VaginalCytology TechniqueFor Vaginal Cytology it will require cotton-tippedapplicators, microscope slides, Diff-Quik stain, and a microscope. It is important that cellsobtained are indeed representative of hormonal changes. An assistant will have to restrain the animal in a standingposition as well as hold the tail up. The vulva has to be cleaned with dilutedchlorhexidine and rinsed. Lubricate a speculum and insert it in the vagina upto a point just cranial to the urethral orifice. Then, usinga sterile cotton-tipped applicator pass it into the vulva in a dorsaldirection into the vagina.
After gently swabbing the wall of the vagina, removethe swab and roll it onto a slide. Label it with name and date. Stain it withDiff Quik and allow it to air dry. When dry scan it under the microscope at 10Xand increase it to 40X. Student Log Book 1Name: MollySpecies: CanineBreed: Siberian HuskySex: FemaleAge: 6YODate of procedure: 01/08/18Description of procedureincluding what part student played in the treatment of the animal:Molly, A 6-year-old,intact female Siberian Husky was presented for breeding management prior toartificial insemination with fresh semen. She had first been observed to be inheat, considered by swelling of the vulva and bloody vaginal discharge 8 daysprior to presentation. Vaginal cytology on the day of presentation wascompletely non-cornified and white blood cells were present. The stage cyclecould be early in proestrus or just out of heat, in diestrus.
To differentiateearly proestrus from diestrus in this dog, the hormone progesterone wasmeasured in blood. Progesterone concentrations are low in proestrus, rise justbefore or during estrus, and remain high throughout diestrus. Serumprogesterone concentration in this dog was 0.4 ng/ml; she was in earlyproestrus on the day she was seen. The serum progesterone concentration wasrechecked five days later and it was 5.7 ng/ml, which indicated ovulation.
Shewas artificially inseminated with good quality semen two days after ovulation.Vaginal cytology was completely cornified, with the majority of cells appearinganuclear, at the time of artificial insemination. Student Log Book 2Name: LolaSpecies: CanineBreed: Labrador RetieverSex: FemaleAge: 3YODate of procedure: 01/03/18Description of procedureincluding what part student played in the treatment of the animal:Lola is 3 year-old Labrador retrievernoticed to have a swollen vulva and to be spotting blood. A series of vaginalsmears were taken as it follows: With Lola standing, the vulvar lips were gently parted and Iinserted a cotton-tipped moistened swab several inches past the vulva atan angle greater than 45 degrees. When the swab stopped forwardprogression, it indicates it is at the vestibular sphincter. I elevated theswab and applied gentle pressure to allow passage through the sphincter and progression to the anterior vagina. Then I rotated swab in the vagina to collect cells and it was subsequently withdrawn.
I rolled the swab across a clean microscope slide, forming three parallel lines on the slide.The slide was alcohol-fixed and stained. The firstlabeled as Day 1 I identified as Proestrus stage since all Intermediates,superficials and red blood cells were noted.
Day 2 I came to the conclusion itwas Estrus because I only noted superficial and red blood cells. On day 3 itwas still Estrus as again I only observed superficial cells. Day 6 Estrus stillall superficial cells were in the slide prepared. Same thing with day 9. Onday 10 I observed all superficial cells and some red blood cells, still estrus.On day 11 it was the first day of diestrus since my slide contained an abruptchange to intermediate and parabasal cells and I also noted red and white bloodcells. I had the opportunity to identify multiple estrus slides. Student Log Book 3Name: BellaSpecies: CanineBreed: MalteseSex: FemaleAge: 5YODate of procedure: 01/16/18Description of procedureincluding what part student played in the treatment of the animal:Maryis a 5 year old Maltese female unsprayed, that the owners desired to breed.
Ownerswere out of town for a week and, on returning noticed Mary had significantvulvar swelling and, when let outside, strongly attracted to the neighbor’smale. They brought her in. I took a sampleand prepared a vaginal smear and again another one 2 days later.
On the 3rdday owners took Mary to the stud and she was bred without difficulty. It is notimpossible, but Mary was very unlikely to be pregnant. She was clearly incytology estrus on Friday (day 1) 100% superficial cells were observed, but indiestrus on Sunday (day 3) as only one superficial cell, with the remaindermostly small intermediates was observed. Breeding took place on Monday, whichwas either diestrus day 2 or 3 and past the normal period of fertility.