Histologic characterization of the cat middle ear: in sickness and in health.

The purpose of this study was to establish microscopic normal in the middle ear of the cat while concurrently characterizing gross and microscopic lesions reflecting spontaneous otitis media. Both ears from 50 cats were examined grossly and processed for histologic examination of the external, middle, and internal ear on a single slide. Gross lesions of the middle ear were present in 14 of 100 (14%) and included turbid fluid, frank pus, hemorrhage, and fibrous thickening of the auricular mucoperiosteum. Histologically, 48 of 100 (48%) ears had evidence of ongoing or previous inflammatory middle ear disease, including proteinaceous fluid; vascular ectasia; expansion of the auricular mucoperiosteum by neutrophils, lymphocytes, and macrophages; cholesterol clefts; hemorrhage; fibrin; granulation tissue; membranous pseudo-glands; fibrosis; proliferation and/or osteolysis of the tympanic and septum bullae. Histologic lesions were identified in 34 of 100 ears (34%) lacking gross evidence of disease. Ears were classified histologically as either normal (52/100 [52%]) or diseased (48/100 [48%]). Diseased ears were further classified as mild to moderate (37/100 [37%]) or severely (11/100 [11%]) affected. Internal ear involvement was present in 11 of 100 (11%) ears. Histologic evidence of middle ear disease in cats is far greater than gross lesions or clinical literature suggests; further investigation and correlation of clinical and histologic disease are warranted. With minimal additional preparation, diagnostic specimens may be readily prepared and evaluated for this integral sensing organ.

Evaluation of efficacy of mineral oil, charcoal, and smectite in a rat model of equine cantharidin toxicosis.

BACKGROUND: The efficacy of orally administered therapeutics for the treatment of cantharidin intoxication has not been evaluated in controlled studies. OBJECTIVE: To develop a model of acute cantharidin intoxication in laboratory rats and to evaluate in this model the relative efficacy of 3 gastrointestinal therapies used to treat equine cantharidin toxicosis. ANIMALS: Sixty-four male Sprague-Dawley rats. METHODS: A blinded, randomized, controlled study was performed on rats surgically implanted with telemetry transmitters for evaluating heart rate, locomotor activity, and body temperature. Orogastric administration of cantharidin was performed within 15 seconds before administration of mineral oil, activated charcoal, or smectite. Negative control groups received therapeutic agents alone. Urine was collected for cantharidin analysis. Rats were sacrificed 24 hours after intoxication, and tissues were collected for histopathologic evaluation. Data analysis included ANOVA procedures and contingency tables. RESULTS: Six of 8 cantharidin-intoxicated rats treated with mineral oil died; bradycardia and hypothermia developed in the animals of this group 0-8 hours after intoxication. Rats treated with mineral oil had higher urine cantharidin concentrations than rats receiving cantharidin alone or with smectite (P = .04). The most severe hypothermia (30.6°C ± 1.0) developed in rats administered mineral oil at 4-8 hours after intoxication, whereas those treated with charcoal (35.2°C ± 0.8) had mean body temperatures higher than all other treatment groups (P = .03). Survival times in the charcoal (P = .16) and smectite (P = .12) treatment groups were not statistically different from negative controls. CONCLUSIONS AND CLINICAL IMPORTANCE: Mineral oil is often used in the treatment of equine cantharidin toxicosis. Our findings suggest that mineral oil increases cantharidin absorption, worsening morbidity and fatality in rats.

Immunohistochemical expression of c-KIT protein in feline soft tissue fibrosarcomas.

C-KIT is the cellular homolog of the feline sarcoma viral oncogene v-KIT, which encodes the tyrosine kinase receptor protein KIT. Mutations and varied expression of this gene have been demonstrated within multiple neoplasms in people and domestic animals. The purpose of this study was to determine if KIT protein is expressed in feline soft tissue fibrosarcomas (ST FSA) using immunohistochemistry (IHC). The computer database at the Oklahoma Animal Disease Diagnostic Laboratory was searched from January 1, 2006, to December 31, 2007, for any domestic cat with an ST FSA. Routinely stained slides from 46 feline ST FSAs were reviewed and graded based on the scale outlined by Kuntz et al. Immunohistochemistry for KIT protein was performed on one representative section from each cat. There were a total of 12/46 (26%) cats that were immunoreactive for KIT. Immunoreactivity was detected in greater than 80% of the neoplastic cells in 4/46 (9%) cats. Immunoreactivity was detected in less than 10% of the neoplastic cells in 8/46 (17%) cats. Immunoreactivity was characterized by evenly distributed cytoplasmic stippling within the neoplastic spindle-shaped cells and/or multinucleated giant cells. Based on these results, KIT immunoreactivity can be detected within feline ST FSAs using IHC. The results of this study also indicate that KIT immunoreactivity in feline ST FSA does not correlate with the histologic grade (P = .141, X(2) = 2.166), survivability (P = .241, X(2) = 1.373), or whether the neoplasm was a spontaneous or an injection site FSA (P = .074, X(2) = 3.184).

inlA premature stop codons are common among Listeria monocytogenes isolates from foods and yield virulence-attenuated strains that confer protection against fully virulent strains.

Previous studies showed that a considerable proportion of Listeria monocytogenes isolates obtained from foods carry a premature stop codon (PMSC) mutation in inlA that leads to production of a truncated and secreted InlA. To further elucidate the role these mutations play in virulence of L. monocytogenes, we created isogenic mutants, including (i) natural isolates where an inlA PMSC was reverted to a wild-type inlA allele (without a PMSC) and (ii) natural isolates where a PMSC mutation was introduced into a wild-type inlA allele; isogenic mutant sets were constructed to represent two distinct inlA PMSC mutations. Phenotypical and transcriptional analysis data showed that inlA PMSC mutations do not have a polar effect on the downstream inlB. Isogenic and natural strains carrying an inlA PMSC showed significantly reduced invasion efficiencies in Caco-2 and HepG2 cell lines as well as reduced virulence in oral guinea pig infections. Guinea pigs were also orally infected with a natural strain carrying the most common inlA PMSC mutation (vaccinated group), followed by challenge with a fully virulent L. monocytogenes strain 15 days postvaccination to probe potentially immunizing effects of exposure to L. monocytogenes with inlA PMSC mutations. Vaccinated guinea pigs showed reduced bacterial loads in internal organs and improved weight gain postchallenge, indicating reduced severity of infections in guinea pigs exposed to natural strains with inlA PMSC mutations. Our data support that (i) inlA PMSC mutations are causally associated with attenuated virulence in mammalian hosts and (ii) naturally occurring virulence-attenuated L. monocytogenes strains commonly found in food confer protective immunity.

Search for Borrelia burgdorferi in kidneys of dogs with suspected "Lyme nephritis".

BACKGROUND: "Lyme nephritis" is a poorly characterized condition associated with proteinuria and often fatal renal failure in dogs with serological evidence of infection with Borrelia burgdorferi. OBJECTIVE: The aim of this study was to determine if intact B. burgdorferi organisms were present in the kidneys of serologically Lyme-positive dogs with histopathologic features of Lyme nephritis. ANIMALS: Twenty-six affected and 10 control dogs were identified over an 8-year period (1996-2004) in databases at Cornell University's College of Veterinary Medicine. Case inclusion required serologic evidence of natural exposure to B. burgdorferi and availability of renal tissue (frozen or paraffin embedded) exhibiting pathology consistent with Lyme nephritis. METHODS: Renal tissue samples were assessed using modified Steiner (silver) (MS) staining, immunohistochemistry (IHC), polymerase chain reaction (PCR) using 4 primer sets (eubacterial, B. burgdorferi, Bartonella, and canine genomic DNA), and fluorescence in situ hybridization (FISH) using a 5'-cy3-eubacterial probe for 16S rRNA. RESULTS: MS stain was positive in 1 case; IHC was negative in all cases. None of the B. burgdorferi or Bartonella PCR reactions was positive. Two of the B. burgdorferi FISH analyses were positive. CONCLUSIONS AND CLINICAL IMPORTANCE: Minimal evidence of the presence of intact B. burgdorferi or any other bacterial organism was found in the renal tissue of dogs with suspected Lyme nephritis. Direct renal invasion by B. burgdorferi organisms does not appear to be responsible for this syndrome.

Ovarian and intestinal angiomatosis in a horse.

Dozens of red, raised nodules scattered along the serosal surface of the small intestine and the right and left ovaries were observed as incidental findings on gross examination in a 21-year-old Thoroughbred mare euthanatized for severe lameness. Histologically, these nodules were composed of numerous, variably sized, redundant vascular profiles filled with red blood cells and fibrin thrombi. Based on the presence of multiple nodules composed of benign vascular channels scattered within the small intestine and ovary, a diagnosis of angiomatosis is proposed. To the authors' knowledge, this is the first report of small intestinal and ovarian angiomatosis in a horse.

Sigma B contributes to Listeria monocytogenes gastrointestinal infection but not to systemic spread in the guinea pig infection model.

Contributions of the alternative sigma factor sigmaB to Listeria monocytogenes infection were investigated using strains bearing null mutations in sigB, prfA, or inlA or in selected inlA or prfA promoter regions. The DeltaP4inlA strain, which has a deletion in the sigmaB-dependent P4inlA promoter, and the DeltasigB strain had significantly reduced invasion efficiencies relative to that of the wild-type strain in the Caco-2 human colorectal epithelial cell line, while the invasion efficiency of a strain bearing a deletion in the partially sigmaB dependent P2prfA promoter region did not differ from that of the wild type. The virulence of the DeltasigB and DeltaP4inlA strains was attenuated in intragastrically inoculated guinea pigs, with the DeltasigB strain showing greater attenuation, while the virulence capacity of the DeltaP2prfA strain was similar to that of the wild-type strain, suggesting that attenuation of virulence due to the DeltasigB mutation does not result from loss of sigmaB-dependent prfA transcription. Our results show that sigmaB-dependent activation of inlA is important for cell invasion and gastrointestinal infection and suggest that sigmaB-regulated genes in addition to inlA appear to contribute to gastrointestinal infection. Interestingly, the virulence of the DeltasigB strain was not attenuated in intravenously infected guinea pigs. We conclude that (i) L. monocytogenes sigmaB plays a critical role in invasion of human host cells, (ii) sigmaB-mediated contributions to invasion are, in part, due to direct effects on inlA transcription but not on prfA transcription, and (iii) sigmaB plays a critical role during the gastrointestinal stage of listeriosis in the guinea pig but is not important for systemic spread of the organism.

Slaughterhouse survey of pyelonephritis in dairy cows.

Twenty-one rejected kidneys from 2426 slaughtered dairy cows (0.87 per cent) had gross signs of pyelonephritis that were confirmed by histopathology. In all the kidneys the findings were consistent with a chronic rather than an acute infection. One species of bacteria was cultured from 12 of the kidneys and two species of bacteria were cultured from six. The most commonly isolated bacteria were Escherichia coli, from eight kidneys, Arcanobacterium pyogenes, from seven kidneys and Corynebacterium renale, from five kidneys. The other bacteria cultured were Corynebacterium cystitidis, Corynebacterium species, Streptococcus species group G and Enterococcus faecalis. E. coli was cultured from all the kidneys from which two species were isolated; the accompanying bacteria were A. pyogenes in three kidneys, C. renale in two and C. cystitidis in one. No bacteria were cultured from two of the kidneys and no significant bacteria were cultured from another. The kidneys with pyelonephritis were slightly larger than a comparison group of 72 kidneys without nephritis.

Incidence of polysaccharide storage myopathy in draft horse-related breeds: a necropsy study of 37 horses and a mule.

Skeletal muscle samples from 38 draft horse-related animals 1-23 years of age were evaluated for evidence of aggregates of glycogen and complex polysaccharide characteristic of equine polysaccharide storage myopathy (EPSSM). Cardiac muscle from 12 of these horses was also examined. Antemortem serum levels of creatine kinase (CK) and aspartate aminotransferase (AST) from 9 horses with EPSSM and 5 horses without EPSSM were compared. Skeletal muscle from 17 horses contained inclusions of periodic acid-Schiff (PAS)-positive, amylase-resistant complex polysaccharide. Similar inclusions were also present in the cardiac muscle of 1 horse. A vacuolar myopathy with aggregates of PAS-positive, amylase-sensitive glycogen was seen in 8 other horses, and these findings are also considered diagnostic for EPSSM. Antemortem serum activities of CK and AST were often higher in EPSSM horses than in horses without EPSSM. Using the presence of amylase-resistant complex polysaccharide as the criterion for diagnosis of EPSSM, the incidence in this population was 45%. Inclusion of horses with aggregates of glycogen but no amylase-resistant complex polysaccharide as representative of the range of pathologic findings in horses with EPSSM resulted in a 66% incidence in this population.

Diagnosis of persistent bovine viral diarrhea virus infection by immunohistochemical staining of formalin-fixed skin biopsy specimens.

The objective of this study was to evaluate the efficacy of immunohistochemical (IHC) staining for diagnosis of persistent bovine viral diarrhea virus (BVDV) infection using formalin-fixed, paraffin-embedded skin biopsy specimens. Skin from 41 of 42 calves shown to be persistently infected (PI) with BVDV by repeated virus isolation more than 3 weeks apart were immunohistochemically positive for BVDV antigen. Positive IHC staining was most pronounced in the keratinocytes and in hair follicle epithelium, hair matrix cells of the hair bulb, and the dermal papilla. All of the skin sections from 10 calves experimentally infected postnatally with BVDV (10(5) median tissue culture infective doses [TCID50]) and biopsied on days 0, 5, 7, and 9 postinfection were negative for viral antigen. Ten calves from a second group experimentally infected with a higher dose of BVDV (10(8) TCID50) were biopsied when viremic between 10 and 14 days postinfection and 4 calves exhibited positive IHC staining for BVDV; however, staining in these skin biopsies was confined to small foci in the nonfollicular epidermis and follicular ostia. This staining was distinct from that observed in skin obtained from PI cattle. Skin biopsy represents an effective method for identifying animals PI with BVDV.

Ulcerative keratitis associated with qualitative tear film abnormalities in cats.

Three cats with indolent corneal ulcers and one cat with bilateral corneal sequestration and normal aqueous tear production were found to have rapid tear break-up times (BUTs). Tear BUTs in clinically affected cats averaged 2.5 +/- 1.29 s and 2.33 +/- 0.58 s for the right and left eyes, respectively. Palpebral conjunctival biopsies were harvested from consistent sites from each eye of affected cats (n = 7 affected eyes), and age-and breed-matched controls (n = 2 unaffected eyes). Light microscopy revealed a marked decrease to complete absence of conjunctival goblet cells (average goblet cell (GC):epithelial cell (EC) density = 18:50), conjunctival epithelial dysplasia, squamous metaplasia, and neutrophilic and mononuclear cell submucosal infiltration in affected cats. Specimens from the control cats had an average GC:EC density of 34:50, and minimal submucosal inflammatory infiltrate. The corneas (n = 7 eyes) healed following surgical keratectomy with (n = 2 eyes) or without (n = 1 eye) conjunctival pedicle flaps, superficial keratectomy and striate keratotomy with (n = 2 eyes) or without (n = 2 eyes) third eyelid flaps, and mucinomimetic tear supplementation (n = 5 eyes). Goblet cell regeneration was confirmed after 5 months of mucinomimetic supplementation (n = 2 eyes). The etiology for these mucin deficiencies remains unknown.