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British Poultry Science Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/cbps20
Pharmacotoxicological aspects of nitrate and nitrite in domestic fowls a
a
a
M. Atef , M. A. M. Abo‐Norage , M. S. M. Hanafy & A. E. Agag
a
a
Department of Pharmacology, Faculty of Veterinary Medicine , Cairo University and Animal Health Research Institute , Cairo, Giza, Egypt Published online: 08 Nov 2007.
To cite this article: M. Atef , M. A. M. Abo‐Norage , M. S. M. Hanafy & A. E. Agag (1991) Pharmacotoxicological aspects of nitrate and nitrite in domestic fowls, British Poultry Science, 32:2, 399-404, DOI: 10.1080/00071669108417365 To link to this article: http://dx.doi.org/10.1080/00071669108417365
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/ terms-and-conditions
British Poultry Science (1991) 32: 399-404
PHARMACOTOXICOLOGICAL ASPECTS OF NITRATE AND NITRITE IN DOMESTIC FOWLS M. ATEF, M. A. M. ABO-NORAGE, M. S. M. HANAFY AND A. E. AGAG Department of Pharmacology, Faculty of Veterinary Medicine, Cairo University and Animal Health Research Institute, Cairo, Giza, Egypt
Downloaded by [Purdue University] at 20:23 13 March 2015
Received for publication 19th April 1990
Abstract 1. The effects of nitrates and nitrites on growth, erythrocytic count, liver and kidney functions, humoral and cell mediated immune responses in cockerels were investigated. 2. Sodium nitrate (4.2 g/kg diet) and sodium nitrite (1.7 g/kg) retarded growth, caused methaemoglobinaemia and changes in erythrocytic count, serum concentrations of glutamic-pyruvic transminase, creatinine and urea. 3. Cockerels given nitrate or nitrite in the food had reduced haemagglutination responses after injection of sheep erythrocytes and a reduced delayed hypersensitivity reaction to purified protein derivative tuberculin following sensitisation to Bacille Calmette Guerin. 4. Nitrates and nitrites are environmental pollutants present in food and water and they may contribute to the aetiology of liver and kidney diseases and problems related to failure of immunity in domestic fowls. INTRODUCTION
The continuous use of nitrogenous fertilisers in agriculture is the major source of nitrates and nitrites which have been shown to be present in a wide variety of food plants and drinking water in relatively high concentrations (Walker, 1975). Nitrates become converted into the much more toxic nitrites in the gastrointestinal tract (Swann, 1975; Mueller et al., 1986). Nitrates react with amines in the food to form potentially cytotoxic and carcinogenic nitrosamines which mediate many of the toxic effects attributed to nitrites (Gilbert et al., 1975; Choi, 1985). Toxicological effects of nitrates and nitrites in different mammalian species are well-documented and include: carcinogenesis, hepatotoxicity, impairment of reproductive functions, endocrine disturbances, growth retardation, destruction of vitamin A, methaemoglobinaemia and impairment of certain defence mechanisms linked to the inflammatory response and tissue injury (Sleight et al, 1972; Swann, 1975; De Saint Blanquat et al, 1983; Prasad, 1983; Choi, 1985; Jahries et al, 1986; Slepchenko and KhnerNitskh, 399
400
M. ATEF, M. A. M. ABO-NORAGE, M. S. M. HANAFY AND A. E. AGAG
1988). Previous investigations in domestic fowls and other avian species were primarily concerned with the effects of nitrates and nitrites on growth and survival (Adams et al, 1966; Arends et al., 1967; Marrett and Sunde, 1968; Adams et al., 1969; Adams, 1974). Thus, while the effects of nitrates and nitrites on mammals have been extensively investigated little is known about their toxic effects in domestic fowls. Reports on field cases of nitrate and nitrite toxicity in poultry farms (Litjens and Eijkelenburg, 1987) and their occurrence in poultry diets (Marrett and Sunde, 1968) have encouraged us to investigate some of the effects of nitrates and nitrites on blood constituents and immunological responses in the domestic fowls.
Downloaded by [Purdue University] at 20:23 13 March 2015
MATERIALS AND METHODS
A slow growing native breed (Balady) was used. Six-month-old cockerels with well-developed wattles were used in the immunological tests. In the other experiments two-months-old cockerels were used. A commercial balanced broiler diet was mixed with 0-05 M sodium nitrate/kg (4-2 g/kg) (NO3 diet) or 0-025 M sodium nitrite/kg (1-7 g/kg) (NO2 diet) or 0-05 M sodium chloride/kg (2-9 g/kg) (control, Cl diet). The different groups of birds were given these diets as described below. Forty-five cockerels were divided into three equal groups and were maintained on the NO3 diet, the NO2 diet or the Cl diet for 4 weeks. Blood samples were obtained from 5 birds from each group by heart puncture at weekly intervals. The blood was collected in heparinised tubes for counting erythrocytes (Natt and Herrick, 1952) and determination of methaemoglobin (Varley et al., 1980) and in plain tubes for separation of serum to determine creatinine, glutamic-pyruvic transaminase (GPT) and urea (Diagnostic kits, BioMerieux, France). Ten birds in each group (mean body weight ± SD = 312 ± 7 g) were weighed at weekly intervals. At the end of the experiment birds were killed and the spleen, bursa of Fabricius, liver and heart were weighed. To establish immune responses to injection with sheep erythrocytes three groups of cockerels (5 birds each) were maintained on the NO3, NO2 or Cl diets for two weeks. The birds were each intravenously inoculated with 0-5 ml of a suspension of 10% loosely packed, washed sheep red blood cells (SRBC) in saline. Feeding of the diets was continued for a further two weeks. Blood samples were obtained, sera were separated and heated at 56°C for 30 min to inactivate complement. Serial double dilutions of serum (25 ß\) were made with barbitone buffer (pH 7-4) in a microtitre plate. Aliquots of 25 fú of a suspension of 10% loosely packed, washed SRBC were added to the dilutions of serum. Plates were left undisturbed for 4 h, then direct haemagglutinin titres were determined (Prescott et al., 1982). Titres were measured as log2 values. To measure delayed hypersensitivity to purified protein derivative three groups each consisting of 8 cockerels were maintained on the NO3, NO2 or Cl diets for two weeks. Five birds in each group were then inoculated intradermally (sensitised) with 0-05 ml of Bacille Calmette Guerin (BCG, 5 mg/ml) (Japan BCG Laboratory, Japan) at the dorsum of the neck. The remaining
401
NITRATE AND NITRITE IN FOWLS
three birds in each group (non-sensitised) were similarly treated but given saline instead of BCG. Feeding of the diets was continued for a further three weeks. The thickness of both wattles was measured using a calliper and each bird was injected with 0-1 ml of tuberculin purified protein derivative (PPD, Egyptian Organisation for Biological Products and Vaccines) into the left wattle and an equal volume of sterile saline was injected into the right wattle. Birds were left for 24 h, then wattle thickness was measured. The proportional increase in the thickness of the right wattle because of the injection of saline (non-specific) was subtracted from that of the left wattle (Prescott et al., 1982).
Downloaded by [Purdue University] at 20:23 13 March 2015
RESULTS
A highly significant reduction (P
British Poultry Science Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/cbps20
Pharmacotoxicological aspects of nitrate and nitrite in domestic fowls a
a
a
M. Atef , M. A. M. Abo‐Norage , M. S. M. Hanafy & A. E. Agag
a
a
Department of Pharmacology, Faculty of Veterinary Medicine , Cairo University and Animal Health Research Institute , Cairo, Giza, Egypt Published online: 08 Nov 2007.
To cite this article: M. Atef , M. A. M. Abo‐Norage , M. S. M. Hanafy & A. E. Agag (1991) Pharmacotoxicological aspects of nitrate and nitrite in domestic fowls, British Poultry Science, 32:2, 399-404, DOI: 10.1080/00071669108417365 To link to this article: http://dx.doi.org/10.1080/00071669108417365
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/ terms-and-conditions
British Poultry Science (1991) 32: 399-404
PHARMACOTOXICOLOGICAL ASPECTS OF NITRATE AND NITRITE IN DOMESTIC FOWLS M. ATEF, M. A. M. ABO-NORAGE, M. S. M. HANAFY AND A. E. AGAG Department of Pharmacology, Faculty of Veterinary Medicine, Cairo University and Animal Health Research Institute, Cairo, Giza, Egypt
Downloaded by [Purdue University] at 20:23 13 March 2015
Received for publication 19th April 1990
Abstract 1. The effects of nitrates and nitrites on growth, erythrocytic count, liver and kidney functions, humoral and cell mediated immune responses in cockerels were investigated. 2. Sodium nitrate (4.2 g/kg diet) and sodium nitrite (1.7 g/kg) retarded growth, caused methaemoglobinaemia and changes in erythrocytic count, serum concentrations of glutamic-pyruvic transminase, creatinine and urea. 3. Cockerels given nitrate or nitrite in the food had reduced haemagglutination responses after injection of sheep erythrocytes and a reduced delayed hypersensitivity reaction to purified protein derivative tuberculin following sensitisation to Bacille Calmette Guerin. 4. Nitrates and nitrites are environmental pollutants present in food and water and they may contribute to the aetiology of liver and kidney diseases and problems related to failure of immunity in domestic fowls. INTRODUCTION
The continuous use of nitrogenous fertilisers in agriculture is the major source of nitrates and nitrites which have been shown to be present in a wide variety of food plants and drinking water in relatively high concentrations (Walker, 1975). Nitrates become converted into the much more toxic nitrites in the gastrointestinal tract (Swann, 1975; Mueller et al., 1986). Nitrates react with amines in the food to form potentially cytotoxic and carcinogenic nitrosamines which mediate many of the toxic effects attributed to nitrites (Gilbert et al., 1975; Choi, 1985). Toxicological effects of nitrates and nitrites in different mammalian species are well-documented and include: carcinogenesis, hepatotoxicity, impairment of reproductive functions, endocrine disturbances, growth retardation, destruction of vitamin A, methaemoglobinaemia and impairment of certain defence mechanisms linked to the inflammatory response and tissue injury (Sleight et al, 1972; Swann, 1975; De Saint Blanquat et al, 1983; Prasad, 1983; Choi, 1985; Jahries et al, 1986; Slepchenko and KhnerNitskh, 399
400
M. ATEF, M. A. M. ABO-NORAGE, M. S. M. HANAFY AND A. E. AGAG
1988). Previous investigations in domestic fowls and other avian species were primarily concerned with the effects of nitrates and nitrites on growth and survival (Adams et al, 1966; Arends et al., 1967; Marrett and Sunde, 1968; Adams et al., 1969; Adams, 1974). Thus, while the effects of nitrates and nitrites on mammals have been extensively investigated little is known about their toxic effects in domestic fowls. Reports on field cases of nitrate and nitrite toxicity in poultry farms (Litjens and Eijkelenburg, 1987) and their occurrence in poultry diets (Marrett and Sunde, 1968) have encouraged us to investigate some of the effects of nitrates and nitrites on blood constituents and immunological responses in the domestic fowls.
Downloaded by [Purdue University] at 20:23 13 March 2015
MATERIALS AND METHODS
A slow growing native breed (Balady) was used. Six-month-old cockerels with well-developed wattles were used in the immunological tests. In the other experiments two-months-old cockerels were used. A commercial balanced broiler diet was mixed with 0-05 M sodium nitrate/kg (4-2 g/kg) (NO3 diet) or 0-025 M sodium nitrite/kg (1-7 g/kg) (NO2 diet) or 0-05 M sodium chloride/kg (2-9 g/kg) (control, Cl diet). The different groups of birds were given these diets as described below. Forty-five cockerels were divided into three equal groups and were maintained on the NO3 diet, the NO2 diet or the Cl diet for 4 weeks. Blood samples were obtained from 5 birds from each group by heart puncture at weekly intervals. The blood was collected in heparinised tubes for counting erythrocytes (Natt and Herrick, 1952) and determination of methaemoglobin (Varley et al., 1980) and in plain tubes for separation of serum to determine creatinine, glutamic-pyruvic transaminase (GPT) and urea (Diagnostic kits, BioMerieux, France). Ten birds in each group (mean body weight ± SD = 312 ± 7 g) were weighed at weekly intervals. At the end of the experiment birds were killed and the spleen, bursa of Fabricius, liver and heart were weighed. To establish immune responses to injection with sheep erythrocytes three groups of cockerels (5 birds each) were maintained on the NO3, NO2 or Cl diets for two weeks. The birds were each intravenously inoculated with 0-5 ml of a suspension of 10% loosely packed, washed sheep red blood cells (SRBC) in saline. Feeding of the diets was continued for a further two weeks. Blood samples were obtained, sera were separated and heated at 56°C for 30 min to inactivate complement. Serial double dilutions of serum (25 ß\) were made with barbitone buffer (pH 7-4) in a microtitre plate. Aliquots of 25 fú of a suspension of 10% loosely packed, washed SRBC were added to the dilutions of serum. Plates were left undisturbed for 4 h, then direct haemagglutinin titres were determined (Prescott et al., 1982). Titres were measured as log2 values. To measure delayed hypersensitivity to purified protein derivative three groups each consisting of 8 cockerels were maintained on the NO3, NO2 or Cl diets for two weeks. Five birds in each group were then inoculated intradermally (sensitised) with 0-05 ml of Bacille Calmette Guerin (BCG, 5 mg/ml) (Japan BCG Laboratory, Japan) at the dorsum of the neck. The remaining
401
NITRATE AND NITRITE IN FOWLS
three birds in each group (non-sensitised) were similarly treated but given saline instead of BCG. Feeding of the diets was continued for a further three weeks. The thickness of both wattles was measured using a calliper and each bird was injected with 0-1 ml of tuberculin purified protein derivative (PPD, Egyptian Organisation for Biological Products and Vaccines) into the left wattle and an equal volume of sterile saline was injected into the right wattle. Birds were left for 24 h, then wattle thickness was measured. The proportional increase in the thickness of the right wattle because of the injection of saline (non-specific) was subtracted from that of the left wattle (Prescott et al., 1982).
Downloaded by [Purdue University] at 20:23 13 March 2015
RESULTS
A highly significant reduction (P
