نوع مقاله : مقاله پژوهشی
نویسندگان
1 جهاد کشاورزی -خرم آباد - ایران
2 گروه علوم دامی، دانشکده کشاورزی، دانشگاه لرستان. خرم آباد. ایران
3 گروه علوم دامی دانشکده کشاورزی دانشگاه لرستان- استان لرستان -خرم آباد
چکیده
کلیدواژه‌ها
موضوعات
عنوان مقاله [English]
نویسندگان [English]
L-carnitine has antioxidant properties and protect the sperm membrane from reactive oxygen substance (ROS). This component reduces the availability of lipids from oxidation. To evaluate the administration of L-carnitine on semen and blood parameters and testicular tissue in aged Ross strain roosters. Forty aged roosters with same weight and age were distributed in four treatments with 10 repetitions in standard cages. The roosters were divided into experimental treatments including control, 100, 250 and 500 mg of L-carnitine per kg of diet. The duration of the experiment was 10 weeks. The length. For habituation to sperm collection used back-abdominal rubbing method. Semen samples were transferred to the laboratory at temperature of 37°C and after initial evaluation, parameters of motility, viability, integrity of sperm plasma membrane and DNA health were evaluated. The serum samples were collected to determine parameters and testicular tissue for histological studies. The rate of motility, viability, membrane integrity and DNA integrity increased in treatments fed with L-carnitine. The maximum integrity was observed in the treatment of 500 mg/kg. The amount of cholesterol and HDL in treatment 4 (500mg) was minimum and maximum respectively, while liver enzyme alanine aminotransferase showed no significant difference in all treatments. The thickness of the germinal layer in seminiferous tubules increased significantly with the addition of 500 and 250 mg of L-carnitine. L-carnitine is able to reduce the effects of oxidative stress in aged roosters. This result can be suggested and used for important breeds that are kept for unusual period.
کلیدواژه‌ها [English]
Extended Abstract
Introduction
Carnitine has an antioxidant property that protects the sperm membrane from free oxygen damage. In addition, carnitine reduces the availability of reactive oxygen species by transferring lipids to mitochondria for beta-oxidation. With this activity, lipids are consumed for production of ATP. The effects of L-carnitine on the reproductive performance of roosters are not precisely known. The effects of L-carnitine on reproductive parameters in humans and pigs were evaluated. In the semen sample of infertile people, the concentration of carnitine is low. It is used as a marker in the epididymal fluid to distinguish between fertile and non-fertile people. The volume of semen in pigs whose diet was supplemented with 720 mg per day was significantly higher than the control group. Although the effects of administration of carnitine to the diet on carcass composition, egg production and chick hatching were evaluated in poultry, the effects of this vitamin on reproductive performance in layer hens, especially aged layers are still unknown. Free radicals have destructive effects on the plasma membrane, so when the cell membrane exposes to reactive oxygen species through the peroxidation of its lipids, this membrane is broken and its activity decreases. Lipid peroxidation occurs when reactive oxygen species overcome or dominate the sperm's antioxidant system. Somatic cells defend against lipid peroxidation with their cytoplasmic enzymes such as superoxide dismutase (SOD) and glutathione peroxidase, while sperm cells release these enzymes during spermatogenesis. Therefore, after being released from the spermatozoa, it suffers from the lack of these enzymes in the reproductive system. Compared to mammals, poultry sperm is rich in unsaturated fatty acids and becomes very vulnerable during processing (transportation and storage). Therefore, lipid peroxidation of semen is a biochemical indicator.The antioxidant effect of carnitine was evaluated in plasma, liver and kidneys of young rats. Adding carnitine to the diet of old rats increased vitamin E and ascorbic acid, as well as glutathione peroxidase and superoxide dismutase activity. The lipid peroxidation in rats treated with carnitine significantly decreased compared to the control group. The increase of ascorbic acid by administration of carnitine in diet of rats can be due to its biosynthesis in the tissue. With the using of L-carnitine ascorbic acid concentration increased and lipid peroxidation decreased. It is assumed that the semen of old roosters will be of better quality with the consumption of carnitine and will be protected from the oxidation of the sperm membrane. The purpose of the present study is to evaluate the effects of oral L-carnitine on the parameters of semen, blood and testicular tissue in aged Ross roosters. Materials and methods In order to carry out this research, 40 cocks were selected. Roosters were placed in a light regime of 16 hours of light and 8 hours of darkness. Roosters were placed in four treatments and ten repetitions. The treatments included the control of other treatments consisted 100, 250 and 500 mg/kg L-carnitine in kg diet. A experimental period was 11 weeks. Back-abdominal rubbing method was used for semen collection. In this study, to evaluate the motility, 10 microliters of each treatment's sperm sample was added to 190 microliters of DMEM culture medium.
Results and Discussion
Sperm motility rate was measured with a CASA system. Eosin-nigrosine staining was used to check sperm viability. Sperm membrane integrity test was performed with the help hypo osmotic solution (Mohammadi, Sharifi, Sharafi, Mohammadi- Sangcheshmeh, et al.). In this experiment, DNA damage was determined by DNA fragmentation kit. In this method DNA sperm was denatured in a micro gel bed under the influence of an acid treatment. In order to evaluate blood parameters, feed was stopped and after than blood samples were collected. Serum was separated and stored at -20◦C. The glucose, creatinine, cholesterol, triglyceride, total protein, ALP, ALT, AST, LDL, HDL were analyzed. To study of the testicular tissue, testis samples were taken and fixed in formalin 10%. Data were analyzed by SAS software, complete random design, and the averages were compared by Duncan's multiple range test. The results of this study showed that the sperm motility in diet supplemented 100 and 250 mg of l-carnitine in old roosters increased significantly compared to the control. Also, in all treatments, the use of L-carnitine (100, 250 and 500 mg/kg of feed) increased the progressive motile sperm, which was consistent with the results of Fattah et al. The amount of 100 and 250 mg decreased the rate of dead sperm. The concentration of HDL and LDL increased and decreased respectively, in roosters whose diets were supplemented with L-carnitine. Probably, by production of steroids in the testis through the increase of HDL, the structural changes of the testicular tissue (increasing the diameter of the seminiferous tubules and the thickness of the germinal layer) can be justified. Carnitine palmitoyl transferase enzyme is involved in beta oxidation of fatty acids and L-carnitine is able to increase the activity of the enzyme.
Conclusion
In this research, the concentration of triglyceride and cholesterol decreased in aged roosters that used L-carnitine. It seems that the decreasing is due to the increase in the activity of carnitine palmitoyl transferase enzyme. The present study showed that L-carnitine in concentrations of 250 and 500 mg/kg in diet can improve sexual characteristics (testis tissue and sperm motility characteristics) in aged roosters. in this regard more research on hormone or gene expression or proteins synthesis is recommended.
Author Contributions
For research articles with several authors, a short paragraph specifying their individual contributions must be provided. The following statements should be used “Conceptualization, X.X. and Y.Y.; methodology, X.X.; software, X.X.; validation, X.X., Y.Y. and Z.Z.; formal analysis, X.X.; investigation, X.X.; resources, X.X.; data curation, X.X.; writing—original draft preparation, X.X.; writing—review and editing, X.X.; visualization, X.X.; supervision, X.X.; project administration, X.X.; funding acquisition, Y.Y. All authors have read and agreed to the published version of the manuscript.” Please turn to the CRediT taxonomy for the term explanation. Authorship must be limited to those who have contributed substantially to the work re-ported.
All authors contributed equally to the conceptualization of the article and writing of the original and subsequent drafts.
In this section, please provide details regarding where data supporting reported results can be found, including links to publicly archived datasets analyzed or generated during the study (see examples). Data available on request from the authors.
If the study did not report any data, you might add “Not applicable” here.
The Acknowledgments section should be a few sentences at the end, but it is important to recognize those people (organizations and individuals) who made considerable impact on the research, provided significant help to the author to formulate and complete the experiment, and improved the research at any stage (from providing access to equipment or field sites to editing the manuscript). However, this is an optional section.
In this section, you can acknowledge any support given which is not covered by the author contribution or funding sections. This may include administrative and technical support, or donations in kind (e.g., materials used for experiments).
The authors would like to thank all participants of the present study.
The study was approved by the Ethics Committee of the University of ABCD (Ethical code: IR.UT.RES.2024.500). The authors avoided data fabrication, falsification, plagiarism, and misconduct.
The author declares no conflict of interest.
The author declares no conflict of interest.
Extended Abstract
Introduction
Carnitine has an antioxidant property that protects the sperm membrane from free oxygen damage. In addition, carnitine reduces the availability of reactive oxygen species by transferring lipids to mitochondria for beta-oxidation. With this activity, lipids are consumed for production of ATP. The effects of L-carnitine on the reproductive performance of roosters are not precisely known. The effects of L-carnitine on reproductive parameters in humans and pigs were evaluated. In the semen sample of infertile people, the concentration of carnitine is low. It is used as a marker in the epididymal fluid to distinguish between fertile and non-fertile people. The volume of semen in pigs whose diet was supplemented with 720 mg per day was significantly higher than the control group. Although the effects of administration of carnitine to the diet on carcass composition, egg production and chick hatching were evaluated in poultry, the effects of this vitamin on reproductive performance in layer hens, especially aged layers are still unknown. Free radicals have destructive effects on the plasma membrane, so when the cell membrane exposes to reactive oxygen species through the peroxidation of its lipids, this membrane is broken and its activity decreases. Lipid peroxidation occurs when reactive oxygen species overcome or dominate the sperm's antioxidant system. Somatic cells defend against lipid peroxidation with their cytoplasmic enzymes such as superoxide dismutase (SOD) and glutathione peroxidase, while sperm cells release these enzymes during spermatogenesis. Therefore, after being released from the spermatozoa, it suffers from the lack of these enzymes in the reproductive system. Compared to mammals, poultry sperm is rich in unsaturated fatty acids and becomes very vulnerable during processing (transportation and storage). Therefore, lipid peroxidation of semen is a biochemical indicator.The antioxidant effect of carnitine was evaluated in plasma, liver and kidneys of young rats. Adding carnitine to the diet of old rats increased vitamin E and ascorbic acid, as well as glutathione peroxidase and superoxide dismutase activity. The lipid peroxidation in rats treated with carnitine significantly decreased compared to the control group. The increase of ascorbic acid by administration of carnitine in diet of rats can be due to its biosynthesis in the tissue. With the using of L-carnitine ascorbic acid concentration increased and lipid peroxidation decreased. It is assumed that the semen of old roosters will be of better quality with the consumption of carnitine and will be protected from the oxidation of the sperm membrane. The purpose of the present study is to evaluate the effects of oral L-carnitine on the parameters of semen, blood and testicular tissue in aged Ross roosters. Materials and methods In order to carry out this research, 40 cocks were selected. Roosters were placed in a light regime of 16 hours of light and 8 hours of darkness. Roosters were placed in four treatments and ten repetitions. The treatments included the control of other treatments consisted 100, 250 and 500 mg/kg L-carnitine in kg diet. A experimental period was 11 weeks. Back-abdominal rubbing method was used for semen collection. In this study, to evaluate the motility, 10 microliters of each treatment's sperm sample was added to 190 microliters of DMEM culture medium.
Results and Discussion
Sperm motility rate was measured with a CASA system. Eosin-nigrosine staining was used to check sperm viability. Sperm membrane integrity test was performed with the help hypo osmotic solution (Mohammadi, Sharifi, Sharafi, Mohammadi- Sangcheshmeh, et al.). In this experiment, DNA damage was determined by DNA fragmentation kit. In this method DNA sperm was denatured in a micro gel bed under the influence of an acid treatment. In order to evaluate blood parameters, feed was stopped and after than blood samples were collected. Serum was separated and stored at -20◦C. The glucose, creatinine, cholesterol, triglyceride, total protein, ALP, ALT, AST, LDL, HDL were analyzed. To study of the testicular tissue, testis samples were taken and fixed in formalin 10%. Data were analyzed by SAS software, complete random design, and the averages were compared by Duncan's multiple range test. The results of this study showed that the sperm motility in diet supplemented 100 and 250 mg of l-carnitine in old roosters increased significantly compared to the control. Also, in all treatments, the use of L-carnitine (100, 250 and 500 mg/kg of feed) increased the progressive motile sperm, which was consistent with the results of Fattah et al. The amount of 100 and 250 mg decreased the rate of dead sperm. The concentration of HDL and LDL increased and decreased respectively, in roosters whose diets were supplemented with L-carnitine. Probably, by production of steroids in the testis through the increase of HDL, the structural changes of the testicular tissue (increasing the diameter of the seminiferous tubules and the thickness of the germinal layer) can be justified. Carnitine palmitoyl transferase enzyme is involved in beta oxidation of fatty acids and L-carnitine is able to increase the activity of the enzyme.
Conclusion
In this research, the concentration of triglyceride and cholesterol decreased in aged roosters that used L-carnitine. It seems that the decreasing is due to the increase in the activity of carnitine palmitoyl transferase enzyme. The present study showed that L-carnitine in concentrations of 250 and 500 mg/kg in diet can improve sexual characteristics (testis tissue and sperm motility characteristics) in aged roosters. in this regard more research on hormone or gene expression or proteins synthesis is recommended.
Author Contributions
For research articles with several authors, a short paragraph specifying their individual contributions must be provided. The following statements should be used “Conceptualization, X.X. and Y.Y.; methodology, X.X.; software, X.X.; validation, X.X., Y.Y. and Z.Z.; formal analysis, X.X.; investigation, X.X.; resources, X.X.; data curation, X.X.; writing—original draft preparation, X.X.; writing—review and editing, X.X.; visualization, X.X.; supervision, X.X.; project administration, X.X.; funding acquisition, Y.Y. All authors have read and agreed to the published version of the manuscript.” Please turn to the CRediT taxonomy for the term explanation. Authorship must be limited to those who have contributed substantially to the work re-ported.
All authors contributed equally to the conceptualization of the article and writing of the original and subsequent drafts.
In this section, please provide details regarding where data supporting reported results can be found, including links to publicly archived datasets analyzed or generated during the study (see examples). Data available on request from the authors.
If the study did not report any data, you might add “Not applicable” here.
The Acknowledgments section should be a few sentences at the end, but it is important to recognize those people (organizations and individuals) who made considerable impact on the research, provided significant help to the author to formulate and complete the experiment, and improved the research at any stage (from providing access to equipment or field sites to editing the manuscript). However, this is an optional section.
In this section, you can acknowledge any support given which is not covered by the author contribution or funding sections. This may include administrative and technical support, or donations in kind (e.g., materials used for experiments).
The authors would like to thank all participants of the present study.
The study was approved by the Ethics Committee of the University of ABCD (Ethical code: IR.UT.RES.2024.500). The authors avoided data fabrication, falsification, plagiarism, and misconduct.
The author declares no conflict of interest.
The author declares no conflict of interest.
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