Feline sporotrichosis: Characterization of cutaneous and extracutaneous lesions using different diagnostic methods.

Sporotrichosis is a mycotic infection of the cutaneous and subcutaneous tissues caused by Sporothrix spp. that can also cause extracutaneous manifestations. This study aimed to characterize cutaneous and extracutaneous sporotrichosis lesions in cats. Over 1 year, 102 cats rescued by the Zoonoses Control Center of Belo Horizonte, Brazil, euthanized with clinical suspicion of feline sporotrichosis were evaluated. After euthanasia, the animals were evaluated by macroscopic, cytological, histopathological, and immunohistochemistry (IHC) examinations; fungal culture; and polymerase chain reaction (PCR). Sporothrix infection was identified by at least one diagnostic technique in all cats (n = 102) evaluated by postmortem examination, including 26/28 cases (93%) evaluated by IHC, 66/90 cases (73%) evaluated by cytology, 70/102 cases (68.6%) evaluated by histopathology, and 62/74 cases (84%) evaluated by fungal culture. Two cats had positive results only by fungal culture. Cytology and histopathology examinations were effective in diagnosing sporotrichosis, although IHC was needed to confirm the diagnosis in cats with low fungal loads. Sporothrix brasiliensis was confirmed by the sequencing of 3 samples. Skin lesions were characterized mainly by pyogranulomatous to granulomatous dermatitis (frequently with subcutaneous inflammation) with different intensities of Sporothrix spp. yeast. Extracutaneous findings associated with sporotrichosis included rhinitis or rhinosinusitis, lymphadenitis, pneumonia, meningitis, periorchitis, conjunctivitis, and glossitis. Extracutaneous infections were observed in 74/102 cases, and a possible association between the chronicity of the disease and the higher pathogenicity of this fungal species in cats requires further investigation.

An immersive simulation strategy to teach canine vaginal cytology: A multi-centre study.

Vaginal cytology (VC) is an essential technique for monitoring the bitch's estrus cycle. Currently, animal-free teaching methodologies have not been investigated for VC. Hence, this study aimed to evaluate an immersive simulation with a VC model and augmented reality tools. Students (n = 219) from four universities were enrolled, having learning stations with models for practising VC that provided immediate feedback on the technique. Augmented reality tools comprised QR codes that endorsed students to short videos of owners' avatars reporting the clinical reproductive story of the simulated animals and slides with QR codes leading to microscopy slide navigation videos. Proestrus, estrus, diestrus, anestrus and vaginitis were identified in the learning stations. The students' perceptions were evaluated through questionnaires assessing satisfaction, motivation, confidence, impact on learning and diagnostic accuracy. Before the immersive simulation, students had no experience with VC, being afraid of doing a VC with a live dog. Almost all the students considered practicing VC as essential and 94% reported that repeating the procedure (>2 times) was the most important parameter for learning. The simulation activity lasted ≈3 h and significantly improved the confidence of students, being less afraid of doing a VC in a live animal. Slide navigation videos improved the diagnostic accuracy of the estrus cycle stage, and students diagnosed the estrus and vaginitis cases more accurately. The immersive simulation strategy allowed repeated practice in a safe, motivated and standardized environment, being appraised by students as an essential strategy for learning VC.

A novel cytological technique for bacterial detection on canine skin.

BACKGROUND: Cytological examination of the skin is an important diagnostic technique in dermatology for disease diagnosis and monitoring of treatment. Impression smear and tape-strip preparation are gold standard cytological techniques. This study introduces a novel cytological method, slurry preparation, in which debris collected from the skin surface is macerated in warm sterile water and dried on the slide. HYPOTHESIS/OBJECTIVES: To compare organism yield between a novel and two standard cytological collection techniques by evaluating bacteria and Malassezia yeast counts. ANIMALS: Thirty client-owned dogs diagnosed with atopic dermatitis and lesions consistent with pyoderma and/or Malassezia dermatitis. MATERIALS AND METHODS: In a prospective, blinded comparison study, dermatological lesions from each dog were sampled using impression smear, tape-strip and slurry preparation methodologies. Ten random reticle fields per lesion, accounting for a total area of 6.25 × 10-3 mm2 (equivalent to ˜5% of a typical ×40 field), were evaluated by light microscopy for each cytological method. Bacterial and Malassezia organisms were measured as counts/sample. The slurry preparation was compared to each of the standard methodologies separately using paired Student's t-tests. RESULTS: The slurry preparation was demonstrated to be more sensitive than gold standard techniques in detecting bacteria, with mean differences of 12.7 and 13.5 additional bacteria per cytological sample, compared to the impression and tape-strip preparations, respectively. The slurry preparation did not differ significantly in detecting Malassezia organisms compared with impression smear preparation. The tape-strip preparation was demonstrated to be more sensitive than the slurry preparation in the detection of Malassezia organisms, finding a mean difference of 0.22 more organisms per sample. CONCLUSION: Slurry preparation presents advantages over the gold standard techniques in the detection of bacteria which can help to manage canine pyoderma more appropriately. Performing a slurry preparation in cases of suspected canine pyoderma could be considered as an alternative to the gold standard techniques.

Repeatability and reproducibility of microscopic examination of adhesive tape strip cytology slides for the quantification of Malassezia spp. in canine skin.

BACKGROUND: The optimal microscopic magnification and number of optical fields of adhesive tape strip cytological slides that should be examined when searching for Malassezia yeasts on canine skin are unknown. OBJECTIVES: To determine the optimal magnification and the minimum number of optical fields that should be examined to maximise intraobserver repeatability and interobserver reproducibility. MATERIALS AND METHODS: Seven experienced examiners counted, twice, the number of yeasts in 10, 20, 30, 40 and 50 optical fields of 40 slides at ×400 and ×1000 magnification. RESULTS: The number of yeasts per unit surface area was significantly higher at ×1000 compared to ×400 magnification. Repeatability and reproducibility for counting the yeasts was very poor. CONCLUSIONS AND CLINICAL RELEVANCE: Adhesive tape strip cytological slides should be examined microscopically for Malassezia spp. at ×1000 magnification. The repeatability of this examination for counting the yeasts is poor.

Contexte - Le grossissement microscopique optimal et le nombre de champs optiques des lames cytologiques de bandes adhésives à examiner lors de la recherche de levures Malassezia sur la peau de chien sont inconnus. Objectifs - Déterminer le grossissement optimal et le nombre minimal de champs à examiner pour maximiser la répétabilité intra-observateur et la reproductibilité inter-observateur. Matériels et méthodes - Sept examinateurs expérimentés ont compté, deux fois, le nombre de levures dans 10, 20, 30, 40 et 50 champs de 40 lames aux grossissements ×400 et ×1 000. Résultats - Le nombre de levures par unité de surface était significativement plus élevé au grossissement ×1 000 par rapport au grossissement ×400. La répétabilité et la reproductibilité du comptage des levures étaient très médiocres. Conclusions et pertinence clinique - Les lames cytologiques de bandes adhésives doivent être examinées au microscope pour Malassezia spp. à un grossissement ×1 000. La répétabilité de cet examen de comptage des levures est faible.

Introducción- se desconoce el aumento microscópico óptimo y el número de campos ópticos de los portaobjetos citológicos en tiras de cinta adhesiva que deben examinarse al buscar levaduras Malassezia en la piel canina. Objetivos- determinar el aumento óptimo y el número mínimo de campos ópticos que deben examinarse para maximizar la repetibilidad intraobservador y la reproducibilidad interobservador. Materiales y métodos- siete examinadores experimentados contaron dos veces el número de levaduras en campos ópticos de 10, 20, 30, 40 y 50 de 40 portaobjetos con aumentos de x ×400 y ×1000. Resultados- el número de levaduras por unidad de superficie fue significativamente mayor con un aumento de ×1000 en comparación con un aumento de ×400. La repetibilidad y reproducibilidad para contar las levaduras fue muy pobre. Conclusiones y relevancia clínica - Los portaobjetos citológicos en tiras de cinta adhesiva deben examinarse microscópicamente para detectar Malassezia spp. con un aumento de ×1.000. La repetibilidad de este examen para contar las levaduras es pobre.

Contexto - A ampliação microscópica ideal e o número de campos ópticos das lâminas citológicas de fita adesiva que devem ser examinados nas pesquisas de leveduras do gênero Malassezia em cães são desconhecidos. Objetivos - Determinar a magnificação ideal e o número mínimo de campos ópticos que devem ser examinados para maximizar a repetibilidade intraobservador e a reproducibilidade interobservador. Materiais e métodos - Sete examinadores experientes contaram duas vezes o número de leveduras em 10, 20, 30, 40 e 50 campos ópticos de 40 lâminas nas magnificações de x400 e x1000. Resultados - O número de leveduras por unidade de área de superfície foi significativamente maior em x1000 em comparação com a ampliação de x400. A repetibilidade e a reprodutibilidade para a contagem de leveduras foi muito pobre. Conclusões e relevância clínica - Lâminas de citologia por fica adesiva devem ser examinadas microscopicamente para Malassezia spp a uma magnificação de x1.000. A repetibilidade deste exame para contagem de leveduras foi pobre.

Vaginal swab cytology as a diagnostic tool for neoplasia of the lower urinary tract in 5 dogs.

Urothelial carcinoma (UC), or transitional cell carcinoma, is the most common canine urothelial malignancy in dogs. Females are predisposed and provide a challenge for diagnostic tumor sampling. The objectives of this study were to investigate the use and tolerability of vaginal swab cytology for UC diagnosis. Five dogs were identified with non-diagnostic urine sediment cytology and UC diagnosed on vaginal cytology with confirmation by another means. All patients tolerated the vaginal swab with minimal restraint. This study confirms the potential of vaginal swab cytology as a simple, inexpensive, and well-tolerated means for lower urinary tract UC diagnosis in female dogs. Key clinical message: Vaginal swab cytology is a non-invasive, low-cost method of obtaining a sample for cytological assessment for UC.

La cytologie vaginal par écouvillonnage comme outil de diagnostic de la néoplasie des voies urinaires inférieures chez cinq chiennes. Le carcinome urothélial (CU), ou carcinome à cellules transitionnelles, est la tumeur maligne urothéliale canine la plus courante. Les femelles sont prédisposées et constituent un défi pour l'échantillonnage diagnostique des tumeurs. Les objectifs de cette étude étaient d'étudier l'utilisation et la tolérabilité de la cytologie vaginale par écouvillonnage pour le diagnostic du CU. Cinq chiennes ont été identifiées avec une cytologie des sédiments urinaires non diagnostique et un CU diagnostiqué sur la cytologie vaginale avec confirmation par un autre moyen. Toutes les patientes ont toléré le prélèvement vaginal avec un minimum de contention. Cette étude confirme le potentiel de la cytologie vaginale par écouvillonnage en tant que moyen simple, peu coûteux et bien toléré pour le diagnostic du CU des voies urinaires inférieures chez les chiennes.Message clinique clé :La cytologie vaginale par écouvillonnage est une méthode non-invasive et peu coûteuse pour obtenir un échantillon pour évaluation cytologique du CU.(Traduit par les auteurs).

Precision-cut bovine udder slices (PCBUS) as an in-vitro-model of an early phase of infection of bovine mastitis.

BACKGROUND: The aim of this study was to establish precision-cut bovine udder slices (PCBUS) as an in-vitro-model to investigate pathophysiological processes in the early phase of mastitis in order to have the possibility to investigate new therapeutic approaches for the treatment of such udder inflammation in later studies. Furthermore, this model should contribute to substitute in-vivo-experiments. Bovine mastitis is one of the most common and costly infectious diseases in the dairy industry, which is largely associated with the use of antimicrobial agents. Given this problem of antimicrobial resistance, it is essential to step up research into bacterial infectious diseases. Thus, the transfer of the in-vitro-model of precision-cut tissue slices to the bovine udder enables broad research into new therapeutic approaches in this area and can also be used to address issues in basic research or the characterisation of complex pathophysiological processes. RESULTS: A stimulation with LPS, PGN or the combination of both substances (LPS:PGN) demonstrates the ability of the PCBUS to react with a significant secretion of IL-1ß, TNF-α and PGE2. CONCLUSION: The slices represent an instrument for investigating pharmacological interactions with udder tissue, which can be useful for studies on pharmacological questions and the understanding of complex pathophysiological processes of infection and inflammation.

Diagnostic Accuracy of a Direct Panfungal Polymerase Chain Reaction Assay Performed on Stained Cytology Slides.

Molecular techniques are increasingly being applied to stained cytology slides for the diagnosis of neoplastic and infectious diseases. Such techniques for the identification of fungi from stained cytology slides have not yet been evaluated. This study aimed to assess the diagnostic accuracy of direct (without nucleic acid isolation) panfungal polymerase chain reaction (PCR) followed by sequencing for identification of fungi and oomycetes on stained cytology slides from dogs, cats, horses, and other species. Thirty-six cases were identified with cytologically identifiable fungi/oomycetes and concurrent identification via fungal culture or immunoassay. Twenty-nine controls were identified with no cytologically or histologically visible organisms and a concurrent negative fungal culture. Direct PCR targeting the internal transcribed spacer region followed by sequencing was performed on one cytology slide from each case and control, and the sensitivity and specificity of the assay were calculated. The sensitivity of the panfungal PCR assay performed on stained cytology slides was 67% overall, 73% excluding cases with oomycetes, and 86% when considering only slides with abundant fungi. The specificity was 62%, which was attributed to amplification of fungal DNA from control slides with no visible fungus and negative culture results. Direct panfungal PCR is capable of providing genus- or species-level identification of fungi from stained cytology slides. Given the potential of panfungal PCR to amplify contaminant fungal DNA, this assay should be performed on slides with visible fungi and interpreted in conjunction with morphologic assessment by a clinical pathologist.

Correlations between clinical signs and corneal cytology in feline eosinophilic keratoconjunctivitis.

OBJECTIVE: To assess correlations between clinical and cytological features of feline eosinophilic keratoconjunctivitis at the time of cytological diagnosis. ANIMALS STUDIED: Fifteen client-owned, domestic breed cats (18 eyes) examined between 2007 and 2019. PROCEDURES: An electronic search and medical record review of cats diagnosed with feline eosinophilic keratitis or keratoconjunctivitis (FEK) based on clinical examination findings and eosinophils detected on corneal cytology were conducted. Clinical severity was graded using a modified version of a previously validated semiquantitative preclinical ocular toxicology scoring (SPOTS) system. Clinical grades were assigned following review of clinical images and medical record descriptions, and cytological grades were assigned following review of archived corneal cytology slides. Correlations were analyzed for significance using Spearman's rank correlation coefficient. RESULTS: Higher total corneal scores correlated with higher total conjunctival scores, but not with total fluorescein scores. Small lymphocyte scores correlated negatively with scores for collagen degeneration or mineralization. Globule leukocytes, a unique cell type not previously described in ocular cytology, were identified in 4 of 18 cytological samples. Higher globule leukocyte scores were correlated with higher scores for mast cells or plasma cells. Specimens with lower eosinophil scores had higher globule leukocyte scores. CONCLUSIONS: Large variability was detected in the cytological characteristics and clinical features of FEK-affected cats. This is the first report of globule leukocytes being identified in ocular cytology from any species. The role of globule leukocytes in the etiopathogenesis and progression of FEK remains unknown and warrants further investigation.

Influence of locoregional lymph node aspiration cytology vs sentinel lymph node mapping and biopsy on disease stage assignment in dogs with integumentary mast cell tumors.

OBJECTIVE: To compare the effect of sentinel lymph node (SLN) histology vs locoregional lymph node (LRLN) fine needle aspiration (FNA) cytology on assigned disease stage and adjunctive treatment recommendations and describe the incidence of anatomic disparity between the LRLN and SLN. STUDY DESIGN: A pre-post study refers to a study design type in which subjects are compared pre and post the intervention of interest. ANIMALS: Seventeen dogs undergoing primary excision of 20 cutaneous and subcutaneous mast cell tumors (MCT). METHODS: Client-owned dogs presenting to the Cornell University Hospital for Animals for surgical removal of a cytologically confirmed cutaneous or subcutaneous MCT >1 cm in diameter were enrolled. Cytological examination of FNA from the LRLN was compared with histology of the SLN. The SLN was identified by indirect computed tomographic lymphangiography (ICTL) after peritumoral injection of iopamidol and scanning at 1, 3, 5, 10, and 15 minutes. Histopathologic node score > 1 was considered metastatic. After case review by an oncologist, LRLN FNA cytology was compared with SLN histology for effect on changes in stage assignment and adjunctive treatment recommendations. RESULTS: Mast cell tumors were graded as 2 low (n = 11), 2 high (n = 2), and subcutaneous (n = 7). Optimal scan timing was 10 minutes after injection of iopamidol. Sentinel lymph node differed anatomically from LRLN in 5 of 18 scans. Metastases were detected by histology in 9 of 20 SLN compared with in 1 of 20 FNA of LRLN (P = .001), changing stage and adjunctive treatment recommendations 8 of 20 tumors. Only 6 of 19 LRLN FNA samples were diagnostic. CONCLUSION: Sentinel lymph nodes were consistently identified with ICTL and differed from LRLN in one-quarter of tumors. Histopathological examination of SLN altered recommendations in half of the dogs compared with the previous standard of care. CLINICAL SIGNIFICANCE: Indirect computed tomographic lymphangiography and SLN excision should be considered as a new standard for dogs with MCT.

Evaluation of cytology and histopathology for the diagnosis of feline orbital neoplasia: 81 cases (2004-2019) and review of the literature.

OBJECTIVE: To provide an updated overview of feline orbital neoplasia, to compare diagnostic utility of cytology and histopathology, and to evaluate minimally invasive sampling modalities. PROCEDURES: A medical records search was performed to identify cats with orbital neoplasia. Data were collected regarding signalment, diagnosis, vision status, imaging modalities, and sample collection methods. A reference population with orbital neoplasia was also identified via literature search for comparison with regard to final diagnosis. RESULTS: Eighty-one cats met selection criteria and 140 cases were identified in the literature. In the study and reference populations, respectively, diagnoses were grouped as follows: round cell tumors 47% and 24%, epithelial tumors 38% and 40%, mesenchymal tumors 14% and 34%, and neurologic origin tumors 1% and 2%. The most common diagnoses in both groups were lymphoma and squamous cell carcinoma (SCC). Feline restrictive orbital myofibroblastic sarcoma (FROMS) was common in the reference population but not diagnosed in the study population. Cytology results were available for 41 cats; histopathology results were available for 65 cats. Both cytology and histopathology results were available for 25 cats, in 44% of which cytologic results were overturned. No significant complications were associated with any sampling method. Lack of cats with multiple samples available for histopathology limited comparison between tissue sampling methods. CONCLUSIONS: Orbital neoplasia is common in cats, with round cell and epithelial tumors diagnosed most commonly in the study population. Histopathology is superior to cytology in providing a definitive diagnosis. Minimally invasive tissue biopsy techniques appear to be safe and effective.

Hematology Red Flags: The Value of Blood Smear Examination in Horses.

This article uses a case-based approach, complemented with diagnostic algorithms and images, to highlight hematologic changes of pathologic relevance in horses, namely, marked erythrocytosis, anemia or leukocytosis, inflammatory leukograms, lymphocytosis in adult horses, thrombocytopenia, and pancytopenia. These hematologic abnormalities occur with certain diseases and their identification can help clinicians narrow to down differential diagnostic lists. This article highlights the importance of blood smear examination, particularly, but not only, when numerical red flags are identified on automated blood counts.

Practical Tips on Sample Handling for Hematology, Chemistry, and Cytology Testing for Equine Patients:: Getting More Bang for your Buck.

Clinical pathology results are only as good as the quality of samples and accompanying information submitted to the diagnostic laboratory. The frustration of nondiagnostic or equivocal test results can often be avoided by taking the time to follow sample handling and submission guidelines. This article discusses preanalytical errors that commonly affect the accuracy of hematology, chemistry, and cytology testing, and offers practical tips for preventing these errors and maximizing diagnostic yield.

A high-throughput method for unbiased quantitation and categorization of nuclear morphology†.

The physical arrangement of chromatin in the nucleus is cell type and species-specific, a fact particularly evident in sperm, in which most of the cytoplasm has been lost. Analysis of the characteristic falciform ("hook shaped") sperm in mice is important in studies of sperm development, hybrid sterility, infertility, and toxicology. However, quantification of sperm shape differences typically relies on subjective manual assessment, rendering comparisons within and between samples difficult. We have developed an analysis program for morphometric analysis of asymmetric nuclei and characterized the sperm of mice from a range of inbred, outbred, and wild-derived mouse strains. We find that laboratory strains have elevated sperm shape variability both within and between samples in comparison to wild-derived inbred strains, and that sperm shape in F1 offspring from a cross between CBA and C57Bl6J strains is subtly affected by the direction of the cross. We further show that hierarchical clustering can discriminate distinct sperm shapes with greater efficiency and reproducibility than even experienced manual assessors, and is useful both to distinguish between samples and also to identify different morphological classes within a single sample. Our approach allows for the analysis of nuclear shape with unprecedented precision and scale and will be widely applicable to different species and different areas of biology.

Diagnostic Validation of a Whole-Slide Imaging Scanner in Cytological Samples: Diagnostic Accuracy and Comparison With Light Microscopy.

Digital slides created by whole-slide imaging scanners can be evaluated by pathologists located in remote sites, but the process must be validated before this technology can be applied to routine cytological diagnosis. The aim of this study was to validate a whole-slide imaging scanner for cytological samples. Sixty cytological samples, whose diagnoses were confirmed by gold-standard examinations (histology or flow cytometry), were digitalized using a whole-slide imaging scanner. Digital slides and glass slides were examined by 3 observers with different levels of cytopathological expertise. No significant differences were noted between digital and glass slides in regard to the number of cases correctly diagnosed, or the sensitivity, specificity, or diagnostic accuracy, irrespective of the observers' expertise. The agreements between the digital slides and the gold-standard examinations were moderate to substantial, while the agreements between the glass slides and the gold-standard examinations were substantial for all 3 observers. The intraobserver agreements between digital and glass slides were substantial to almost perfect. The interobserver agreements when evaluating digital slides were moderate between observers 1 and 2 and between observers 1 and 3 while they were substantial between observers 2 and 3. In conclusion, our study demonstrated that the digital slides produced by the whole-slide imaging scanner are adequate to diagnose cytological samples and are similar among clinical pathologists with differing levels of expertise.

Comparison of 3 corneal cytology collection methods for evaluating equine ulcerative keratitis: Cytobrush, kimura platinum spatula, and handle edge of scalpel blade.

PURPOSE: To compare corneal cytology samples from three common sampling techniques: cytobrush (CB), Kimura platinum spatula (KS), and the handle edge of a scalpel blade (SB). METHODS: Equine patients presenting to the University of Florida College of Veterinary Medicine with ulcerative keratitis were included. Following diagnosis of corneal ulcer and sampling for microbial culture, two cytology samples per technique were collected with sterile CB, KS, and SB in a randomized order. Cytologic evaluation was performed by two observers masked to collection method. Objective measures of sample cellularity, quality, distribution, and identification of infectious organisms were recorded per 10 monolayer cell populations using 50× magnification with oil immersion which were compared to culture results. Variables were compared using ANOVA with Student's t test when appropriate and Cohen's kappa (k) to evaluate inter- and intra-observer agreement (IOA) between observers and techniques. RESULTS: Twenty equine patients (120 samples) were included. The IOA between observers was substantial (k = 0.75 ± 0.06) for cytological parameters. SB provided the most cellular samples (P < .01). There was a trend toward agreement (k = 0.12 ± 0.16) in technique for sample quality (P = .08). CB and SB had significantly poorer cell distribution than KS (P < .05). Infection was confirmed in 12 of 20 patients with SB and CB techniques having a significantly higher diagnostic yield than KS (P < .05) and was most consistent with infection confirmed on culture. CONCLUSIONS: The SB provided the most diagnostic samples but all three techniques are clinically useful in evaluating equine ulcerative keratitis.

External ear cytological results and resident flora of clinically normal alpacas (Vicugna pacos).

BACKGROUND: Otitis is common in alpacas. Suppurative otitis media/interna can be an extension from the external ear canal or from a respiratory infection. Cytological evaluation provides rapid and inexpensive information to assist in therapeutic decision; to date, there is no published information regarding the normal cytological results and flora of the alpaca external ear canal. HYPOTHESIS/OBJECTIVES: To describe normal resident cytological findings and flora and possible variation over time, we sampled clinically normal alpaca external ear canals during two different seasons. ANIMALS: Fifty privately owned, healthy alpacas of different ages and sexes in two northeastern United States flocks. METHODS AND MATERIALS: One ear per alpaca had both cytological swabs (ectoparasites, inflammatory and epithelial cells, bacteria and yeast) and sterile swabs (bacterial and fungal cultures) taken. This was done in August 2017 and repeated in January 2018. RESULTS: Yeast organisms were noted cytologically in 2-4% of the samples. Prevalence of total yeast genera was 6% in August and 30% in January. Cytologically, rod-shaped bacteria [maximum 4-10/high power field (HPF); median 0-0.5/HPF] were seen in 50% of alpacas in August and 26% in January. Coccal bacteria (maximum 6-10/HPF; median 0/HPF) were seen in 32% of alpacas in August and 16% in January. No statistically significant findings were noted between sampling months. Common bacterial genera isolated in August were Bacillus (44%), Arthrobacter (40%) and nonhaemolytic Staphylococcus (26%), and in January were Bacillus (42%) and Pantoea (38%). CONCLUSIONS AND CLINICAL IMPORTANCE: This information may be useful when evaluating alpaca external ear canal samples, which subsequently may help dictate empirical therapy.

Diagnostic performance of cytology for assessment of hepatic lipid content in dairy cattle.

The objective of our study was to characterize the diagnostic performance of cytology for assessing hepatic lipid content (HLC) in dairy cows by comparing microscopic evaluation of lipid vacuolation in touch imprint slide preparations of liver biopsies with quantitative measurement of triglyceride concentration ([TG]; mg/mg of wet weight) in paired biopsy samples. Our study also sought to compare the diagnostic performance of liver cytology, plasma nonesterified fatty acid concentration ([NEFA]), and plasma ß-hydroxybutyrate concentration ([BHB]) derived from a measurement performed on whole blood, for assessing HLC. Chemical extraction of TG from liver tissue remains the gold standard for quantifying HLC, largely because available blood tests, although useful for detecting some types of pathology, such as increased lipid mobilization, ketosis, or hepatocellular injury, are nonspecific as to etiology. Veterinary practitioners can sample bovine liver for cytological evaluation in a fast, minimally invasive, and inexpensive manner. Thus, if highly predictive of HLC, cytology would be a practical diagnostic tool for dairy veterinarians. In our study, liver biopsy samples from Holstein cows (219 samples from 105 cows: 52 from cows 2 to 20 d prepartum, 105 from cows 0 to 10 d in milk, 62 from cows 18 to 25 d in milk) were used to prepare cytology slides and to quantify [TG] using the Folch extraction method followed by the Hantzch condensation reaction and spectrophotometric measurement. An ordinal scale (0-4) based on amount of hepatocellular cytoplasm occupied by discrete clear vacuoles was used by 3 blinded, independent observers to rank HLC in Wright-Giemsa-stained slides. Interobserver agreement in cytology scoring was good. Corresponding plasma [NEFA] and [BHB] measurements were available for 187 and 195 of the 219 samples, respectively. Liver [TG] correlated more strongly with cytology score than with NEFA or BHB, and receiver operating characteristic curve analysis showed that cytology had better diagnostic performance than either NEFA or BHB for correctly categorizing [TG] at thresholds of 5, 10, and 15%. Hepatic lipidosis in high-producing dairy cows is of major clinical and economic importance, and this study demonstrates that cytology is an accurate means of assessing HLC. Additional work is indicated to evaluate the diagnostic utility of liver cytology.

Accuracy of ultrasonography and fine-needle aspiration cytology in the diagnosis of prostate diseases in dogs.

Clinical signs of prostatic diseases in dogs are often non-specific. Appropriate treatment should be based on a detailed investigation using reliable diagnostic tools. The aim of our study was to evaluate the diagnostic value of ultrasonography (US) and fine-needle aspiration (FNA) cytology in dogs' prostate diseases. The mean accuracy of FNA cytology and US were 0.72 and 0.88 (n = 13), respectively. US gland size measurements and actual gland dimensions were highly concordant. Obtained results confirm the high diagnostic value of US and FNA biopsy and in prostatic diseases. Diagnosis based on US is highly reliable; however, it should be combined with clinical signs. Therefore, cytological evaluation of prostate gland material may be performed to differentiate or confirm presumptive diagnosis.

Comparison of two ear cytological collection techniques in dogs with otitis externa.

BACKGROUND: Canine otitis externa is a common disease. Cytological evaluation of otic exudate is a useful diagnostic test to direct and monitor treatment for otitis externa. One method of collecting ear cytological specimens utilizes a cotton-tipped swab inserted into the vertical ear canal. A proposed alternative method is to aspirate exudate from the deep horizontal canal using a rubber tube. HYPOTHESIS/OBJECTIVES: To compare cytological findings between two different collection techniques by evaluating the numbers of bacteria, Malassezia yeast and inflammatory cells. ANIMALS: Thirty client-owned dogs with otitis externa. METHODS AND MATERIALS: A prospective, randomized, blinded comparison study. Ear canals from each dog were sampled using cotton swab and rubber tube. Ear cytological preparations were evaluated independently by two investigators for polymorphonuclear leucocytes (PMN), monocytes/lymphocytes, macrophages, yeast, intracellular (IC) cocci, extracellular (EC) cocci, IC bacilli and EC bacilli. RESULTS: A paired Student's t-test was used to compare the two techniques. The inter-investigator reliability for PMN, EC bacilli and yeast was good, and for EC cocci was moderate. There were significantly higher numbers of PMNs obtained by the tube method (P = 0.0024) than by the cotton swab method. There were no statistically significant differences between cotton swab and rubber tube methods for monocytes/lymphocytes (P = 0.7780), macrophages (P = 0.1751), EC cocci (P = 0.1262), EC bacilli (P = 0.1162), yeast (P = 0.5371), IC cocci (P = 0.6606) or IC bacilli (P = 0.6761). The technique was well tolerated. CONCLUSION: An alternative ear cytological collection technique was identified which enables sampling of the deep horizontal canal.

Cytomorphometry of serosal effusion in dogs.

Cytomorphometry made on cytological slides is the quantitative method of precise analysis of cellular structures, including both cytoplasm and nucleus. The aim of this study was to describe cytomorphometric parameters of mesothelial cells in the course of benign reactive and malignant proliferation and to compare them to carcinomas and adenocarcinomas located within serosal cavities in dogs. The second aim was to evaluate applicability of cytomorphometry to diagnostics of diseases causing accumulation of effusion in serosal cavities. Cytological samples of normal and non-malignant mesothelium, mesothelioma and various carcinomas were collected from dogs. Cytomorphometry was made on the smears stained with Giemsa solution. Mean nuclear and cellular perimeter, mean nuclear and cellular area, mean nuclear and cellular diameter, and mean nuclear and cellular roundness were determined. Moreover, nuclear to cytoplasmic ratio (N/C) was calculated. The data revealed statistically significant differences for all parameters, excluding mean nuclear perimeter, between compared groups. Normal mesothelium cells and their nuclei were significantly smaller and more elongated than cells and nuclei of both benign reactive and malignant neoplastic mesothelium. Only a few differences were observed between benign reactive mesothelium cells and mesothelioma cells - mean nuclear area and mean nuclear diameter of benign reactive mesothelium cells were significantly larger and N/C ratio was higher in comparison to mesothelioma cells. Even though some significant differences were observed, considerable overlap of these cytomorphometric parameters in animals with different diseases limited practical role of these observations. Cytomorphometric analysis of cellular samples collected from dogs with proliferative processes affecting serosal cavities can be only an auxiliary method increasing accuracy of preoperative diagnosis.