The effects of active and inactivate Saccharomyces cerevisiae and their combination on performance, antioxidant levels and lipid peroxidation index in the blood of Japanese quail
Subject Areas : Veterinary Clinical Pathologyحسین نیک پیران 1 , حامد منافی 2 , توحید وحدت پور 3
1 - گروه علوم درمانگاهی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران
2 - فارغ‌التحصیل دکتری عمومی دامپزشکی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران
3 - گروه علوم دامی، دانشکده کشاورزی و دامپزشکی، واحد شبستر، دانشگاه آزاد اسلامی، شبستر، ایران
Keywords: Performance, Malondialdehyde, total antioxidant capacity, Japanese quails, Peroxidation,
Abstract :
Considering that various effects of probiotics, prebiotics, and symbiotic on production and growth of broiler chicks has been reported previously, the aim of this study was to investigate the effects of probiotics and prebiotics and their mixture, on performance, antioxidant levels and lipid peroxidation index in the blood of Japanese quails. Thus, 192 day- old- Japanese quails were distributed randomly to 4 groups with 4 replicates of 12 male and female and evaluated for 42 days. The control group only received basal diet, but in the 2nd group (probiotic group) active saccharomyces cerevisiae 1 gr/Kg of feed, in the 3rd group (prebiotic group) inactivated saccharomyces cerevisiae 0.5 gr/Kg-feed of feed, and in the 4th group (synbiotic group) 0.5 gr/Kg saccharomyces cerevisiae and 250 mg/Kg-feed inactivated saccharomyces cerevisiae was added to the basal diet. Results indicated that mean serum levels of Malondialdehyde (MDA) was statistically different among males of different groups (p<0.05). Also the highest and lowest MDA levels were observed in the 4th and 3rd groups respectively with a statistically significant difference (p<0.05). Total antioxidant capacity (TAC) of plasma was not different statistically in the studied groups. On the other hand, mean feed consumption and body weight in the 2nd and 3rd groups was increased in comparison to control group and FCR was decreased significantly in comparison to control and synbiotic group. The best carcass efficiency was observed in the prebiotic group. The results showed that the employed probiotics and prebiotics were only effective in performance of Japanese quails, but did not have a specific effect on serum-MDA and plasma-TCA levels.
References:
Aluwong, T., Kawu, M., Raji, M., Dzenda, T., Govwang, F., Sinkalu, V., et al. (2013). Effect of yeast probiotic on growth, antioxidant enzyme activities and malondialdehyde concentration of broiler chickens. Antioxidants, 2(4): 326-339.
Ao, X., Yoo, J., Zhou, T., Wang, J., Meng, Q., Yan, L., et al. (2011). Effects of fermented garlic powder supplementation on growth performance, blood profiles and breast meat quality in broilers. Livestock Science, 141(1): 85-89.
Asadi, G., Ebrahimnejad, Y., Nazeradl, K. and Ahmadzade, A. (2010). Impact of the size of corn on the weight and some internal organs of japanese quail. Fifth Animal Science Congress, pp: 57-64.
Awad, W., Ghareeb, K., Abdel-Raheem, S. and Böhm, J. (2009). Effects of dietary inclusion of probiotic and synbiotic on growth performance, organ weights, and intestinal histomorphology of broiler chickens. Poultry Science, 88(1): 49-56.
Bonos, E., Christaki, E. and Florou-Paneri, P. (2010). Performance and carcass characteristics of japanese quail as affected by sex or mannan oligosaccharides and calcium propionate. South African Journal of Animal Science, 40(3): 173-184.
Bonos, E.M., Christaki, E.V. and Florou-Paneri, P.C. (2010). Effect of dietary supplementation of mannan oligosaccharides and acidifier calcium propionate on the performance and carcass quality of japanese quail (Coturnix japonica). International Journal of Poultry Science, 9(3): 264-272.
Burgat, V. (1991). Residues of drugs of veterinary use in food. La Revue du Praticien, 41(11): 985-990.
Cakir, S., Midilli, M., Erol, H., Simsek, N., Cinar, M., Altintas, A., et al. (2008). Use of combined probiotic-prebiotic, organic acid and avilamycin in diets of japanese quails. Revue de Médecine Vétérinaire, 159(11): 565-569.
Capcarova, M., Weiss, J., Hrncar, C., Kolesarova, A. and Pal, G. (2010). Effect of Lactobacillus fermentum and Enterococcus faecium strains on internal milieu, antioxidant status and body weight of broiler chickens. Journal of Animal Physiology and Animal Nutrition, 94(5): e215-e224.
Casewell, M., Friis, C., Marco, E., Mcmullin, P. and Phillips, I. (2003). The european ban on growth-promoting antibiotics and emerging consequences for human and animal health. Journal of Antimicrobial Chemotherapy, 52(2): 159-161.
Chichlowski, M., Croom, J., Mcbride, B., Daniel, L., Davis, G. and Koci, M. (2007). Direct-fed microbial primalac and salinomycin modulate whole-body and intestinal oxygen consumption and intestinal mucosal cytokine production in the broiler chick. Poultry Science, 86(6): 1100-1106.
Choudhury, K., Das, J., Saikia, S., Sengupta, S. and Choudhury, S. (1998). Short communication-supplementation of broiler diets with antibiotic and probiotic fed muga silk worm pupae meal. Indian Journal of Poultry Science, 33(3): 339-342.
Coates, M.E. and Fuller, R. (1977). The gnoto animal in the study of gut microbiology. In: Microbial Ecology of the Gut. Clarke, R. and Bauchop, T. editors. London: Academic Press Inc.(London) Ltd., pp: 311-346.
Duval-Iflah, Y. (2001). Comparison of yogurt, heat treated yogurt, milk and lactose effects on plasmid dissemination in antibiotic mice. International Journal of Genetics and Molecular Biology, 79: 199.
Fooks, L. and Gibson, G. (2002). Probiotics as modulators of the gut flora. British Journal of Nutrition, 88(S1): s39-s49.
Frost, A. and Woolcock, J. (1991). Antibiotics and animal production. In: Microbiology of Animals and Animal Products. Gill, C. and Woolcock, J. editors. New York, NY: Elsevier, Amsterdam, pp: 181-194.
Fuller, R. (1989). A Review: Probiotics in man and animals. Journal of Applied Bacteriology, 66: 365-378.
Garcia, V., Catala-Gregori, P., Hernandez, F., Megias, M. and Madrid, J. (2007). Effect of formic acid and plant extracts on growth, nutrient digestibility, intestine mucosa morphology, and meat yield of broilers. The Journal of Applied Poultry Research, 16(4): 555-562.
Genchev, A., Mihaylova, G., Ribarski, S., Pavlov, A. and Kabakchiev, M. (2008). Meat quality and composition in japanese quails. Trakia Journal of Sciences, 6(4): 72-82.
Ghosh, H., Halder, G., Samanta, G., Paul, S. and Pyne, S. (2007). Effect of dietary supplementation of organic acid and mannan oligosaccharide on the performance and gut health of japanese quail (Coturnix coturnix japonica). Asian Journal of Poultry Sciences, 1(1): 1-7.
Gibson, G.R. and Roberfroid, M.B. (1995). Dietary modulation of the human. colonic microbiota. introducing the concept of prebiotics. Journal of Nutrition, 125: 1401-1412.
Glenn, G. and Roberfroid, M. (1995). Dietary modulation of the human colonic microbiota: Introducing the concept of prebiotics. The Journal of Nutrition, 125(6): 1401-1412.
Guclu, B. (2003). The effect of mannanoligosaccharides on fattening performance of quails. Indian Veterinary Journal, 80(10): 1018-1021.
Homma, H. and Shinohara, T. (2004). Effects of probiotic Bacillus cereus toyoi on abdominal fat accumulation in the japanese quail (Coturnix japonica). Animal Science Journal, 75(1): 37-41.
Khosravi, A., Boldaji, F., Dastar, B. and Hasani, S. (2008). The use of some feed additives as growth promoter in broilers nutrition. International Journal of Poultry Science, 7(11): 1095-1099.
ŁperňÊkovÊ, D., MÊtÉ, D., RÌżaňska, H. and KovÊč, G. (2007). Effects of dietary rosemary extract and"-tocopherol on the performance of chickens, meat quality, and lipid oxidation in meat storaged under chilling conditions. Bulletin of the Veterinary Institute in Pulawy, 51: 585-589.
Marcinčák, S., Cabadaj, R., Popelka, P. and Šoltýsová, L. (2008). Antioxidative effect of oregano supplemented to broilers on oxidative stability of poultry meat. Slovenian Veterinary Research, 45: 61-66.
Mehdipour, Z., Afsharmanesh, M. and Sami, M. (2013). Effects of dietary synbiotic and cinnamon (cinnamomum verum) supplementation on growth performance and meat quality in japanese quail. Livestock Science, 154(1): 152-157.
Mirbabaie, N., Mohamadi, M. and Rostaei, M. (2012). Effect of probiotic protexin and formic acid on broiler performance. Animal Production Research, 3: 9-16. [In Persian]
Mirbabaie, N., Mohamadi, M. and Rostaei, M. (2012). Effect of probiotic protexin and formic acid on safety systems. Iranian Journal of Animal Science, 4: 449-456.
Nasehi, B., Chaji, M., Ghodsi, M. and Poranian, M. (2014). Effect of probiotics addition in feed of japanese quail on the chemical and microbial properties of its meat during storage. Iranian Journal of Nutrition Sciences & Food Technology, 9(4): 77-86. [In Persian]
Oguz, H. and Parlat, S. (2004). Effects of dietary mannanoligosaccharide on performance of japanese quail affected by aflatoxicosis. South African Journal of Animal Science, 34(3): 144-148.
Panda, A.K., Savaram, V., Rao, R., Mantena, V.L.N., Raju, S. and Sharma, R. (2006). Dietary supplementation of lactobacillus sporogenes on performance and serum biochemio-lipid profile of broiler chickens. Journal of Poultry Science, 43: 235-240.
Panda, B. and Singh, R. (1990). Developments in Processing quail meat and eggs. World's Poultry Science Journal, 46(03): 219-234.
Parizadian, K., Jafary, A., Shams, S. and Sardarzade, A. (2013). Investigation of carcass characteristics, meat quality and blood parameters of male japanese quail fed dietary supplements of L-Carnitine. Journal of Animal Science, 99: 16-25. [In Persian]
Parlat, S., Yildiz, A. and Yazgan, O. (2003). Effect of dietary addition of probiotics (mannanoligosaccharides) or antibiotics (virginiamycin) on performance of japanese quail (Coturnix coturnix japonica). Proceedings of Balkan Animal Science Conference, Bucharest, Romania, pp:119-126.
Rolfe, R.D. (2000). The role of probiotic cultures in the control of gastrointestinal health. The Journal of Nutrition, 130(2): 396S-402S.
Sarica, S., Corduk, M., Ensoy, U., Basmacioglu, H. and Karatas, U. (2007). Effects of dietary supplementation of L-Carnitine on performance, carcass and meat characteristics of quails. South African Journal of Animal Science, 37(3): 189-201.
Sarica, S., Corduk, M., Yarim, G., Yenisehirli, G. and Karatas, U. (2009). Effects of novel feed additives in wheat based diets on performance, carcass and intestinal tract characteristics of quail. South African Journal of Animal Science, 39(2): 144-157.
Sorum, H. and Sunde, M. (2001). Resistance to antibiotics in the normal flora of animals. Veterinary Research, 32(3-4): 227-241.
Spring, P., Wenk, C., Dawson, K. and Newman, K. (2000). The effects of dietary mannaoligosaccharides on cecal parameters and the concentrations of enteric bacteria in the ceca of salmonella-challenged broiler chicks. Poultry Science, 79(2): 205-211.
Taherpour, K., Moravej, H., Shivazad, M., Adibmoradi, M. and Yakhchali, B. (2009). Effects of dietary probiotic, prebiotic and butyric acid glycerides on performance and serum composition in broiler chickens. African Journal of Biotechnology, 8(10): 2329-2334.
Talebi, A., Amirzadeh, B., Mokhtari, B. and Gahri, H. (2008). Effects of a multi-strain probiotic (Primalac) on performance and antibody responses to newcastle disease virus and infectious bursal disease virus vaccination in broiler chickens. Avian Pathology, 37(5): 509- 512.
Yalcin, S., Oğuz, İ. and Ötleş, S. (1995). Carcase characteristics of quail (Coturnix coturnix japonica) slaughtered at different ages. British Poultry Science, 36(3): 393-399.
Yalçin, S., Özsoy, B. and Erol, H. (2008). Yeast culture supplementation to laying hen diets containing soybean meal or sunflower seed meal and its effect on performance, egg quality traits, and blood chemistry. The Journal of Applied Poultry Research, 17(2): 229-236.
Yildiz, A., Parlat, S. and Yildirim, I. (2004). Effect of dietary addition of live yeast (saccharomyces cerevisiae) on some performance parameters of adult japanese quail (Coturnix coturnix japonica) induced by aflatoxicosis. Revue De Medecine Veterinaire, 155(1): 38-41.
Zhang, A., Lee, B., Lee, S., Lee, K., An, G., Song, K., et al. (2005). Effects of yeast (Saccharomyces cerevisiae) cell components on growth performance, meat quality, and ileal mucosa development of broiler chicks. Poultry Science, 84(7): 1015-1021.
Zhang, Z., Zhou, T., Ao, X. and Kim, I. (2012). Effects of Β-glucan and bacillus subtilis on growth performance, blood profiles, relative organ weight and meat quality in broilers fed maize–soybean meal based diets. Livestock Science, 150(1): 419-424.
Aluwong, T., Kawu, M., Raji, M., Dzenda, T., Govwang, F., Sinkalu, V., et al. (2013). Effect of yeast probiotic on growth, antioxidant enzyme activities and malondialdehyde concentration of broiler chickens. Antioxidants, 2(4): 326-339.
Ao, X., Yoo, J., Zhou, T., Wang, J., Meng, Q., Yan, L., et al. (2011). Effects of fermented garlic powder supplementation on growth performance, blood profiles and breast meat quality in broilers. Livestock Science, 141(1): 85-89.
Asadi, G., Ebrahimnejad, Y., Nazeradl, K. and Ahmadzade, A. (2010). Impact of the size of corn on the weight and some internal organs of japanese quail. Fifth Animal Science Congress, pp: 57-64.
Awad, W., Ghareeb, K., Abdel-Raheem, S. and Böhm, J. (2009). Effects of dietary inclusion of probiotic and synbiotic on growth performance, organ weights, and intestinal histomorphology of broiler chickens. Poultry Science, 88(1): 49-56.
Bonos, E., Christaki, E. and Florou-Paneri, P. (2010). Performance and carcass characteristics of japanese quail as affected by sex or mannan oligosaccharides and calcium propionate. South African Journal of Animal Science, 40(3): 173-184.
Bonos, E.M., Christaki, E.V. and Florou-Paneri, P.C. (2010). Effect of dietary supplementation of mannan oligosaccharides and acidifier calcium propionate on the performance and carcass quality of japanese quail (Coturnix japonica). International Journal of Poultry Science, 9(3): 264-272.
Burgat, V. (1991). Residues of drugs of veterinary use in food. La Revue du Praticien, 41(11): 985-990.
Cakir, S., Midilli, M., Erol, H., Simsek, N., Cinar, M., Altintas, A., et al. (2008). Use of combined probiotic-prebiotic, organic acid and avilamycin in diets of japanese quails. Revue de Médecine Vétérinaire, 159(11): 565-569.
Capcarova, M., Weiss, J., Hrncar, C., Kolesarova, A. and Pal, G. (2010). Effect of Lactobacillus fermentum and Enterococcus faecium strains on internal milieu, antioxidant status and body weight of broiler chickens. Journal of Animal Physiology and Animal Nutrition, 94(5): e215-e224.
Casewell, M., Friis, C., Marco, E., Mcmullin, P. and Phillips, I. (2003). The european ban on growth-promoting antibiotics and emerging consequences for human and animal health. Journal of Antimicrobial Chemotherapy, 52(2): 159-161.
Chichlowski, M., Croom, J., Mcbride, B., Daniel, L., Davis, G. and Koci, M. (2007). Direct-fed microbial primalac and salinomycin modulate whole-body and intestinal oxygen consumption and intestinal mucosal cytokine production in the broiler chick. Poultry Science, 86(6): 1100-1106.
Choudhury, K., Das, J., Saikia, S., Sengupta, S. and Choudhury, S. (1998). Short communication-supplementation of broiler diets with antibiotic and probiotic fed muga silk worm pupae meal. Indian Journal of Poultry Science, 33(3): 339-342.
Coates, M.E. and Fuller, R. (1977). The gnoto animal in the study of gut microbiology. In: Microbial Ecology of the Gut. Clarke, R. and Bauchop, T. editors. London: Academic Press Inc.(London) Ltd., pp: 311-346.
Duval-Iflah, Y. (2001). Comparison of yogurt, heat treated yogurt, milk and lactose effects on plasmid dissemination in antibiotic mice. International Journal of Genetics and Molecular Biology, 79: 199.
Fooks, L. and Gibson, G. (2002). Probiotics as modulators of the gut flora. British Journal of Nutrition, 88(S1): s39-s49.
Frost, A. and Woolcock, J. (1991). Antibiotics and animal production. In: Microbiology of Animals and Animal Products. Gill, C. and Woolcock, J. editors. New York, NY: Elsevier, Amsterdam, pp: 181-194.
Fuller, R. (1989). A Review: Probiotics in man and animals. Journal of Applied Bacteriology, 66: 365-378.
Garcia, V., Catala-Gregori, P., Hernandez, F., Megias, M. and Madrid, J. (2007). Effect of formic acid and plant extracts on growth, nutrient digestibility, intestine mucosa morphology, and meat yield of broilers. The Journal of Applied Poultry Research, 16(4): 555-562.
Genchev, A., Mihaylova, G., Ribarski, S., Pavlov, A. and Kabakchiev, M. (2008). Meat quality and composition in japanese quails. Trakia Journal of Sciences, 6(4): 72-82.
Ghosh, H., Halder, G., Samanta, G., Paul, S. and Pyne, S. (2007). Effect of dietary supplementation of organic acid and mannan oligosaccharide on the performance and gut health of japanese quail (Coturnix coturnix japonica). Asian Journal of Poultry Sciences, 1(1): 1-7.
Gibson, G.R. and Roberfroid, M.B. (1995). Dietary modulation of the human. colonic microbiota. introducing the concept of prebiotics. Journal of Nutrition, 125: 1401-1412.
Glenn, G. and Roberfroid, M. (1995). Dietary modulation of the human colonic microbiota: Introducing the concept of prebiotics. The Journal of Nutrition, 125(6): 1401-1412.
Guclu, B. (2003). The effect of mannanoligosaccharides on fattening performance of quails. Indian Veterinary Journal, 80(10): 1018-1021.
Homma, H. and Shinohara, T. (2004). Effects of probiotic Bacillus cereus toyoi on abdominal fat accumulation in the japanese quail (Coturnix japonica). Animal Science Journal, 75(1): 37-41.
Khosravi, A., Boldaji, F., Dastar, B. and Hasani, S. (2008). The use of some feed additives as growth promoter in broilers nutrition. International Journal of Poultry Science, 7(11): 1095-1099.
ŁperňÊkovÊ, D., MÊtÉ, D., RÌżaňska, H. and KovÊč, G. (2007). Effects of dietary rosemary extract and"-tocopherol on the performance of chickens, meat quality, and lipid oxidation in meat storaged under chilling conditions. Bulletin of the Veterinary Institute in Pulawy, 51: 585-589.
Marcinčák, S., Cabadaj, R., Popelka, P. and Šoltýsová, L. (2008). Antioxidative effect of oregano supplemented to broilers on oxidative stability of poultry meat. Slovenian Veterinary Research, 45: 61-66.
Mehdipour, Z., Afsharmanesh, M. and Sami, M. (2013). Effects of dietary synbiotic and cinnamon (cinnamomum verum) supplementation on growth performance and meat quality in japanese quail. Livestock Science, 154(1): 152-157.
Mirbabaie, N., Mohamadi, M. and Rostaei, M. (2012). Effect of probiotic protexin and formic acid on broiler performance. Animal Production Research, 3: 9-16. [In Persian]
Mirbabaie, N., Mohamadi, M. and Rostaei, M. (2012). Effect of probiotic protexin and formic acid on safety systems. Iranian Journal of Animal Science, 4: 449-456.
Nasehi, B., Chaji, M., Ghodsi, M. and Poranian, M. (2014). Effect of probiotics addition in feed of japanese quail on the chemical and microbial properties of its meat during storage. Iranian Journal of Nutrition Sciences & Food Technology, 9(4): 77-86. [In Persian]
Oguz, H. and Parlat, S. (2004). Effects of dietary mannanoligosaccharide on performance of japanese quail affected by aflatoxicosis. South African Journal of Animal Science, 34(3): 144-148.
Panda, A.K., Savaram, V., Rao, R., Mantena, V.L.N., Raju, S. and Sharma, R. (2006). Dietary supplementation of lactobacillus sporogenes on performance and serum biochemio-lipid profile of broiler chickens. Journal of Poultry Science, 43: 235-240.
Panda, B. and Singh, R. (1990). Developments in Processing quail meat and eggs. World's Poultry Science Journal, 46(03): 219-234.
Parizadian, K., Jafary, A., Shams, S. and Sardarzade, A. (2013). Investigation of carcass characteristics, meat quality and blood parameters of male japanese quail fed dietary supplements of L-Carnitine. Journal of Animal Science, 99: 16-25. [In Persian]
Parlat, S., Yildiz, A. and Yazgan, O. (2003). Effect of dietary addition of probiotics (mannanoligosaccharides) or antibiotics (virginiamycin) on performance of japanese quail (Coturnix coturnix japonica). Proceedings of Balkan Animal Science Conference, Bucharest, Romania, pp:119-126.
Rolfe, R.D. (2000). The role of probiotic cultures in the control of gastrointestinal health. The Journal of Nutrition, 130(2): 396S-402S.
Sarica, S., Corduk, M., Ensoy, U., Basmacioglu, H. and Karatas, U. (2007). Effects of dietary supplementation of L-Carnitine on performance, carcass and meat characteristics of quails. South African Journal of Animal Science, 37(3): 189-201.
Sarica, S., Corduk, M., Yarim, G., Yenisehirli, G. and Karatas, U. (2009). Effects of novel feed additives in wheat based diets on performance, carcass and intestinal tract characteristics of quail. South African Journal of Animal Science, 39(2): 144-157.
Sorum, H. and Sunde, M. (2001). Resistance to antibiotics in the normal flora of animals. Veterinary Research, 32(3-4): 227-241.
Spring, P., Wenk, C., Dawson, K. and Newman, K. (2000). The effects of dietary mannaoligosaccharides on cecal parameters and the concentrations of enteric bacteria in the ceca of salmonella-challenged broiler chicks. Poultry Science, 79(2): 205-211.
Taherpour, K., Moravej, H., Shivazad, M., Adibmoradi, M. and Yakhchali, B. (2009). Effects of dietary probiotic, prebiotic and butyric acid glycerides on performance and serum composition in broiler chickens. African Journal of Biotechnology, 8(10): 2329-2334.
Talebi, A., Amirzadeh, B., Mokhtari, B. and Gahri, H. (2008). Effects of a multi-strain probiotic (Primalac) on performance and antibody responses to newcastle disease virus and infectious bursal disease virus vaccination in broiler chickens. Avian Pathology, 37(5): 509- 512.
Yalcin, S., Oğuz, İ. and Ötleş, S. (1995). Carcase characteristics of quail (Coturnix coturnix japonica) slaughtered at different ages. British Poultry Science, 36(3): 393-399.
Yalçin, S., Özsoy, B. and Erol, H. (2008). Yeast culture supplementation to laying hen diets containing soybean meal or sunflower seed meal and its effect on performance, egg quality traits, and blood chemistry. The Journal of Applied Poultry Research, 17(2): 229-236.
Yildiz, A., Parlat, S. and Yildirim, I. (2004). Effect of dietary addition of live yeast (saccharomyces cerevisiae) on some performance parameters of adult japanese quail (Coturnix coturnix japonica) induced by aflatoxicosis. Revue De Medecine Veterinaire, 155(1): 38-41.
Zhang, A., Lee, B., Lee, S., Lee, K., An, G., Song, K., et al. (2005). Effects of yeast (Saccharomyces cerevisiae) cell components on growth performance, meat quality, and ileal mucosa development of broiler chicks. Poultry Science, 84(7): 1015-1021.
Zhang, Z., Zhou, T., Ao, X. and Kim, I. (2012). Effects of Β-glucan and bacillus subtilis on growth performance, blood profiles, relative organ weight and meat quality in broilers fed maize–soybean meal based diets. Livestock Science, 150(1): 419-424.
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