5. Collection and Investigation of the Preputial Flushing

How can samples of smegma be collected for investigation?

Samples of smegma, a mixture of exfoliated epithelial cells and sebaceous gland secretions, can be collected using two methods: pipette and douche.

Describe the Pipette method for collection of smegma

* Dry pipette: A dry AI (artificial insemination) pipette with a 10-15 ml syringe attached is introduced into the fornix of the bull's prepuce. Negative pressure is applied while the pipette is scraped and slid back and forth in the sheath. At least 1 ml of milky white smegma at the pipette's tip is mixed with 3-5 ml of transport medium.
* Wet pipette: Similar to the dry technique, but involves infusing 5-10 ml of physiological saline or transport medium into the prepuce with the pipette. The prepuce is then held to prevent fluid from escaping, and vigorous massage is performed. The fluid is then drawn back into the pipette.

Describe the douche method for collection of smegma

The sheath is infused with 100-200 ml of physiological saline solution (PSS) or transport medium. The preputial opening is occluded, and the trapped fluid is vigorously massaged before being recollected.

How should collected samples be handled for investigation?

Collected samples should be handled with cleanliness and minimal exposure to air and sunlight. They should also be protected from extreme temperature changes and placed into a suitable culture medium, such as lactated Ringer's solution or buffered physiological saline, as soon as possible. Proper handling and transport of the samples are crucial to maintain their viability and integrity for further investigation.

What are the methods of investigation for Trichomonas?

* Microscopic observation: The collected mucus can be observed microscopically for the presence of Trichomonas protozoa. Morphological criteria such as an undulating membrane, three anterior flagella, irregular and jerky movement, and continuous rolling can aid in identifying Trichomonas.
* Culture: A small aliquot of the preputial smegma in transport medium is layered on the surface of a culture medium and incubated at 37°C. To determine if a culture is positive, a sample is placed on a microscope slide with a coverslip and viewed under a microscope, preferably with phase or dark-field illumination.

What are the methods of investigation for Bovine genital campylobacteriosis?

Fluorescent antibody test: When preputial samples cannot be processed immediately, a selective transport and enriched medium (such as Oxoid CM 391) can be used. The viability of the bacteria in this medium is around 1 week. Fluorescent antibody tests can be performed on the samples to detect the presence of Campylobacter fetus, the causative agent of bovine genital campylobacteriosis.