Histology, prevalence, and environmental sources for pulmonary silicates depositions in domestic and wild animals.

The source and significance of pulmonary silicate crystals in animals and people are poorly understood. To estimate the prevalence and characterize the pulmonary crystalline material in animals from St. Kitts, tissue samples from dogs, horses, cattle, sheep, goats, pigs, chickens, mongooses, and monkeys were examined by light microscopy, scanning electron microscopy with energy-dispersive x-ray analysis (SEM/EDXA), and x-ray diffraction. Crystalline material was seen in 201 of 259 (77.6%) lung samples as perivascular and interstitial accumulations of heterogeneous crystalline particulate material, free or within macrophages (silicate-laden macrophages [SLMs]), mostly lacking evidence of chronic inflammation or fibrosis. The crystalline material was birefringent, basophilic on acid-fast, and composed of silicas on SEM/EDXA. Mongooses (100%) and monkeys (98%) had the highest prevalence of SLM, followed by cattle and chickens. Lesions were graded on a 3-point scale based on the histologic location and extent of silicates and SLM and were significantly more severe in mongooses (median = 3) than in monkeys (median = 2), dogs (median = 2), and chickens (median = 1). On EDXA, the crystalline material from lungs, air, and topsoil was composed of silicon, oxygen, aluminum, and iron, with a particulate matter size between 2.5 and 10 µm. We hypothesize Saharan dust, volcanic ash, topsoil, and rock quarry dust are potential sources of siliceous dust inhalation and SLM accumulations lacking chronic inflammation (silicosis); dust generation may be potentiated by road vehicle or wind suspension. Future investigations are warranted on the role of silicate inhalation and respiratory comorbidities in people, with monkeys, mongooses, or chickens serving as possible sentinels for exposure.

International Guidelines for Veterinary Tumor Pathology: A Call to Action.

Standardization of tumor assessment lays the foundation for validation of grading systems, permits reproducibility of oncologic studies among investigators, and increases confidence in the significance of study results. Currently, there is minimal methodological standardization for assessing tumors in veterinary medicine, with few attempts to validate published protocols and grading schemes. The current article attempts to address these shortcomings by providing standard guidelines for tumor assessment parameters and protocols for evaluating specific tumor types. More detailed information is available in the Supplemental Files, the intention of which is 2-fold: publication as part of this commentary, but more importantly, these will be available as "living documents" on a website (www.vetcancerprotocols.org), which will be updated as new information is presented in the peer-reviewed literature. Our hope is that veterinary pathologists will agree that this initiative is needed, and will contribute to and utilize this information for routine diagnostic work and oncologic studies. Journal editors and reviewers can utilize checklists to ensure publications include sufficient detail and standardized methods of tumor assessment. To maintain the relevance of the guidelines and protocols, it is critical that the information is periodically updated and revised as new studies are published and validated with the intent of providing a repository of this information. Our hope is that this initiative (a continuation of efforts published in this journal in 2011) will facilitate collaboration and reproducibility between pathologists and institutions, increase case numbers, and strengthen clinical research findings, thus ensuring continued progress in veterinary oncologic pathology and improving patient care.

Testicular Teratocarcinoma in a Ferret (Mustela putorius furo).

A 7-month-old male domestic ferret (Mustela putorius furo) was presented for evaluation of unilateral testicular enlargement. Microscopic examination of the left testicle revealed a neoplasm with differentiation along multiple cell lines (ectoderm, endoderm, mesoderm) including respiratory epithelium, bone and haired skin. A poorly differentiated epithelial component was dispersed throughout the neoplasm with invasion of testicular lymphatics. The animal developed progressive dysuria and was euthanized. At necropsy, metastasis of the poorly differentiated epithelial component was present in the urinary bladder, ureters, prostate gland, pelvic fat, abdominal and thoracic lymph nodes, kidney and lung. This is the first report of a malignant testicular teratoma with widespread metastasis in this species.

Filarial dermatitis caused by Filaria taxideae in domestic ferrets (Mustela putorius furo) from the western United States.

Filaria taxideae is a common subcutaneous filarial parasite of American badgers (Taxidea taxus) that has also been reported in skunks in the Western USA. Dermatitis associated with this parasite has been reported in badgers and skunks; however, many individuals have no gross lesions. With the exception of a captive red panda in California, there have been no reports of F. taxidae infection in domestic or exotic/pet animals. In this case series, we document F. taxideae in domestic ferrets from California, Texas, and Colorado and describe the clinical presentation, gross and histologic lesions, and the molecular characterization of this parasite. In two cases, ferrets were positive on Dirofilaria immitis antigen tests but had no evidence of heartworm infection suggesting that F. taxideae antigens can cross-react. These findings indicate that F. taxideae should be considered in cases of dermatitis in ferrets in areas where this parasite may occur.

Management of severe craniofacial vascular malformation operated under bypass: evolution from a "no-flow" to a "low-flow".

BACKGROUND: Cardiopulmonary bypass (CBP) and circulatory arrest as an assist in the surgical excision of a severe facial vascular malformation were first described by Mulliken et al in 1979. Later on, its use had expanded for resection of intracranial vascular malformations. However, to date, there have not been any published series of these procedures being used in the resection of craniofacial vascular malformations. We sought to review the first 10 surgical procedures performed at McGill University Health Centre for large vascular malformations resection using hypothermic CBP with or without circulatory arrest. METHODS: All consecutive patients at the McGill University Health Centre who had a craniofacial vascular malformation resected with the aid of CBP were reviewed. A comparison of the classic midline sternotomy with cardiac arrest to percutaneous femoral bypass with hypothermic "low flow" was performed. Charts were reviewed for the operative intervention including bypass parameters and short- and long-term complications of the procedure. RESULTS: Cardiopulmonary bypass was used in 9 patients for 10 surgical procedures for the resection of a variety of craniofacial vascular malformations from 1987 to 2001. All lesions had sclerotherapy and embolization of the feeding vessels 72 to 96 hours preoperatively. The average age of our patients was 21 ± 13.4 years (2-37 years). Procedures were conducted via either an open bypass or a closed femoral approach. There were no mortalities. There were 2 major cardiac intraoperative complications and 1 major postoperative complication, which were managed with no sequelae. The average length of postoperative hospital stay was 10 days. All patients went on to full recovery. The blood transfusions varied from 10 U to 0 U for our last patient. CONCLUSIONS: The assistance and adjunct of CBP are a useful procedure in the resection of very large vascular malformations, in selected cases. There were no major long-term complications in this series. With the evolution of our approach, the use of complete circulatory arrest was not required in the majority of cases, and an adequate resection was usually possible with the low-flow state alone as we developed this technique with more experience through the process.

Preoperative diplopia: the most important prognostic factor for diplopia after surgical repair of pure orbital blowout fracture.

BACKGROUND: No consensus exists in the literature regarding the risk factors associated with new or residual diplopia after pure orbital blowout fracture (BOF) repair. OBJECTIVE: To assess and evaluate the risk factors associated with diplopia after surgical repair of pure BOF. METHODS: Patients with pure BOF who were managed surgically were identified in an 11-year period at the McGill University Health Center. The association between new or residual diplopia postsurgical repair and various risk factors was assessed using chi and Fisher exact tests, and multivariate analysis was conducted using logistic regression. RESULTS: A total of 61 patient charts were reviewed. Results demonstrated the presence of preoperative diplopia and radiologic evidence of extraocular muscle (EOM) swelling to be strongly associated with diplopia at 6 months after repair (P < 0.05). Patients who presented preoperatively with diplopia had a 9.91 times greater probability of developing diplopia postoperatively (P = 0.035; 95% confidence interval, 1.17-83.80). CONCLUSIONS: Preoperative diplopia is the best predictor of the presence of postoperative diplopia after BOF repair. Initial injury to the EOM leading to EOM swelling and preoperative diplopia seems to be the origin of diplopia after surgical repair of pure BOF.

Multiple urogenital abnormalities in a Persian cat.

A 1.5-year-old female Persian cat was presented for inappetence and azotemia. Ultrasonography and urography revealed multiple abnormalities involving the genitourinary tract, including agenesis of the right kidney and ureter. Gross examination of the abnormal uterus revealed segmental aplasia of right caudal uterine horn causing cranial distension with fluid, a normal left uterine horn, and both normal ovaries. Microscopically, endometrial glands of the right uterine horn were markedly decreased in number. The right uterine horn was hemorrhagic suggesting estrus. This is the first report of this combination of urinary and uterus abnormalities in the veterinary literature.

Discoid lupus erythematosus (DLE) in a Spitz dog.

A 3-year-old, female Spitz, was presented due to lack of response to therapies with a 6-month history of skin lesions characterized by diffuse erythema and scaling on the dorsal trunk. Physical examination revealed the dog was active and healthy. Skin culture isolated no fungus. Histological examination of skin biopsy specimens revealed interface dermatitis with hydropic degeneration of the basal layers, predominant plasmacytic perivascular accumulation in the dermis, and intensive plasma cell-rich interface mural folliculitis. Moderate CD3-positive lymphocytes infiltrated the superficial dermis. This report may provide unique information of canine discoid lupus erythematosus in an unusual breed with atypical cutaneous lesions.

Calcinosis circumscripta on lingual muscles and dermis in a dog.

A 10-month old male Shi-tzu was presented with a week history of scurfy skin and whitish masses located in the ventral portion of tongue. Grossly, two whitish prominences with diameters of 3 to 4 mm were found on the underside of the tongue. Microscopically, in ulcerative epidermis, deposition of dense, amorphous granular basophilic calcium salts was separated by thin fibrous connective tissue containing mild inflammation. Many fascicles were characterized by replacement of degenerating myofibers with calcification. The precise cause of calcification could not be determined; however, it is interesting that two different lesions shown in calcinosis circumscripta and slight nutritional myopathy were also observed simultaneously only in the tongue.

An array microscope for ultrarapid virtual slide processing and telepathology. Design, fabrication, and validation study.

This paper describes the design and fabrication of a novel array microscope for the first ultrarapid virtual slide processor (DMetrix DX-40 digital slide scanner). The array microscope optics consists of a stack of three 80-element 10 x 8-lenslet arrays, constituting a "lenslet array ensemble." The lenslet array ensemble is positioned over a glass slide. Uniquely shaped lenses in each of the lenslet arrays, arranged perpendicular to the glass slide constitute a single "miniaturized microscope." A high-pixel-density image sensor is attached to the top of the lenslet array ensemble. In operation, the lenslet array ensemble is transported by a motorized mechanism relative to the long axis of a glass slide. Each of the 80 miniaturized microscopes has a lateral field of view of 250 microns. The microscopes of each row of the array are offset from the microscopes in other rows. Scanning a glass slide with the array microscope produces seamless two-dimensional image data of the entire slide, that is, a virtual slide. The optical system has a numerical aperture of N.A.= 0.65, scans slides at a rate of 3 mm per second, and accrues up to 3,000 images per second from each of the 80 miniaturized microscopes. In the ultrarapid virtual slide processing cycle, the time for image acquisition takes 58 seconds for a 2.25 cm2 tissue section. An automatic slide loader enables the scanner to process up to 40 slides per hour without operator intervention. Slide scanning and image processing are done concurrently so that post-scan processing is eliminated. A virtual slide can be viewed over the Internet immediately after the scanning is complete. A validation study compared the diagnostic accuracy of pathologist case readers using array microscopy (with images viewed as virtual slides) and conventional light microscopy. Four senior pathologists diagnosed 30 breast surgical pathology cases each using both imaging modes, but on separate occasions. Of 120 case reads by array microscopy, there were 3 incorrect diagnoses, all of which were made on difficult cases with equivocal diagnoses by light microscopy. There was a strong correlation between array microscopy vs. "truth" diagnoses based on surgical pathology reports. The kappa statistic for the array microscopy vs. truth was 0.96, which is highly significant (z=10.33, p <0.001). There was no statistically significant difference between rates of agreement with truth between array microscopy and light microscopy (z=0.134, p >0.05). Array microscopy and light microscopy did not differ significantly with respect to the number/percent of correct decisions rendered (t=0.552, p=0.6376) or equivocal decisions rendered (t=2.449, p=0.0917). Pathologists rated 95.8% of array microscopy virtual slide images as good or excellent. None were rated as poor. The mean viewing time for a DMetrix virtual slide was 1.16 minutes. The DMetrix virtual slide processor has been found to reduce the virtual slide processing cycle more than 10 fold, as compared with other virtual slide systems reported to date. The virtual slide images are of high quality and suitable for diagnostic pathology, second opinions, expert opinions, clinical trials, education, and research.

Image quality issues in a static image-based telepathology consultation practice.

Field selection and image quality have often been identified as impediments in the successful employment of static-image telepathology. One thousand seven hundred fifty-three electronic consultations using static images were performed at the Department of Telemedicine, Armed Forces Institute of Pathology (AFIP) between November 1994 and September 2001, with 98.3% receiving a telepathology diagnosis. In 47.9% of cases, imagery was considered good by AFIP consultants, 38.5% were considered adequate, and 14.6% of cases were considered to have poor-quality imagery. Deficiencies in image quality were recorded for each case. Cases with imagery rated as good averaged significantly fewer deficiencies per case (0.45, range: 0 to 3) than cases with imagery rated adequate (0.95, range: 0 to 6) or poor (2.4, range: 0 to 7). Deficiencies in focus were most commonly identified in this series of cases (28.1%), followed by improper white balancing of the capture device (14.1%) and inadequate resolution (10%). Cases in which images were of inadequate resolution showed an increased likelihood for discordance between the telepathology diagnosis and the diagnosis rendered on follow-up material ("truth diagnosis"). Inadequate field selection, although only cited in 6.7% of cases overall, was seen with a significantly higher frequency in cases in which there was discordance between the telepathology and truth diagnosis. A review of common image deficiencies in static-image telepathology and possible causes is presented.

Hepatic cirrhosis occurring in a young woodchuck (Marmota monax) due to vertical transmission of woodchuck hepatitis virus (WHV).

Histologic and clinicopathologic findings of a woodchuck (Marmota monax) vertically infected with woodchuck hepatitis virus (WHV) are presented. The liver exhibits marked cirrhotic changes, which is characteristic of the pre-transformation phase of WHV. At necropsy, the woodchuck exhibited ascites and the liver had a grossly nodular appearance. Microscopically, focal hepatocyte necrosis and inflammatory cells were observed in midzonal and periportal areas in the liver. In Macchiavellos stained sections, cytoplasmic inclusion bodies appeared reddish granular materials. We believe that this may represent a new suitable and cost-effective cirrhotic model for the disease processes associated with hepadnaviruses in a number of other species, most notably Hepatitis B virus infection in man.

Fetal bladder outlet obstruction in a stillborn bovine fetus.

A stillborn bovine male fetus with abdominal distention, arthrogryposis and atresia ani was presented for diagnostic evaluation. At necropsy, this fetus had a large amount of ascites, urachal obstruction and marked bladder distention. The ventral surface of the bladder had ruptured and attached to the abdominal wall by fibrinous adhesions. There was bilateral hydronephrosis with moderate pelvic dilatation and cortical attenuation. The rectum was filled with meconium but the anus was imperforate. The right forelimb was contracted. The cause(s) of these abnormalities could not be determined; however, we believe that developmental abnormalities during embryogenesis may be the result of chromosomal abnormalities.

Structural characterization of lipopeptide biomarkers isolated from Bacillus globigii.

Spectra obtained using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry of Bacillus globigii (Bacillus subtilis niger) spores, vegetative cells and the culture supernatant show a cluster of biomarkers centered at a molecular mass of 1478 Da. Three biomarkers were isolated from the cell-free culture supernatant by solid-phase extraction and reversed-phase high-performance liquid chromatography, and characterized using various kinds of mass spectrometry. A Fourier transform mass spectrometer with a MALDI source was used to determine the monoisotopic protonated masses at 1463.8, 1477.8, and 1505.8 Da in order of elution. The mass differences of 14 and 28 Da suggest that they are homologous molecules. Alkaline hydrolysis of each species showed that it contained a lactone linkage. Strong acid hydrolysis released a fatty acid from an amide bond, consistent with a lipopeptide. A quadrupole time-of-flight instrument with a nanospray source was used to sequence the hydrolyzed forms of the three biomarkers. The cyclic lipopeptides were found to have amino acid sequences identical with those in fengycins and plipastatins, antimicrobial compounds with phospholipase inhibitor activity, previously identified in related species of Bacillus subtilis and Bacillus cereus.