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Blood of the birds of all the groups was taken on day 17th of the experiment after slaughtering the birds. In this study, Haemoglobin, HCT, MCV, MCH, MCHC, TLC, Heterophil, Lymphocytes, Mononcytes, Eosinophils and Basophils were measured to determine the effect of various doses of ivermectin in broilers on the haematological parameters which is depicted in table-1.
These results revealed, non significant changes in the birds of all the groups indicating that Ivermectin did not affect the blood functioning in the broilers of non medicated and medicated (therapeutic or higher doses) groups. There were no significant differences in the haematological values between the control and experimental groups.
Total leucocyte count (TLC/Âµl) increased non-significantly in Groups B, D, E and F from the control (Group A) after i/v administration of ivermectin. These findings are different from the earlier findings of Kelawala etal.,(1996), who reported non-significant decrease in (TLC) after epidural administration of xylazine in dogs. In this case drug and specie were different from ours.
Our results are in accordance with Sharma et al., (1990), who studied the efficacy of ivermectin againstÂ Ascaridia galliÂ infection in chickens under controlled laboratory conditions in the experiments. The chicks in the treated group were subcutaneously injected with ivermectin at a dose of 0.3 mg kgâˆ’1Â body weight on Day 10. The lower lesion score and post-treatment near-normal haematobiochemical picture in treated birds confirmed these observations. The treated birds also had a better growth rate than the untreated chickens.
This study Talebi et al., (2005) has contributed to the blood profiles of four main broiler strains (Ross, Cobb, Arbor-Acres, and Arian) during 8 weeks husbandry period. Study showed that age affects significantly the haematological profiles of the broiler strains. With increasing of age, the erythrocytic parameters and leukocytic parameters were significantly increased, but MCV, MCH and absolute count of heterophils as well as H/L ratio were significantly decreased. As this study was conducted on various strains of broiler chicks and at 8 weeks of age and we used single strain and less than 8 weeks of age so we have observed no significant change in these parameters after using different concentrations of ivermectin in broilers.
Available information indicates that haematological values of avian species are also significantly influenced by poultry diseases including fowl typhoid, mycoplasmosis, avian coccidiosis, infectious bursal disease, Newcastle disease and toxoplasmosis. (Kokosharov and Todorova, 1987; Branton et al., 1997; Burnham et al., 2003; Koinarski et al., 2001; Panigraphy et al., 1986; Juranova et al., 2001; Galindo-Muniz et al., 2001 and Kaneto et al., 1997). Our data showed no above mentioned diseases in the broilers indicating healthy birds in all the groups.
Rabbits were treated with the Â ivermectinÂ at subcutaneous doses of 0.2, 1 and 2.5 mg/kg for four weeks for measuring someÂ bloodÂ parameters by Ali (1990). At a dose of 0.2 or 1 mg/kg the drug had no significant effect on any of the parameters measured. At a dose of 2.5 mg/kg, the drug reduced significantly theÂ haemoglobinÂ concentration,Â haematocrit,Â mean corpuscularÂ haemoglobinÂ concentration and mean corpuscularÂ haemoglobinÂ on weeks 1, 2, 3 and 4. Platelet counts were not significantly affected. This study is not in agreement with our research as we used single injection of ivermectin in different doses in the broilers in our study instead of rabbits so specie variation may be there and we used treatments once.
EFFECT OF IVERMECTIN ON BLOOD CHEMISTRY IN BROILER CHICKENS
Blood of the birds of all the groups was taken on day 17th of the experiment after slaughtering the birds.
In the this study, Bilirubin, SGPT, SGOT, Total Protein, Albumin, Globulin and A/G were measured to determine the effect of various doses of ivermectin in broilers on the liver function tests.
The results for liver function tests (Bilirubin, SGPT, SGOT, Total Protein, Albumin, Globulin and A/G) revealed, non significant changes in the birds of all the groups indicating that Ivermectin did not affect the liver functioning in the broilers of non medicated and medicated (therapeutic or higher doses) groups. There were no significant differences in serum biochemistry values between the control and experimental groups.
There are two general categories of "liver enzymes." The first group includes the alanine aminotransferase (ALT) and the aspartate aminotransferase (AST), formerly referred to as the SGPT and SGOT. These are enzymes that are indicators of liver cell damage.
The ALT (SGPT) and AST (SGOT) are enzymes that are located in liver cells and leak out and make their way into the general circulation when liver cells are injured. The ALT is thought to be a more specific indicator of liver inflammation, since the AST may be elevated in diseases of other organs such as the heart or muscle. ALT and AST are often used to monitor the course of chronic hepatitis and the response to treatments.
Bilirubin is formed primarily from the breakdown of a substance in red blood cells called "heme." It is taken up from blood processed through the liver, and then secreted into the bile by the liver. Conditions which cause increased formation of bilirubin, such as destruction of red blood cells, or decrease of its removal from the blood stream, such as liver disease may result in an increase in the level of serum bilirubin.Â However, serum bilirubin is generally considered a true test of liver function (LFT), since it reflects the liver's ability to take up, process, and secrete bilirubin into the bile.
The other used indicator of liver function is the serum albumin. Albumin is a major protein which is formed by the liver, and chronic liver disease causes a decrease in the amount of albumin produced. Therefore, in more advanced liver disease, the level of the serum albumin is reduced. (Peter W. Gardner and Stuart Waldstreicher (1995). Gastroenterology Consultants, Report of The American Liver Foundation, 1425 Pompton Avenue, Cedar Grove, NJ).
At therapeutic dose of ivermectin in healthy and diseased dogs, a no significant increase in the levels of alkaline phosphatase and alanine aminotransferase was observed by Qayyum, (2002). These results are in accordance with the results of our study which showed no significant changes in the values of serum biochemistry at different doses of ivermectin in the healthy broilers.
The results are also in line with the results of Uysal and Mahzunlar (1989) who reported that the enzymes (alkaline phosphatase and alanine aminotransferase) activity did not differ significantly when the healthy dogs were treated with therapeutic doses of ivermectin.
Our results are quite similar to the results of Lierz (2001) who injected single dose of ivermectin intramuscularly ranging from 0.2 mg/kg to 11 mg/kg bodyweight in different groups of falcon hybrids (Falco rusticolusÂ xÂ Falco peregrinus). Doses of ivermectin between 0.2 and 5 mg/kg failed to produce clinical signs of illness in the birds.
Light changes in the mean plasma activities of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase (AP) were detected in the group dosed with 5 mg/kg, and higher dosages caused marked changes in these enzymes . These results differ with our results although we have also used avian species in this study but there was no change in the values on different doses which we have given to the birds. This may be the reason that higher doses than those which we have used in our experiment can alter liver functioning in broilers too.
The healthy dogs which were given ivermectin at 10 times higher doses one of the ten dogs had subdermal necrosis which can be compared with the study of some researchers who reported subdermal necrosis at the injection site. One of the dogs which were treated with ivermectin in higher doses (2 mg/kg) showed the allergic reactions. The results of the present study did not coincide with our results although we have used seventeen times more doses of ivermectin and did not see any sub dermal lesions in the birds. It may be due to specie variation
Herrera et al (2002) demonstrated that during the course of experimentalÂ TrypanosomaÂ evansiÂ infection, a gradual decline inÂ serumÂ alkaline phosphatase andÂ albumin, in the infectedÂ coatis (raccoonÂ family), suggested hepatic damage, while the increase in globulin levels might have been due to enhanced antibody production. The high values forÂ ALTÂ andÂ ASTÂ shown by the infectedÂ coatis, in this study, may be related to the hepatic and cardiac lesions demonstrated. As such changes were not observed in the values of serum biochemistry in our study, so we can say that at our doses of ivermectin did not cause any hepatic and cardiac lesions in the broilers.
To the authors' knowledge, no studies have been carried out to investigate the effect of ivermectin on Serum Bilirubin, SGPT, SGOT, Total Protein, Albumin, Globulin and A/G activities in broilers.
The results of the present study did not coincide with these results, because damage to the hepatocytes was not there to become statistically different, which can be due to the breed difference or environmental factor. It is further indicated that ivermectin is a safe choice as it does not cause any significant change in serum biochemistry. It is concluded that ivermectin may be safely used at 0.3, 1, 3 and 5 mg/kg body weight in broilers.
Bilirubin is produced from breakdown of haem in red blood cells. once levels are increased than normal, jaundice can be seen. Causes of a raised plasma bilirubin can be classified as Pre-hepatic - haemolysis, Hepatic viruses, drugs, cirrhosis and Post-hepatic gallstones in common bile duct
ALT is enzyme located in cytosol of liver cells. The levels increased by any condition that damages the liver cells and results in the enzyme leaking out. ALT is the most sensitive marker for liver cell damage.
AST, enzyme is located in cytosol and mitochondria of liver cells. This is less sensitive for liver damage than ALT. The levels elevated in liver disease, myocardial infarction, skeletal muscle damageÂ
Albumin is synthesized in the liver. Low albumin may be due to liver disease, nephrotic syndrome, malabsorption, malnutrition. High albumin: due to dehydrationÂ
In a study done by Awan (2009), different toxic levels (at the dosage rate of 1 mg/kg and 2 mg/kg) of dipyrone were administered intra-muscularly to broiler chicks for four days. The biochemical analysis showed that there was no change in serum uric acid, creatinine, ALT and AST levels in the samples collected at different times during experiment.
Our results are in agreement of the study of Silva (2007). They analyzed blood serum samples of HYBRO PG broilers, collected from 21, 35, and 42-day-old birds, with the aim of establishing normal values of some blood serum parameters. There was no influence of age on total bilirubin and albumin levels. Serum activity of all enzymes changed as a function of bird age. AST serum activity progressively increased with age, which is consistent with the observations of other studies. This is possibly due to the increase of liver metabolism and to the significant muscle development. The differences detected among groups relative to serum enzymes are probably due to physiological changes that are normal in these different ages.
The rabbits were fed raw or processed pigeon pea seed based diets. Values for conjugated and total bilirubin (mg/dl) did not show significant differences (P> 0.05) among rabbits of different treatment groups. Total and conjugated bilirubins are indicators of protein adequacy. An increase in serum glutamic oxalo-acetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT) signal necrosis and myocardial infarctions, which are indicators of poor quality protein of diets fed. As no changes occur in these values in our research study so we can say that there might be no necrosis and myocardial infarctions due to the different doses of ivermectin in all the groups.
The results of our study revealed immunoenhancement in highly medicated birds of Group F. Lymphocyte hypersensitivity response, in terms of skin thickness was significantly high in group F. The inflammatory response against DNCB was much higher at highest dose of ivermectin after 24 and even after 72 hours of sensitization. The response of DNCB remained elevated in birds of highest dose of ivermectin after 24 and 72 hours may be due to its immunopotentiating action, because the resolution of inflammation is directly proportional to the reduced number and activity of immune cells. An other reason that may explain the sustained skin thickness during 24-72 hours in the highest dose of DNCB inoculation may be that, heterophils have proteases and hydrolases within granules and if released, would cause damage to the extracellular matrix, promote chemotaxis, the apoptosis of down-regulated and switched off heterophils might have increased inflammation in that area.
The mean skin thickness values in birds of Group F (Highly medicated) was significantly (P<0.05) higher than in birds of all the groups. Skin sensitivity at 24 and 48 hrs. was highest in the birds administered with 5 mg/kg body weight (the highest dose) of ivermectin.
At 16th day (48 hrs.) postchallenge, the birds of Group F showed prolonged effect of drug on skin thickness.
At 17th day (72 hrs.) postchallenge, the skin thickness was more as compared to the other groups but less as compared to the previous values in the same group, indicating that the effect of drug was decreased but still present as compared to the other groups.
Our results are in accordance with Tiwary and Goel 1985. They reported that cutaneous hypersensitivity developed slowly, reaching its maximum manifestation after 24 h of challenge and gradually declining thereafter.Â The development of a skin reaction was typical of delayed type hypersensitivity and was characterized histopathologically by congestion, oedema, mononuclear and heterophilic cell infiltration in the dermal layer and lymphocytic perivascular cuffing.
IvermectinÂ has a dose-dependent pharmacokinetics inÂ humans,Â dogsÂ and redÂ deerÂ resulting in linear increase inÂ blood plasmaÂ concentrations with increase in dose (Krishna and Klotz, 1993). So, theÂ immune responseÂ to antigen may also vary with variableÂ blood plasmaÂ concentrations ofÂ ivermectin. The ability of an individual to develop contact sensitivityÂ is a measure of cellular immunity to a new antigen to which it has not been exposed previously (Stites, 1978). Delayed-typeÂ hypersensitivity reactionÂ was determined byÂ DNCB, a highly reactive substance which can form dinitrophenylÂ protein complexesÂ with variousÂ skinÂ proteins. Krishna and Klotz, 1993Â D.R. Krishna and U. Klotz, Determination of ivermectin in human plasma by high performance liquid chromatography,Â Arzneimittel-ForschungÂ 43Â (1993), pp. 609-611.Â
Stites, 1978Â Stites, D.P., 1978. Clinical laboratory methods of detecting cellular immune functions. In: Fudengergh, H.H., Stites, D.P., Caldwell, J.L., Wells, J.V. (Eds.), Basic and Clinical Immunology, second ed. Lange Medical Publications, Maruzen Company Ltd., New York, U.S.A., pp. 375-388.
In a study done by Sajid et al 2007, on rabbits, the cell mediated immunity against dinitrochlorobenzene (DNCB) was determined by delayed-type hypersensitivity The skin sensitivity to DNCB at 24 and 48Â h was highest (PÂ >Â 0.05) in rabbits administered with 600Â Î¼g/kg b.w. A graded dose immune response suggested an immunopotentiating effect of ivermectin at higher doses. This is inaccordance with the mean skin thickness values in birds of Group F (Highly medicated) in our study which is significantly (P<0.05) higher than in birds of all the groups. Skin sensitivity at 24 and 48 hrs. was highest in the birds administered with 5 mg/kg body weight (the highest dose) of ivermectin.
Similar results were shown by Munir et al 2009. They reported that skinÂ thickness and lymphoproliferation of salinomycin medicated chicks wereÂ significantly greater (PÂ < 0.05) than those of levamisole, cyclophosphamide and cyclosporine treatedÂ chicks. Thet concluded that salinomycin had beneficialÂ effects on the cell mediated immune responses of broiler chicks against hydropericardium syndrome virusÂ and NewcastleÂ disease.
The results of this study contradict with the findings of Kadian et al 1988. They observed that 0.3 ppm dietry aflatoxin B 1 significantly suppressed the
Cell mediated immunity of chickens as the delayed type hypersensititivity response at 15, 30 and 45 days of age of aflatoxin B 1 fed chickens to Dinitrofluorobenzene.
Weights of all the birds of all the groups were taken on day 1st of the experiment and on day 17th (last day) of the experiment.
The results indicated, non significant difference in all the birds of all the groups indicating that Ivermectin did not cause any negative effect on weight gain on higher doses in the broilers. Although the birds of Groups D, E and F showed less weight gain as compared to the birds of Group A (Normal) but it was non significant. This result of the study contradict with the fidings of Miller R. W. (1990). He reported that if chickens were fed ivermectin at a level of 2 ppm for 5 wk to determine its efficacy against the lesser mealworm, Alphitobius diaperinus, the feeding of ivermectin may reduce body weights, body weight gain and feed efficiency at older ages. But in our study, single application of sub cutaneous injection of ivermectin in each bird was used.
Our results are in accordance with the results of Smith et al (1987) who evaluated the effect of a single injection of ivermectin in mid summer in a herd of cows and calves. Ivermectin treatment icreased lactating cow weight by 23 lbs and improved calf gains by 21 lbs. Weight change in dry cows was not affected by treatment. Body condition score was significantly improved in lactating cows that were treated with ivermectin.
Quiroza et al (2003) determined the difference on the weight gain and the reduction of eggs of gastrointestinal nematodes (GIN)) in grazing weaning calves treated with ivermectin. Group 1, was treated with ivermectin at 200 mcg/kg/sc, and the Group 2, was the control, treated only once with albendazole at the beginning of the study. The difference on the average
weight gain was 30.8 kg per calf (more) versus the control (Pâ‰¤0.05). But in our case the specie variation was there, as they used calves and we used broilers in our study.