na' and K' and hene, to equilibrate the eletrial potential onsiderable amounts of negatively harged ions entered the ell whih may explain the stimulation in the enzyme responsible. Results of the statistial analysis of the ativity of the enzymes studied revealed a predominant exposure duration dependent effet (Table 4). The overall results of r(vi) intoxiation revealed a drasti impairment in the ellular ative transport of different vital ions and metabolites in liver, brain and musle tissues. The hampered uptake and transport of important ions suh as a++ and Mg++ may ause failure of the ontratile property of the musle. The Table 4- F' ratios derived from the two way nested ANOV A test of the speifi ativity of various enzymes studied between different treatments and exposure durations. alulated "F" values are given in the Table. "Degrees of Freedom" (dt) are (a) among doses = 3, 8 and (b) among durations = 8, 24. ritial values of F for these df are also given Liver Among durat ions Brain Among durations Musle Na-K a i nzymes Mg a-ho) Mg-HO J Among durations ritial values of "F" at various df : Probability F [3,8] F [8,24] P = 5 P = 1 p =

THAKR et af. : r fndd HANGS fn P. dipes 49 observed depolarization was probably due to inreased intraellular Na+ resulting from inhibition of membrane (Na+K+)-A TPase ativity.

5 THAKR et af. : r fndd HANGS fn P. dipes 49 observed depolarization was probably due to inreased intraellular Na+ resulting from inhibition of membrane (Na+K+)-A TPase ativity. This leads to rise in the intraellular a++ as is evident from the stimulation in the ativity of (a++)-a TPase. This ultimately effeted the ontratility of musular tissue 17. The fatal onseqqenes of the blokade in the ion pumps and metabolite transport may be oupled with the behavioural observations where a failure in musular mobility followed by paralysis was evident. Aknowledgement Authors are thankful to Dr. Harish Dave, Deputy Diretor of Fisheries, Govt. of Gujarat, India for tehnial assistane. This work was fmanially supported by Ministry of nvironment & Forests, Govt. of India, Grant No. 19/17/91-R of Referenes I Venko V, J Hyg pidemiol Mirobiol Immunol (Prague) 29 (1985) Patel R & Saxena A B, Indian J Zool, II (1983) Renfro J L, Shmidt-Nielson B, Miller D, Benos D & Allen J, in Pollution and physiology of marine organisms, edited by F J Vernberg & W B Vern berg (Aademi Press, New York) 1973,11. 4 Lakshmi R, Kundu R & Mansuri A P, Ata Hydrohim Hydrobiol, 18 (199) Thaker J, hhaya J, Nuzhat S, Mittal R, Mansuri A P & Kundu R, Toxiology, 112 (1996) Kundu R, Lakshmi R & Mansuri A P, Pro Aad nviron Bioi, I (1992) I. 7 Kundu R, Lakshmi R & Mansuri A P, Bull nviron ontam Toxiol, 55 (1995) Sokal R R & Rohlf F J, Biometry, (W H Freeman and ompany, San Franiso) 1969, pp 26 9 Gilles R, in Mehanisms of osmoregulation in animals, edited by R. Gilles (Wiley, New York) 1979, Marty L S & Houston A H, J Fish Bioi, 9 (1976), II. II Strik J J TWA, De longh H H, Van Rijn Van Alkemade J W A & Wuite T P, in Sublethal effets of toxi hemial on aquati animals, edited by J H Koeman & J J TWA Strik (lsevier, New York) 1975, Sahana S S, Jana S S, houdhuri N A & houdhury D K, Physiol Bohemoslovaa, 35 (1986), Verma S R, Jain M & Tonk I P, Toxiol Lett (Arnst), 16 (1983), hhaya J, Thaker J, Mittal R, Nuzhat S, Mansuri A P & Kundu R, Indian J Mar Si, 26 (1997) Hseih K, Sue Y, Yau-Huai W, heih-fu & Ru-Dong W, Toxio!., 24 (1986) Needham D M, in The Struture and funtion of musle, edited by G H Bourne, (Aademi Press, New York) 1973, hen Z, oan, Fielding L & assafer G, J Bioi hem, 266 (1991),

EFFECT OF DIFFERENT METHODS OF PRESERVATION ON THE QUALITY OF CATTLE AND GOAT MEAT. Abstract

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