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Haematobiochemical Alterations of Oral Acute Profenofos Toxicity in Broiler Birds

Arjun Kafle Dulal C Roy Jadav Sarma Satya Sarma Rajkishor Gogoi
Vol 7(3), 123-128
DOI- http://dx.doi.org/10.5455/ijlr.20170303025518

The present study was carried out to evaluate the acute effect of profenofos, in broiler chicken in relation to haematological and biochemical alterations. Twenty day old chicks were divided into two groups (Group I and Group II), ten in each group. Group I was kept as control, while Group II was treated with 16 mg profenofos (LD50) once orally. Blood was collected periodically from 0 to 36 hours to estimate haematological (Hb, TEC, TLC, Lymphocyte and heterophil count) and biochemical (ALT, AST, ALP, Uric acid, Total cholesterol and cholinesterase) changes. The level of all haematological parameters tested was found to be significantly higher except the lymphocyte count. Under biochemical estimation the level of cholinesterase was significantly lower while the rest showed significant elevation. The results indicated that the profenofos cause significant changes in both haematological and biochemical parameters of the birds under study.


Keywords : Acute Profenofos Broiler Haematological Biochemical

Introduction

Organophosphate (OP) group of insecticides are widely used for the control of agriculture and household pests (IPCS, 1990). Organophosphate ranks top in the list of pesticides. They act by inhibiting cholinesterase, an enzyme required for the destruction of acetylcholine and hence termination of action. Profenofos is one of the recommended insecticides and preferred over the conventionally employed toxic chemical pesticides (Venkateswara Rao et al., 2003). Pesticides used against agricultural pests and ecto- parasite infestation in animals may cause adverse side effects in farm animals (Maffini et al., 2006). Irrational use of such pesticide can make its entry into the food chain. Profenofos has been registered to be use in one of the most common feed ingredient in broiler farms i.e., maize. The studies on effect of profenofos is limited to mammals and fishes mostly and since it forms the main ingredient in broiler ration the present study was designed to determine the effect of profenofos on the haematology and biochemical parameters of the said subject.

Material and Methods

Subjects (Birds)

Twenty unsexed day old chicks, divided into two groups, Group I as control and Group II as treatment group. The birds were procured from poultry corner, Khanapara, Guwahati- 781022. Wing band was fixed for ease of identification and reared in Instructional Poultry Farm (IPF), CVSc, Khanapara. They were housed in deep litter system, with ad libitum supply of feed and water. The birds were kept in the farm for 7 days before the experimentation for acclimatisation. The experimental trials were approved by the Institutional Animal Ethics Committee (No.770/ac/CPCSEA/FVSc/AAU/IAEC/15-16/356).

Chemical

Profenofos under the brand name, current (89 % purity), procured from Hazipur, India was used for this study.

Experimental Design

The test compound was administered to birds of group II @ 16 mg/kg orally using oral gavage needle. The dose selected was the LD 50 which was obtained after conducting pilot study. Blood was collected from jugular and wing vein at 0 hr, 3 hrs, 6 hrs, 12 hrs, 24 hrs and at 36 hrs. One part of the collected blood was used for estimation of haematological parameters (Haemoglobin, Total Erythrocyte Count , Total Leucocyte Count, Lymphocyte Count and Heterophil Count) using automated haematology cell counter (Model : MS4S, Sartorius) while the other part was used for serum biochemical analysis namely, aminotransferase (alanine amino transferase and aspartate amino transferase), alkaline phosphates, total cholesterol, uric acid and cholinesterase as per the protocol mentioned in the kit provided by Keediagnostics.

Statistical Analysis

Statistical analysis for the experimental data was performed by one way ANOVA using SPSS 16.0. Values were described as mean ± SE. P value less than 0.05 was considered statistically significant.

Result

Haematological Alterations

The effect of profenofos on haematological profile is shown in Table 1 and Table 2. The Hb level and heterophil count was found to be increased from 6 hrs onwards till 36 hrs in treated group as compared to the control. Significant (P<0.01) increased was observed from 12 hrs onwards in treated group in relation to the control. Data in Table 1 depicts the TEC level to be subsequently increased from 3 hrs onwards till 36 hrs in group II. The statistical analysis revealed that there was significant (P <0.05, P<0.01) increase of TEC from 12 hrs in treated group while comparing to the control. The TLC shows increasing trend than the control group (Table 1). Significant (P <0.05,) increase of TLC from 6 hrs onwards between control and treated group was noticed on statistical analysis. Lymphocyte percent, however showed decreasing trend than the control. The reduction in the percent of lymphocyte was found to be highly significant (P<0.01) from 6 hrs onwards in treated group as compared to the control.

Table 1:

Haematological

Parameters

Time of blood collection (hours)

Group I (Control)

0 3 6 12 24 36
Hb (g/dl) 9 .67±0.22 9.70±0.34 9.80±0.13 9.73±0.31 9.77±0.21 10.02±0.35
TEC (106/µl) 1.99±0.25 2.34±0.29 2.39±0.32 2.42±0.32 2.51±0.17 2.52±0.15
TLC (103/µl) 27.91±0.55 27.74±0.54 25.75±0.32 26.73±0.36 25.74±0.18 27.26±0.36
Lymphocyte (%) 85 .85±0.39 85.26±1.37 84.80±0.34 87.13±0.36 85.17±0.36 86.07±0.50
Haterophil (%) 4.41 ± 0.25 4.65 ± 0.31 4.34 ± 0.49 4.23 ± 0.47 4.16 ± 0.42 3.34 ± 0.29

Table 2:

Haematological Parameters Time of blood collection (hours)

Group II (Treatment)

0 3 6 12 24 36
Hb (g/dl) 9.92±0.20 9.67±0.51 10.53±0.51 11.35±0.37** 12.10±0.31** 13.00±0.46**
TEC (106/µl) 2.34±0.49 2.62±0.21 2.83±0.20 2.96±0.25 3.68±0.34* 5.89±0.18**
TLC (103/µl) 26.76±0.45 27.17±0.35 28.64±0.69* 33.11±0.66** 30.16±0.16** 34.52±0.27**
Lymphocyte (%) 84.00±1.03 80.67±0.33 78.11±0.34** 83.97±0.48** 80.56±0.40** 76.45±0.64**
Haterophil (%) 4.2 ± 0.38 4.2 ± 0.34 4.4 ± 0.40 7.4 ± 0.41** 8.8 ± 0.36** 13.7 ± 0.32**

*(P <0.05); ** (P<0.01)

Biochemical Study

The effect of profenofos on biochemical parameters is shown in Table 3and Table 4. The ALT level was subsequently increased from 3 hrs onwards till 36 hrs in treated group relative to the control. Statistical analysis showed highly significant (P <0.01) difference from 3 hrs to 36 hrs between treated and control groups. The AST level was shown to increase from 3 hrs onwards till 36 hrs in treated group as compared to the control. The mean values of AST significantly increased (P <0.01) from 6 hrs onwards till the end of the experiment. The ALP level was found to be increased gradually from 3 hrs onwards till 36 hrs in treated group as compared to the control. As shown in table 4 the mean values of Serum Alkaline Phosphates increased significantly (P <0.05, P<0.01) from 3 hr onwards between control and treated group. The CHE level was subsequently decreased from 3 hrs onwards till 36 hrs in treatment group as compared to the control. The mean values of CHE showed significant (P <0.05, P<0.01) inhibition from 3 hrs to 36 hrs between treated and control groups. TC was found to be increased from 3 hrs onwards till 36 hrs in treated group as compared to the control. Statistical analysis showed significant (P <0.05, P<0.01) increase from 3 hrs to 36 hrs between treated and control groups. The uric acid level was gradually increased from 3 hrs onwards till 36 hrs in treated group as compared to the control. Statistical analysis of the mean uric acid level between the groups showed significant (P <0.05, P<0.01) increase from 12 hrs to 36 hrs between treated and control groups.

Table 3:

Biochemical parameters Time of blood collection (hours)

Group I (Control)

0 3 6 12 24 36
ALT (IU/L) 25.96±0.60 25.49±0.63 27.54±0.60 26.46±0.77 26.51±0.76 27.48±0.61
AST (IU/L) 51.29±0.32 52.10±0.23 52.19±0.30 52.64±0.61 53.08±1.71 52.47±0.68
ALP (IU/L) 108.56±2.04 109.11±2.63 108.62±1.13 108.98±1.58 108.43±1.30 108.77±2.74
CHE (IU/L) 453.92±1.53 453.65±0.58 453.06±0.52 453.06±0.57 453.62±0.56 453.69±0.63
TC (IU/L) 148.65±0.26 148.28±0.56 148.22±0.67 148.74±1.21 148.88±1.21 147.65±0.47
UA (IU/L) 8 .61±0.17 7.87±0.27 6.54±0.55 7.43±0.17 6.28±0.16 5.54±0.17

Table 4:

Biochemical parameters Time of blood collection (hours)

Group II (Treatment)

0 3 6 12 24 36
ALT (IU/L) 25.21±0.59 32.83±1.01** 37.18±1.25** 48.74±0.89** 60.50±0.90** 83.82±0.75**
AST (IU/L) 51.36±0.29 54.79±3.64 64.98±0.32** 79.56±0.33** 97.08±1.25** 124.85±0.37**
ALP (IU/L) 100.69±3.21 154.67±2.23* 205.45±0.77* 278.05±1.81** 354.23±0.96** 398.55±1.81**
CHE (IU/L) 455.27±0.87 443.00±2.97* 436.81±4.05* 434.22±4.68* 400.46±0.61** 350.79±0.53**
TC (IU/L) 149.01±0.71 152.87±1.51* 169.38±3.42** 176.99±1.31** 175.20±1.22** 196.87±0.89**
UA (IU/L) 8.12±0.28 9.73±1.08 13.53±1.86* 14.94±0.76** 20.90±0.15** 23.22±0.28**

*(P <0.05); ** (P<0.01)

Discussion

The effect of profenofos on both haematological and biochemical parameters was found to be progressing over time. The level of Haemoglobin and TEC was found to be increased in treated group relative to the control. On the contrary, some author (Ezhilmathy et al., 2014) reported the Hb concentration to be reduced (due to anaemia) from 6.442±0.052 to 3.386±0.069 g/dl at 96 hrs while studying the toxicity of mixture of heavy metal (lead nitrate) and profenofos against the estuarine fish Lates calcarifer. The increased in Hb and TEC in the present study was might be due to diarrhoea causing dehydration leading to haemoconcentration (Yadav et al., 2003). The level of TLC was also found to be significantly higher in treated group suggesting similar findings while studying the impact of profenofos in fishes studied for duration of 96 hrs which showed the increase in total leukocyte count from 682.1 ± 5.1 in control to 937.4 ± 3.7 103/mm 3in treatment group (Khalid et al., 2015). Toxicants are found to cause stress in birds and it could be the reason for low lymphocyte count in this study. Organophosphate exerts its toxic effect on lymphoid tissue resulting in lymphopenia (Coles, 1980). Increase in the levels of aminotransferase (ALT and AST), ALP and total cholesterol in the serum were observed which could be the major diagnostic symptoms of liver diseases (Chatterjea and Shinde, 2005). Any changes in the hepatocytes which cause disruption of the cell membrane leads to release of aminotransferase in the blood (Manal et al., 2008). This increment of enzymes may be an indicator of liver damage. Our result is in agreement with many workers (Rahman and Siddiqui, 2003; Rezg et al., 2008). Liver helps to take up the excess cholesterol and its dysfunction results in the elevation of the latter in the circulating blood. The elevation of uric acid in the current study could be suggestive of renal function impairment as uric acid is an end product of metabolic breakdown of purine nucleotides, which is filtered through kidneys and is finally excreted as a normal component in urine of birds. Any factor that reduce glomerular filtration rate (GFR) will cause an increase in serum uric acid level. The test compound, profenofos causes irreversible inhibition of cholinesterase enzyme which indirectly increases the level of acetylcholine in the nerve endings, Hence the level of cholinesterase in the serum was found to be reduced in the present study. The significant inhibition of serum cholinesterase in the present study might be attributed to the irreversible phosphorylation of the hydroxyl group located in the active site of the molecule (Kousba et al., 2004). Profenofos cause significant decrease in the level of cholinesterase in serum when fed chlorpyrifos (32 mg/kg) to indigenous birds (Begum et al., 2015).

Conclusion

It can be concluded from the present study that profenofos at the dose rate of 16 mg/kg body has the potential to cause toxicity in broiler birds which was shown by significant changes in haematological as well as biochemical parameters so public awareness should be raised to prevent irrational use of profenofos in agricultural field.

Acknowledgement

The authors acknowledges the help received from the Dean, College of Veterinary Science, Assam Agricultural University, Khanapara for providing necessary facilities to carry out the research work successfully.

References

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