Cytology 1.docx

Cytology 1 Vaginal cytology Ear Cytology Fecal Cytology - Objectives - Technique and evaluation of exfoliative vaginal cytology - Stages of estrus-clinical signs and cytological findings - Common causes of otitis externa - Cytological examination of the external ear canal - Di...

Cytology 1 Vaginal cytology Ear Cytology Fecal Cytology - Objectives - Technique and evaluation of exfoliative vaginal cytology - Stages of estrus-clinical signs and cytological findings - Common causes of otitis externa - Cytological examination of the external ear canal - Discuss fecal cytology Vaginal Cytology - Exfoliative vaginal cytology is useful adjunct to the history and clinical examination in determining the stage of the estrous cycle in bitches and queens - Assists with optimal timing of mating or artificial insemination in small animals - Assist in diagnosing vaginitis Technique for obtaining sample - Animal is restrained in a standing position - Vulva is cleaned and rinsed - Place a lubricated speculum into the vulva - Using a sterile premoistened (saline) swab introduce the swab into the vagina - Rotate the swab - Remove swab and gently roll on 2 or 3 clean microscope slides - Dry and stain slides with diff quik - Dove video - Evaluation of Smear - always cytologic findings must be interpreted along with history and clinical signs - Four stages of estrus are: - Anestrus - Proestrus - Estrus - Diestrus Working Like a Dog \| Flickr - Photo Sharing! Why Vaginal Smears?? - Rising levels of estrogen cause the vaginal epithelium to become \"cornified\" - the surface cells become large and flattened, with small or absent nuclei. - Tracking changes in the morphology of desquamated vaginal epithelial cells provides a convenient means of assaying changes in estrogen levels.  NonCornified Epithelial cell  NonCornified with pyknotic nucleus CornifiedPykNuc NonCornified  NonCornified  Cornified CornifiedNoNucl Cornified  Anestrus - There is no vulvar swelling and does not attract male dogs - Vaginal smear - Noncornified squamous epithelium - Large cells with rounded border, abundant basophilic cytoplasm, and a large rounded nucleus - Based on size may be categorized as intermediate (larger) or parabasal (smaller) epithelial cells - Neutrophils may be present in small number - RBCs are seen Proestrus - Swollen vulva - Reddish discharge - Attract but not will not accept male in attempts to breed - Microscopically you will see: - proestrus progresses towards estrus the noncornified cells become Cornified squamous epithelium also called superficial cells - Cells with angular, jagged borders, homogeneous, pyknotic nucleus - Neutrophils - RBCs are present Proestrus Proestrus 2 Estrus - History of proestrus - Discharge has changed from red to pinkish and becoming whiter later in estrus more towards metestrus - Accepts male - Microscopically will see: - Vaginal smear reveals that all the squamous epithelial cells are cornified - Can appear to have no nucleus - neutrophils are seen - RBCs are present but decreased Estrus Estrus2 Diestrus - Has a recent history of estrus - Vulvar swelling and discharge have decreased - No longer attract males - Microscopically you will see: - Cornified epithelium are replaced by noncornified squamous epithelium - Neutrophils are present again - RBCs are absent Diestrus - Longest stage of the canine estrous cycle - 60days - No vulvar discharge - No longer attract males - Vulvar swelling and discharge is gone - Predominantly noncornified epithelial cells with a few WBC's - Progesterone secretion is at its peak - Pseudocyesis  - Ear Cytology - Otitis externa is commonly seen in the veterinary practice - Acute - Chronic - Bilateral - Unilateral - Signs: - Shaking head-mal odor - Pain pruritus -alopecia around the ear - tic discharge-excoriation Sounds Only Dogs Can Hear -- The Moderate Voice Conformation of the Ear Canal - Long , narrow ear canal - Traps moisture, foreign debris, and glandular secretions - Excessive hair in the external ear canal will also inhibit ventilation and increase the retention of cerumen - Pendulous ears - This kind of environment can destroy the protective barrier of the epidermis and allow for infections to occur - Animal's habits can lead to a predisposition for otitis - Hunting dogs-foreign objects - Swimmers- stimulates the production of cerumen and also traps moisture in the ear - Allergies (immune-mediated, contact) - Parasites - Bacteria - Yeast - Tumors - Trauma Collection of Sample - Cotton swab - Obtain sample after otoscope examine has been performed - Roll swab on a clean slide - not smear!!! - Collect more than one swab if you are to examine it for parasites - Stain it with diff quick - Examine with oil immersion lens looking for bacteria and yeast (100X) - Dove video - Cytological Examination - Should be evaluated for: - Bacteria (rods, cocci) - Yeast (Malassezia spp) - Fungal hyphae - Mites - Neutrophils - Cerumen - Neoplastic cells Recording Findings - Bacteria- - type/morphology - Grouping - Quantity - Always include the FOV!! - Yeast - Quantity & FOV - Record anything else seen and quantify - RBC or WBC - Increased \# of epithelial cells - Any other pertitant information - Bacteria (rods, cocci) - Normal flora of the ear canal can contain a small number of bacteria - Microscopically you should see none to occasional bacteria - Many of the bacteria are potentially pathogenic and when the conditions are optimal can colonize in the ear canal - In bacterial infections the cytological examine can reveal large numbers of bacteria along with neutrophils (PMNs) - Cocci shaped bacteria are usually Staphylococcus and Streptococcus spp - Rod shaped bacteria are usually Pseudomonas aeruginosa or Proteus mirabilis - Yeast (Malassezia spp) - Formerly known as Pityrosporum spp - Budding, gram positive yeast - Normal inhabitant of the external ear canal - Most common cause of mycotic otitis externa - Cytologic examine: - Organisms that are "peanut shaped" (round or elliptical shaped), dark blue staining, budding - hpf suggest overgrowth  - Concurrent bacterial infections are common - Gram positive cocci - Other fungi are possible but are rare - When unidentifiable yeast or hyphae are observed a culture should be done  Mites - Common problem in dogs and cats - Otodectes cynotis - Reddish brown or black granular discharge is seen - Secondary bacteria and/or yeast can exist - Slide preparation - Mineral oil - Swab (with or without oil on it) - Slide - 10x observation - Fecal Cytology - Clostridium perfringens (CP) - Enteric anaerobic gram positive rod - Normal enteric bacteria - Sporulation - Certain factors may cause CP organism to undergo enteric sporulation - During the sporulation process that the enterotoxin is released - Enterotoxin production causes intestinal damage and fluid accumulation in the intestines - Not very specific - alterations in bacterial micorflora - antibiotic therapy - intestinal epithelial damage - stress - acquired or nosocomial disease -   Causes - Signs may be acute or chronic - include (maybe intermittent ): - Diarrhea - Gas - Abdominal discomfort - ccasionally hemorrhagic diarrhea - ccasional vomiting Diagnosis - [Must be examined during a clinical episode] - Enterotoxin ELISA test - Common in house test is the fecal cytology - High numbers of CP spores serves as a quick and simple screening test - Identification of numerous spores on fecal cytology in conjunction with typical clinical signs tends In House Test - Fecal Smear - Make a very thin smear of the feces - Heat fix or air dry - Stain with Diff quick - Under oil immersion identify the spores - OIF suggests - characteristic gas "bubble" which can be located on the ends, middle, or encompass a majority of the organism - ften describes as having a safety pin appearance  Review - Technique and evaluation of exfoliative vaginal cytology - Stages of estrus-clinical signs and cytologic findings - Common causes of otitis externa - Cytologic examination of the external ear canal - Fecal Exam - CP