Marcia R. S. Ilha

Clinical, pathological and immunohistochemical study of feline mammary fibroepithelial hyperplasia following a single injection of depot medroxyprogesterone acetate

Feline mammary fibroepithelial hyperplasia (FMFH) following a single injection of depot medroxyprogesterone acetate (MPA) was observed in eight intact young queens. The repository compound is marketed as a veterinary product by a local pharmaceutical company with an indication for contraception in cats. The drug was administered according to the recommended doses and injection frequencies. Serum hormone assays performed immediately before neutering and 3 weeks after neutering detected persistently high levels of progesterone suggesting that depot MPA was still exerting its influence. No corpora lutea were found in those cases ruling out ovaries as the main site of progesterone. Immunohistochemistry performed on the hyperplastic mammary glands detected progesterone receptors in the nuclei of ductal cells, and growth hormone (GH) and insulin-like growth factor-I (IGF-I) in the cytoplasm of ductal epithelium. Overdosing should be considered here as the animals received at least 10 mg/kg of depot MPA in a single injection. Progestin-induced local synthesis of GH and IGF-I in mammary epithelial cells is suggested as one of the pathogenic mechanisms involved in the development of FMFH.

Evaluation of novel oral vaccine candidates and validation of a caprine model of Johne's disease

Johnes disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a major threat to the dairy industry and possibly some cases of Crohns disease in humans. A MAP vaccine that reduced of clinical disease and/or reduced fecal shedding would aid in the control of JD. The objectives of this study were (1) to evaluate the efficacy of 5 attenuated strains of MAP as vaccine candidates compared to a commercial control vaccine using the protocol proposed by the Johnes Disease Integrated Program (JDIP) Animal Model Standardization Committee (AMSC), and (2) to validate the AMSC Johnes disease goat challenge model. Eighty goat kids were vaccinated orally twice at 8 and 10 weeks of age with an experimental vaccine or once subcutaneously at 8 weeks with Silirum® (Zoetis), or a sham control oral vaccine at 8 and 10 weeks. Kids were challenged orally with a total of approximately 1.44 × 10(9) CFU divided in two consecutive daily doses using MAP ATCC-700535 (K10-like bovine isolate). All kids were necropsied at 13 months post-challenge. Results indicated that the AMSC goat challenge model is a highly efficient and valid model for JD challenge studies. None of the experimental or control vaccines evaluated prevented MAP infection or eliminated fecal shedding, although the 329 vaccine lowered the incidence of infection, fecal shedding, tissue colonization and reduced lesion scores, but less than the control vaccine. Based on our results the relative performance ranking of the experimental live-attenuated vaccines evaluated, the 329 vaccine was the best performer, followed by the 318 vaccine, then 316 vaccine, 315 vaccine and finally the 319 vaccine was the worst performer. The subcutaneously injected control vaccine outperformed the orally-delivered mutant vaccine candidates. Two vaccines (329 and 318) do reduce presence of JD gross and microscopic lesions, slow progression of disease, and one vaccine (329) reduced fecal shedding and tissue colonization.

Uterine Lesions in 32 Female Miniature Pet Pigs

Thirty-two 4-month-old to 19-year-old female miniature pet pigs were spayed. Uterine lesions were present in all except 8 pigs. The 24 remaining pigs had diffuse cystic endometrial hyperplasia, of which 14 had smooth muscle tumors, including leiomyomas and leiomyosarcomas, in the uterus or broad ligament. Nodular endometrial lesions—including adenocarcinomas, adenomas, and/or adenomyosis—were present in 10 pigs, 3 of which had concurrent smooth muscle tumors. Pyometra was present in 3 pigs. In uterine sections with cystic endometrial hyperplasia, adenomyosis, or adenomas, approximately 70% of epithelial nuclei expressed estrogen receptor and progesterone receptor immunohistochemically; in adenocarcinomas, expression was 20%. Regardless of malignancy, more than 50% of nuclei in smooth muscle tumors expressed estrogen receptor and progesterone receptor. Aging was associated with the development of uterine lesions in miniature pet pigs.

Intoxicação experimental por monensina em eqüinos

Seven horses were experimentally treated with sodium monensin. Two of them were fed 3-4 kg/horse/day with a commercial ration (containing 180 ppm±20) known to be implicated in natural outbreaks of monensin poisoning in horses. A third horse was fed a single dose of 5 mg/kg and another one received four daily doses of 1 mg/kg of sodium monensin (from a premix source). These four horses died or were euthanitised in extremis 3-8 days after the beginning of administration of the drug. A fifth horse was fed a single dose of 5 mg/kg of monensin, became mildly ill but recovered. Two experimental horses did not develop signs of the toxicosis. One of them was fed 40 daily doses of 0.5 mg/kg of monensin. The other was fed 3 kg/day of the same brand of ration used in farms where outbreaks of monensin poisoning in horses have been reported (but from another batch later determined to have monensin levels of only <5 ppm). The onset of clinical signs varied from 2 to 5 days after administration of the drug, and clinical courses lasted from 24 to 76 hours. Clinical signs included tachycardia and cardiac arythmia, groaning, incoordination, sudoresis, sternal or lateral recumbency, paddling and death. Marked increases in the plasmatic activity of creatine phosphokinase were observed in five poisoned horses, and a mild increase in the aspartate aminotransferase plasmatic activity was detected in one of the animals. Main necropsy findings were in the skeletal muscles and consisted of focal to focally extensive white or yellow areas of discoloration with bilateral symmetric distribution. These lesions were associated with gelatinous transluscent edema in the intermuscular fasciae. More intensively affected muscles included quadriceps femoris, adductor, pectineus, gracilis, semimembranous, supraspinatus, subscapularis and brachycephalic. Histopathological findings consisted of segmentar, multifocal to coalescent, degenerative myopathy. Both gross and microscopic lesions were absent in the skeletal muscles from two of the horses fed the monensin premix [5 mg/kg (single dose) and 4 mg/kg (four daily doses of 1 mg/kg)]. Neither gross nor microscopic lesions were observed in the myocardium of any of these horses.

Cytologic Criteria for Mast Cell Tumor Grading in Dogs With Evaluation of Clinical Outcome

A 2-tiered histologic grading scheme for canine cutaneous mast cell tumors (MCTs) is based on morphologic characteristics of neoplastic cells, including karyomegaly, multinucleation, nuclear pleomorphism, and mitotic figures. Aspirates from MCTs may provide the same information more quickly, inexpensively, and less invasively. This study used these criteria to develop a cytologic grading scheme for canine MCTs to predict outcome. Three anatomic pathologists graded histologic samples from 152 canine MCTs. Three clinical pathologists evaluated aspirates from these masses using similar criteria. A cytologic grading scheme was created based on correlation with histologic grade and evaluated with a kappa statistic. Survival was evaluated with Kaplan-Meier survival curves. Cox proportional hazards regression was used to estimate hazard ratios for tumor grades and individual grading components. Simple logistic regression tested for relationships between risk factors and mortality. The cytologic grading scheme that best correlated with histology (kappa = 0.725 ± 0.085) classified a tumor as high grade if it was poorly granulated or had at least 2 of 4 findings: mitotic figures, binucleated or multinucleated cells, nuclear pleomorphism, or >50% anisokaryosis. The cytologic grading scheme had 88% sensitivity and 94% specificity relative to histologic grading. Dogs with histologic and cytologic high grade MCTs were 39 times and 25 times more likely to die within the 2-year follow-up period, respectively, than dogs with low grade MCTs. High tumor grade was associated with increased probability of additional tumors or tumor regrowth. This study concluded that cytologic grade is a useful predictor for treatment planning and prognostication.

Genetic diversity of porcine circovirus type 2 and implications for detection and control.

Porcine circovirus type 2 (PCV2) is the etiological agent of post- weaning multi-systemic wasting syndrome (PMWS), an economically important disease of swine. Severe wasting and lymphadenopathy are typical signs of PMWS. Effective vaccines against PCV2 and reliable diagnostic tests are available. Since PCV2s discovery in the mid-90s and the introduction of commercial vaccines, several new recombinant strains and variants with genetic mutations have emerged. Two noteworthy changes include; a major type switching event that resulted in the previously predominant PCV2a genotype being replaced by PCV2b, and the recent emergence of a mutant PCV2b with a capsid protein containing an additional lysine. The mutant PCV2b exhibits increased virulence and is spreading rapidly in various regions of the world. This article provides an overview of the recent molecular epidemiology in the context of the current methods for the detection and prevention of PCV2, emphasizing the need for updated PCV2 vaccines.

Canine parvovirus-2b-associated erythema multiforme in a litter of English Setter dogs

Erythema multiforme (EM) was diagnosed in a litter of English Setter puppies. The puppies developed erythematous cutaneous lesions at the age of 2 weeks. Microscopically, there was individual keratinocyte apoptosis associated with lymphocyte exocytosis in all layers of the epidermis. Intranuclear viral inclusions were seen in multiple tissues and organs. Tissues from the tongue, lymph node, spleen, skin, and small intestine were positive for Canine parvovirus-2 (CPV-2) and negative for Canine distemper virus (CDV) and Canid herpesvirus 1 by fluorescent antibody test. Negative-staining electron microscopy detected parvovirus particles in the intestinal contents. The skin and small intestine were positive for CPV-2b and negative for CDV by polymerase chain reaction. The mucocutaneous junctions and small intestines stained positive for CPV by immunohistochemistry. The present report documents CPV-2b–associated EM in a litter of English Setters and substantiates the single previous report associating EM with CPV-2. The finding suggests that CPV should be considered as a possible cause of EM in dogs.

Meningoencephalitis Caused by Mycoplasma Edwardii in a Dog

A 6-week-old, female, mixed-breed dog with a clinical history of sudden onset of neurologic signs was presented for necropsy. The dog was diagnosed with suppurative and histiocytic meningoencephalitis based on necropsy findings and histopathology. Mycoplasma sp. was isolated in pure culture from the brain and meninges and was identified as Mycoplasma edwardii using DNA sequencing.

Detection of asymptomatic renal Leptospira infection in abattoir slaughtered cattle in southeastern Georgia, United States.

Objectives: Leptospirosis is one of the most widespread zoonotic infectious diseases affecting humans and animals. Several animal species, including cattle, can act as potential asymptomatic carriers facilitating zoonotic transmission of Leptospira. This study was conducted to assess the occurrence of asymptomatic renal Leptospira carriers among cattle slaughtered in southeastern Georgia, United States. Methods: A battery of diagnostic tests, including dark field microscopy, direct fluorescent antibody staining, polymerase chain reaction, and culture, were performed on a set of bovine kidneys (n = 37) collected from an abattoir in southeastern Georgia, United States. Virulence of a field isolate obtained from this study was tested in a hamster experimental model. Results: Motile spirochete-like structures were observed by dark field microscopy in 23 (59%) out of 37 kidney samples tested. In all, 29 samples (78%) were positive by direct fluorescent antibody staining. Only 11 (29.7%) samples by polymerase chain reaction and 3 (8.1%) by culture were positive for Leptospira sp. The isolates obtained by culture were confirmed as Leptospira borgpetersenii. Hamsters experimentally infected with one of the Leptospira field isolates obtained from this study did not show clinical signs but developed renal infection with interstitial nephritis and tubular necrosis. Conclusions: This study confirms that asymptomatic Leptospira renal infection is present among cattle in the region. Our findings underscore the need for future studies to assess the potential environmental contamination and transmission to humans in contact with infected cattle.

Ligneous conjunctivitis secondary to a congenital plasminogen deficiency in a dog

CASE DESCRIPTION A 7-month-old 16.6-kg (36.5-lb) sexually intact female Golden Retriever was evaluated because of progressive severe bilateral membranous conjunctivitis, oral lesions, nasal discharge, and cough. CLINICAL FINDINGS Histologic examination of conjunctival biopsy specimens revealed findings consistent with ligneous conjunctivitis. Circulating plasminogen activity was repeatedly low, and congenital plasminogen deficiency was identified as the underlying cause of the ocular, oral, and respiratory lesions. TREATMENT AND OUTCOME Topical and subconjunctival administrations of fresh frozen plasma (FFP), topical administration of cyclosporine, and oral administration of azathioprine had no effect on the conjunctival membranes. Excision of the membranes followed by intensive treatment with topical applications of heparin, tissue plasminogen activator, corticosteroid, and FFP and IV administration of FFP prevented membrane regrowth. Intravenous administration of FFP increased plasma plasminogen activity to within reference limits, improved respiratory and oral lesions, and resulted in weight gain; discontinuation of this treatment resulted in weight loss, signs of depression, and worsening of lesions. After euthanasia because of disease progression, necropsy findings included mild hydrocephalus; multifocal intestinal hemorrhages; and fibrinous plaques in the oral cavity, nasopharynx, trachea, esophagus, and pericardium. Microscopically, the plaques were composed of fibrin and poorly organized granulation tissue. Fibrin thrombi were present within vessels in the lungs, oral cavity, and trachea. CLINICAL RELEVANCE In dogs, congenital plasminogen deficiency can occur and may be the underlying cause of ligneous conjunctivitis. A combination of surgical and medical treatments may improve conjunctival membranes, and administration of FFP IV appears to be effective in treating nonocular signs of plasminogen deficiency.