Amylase activity in cerical mucus and serum during estrus in normal and

Aim: To compare the amylase activity in the cervical mucus and serum during estrus in normal and repeat breeder cattle and find out importance in fertility. Materials and methods: Twenty cyclic crossbred cattle were divided into two groups: normal (n =10) and repeat breeder (n =10). Animals exhibiting spontaneous estrus, were artificially inseminated (AI) twice, as per AM-PM schedule along with simultaneous collection of cervical mucus and blood. Amylase activity was measured by using kits in collected samples. Result and discussion: Cervical amylase activity was significantly (p<0.01) higher in normal compared to repeat breeder animals showing important role of cervical amylase in fertility without significant difference in serum profiles between these


Introduction
postovulatory phases and declining around ovulation [3].The Physical and chemical properties of bovine Assessment of fertility in female by examining cervical mucus has been determined in previous genital tract fluids has been receiving considerable studies, but there is a paucity of literature about the attention.The cervix with its unique anatomical and activity of enzymes [4].Therefore, it is logical to histological structure and secretory function plays an determine and compare the activity of enzymes during important role in sperm survival and transport to estrus in cervical mucus in normal and repeat breeder uterine cavity.The primary function of cervical mucus cattle in order to find out abnormalities in cervical is sperm transport and to act as a mechanical barrier mucus and attempt for correction to improve fertility against microbial infection of the uterus.Cervical in problematic cases.Thus, the aim of the present mucus contain about 92-95% water along with low study was to determine the amylase activity in the molecular weight substances such as electrolytes, cervical mucus and serum during estrus in normal and carbohydrates, amino acids, lipids and soluble repeat breeder cattle.macromolecular compounds such as proteins and polysaccharides dissolved in it [1].During estrus, the Materials and Methods modification of physical and biochemical properties The present investigation was conducted on of cervical mucus takes place under influence of sex twenty clinically healthy crossbred cattle of 4-7 years steroid hormones in order to facilitate the transport of of age, maintained at the Institute Dairy Farm, spermatozoa.Cyclic variation in the alkaline Livestock Production and Management Section, phosphatase, amylase and lysozyme in cervical mucus Indian Veterinary Research Institute, Izatnagar.reported by Schumacher [1] and level of peroxidase in Animals were checked for estrus twice daily using the cattle could be used to detect ovulation [2].
teaser bull along with observing behavioural signs of Amylase enzyme shows cyclic changes in the estrus.The selected animals have normal parturition cervical mucus of women, rising during the pre-and www.veterinaryworld.orgVeterinary World, Vol.5 No.8 August 2012 487 and not have postpartum anestrus and breeding was water (4.0ml) and working iodine solution (0.2ml) done after 45 days of voluntary waiting periods.BCS mixed well and measured optical density (O D) using was 4.5-5 (scale 0-8) and the status of genital tract and red filter at 620nm wavelength against purified water.ovaries was examined by rectal plalpation for proper Statistical analysis: The comparison of amylase in uterine involution, cystic ovaries and found normal at the cervical mucus and serum were performed by days 30 postpartum as well as at time of AI.Animals student's 't' test using the statistic software program were grouped in two groups: Group І (normal cyclic, SPSS (SPSS Inc., Chicago, Illinois, USA).n=10) were inseminated for less than 3 times after the last calving and Group ІІ (repeat breeder, n= 10) which Results and Discussion were inseminated for more than 3 times but failed to The difference in mean values of cervical mucus conceive.The animals were maintained under amylase between two groups was significantly (P isomanagerial conditions with intensive system of <0.01) higher in normal as compared to repeat breeder feeding.They were housed in a well ventilated brick animals.However, Wagh and Hukeri, [5] reported no cemented house with non-slippery floor and offered significant difference between two groups which is standard ration having green and dry fodder along with contradictory to present finding.Serum amylase concentrate.Prior approval for experiments was taken activity did not statistically differ between groups.from Institutional Animal Ethics Committee as per Tsiligianni et al. [6] documented cervical mucus CPCSEA (Govt. of India) norms.
amylase activity in spontaneous and induced estrus in Cervical mucus and blood samples were Friesian cattle with no significant difference between collected from mid to late estrus animals i.e. if the these groups despite the fact that cervical mucus in the animals were detected in estrus during morning, the induced estrus groups was thicker which indicates that sampling and AI was done in same evening and if the water content in the cervical mucus does not influence animals came in heat in the evening then sampling and amylase activity.Cervical mucus amylase concen-AI was done in next morning.Cervical mucus samples tration was significantly higher in normal compared to were collected from animals prior to insemination repeat breeder animals showing important role in using AI gun sheath and syringe from cervix with fertility and was in accordance with the findings of precautions to avoid contamination by proper washing O'Hagan et al.
[7] that addition of amylase in bull of perineal region with diluted potassium semen dilutors improve conception rate in cattle.permanganate solution in water followed by cleaning Similar successful attempts have been made to with spirit swab.Cervical mucus was placed into a 10 improve sperm transport by adding compounds such ml plastic tube and transported to the laboratory in a as amylases and glucosidases to bull semen diluents refrigerated box (4°C).Aliquots of 1.5 to 2 ml of and fertility improved significantly [8].According to cervical mucus were homogenized in a vortex Hawk, [9] mostly failure of fertilization occur due to homogenizer and stored at -20°C for measurement of failure of sperm ova contact rather than from enzymatic activity in future.Blood samples were unfertilizability of gametes.Amylase is involved in collected from jugular vein aseptically and serum was making proper channel in cervical mucus for sperm separated by centrifugation at 3000 rpm for 15 min and transport by hydrolysis of linkage of large polysastored at -20°C until analysis.
ccharides composed of glycogen and making reducing Amylase enzyme activity in the cervical mucus sugars available for cleavage.Lactate and pyruvate are and serum samples was assayed by as per kit (Span important substrate for sustaining spermatozoa Diagnostics Ltd.Surat, India) using spectrophotometabolism during their transit through reproductive meter.For test sample was taken 0.5 ml of buffered tract [10].Amylase significantly increases the lactate substrate, mixed well and incubated at 37°C for 3 min, then added homogenized mucus (50µl) and incubated production of rabbit sperm in vitro and beneficial for for 15 min at 37°C followed by addition of purified maintenance of sperm motility indicating vital role of

Table - 1
. Amylase concentration (IU/L) in cervical mucus and serum of normal and repeat breeder cattle (Mean ± SEM) ** Mean differ Significantly (P <0.01) in column