Importance of Cervical mucus examination in diagnosing bovine infertility

M. Mutha Rao, A.K. Pandey and Sudhir Kumar Division of Animal Reproduction, Gynaecology and obstetrics, Faculty of Veterinary Sciences & Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu RS Pura, Jammu. When a cow/ buffalo is presented in a veterinary clinic for the purpose of AI or for treatment of any genital infection, the foremost bench mark that a veterinarian looks for is the nature of cervical mucus- the colour, odour, quantity, viscosity, elasticity, stringiness, presence of pus, flakes, blood clots, etc. It enables the veterinarian / technician to arrive at the tentative diagnosis of the case presented. Thus cervical mucus examination provides a useful clue in AI management as well as to determine the line of treatment in various reproductive ailments. The chemical composition of cervical mucus reveals that it is made up of macromolecules of mucus of epithelial origin which are composed of glycoproteins that contain 25% aminoacids and 75% carbohydrates. The carbohydrate portion is made up of galactose, glucosamine, fructose, and sialic acid. The proteins of cervical mucus include pre-albumin, albumin, globulin etc. Besides carbohydrates and proteins, it also contains several enzymes like amylase, esterase, glucoronidase etc. The secretion of cervical mucus varies depending on the stage of estrous cycle. At the time of estrus and ovulation, there is profuse secretion of cervical mucus under the influence of estrogens. In a normal healthy cyclic animal, a clear, copious vaginal discharge is evident around the time of estrus. An experienced veterinarian can diagnose the stage of estrus (early, mid, late) based on the characteristics of mucus discharge. Under field conditions, the nature of cervical/vaginal mucus is compared to the white (albumin portion) of an egg a highly transparent, stringy secretion. During early estrus, the discharge is more watery, transparent, less stringy, less elastic and the string breaks almost immediately. During midestrus (standing estrus) which is an ideal time for AI, the discharge is more transparent, less watery, more elastic and above all it is more stringy in nature. Per rectal palpation reveals this stringy nature due to its hanging from vulva up to hock region. In late estrus the discharge is scanty, less transparent, sticky, less stringy and may contain some cell debris. The secretory activity of cervical mucus also varies with breed, season, nutrition and of course, the stage of estrus cycle. Fern Pattern: The cervical mucus of estrus cow exhibits fern pattern of crystallization. Freshly collected cervical mucus is taken on a clean, grease free slide and allowed to spread on the slide on its own. The slide is allowed to dry under sunlight. It is then examined under lower power (10 X) of microscope for fern pattern. The fern pattern, associated with high chloride content of the mucus, does not usually occur during luteal phase or during pregnancy. The phenomena of fern pattern may be of some value, when combined with other observations for early pregnancy diagnosis. The secretion of cervical mucus is stimulated by ovarian estrogen and

inhibited by progesterone. The biophysical properties of cervical mucus such as increase in quantity, fern pattern, decrease in viscosity etc. will facilitate sperm penetration during estrus, thus contributing to higher fertilization rate during mid estrus. In a normal fertile animal, the cervical mucus is clear without any debris. However, during conditions like endometritis and other uterine infections, the colour and odour of mucus is changed. Depending on the severity of endometritis, the mucus may be cloudy, turbid, purulent with inspissated pus clots or reddish yellow. Under such conditions the mucus should be collected aseptically and sent for isolation of infectious agent and antibiotic sensitivity testing. Collection of Cervical mucus: The cervical mucus is collected by rectovaginal technique. A sterile glass pipette or AI sheath passed through vulva is placed at the floor of vagina with a hand in the rectum manipulating the position and placement of sheath. The other end of pipette/ sheath is connected to syringe with adopter and the mucus is aspirated. Caution is taken while passing the sheath into vagina so as to avoid external contamination. The mucus thus collected is taken in a sterile test tube for estimation of bacterial load as well as for antibiogram. The estrus mucus can also be used to diagnose subclinical emdometritis in which case the colour of mucus is turned yellow when it is boiled with equal volume of 5% Na OH solution. The samples with no colour reaction are considered negative for endometritis. Recently, a simple grading system based on the character of vaginal mucus has been developed to evaluate cows with endometritis. The endometritis grade correlates with the presence of pathogenic organisms associated with uterine disease and is prognostic for the likely outcome of treatment. Score 0 Score 1 Score 2 Score 3 = = = = clear and translucent mucus mucus containing flakes of white or off-white pus discharge containing 50% white or off-white mucopurulent material discharge containing 50% purulent material usually white or yellow but occasionally sanguineous

Thus, cervical mucus examination should forma a part of routine investigation of various reproductive problems as well as in AI management. References: Sheldon I.M., Lewis G.S., Le Blanc S, Gilbert R.O. (2006). Defining post partum uterine disease in cattle. Theriogenology 65: 1516-1530. Hafez B and Hafez E.S.E. (2000). Reproduction in farm animals. 7th Williams & Wilkins, Philadelphia, USA. pp 26. edition. Lippincott

Fig 1: Normal fern leaf with typical branching pattern

Fig 2: Cervical mucus sample showing typical fern pattern (20X)

Fig 3: Cervical mucus sample showing typical fern (40 X)

Fig 4: Endometritis Scoring System