The response of blood plasma metabolite concentrations of anestrus lactating buffalo to CIDR-eCG with or without melatonin treatment

Main Article Content

Tamer Awad Ramadan Rakesh Kumar Sharma    Sushil Kumar Phulia Ashok Kumar Balhara Sarvpreet Singh Ghuman Inderjeet Singh


The objective of this study was to compare the efficiency of CIDR-eCG protocol with or without melatonin treatment on blood plasma metabolites in anestrus Murrah buffaloes during out-of-breeding season. Twelve lactating Murrah buffaloes were allotted to control and treated groups each of six animals. Treated buffaloes were implanted with melatonin (18 mg melatonin / 50 kg body weight) for 45 days, then animals of both groups received CIDR for 9 days. All animals received 500 IU eCG intramuscularly one day before CIDR removal and 10 µg GnRH on the day of AI. The animals were subjected to estrus detection daily as per farm routine. At weekly intervals, blood samples were collected to determine the concentration of blood plasma metabolites over 84 days’ period between June and September. All buffaloes exhibited estrus after CIDR removal. CIDR-eCG preceded with melatonin treatment increased plasma total protein, albumin, cholesterol and HDL-cholesterol (P<0.05) in comparison to only CIDR treated control. Buffaloes treated with melatonin plus CIDR exhibited increased (P<0.01) plasma glucose on days 28, 53 and 55 and reduction (P<0.01) in HDL and AST at time of pregnancy diagnosis, while plasma concentration of phosphorus was the highest (P<0.05) at day 84. In conclusion, CIDR-eCG treatment preceded with melatonin implantation improved reproductive performance and altered blood metabolite composition of lactating buffaloes during out-of-breeding season as compared to CIDR-eCG only.


Download data is not yet available.

Article Details

How to Cite
RAMADAN, Tamer Awad et al. The response of blood plasma metabolite concentrations of anestrus lactating buffalo to CIDR-eCG with or without melatonin treatment. Buffalo Bulletin, [S.l.], v. 41, n. 2, p. 351-362, june 2022. ISSN 2539-5696. Available at: <>. Date accessed: 16 aug. 2022.
Original Article


Ali, M.D.M., B.C. Kanjilal, R. Roychoudhury, S.K. Bandopadhyay and B.B. Ghosh. 1991. Total serum protein and hemoglobin content in anestrus rural crossbred heifers. Indian J. Anim. Reprod., 12(2): 159-161.

Antunovic, Z., J. Novoselec, H. Sauerwein, M. Speranda, M. Vegara and V. Pavic. 2011. Blood metabolic profile and some of hormones concentration in ewes during different physiological status. Bulg. J. Agric. Sci., 17(5): 687-695. Available on:

Armstrong, W.D. and C.W. Carr. 1964. Physiological Chemistry Laboratory Directions, 3rd ed. Burges Publishing Co., Minneapolis, USA.

Arshad, H.M., N. Ahmad, Zia-ur-Rahman, H.A. Samad, N. Akhtar and S. Ali. 2005. Studies on some biochemical constituents of ovarian follicular fluid and peripheral blood in buffaloes. Pak. Vet. J., 25(4): 189-193.

Barham, D. and P. Trinder. 1972. An improved colour reagent for the determination of blood glucose by the oxidase system. Analyst, 97(151): 142-145. DOI: 10.1039/an9729700142

Beeler, M.F. and P.G. Catrou. 1983. Disorders of calcium metabolism in Interpretations, p. 33-44. Clinical Chemistry. American Society for Clinical Pathology Press, Chicago, USA.

Bhat, G.R., G.S. Dhaliwal, S.S. Ghuman and M. Honparkhe. 2015. Size of dominant follicle, plasma progesterone and estradiol levels on the day of ovulation and subsequent conception rate in buffalo (Bubalus bubalis) following modified ovsynch + CIDR protocol. J. Appl. Anim. Res., 34(3): 314-317. DOI: 10.1080/09712119.2014.964249

Brantmeier, S.A., R.R. Grummer and R.L. Ax. 1987. Concentrations of high density lipoproteins vary among follicular sizes in the bovine. J. Dairy Sci., 70(10): 2145-2149. DOI: 10.3168/jds.s0022-0302(87)80266-7

Bremmer, D.R., S.J. Bertics, S.A. Besong and R.R. Grummer. 2000. Changes in hepatic microsomal triglyceride transfer protein and triglyceride in periparturient dairy cattle. J. Dairy Sci., 83(10): 2252-2260. DOI: 10.3168/jds.s0022-0302(00)75109-5

Burstein, M., H.R. Scholnick and R. Morfin. 1970. Rapid method for the isolation of lipoproteins from human serum by precipitation with polyanions. J. Lipid. Res., 11(6): 583-595.

Das, G.K. and F.A. Khan. 2010. Summer anoestrus in buffalo - A review. Reprod. Domest. Anim., 45(6): e483-e494. DOI: 10.1111/j.1439-0531.2010.01598.x

Delfino, L.J.B., B.B. Souza, R.M.N. Silva and W.W. Silva. 2012. Efeito do estresse calórico sobre o eritrograma de ruminantes. Agropecuaria Cientifica no Semiárido, 8(2): 1-7. DOI: 10.30969/acsa.v8i2.170

Dhoble, R.L. and S.K. Gupta. 1996. Serum calcium and inorganic phosphorus during postpartum anoestrus in buffaloes. Indian J. Anim. Health, 25(2): 123-126.

Doumas, B.T., R.L. Arends and P.C. Pinto. 1972. Determination of serum albumin. Stand. Meth. Clin. Chem., 7: 175-189. DOI: 10.1016/b978-0-12-609107-6.50022-2

El-Mokadem, M.Y., A.N.M. Nour El-Din, T.A. Ramadan, A.M.A. Rashad, T.A. Taha and M.A. Samak. 2019. Alleviation of reproductive seasonality in Barki ewes using CIDR-eCG with or without melatonin. Small Ruminant Res., 174: 170-178. DOI: 10.1016/j.smallrumres.2019.03.019 

FASS. 2010. Guide for the Care and Use of Agricultural Animals in Research and Teaching, 3rd ed. Federation of Animal Science Societies, Champaign, Illinois, USA.

Guesnet, P.M., M.J. Massoud and Y. Demarne. 1991. Regulation of adipose tissue metabolism during pregnancy and lactation in the ewe: The role of insulin. J. Anim. Sci., 69(5): 2057-2065. DOI: 10.2527/1991.6952057x

Hamra, A.H., J.W. McNally, J.M. Marcek, K.M. Carlson and J.E. Wheaton. 1989. Comparison of progesterone sponges, cronolone sponges and controlled internal drug release dispensers on fertility in anestrous ewes. Anim. Reprod. Sci., 18(1-3): 219-226. DOI: 10.1016/0378-4320(89)90023-7

Hassan, G.A., F.D. El-Nouty, M.A. Samak and M.H. Salem. 1986. Relationship between milk production and some blood constituents in Egyptian Baladi goats. Beitr. Trop. Landwirt., 24(2): 213-219.

Ingvartsen, K.L. and N.C. Friggens. 2005. To what extent do variabilities in hormones, metabolites and energy intake explain variability in milk yield? Domest. Anim. Endocrin., 29(2): 294-304. DOI: 10.1016/j.domaniend.2005.05.001

Jacob, N. and V.P. Vadodaria. 2001. Levels of glucose and cortisol in blood of Patanwadi ewes around parturition. Indian Vet. J. 78(10): 890-892.

Janicek, B.N., P.J. Kononoff, A.M. Gehman, K. Karges and M.L. Gibson. 2007. Effect of increasing levels of corn bran on milk yield and composition. J. Dairy Sci., 90(9): 4313-4316. DOI: 10.3168/jds.2007-0275

Kaneko, J.J., J.W. Harvey and M.L. Bruss. 1997. Clinical Biochemistry of Domestic Animals, 5th ed. Harcourt Brace and Company, Forum Galleria, Singapore.

Karapehlivan, M., E. Atakisi, O. Atakisi, R. Yucayurt and S.M. Pancarci. 2007. Blood biochemical parameters during the lactation and dry period in Tuj ewes. Small Ruminant Res., 73(1-3): 267-271. DOI: 10.1016/j.smallrumres.2006.12.006

Kendall, P.E. and J.R. Webster. 2009. Season and physiological status affects the circadian body temperature rhythm of dairy cows. Livest. Sci., 125(2-3): 155-160. DOI: 10.1016/j.livsci.2009.04.004

Kumar, R., M. Ghosh, S.S. Rawat, P. Kumar and S. Kumar. 2016. Plasma mineral profile of normal cyclic and postpartum anestrous Murrah buffaloes in organized farms. Anim. Sci., 10(2): 12-14. Available on:

Kumar, S., A. Saxena and Ramsagar. 2010. Comparative studies on metabolic profile of anestrous and normal cyclic Murrah buffaloes. Buffalo Bull., 29(1): 7-11.

Lindsay, D.B. 1973. Metabolic changes induced by pregnancy in the ewe. In Payne, E.J.M., K.G. Hibitt and B.F. Sansom (eds.) Production Disease in Farm Animals. Bailliere, Tindal, London, UK.

Madan, M.L. 1988. Status of reproduction in female buffalo. - In Buffalo Production and Health: A Compendium of Latest Research Information Based on Indian Studies, ICAR Publication, New Delhi, India.

Marcos, E., A. Mazur, P. Cardot and Y. Rayssiguier. 1990. The effect of pregnancy and lactation on serum lipid and apolipoprotein B and A-I levels in dairy cows. J. Anim. Physiol. An. N., 64(1-5): 133-138. DOI: 10.1111/j.1439-0396.1990.tb00214.x

Nardone, A., B. Ronchi, N. Lacetera, M.S. Ranieri and U. Bernabucci. 2010. Effects of climate changes on animal production and sustainability of livestock systems. Livest. Sci., 130(1-3): 57-69. DOI: 10.1016/j.livsci.2010.02.011

Nazifi, S., M. Saeb and S.M. Ghavami. 2002. Serum lipid profile in Iranian fat-tailed sheep in late pregnancy, at parturition and during the post-parturition period. J. Vet. Med. A., 49(1): 9-12. DOI: 10.1046/j.1439-0442.2002.00405.x

NRC. 2001. Nutrient Requirement of Dairy Cattle, 7th ed. National Academy of Science, Washington, D.C., USA.

Papachristoforou, C., A. Koumas and C. Photiou. 2007. Initiation of the breeding season in ewe lambs and goat kids with melatonin implants. Small Ruminant Res., 73(1-3): 122-126. DOI: 10.1016/j.smallrumres.2006.12.004

Perera, B.M.A.O. 2011. Reproductive cycles of buffalo. Anim. Reprod. Sci., 124: 194-199. DOI: 10.1016/j.anireprosci.2010.08.022

Ramadan, T.A., R.K. Sharma, S.K. Phulia and I. Singh. 2020. Effect of melatonin implantation on haematological parameters in anestrus lactating buffalo during summer season under tropical conditions. Large Anim. Rev., 26(6): 283-290. Available on:

Ramadan, T.A., D. Kumar, S.S. Ghuman and I. Singh. 2019. Melatonin-improved buffalo semen quality during nonbreeding season under tropical condition. Domest. Anim. Endocrin., 68: 119-125. DOI: 10.1016/j.domaniend.2019.01.010

Ramadan, T.A., R.K. Sharma, S.K. Phulia, A.K. Balhara, S.S. Ghuman and I. Singh. 2014. Effectiveness of melatonin and controlled internal drug release device treatment on reproductive performance of buffalo heifers during out-of-breeding season under tropical conditions. Theriogenology, 82(9): 1296-302. DOI: 10.1016/j.theriogenology.2014.08.014

Ramadan, T.A., R.K. Sharma, S.K. Phulia, A.K. Balhara, S.S. Ghuman and I. Singh. 2016. Manipulation of reproductive performance of lactating buffaloes using melatonin and controlled internal drug release device treatment during out-of-breeding season under tropical conditions. Theriogenology 86(4): 1048-1053. DOI: 10.1016/j.theriogenology.2016.03.034

Ramadan, T.A., R.K. Sharma, S.K. Phulia, A.K. Balhara, S.S. Ghuman and I. Singh. 2015. Effects of melatonin and controlled internal drug release device treatment on blood metabolites of buffalo heifers during out-of-breeding season under tropical conditions. Egyptian Journal of Animal Production, 52(4): 9-17. DOI: 10.21608/ejap.2015.170496

Reitaman, S. and S. Frankel. 1957. A colorimetric method for the determination of serum glutamic oxaloacetic and glutamic pyruvic transaminases. Am. J. Clin. Pathol., 28(1): 56-63. DOI: 10.1093/ajcp/28.1.56

Rigout, S., C. Hurtaud, S. Lemosquet, A. Bach and H. Rulquin. 2003. Lactational effect of propionic acid and duodenal glucose in cows. J. Dairy Sci., 86(1): 243-253. DOI: 10.3168/jds.S0022-0302(03)73603-0

Roginosu, E. 2011. The biochemical profile in cows with reproductive disorders. Cercetari Agronomicein Moldova, 64(2): 75-86. DOI: 10.2478/v10298-012-0035-4

SAS. 2000. SAS User’s Guide: Statistics, Version 8. SAS Inst., Inc., Cary, North Carolina, USA.

Satish, Kumar. 2003. Management of infertility due to mineral deficiency in dairy animals, p. 128-137. In Proceedings of ICAR Summer School on “Advance Diagnostic Techniques and Therapeutic Approaches to Metabolic and Deficiency Diseases in Dairy Animals” Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.

Scholtz, M.M., C. McManus, K.J. Leeuw, H. Louvandini, L. Seixas, C.B. Melo, A. Theunissen and F.W.C. Neser. 2013. The effect of global warming on beef production in developing countries of the southern hemisphere. Natural Science, 5(1A): 1-14. DOI: 10.4236/ns.2013.51A017

Shah, S.N.H. 1988. Comparative studies of seasonal influence on breeding behaviour and conception rate of dairy buffalo and zebu cattle. In Proceedings 11th International Congress on Animal Reproduction and Artificial Insemination. Dublin, Ireland.

Singh, J., S.P.S. Ghuman, D. Dadarwal, M. Honparkhe, G.S. Dhaliwal and A.K. Jain. 2010. Estimations of blood plasma metabolites following melatonin implants treatment for initiation of ovarian cyclicity in true anestrus buffalo heifers. Indian J. Anim. Sci., 80(3): 229-231.

Singh, J., A.S. Nanda and G.P. Adams. 2000. The reproductive pattern and efficiency of female buffaloes. Anim. Reprod. Sci., 2(60-61): 593-604. DOI: 10.1016/s0378-4320(00)00109-3

Singh, R. 1993. Studies on the onset of postpartum ovarian activity in relation to climatic and nutritional status in buffaloes. M.V.Sc. Thesis, Punjab Agriculture, University of Ludhiana, India.

Swenson, M.J. 1977. Blood circulation and the cardiovascular system, p. 14-174. In Dukes, H.H., M.J. Swenson (eds.) Dukes’ Physiology of Domestic Animals, 9th ed. Comstock Publishing Association, Cornell University Press, Ithaca, New York, USA.

Tailor, S.P., L.S. Jain, H.K. Gupta and J.S. Bhatia. 1990. Oestrus and conception rates in buffaloes under village conditions. Indian J. Anim. Sci., 60(8): 1020-1021.

Watson, D. 1960. A simple method for the determination of serum cholesterol. Clin. Chim. Acta, 5: 637-638. DOI: 10.1016/0009-8981(60)90004-8

Zicarelli, L. 1997. Reproductive seasonality in buffalo. In Proceedings of Third Course on Biotechnology of Reproduction in Buffaloes, Caserta, Italy.

Zicarelli, L. 2007. Can we consider buffalo a non precocious and hypofertile species? Ital. J. Anim. Sci., 6: 143-154. DOI: 10.4081/ijas.2007.s2.143

Zilva, J.F. and P.R. Pannall. 1979. Plasma enzymes in diagnosis. In Clinical Chemistry in Diagnosis and Treatment. Lioyd-Luke, London, UK. Chapter, 17: 338-339.