Evaluation of biochemical alterations produced by combined exposure of fenvalerate and nitrate in Bubalus bubalis

Aim: Evaluation of combined effect of fenvalerate and nitrate on biochemical parameters in buffalo calves. Materials and Methods: Sixteen male buffalo calves were divided into four groups of four calves each. Group I receiving no treatment served as the control. Group II and III animals were orally administered with fenvalerate (1.0 mg/kg/day) and sodium nitrate (20 mg/kg/day), respectively, for 21 consecutive days and were kept as positive control. Group IV animals were co-administered with fenvalerate and sodium nitrate at the above dose rates for 21 consecutive days. Biochemical parameters including Aspartate aminotransferase (AST), Alkaline phosphatase (ALP), Gamma-glutamyl transpeptidase (GGT), Lactate dehydrogenase (LDH), Glucose, Total protein, Albumin, Cholesterol, Blood urea nitrogen (BUN) and Creatinine were th determined on 0, 3, 7, 10, 14, 17 and 21 day of treatment. Estimation of these parameters was also done on 7 day of posttreatment period. Results: Co-administration of fenvalerate and sodium nitrate produced significant increase in the plasma levels of AST, ALP, GGT, LDH, glucose, BUN, cholesterol and creatinine while significant decrease in the plasma levels of total proteins was observed. No significant alteration was observed in albumin levels. Extent of organ damage as evidenced by biochemical alterations was more pronounced in calves exposed to combination of fenvalerate and sodium nitrate as compared to their individual exposures. Conclusion: Fenvalerate and sodium nitrate co-administration potentiates the toxicological injury produced, in comparison to their individual exposure.


Introduction
quently lead to ecosystem pollution by the accumulation of nitrates in vegetables and fodder as well as the Pesticides have played a pivotal role in bringing contamination of surface and ground water [4].about green revolution in the world and have evolved Therefore, nitrates are a major threat to environment in as an irreplaceable component for the control of different agricultural situations and a potential health agricultural pests and insects causing public health risk for humans and animals [5].Ruminants are more hazards.Several health ailments and/or outbreaks are susceptible to nitrate poisoning than non-ruminant reported to occur among animals and humans from species.Several clinical studies have documented insecticide toxicity, which mostly occur either from renal and/or hepatotoxicity in humans associated with direct exposure to insecticides or indirectly from excess nitrate intake [6].However, relatively few contaminated feeds or water.Eventhough the average studies have assessed the degree of hazard posed by utilization of pesticides in India is considered to be simultaneous exposure to pesticides and nitrate [7].much lower than many other developed countries, the Humans and animals can be exposed concurrently problem of pesticide residues is relatively high [1].
to more than one chemical in the environment.Such Pyrethroides, derivatives of carbamic acid, represent a interactions may be deleterious, as both the kinetics large variety of compounds which have some field and dynamics of the environmental chemicals can be applications as insecticides, herbicides and fungicides.
modified by their co-occurrence [8].Biochemical Fenvalerate is an insecticide of moderate mammalian biomarkers are increasingly used in ecological risk toxicity which is used against a broad range of pests [2] assessment to identify the incidence and effects of and is reported to cause alterations in the biochemical environmental pollutants [9].In domestic animals and parameters in different tissues of rats [2,3].
humans, simultaneous exposure to fenvalerate and nitrate Excessive use of nitrogenous fertilizers subsecan lead to various health afflictions and tissue/ organ damage which can be assessed by measuring levels of various biochemical enzymes and other parameters indicative of specific organ damage.In India, buffalo is an important dairy animal which contributes about buffalo calves to resemble the levels of the insecticide 70% of the total milk production.
to which dairy animals are likely to be exposed.The The present research investigation was therefore dosage of sodium nitrate was selected on the basis of conducted with the primary aim to elucidate the previous literature as similar dose was used in a study interactive effect of fenvalerate and sodium nitrate on conducted by Shahid Mahboob et al [10] in rabbits for a biochemical parameters in buffalo calves.
period of 40 days to study haematological parameters.

Materials and Methods
Chemicals: Fenvalerate (Reagent®, 20% EC) insecticide was commercially obtained from Bharat Insecticides Ethical approval: The experimental protocol followed Limited, New Delhi.Sodium nitrate (analytical grade) the ethical guidelines on the proper care and use of was procured from Merck Specialiaties Private animals and had been approved by the Institutional Limited, Mumbai.Animal Ethics Committee.

Estimation of biochemical parameters:
To study the Animals: The experiment was performed on sixteen biochemical parameters, blood samples were collected healthy male buffalo calves of 6-12 months age and in heparinized vials from the jugular vein of animals on weighing between 100-130 kg, kept under normal 0, 3, 7, 10, 14, 17, 21 day of treatment and 7 day post ambient conditions in the experimental animal shed of treatment.Plasma was separated by centrifugation at the department.During this period, all animals were 3000 rpm for 15 min.Biochemical parameters including dewormed and subjected to regular clinical examination.
Lactate dehydrogenase (LDH), Glucose, Total protein, Albumin, Cholesterol, Blood urea nitrogen (BUN) and Experimental design: The animals were randomly Creatinine were estimated using Bayer Autopack kits divided into four groups of 4 animals each and their (Bayer Diagnostics India Ltd., India).baseline biochemical values were determined.Group I serving as control, was administered with 50 ml Statistical analysis: All results were subjected to distilled water.Group II animals were orally adminianalysis of variance carried in completely randomized stered with fenvalerate at 1.0 mg/kg/day for 21 consecutive design and the significance was tested using Duncan's days, whereas sodium nitrate at 20 mg/kg/day was multiple range test [11].The significance was assayed administered to group III animals.Group IV animals at 5 % (p<0.05)levels.These statistical calculations were administered both fenvalerate and sodium nitrate were carried out with SPSS 16.0 software.at the same dosages and for the same duration as group

Results and Discussion
II and III, respectively.The requisite amount of insecticide and nitrate was suspended in 50 ml of The results for the combined effect of fenvalerate distilled water and drenched to animals of subsequent and sodium nitrate on AST, ALP and GGTP are groups.The daily oral dose of fenvalerate was selected presented in   [20].Studies relating to the th significant increase in plasma glucose levels from 14 effects of fenvalerate and nitrate in combination have day onwards.Maximum increase of 60.6% was observed not been documented.The liver plays an important role st on 21 day of treatment which was comparatively in metabolism to maintain energy level and structural higher than that observed in fenvalerate (39.9%) and stability of body.It is also the site of detoxification sodium nitrate (53.1%) exposed groups.Fenvalerate where toxic compounds get biotransformed into less and sodium nitrate co-exposure in buffalo calves harmful products to reduce toxicity [21].However, produced significant decrease in plasma protein levels these toxic compounds damage the liver cells and th th on 7 day and then 14 day onwards.Maximum decline produce hepatotoxicity.High plasma concentrations of st of 29.2% was observed on 21 day of treatment which ALT and AST are considered as an index of liver was comparatively higher than fenvalerate (20.5%) damage [22].An increase of aminotransferase activity and sodium nitrate (7.69%) exposed groups.No signiin the extracellular fluid or plasma is a sensitive indicator

ficant alteration was observed in albumin levels in any
present study could either be due to impairment of of the treated groups.
protein synthesis or due to increased protein catabolism The blood sugar level is an important indicator of following combined exposure to toxicants.on-going body homeostatic mechanisms.The blood The effect produced by combined exposure of sugar levels are tightly regulated by the body, but fenvalerate and sodium nitrate on cholesterol, BUN certain metabolic and toxic conditions cause the levels and creatinine are presented in th damage which can also be the contributing factor for An increasing trend in BUN levels from 10 day hyperglycemia since the liver is an important site for onwards was observed following combined exposure metabolism of glucose.Decrease in plasma protein to fenvalerate and sodium nitrate.Maximum increase st levels are usually seen in chronic liver diseases due to of 90% was observed on 21 day of combined treatment impaired synthesis and in renal diseases due to loss of of toxicants which was comparatively higher than that proteins.The declining protein levels of plasma in the of fenvalerate (64.8%) and sodium nitrate (50.4%)Gill et al. [34] reported that combined exposure to assessment of pesticide contaminated environment.Afr.J.

Table -
fodder and average daily consumption of fodder by Maximum increase of 42.8% was observed in AST

Table - 1
[15]fect of [18]acute oral administration of fenvalerate (1mg/kg/day) and sodium nitrate (20mg/kg/day) combination on plasma aspartate aminotransferase, alkaline phosphatase and gamma-glutamyl transpeptidase in buffalo calves.The increase in plasma LDH activity in present Several studies have documented the biochemical investigation reflects damage to a range of tissues alterations on fenvalerate [2,3,12-14] and nitrate exposure including skeletal or cardiac muscles, kidney and liver.[15][16][17][18]indifferent species of animals.Increases in the The effects produced by fenvalerate and sodium activity of phosphatases and aminotransferases have nitrate co-administration on glucose, total protein and been reported with pyrethroids [19].Administration of albumin levels are shown in Table-2.Combined either nitrate or fluoride or their combination also elevated exposure to fenvalerate and sodium nitrate produced phosphatase activity in rats [24]ues given are mean ± SE, n= 4*, *Means with at least one common superscript do not differ significantly (p<0.05).Group I -Control, II -Fenvalerate, III -Sodium nitrate, IV -Fenvalerate and Sodium nitrate combination.st of cellular damage[23].Alkaline phosphate is a zinclevels on 21 day of treatment.Although there was containing enzyme[24]which is secreted in liver, significant increase in AST levels in animal groups biliary tracts, small intestines, bones, lungs and kidney.treated individually with either fenvalerate or nitrate, Elevated levels in blood may indicate damage to the the percent increase following combined exposure of liver and other organs due to non-specific irritation [8].toxicants was comparatively greater than the corres-Although the exact cause of increased GGTP level in ponding values for individual exposures.The levels of the present study could not be ascertained, cholestatic plasma AST in fenvalerate-nitrate interaction group disorders of all species are associated with increased varied significantly from those of fenvalerate and nitrate

Table - 2
. Effect of sub-acute oral administration of fenvalerate (1 mg/kg/day) and sodium nitrate (20 mg/kg/day) combination on plasma glucose, total proteins and albumin in buffalo calves.Values given are mean ± SE, n= 4*, *Means with at least one common superscript do not differ significantly (p<0.05).Group I -Control, II -Fenvalerate, III -Sodium nitrate, IV -Fenvalerate and Sodium nitrate combination.

Table - 3
. Effect of sub-acute oral administration of fenvalerate (1 mg/kg/day) and sodium nitrate (20 mg/kg/day) combination on plasma cholesterol, blood urea nitrogen and creatinine in buffalo calves.
[32]ration[32].Increase in plasma BUN and creatinine 4. Testud, F., (2005) Inorganic fertilizers.EMC Toxicol.levels in present study are indicative of renal damage, Pathol., 1: 21-28.which may be attributed to urinary obstruction that 5. Kaya, S. and Akar, F. (2002) Inorganic Substances In: Toxicology in Veterinary Medicine, Kaya S, Pirincci I and potentiates decreased secretion of urea from the body.Bilgili A. (eds), Second Edition.Ankara Medisan Publishing Greater elevation of BUN and creatinine levels on mixtures at ground water concentrations.Toxicol.Ind. parameters.Consistent to our findings, Dubey et al. [8] fluoride.In another study, Sodium fluoride and delta-