Adriana R. Massone

Histochemical and ultrastructural study of the chicken salivary palatine glands

Salivary glands are a good model to investigate the relationship between cell secretion and glandular structure. Most studies of this organ deal with mammals, but we are interested in a morphofunctional characterization of these glands in poultry in relation with particular feeding habits. For this purpose, conventional and lectin histochemical methods as well as ultrastructural methods have been applied to the chicken lateral and medial palatine salivary glands. It was found that periodic acid-Schiff (PAS)-positive, alcianophilic, and metachromatic or orthochromatic cells were present with a more homogeneous distribution pattern in lateral glands than in medial palatine glands. Lectin staining depended on the lectin type that was applied, but also on the glandular part both in lateral and medial glands. Ultrastructural studies showed cytoplasmic membranous structures with a scattered granular or filamentous content depending on the secretory cell. In conclusion, morphofunctional characteristics of salivary glands of chicken suggest that their products are involved in lubrication and humidification of food ingested, and probably in protection of the oral surface, as has been previously described for other animals showing similar histochemical staining patterns.

A simple method of infecting rabbits with Bovine herpesvirus 1 and 5.

This report describes an alternative technique to inoculate rabbits and to reproduce infection by Bovine herpesvirus 1 and 5. First, the nostrils are anaesthetized by aspersion with local anaesthetic. A few seconds later, and after proving the insensitivity of the zone, the rabbits are put on their back legs with their nostrils upwards and the inoculum is introduced slowly into each nostril by using disposable droppers. Clinical signs, viral isolation from nasal swabs, histological lesions found, positive polymerase chain reaction and antibodies production confirm the infection. This very simple and bloodless technique, where the animals are exposed to minor distress, may be useful for evaluating the virulence of BoHV-1 and BoHV-5 strains, to study the establishment of latent virus infection and to test the potential of experimental vaccines or properties of antiviral drugs. It may be also suitable for experimental infection with other respiratory viruses in this animal model.

Lack of prognostic significance of angiogenesis in canine melanocytic tumours.

The prognostic significance of angiogenesis in some canine tumours has been investigated, but little is known about its relevance in canine melanocytic tumours (MTs). The aim of this study was to evaluate the prognostic significance of angiogenesis in canine MTs. A total of 36 cutaneous melanocytomas (benign MTs), 40 cutaneous melanomas (malignant MTs) and 43 oral melanomas were studied. Survival data were available for a subset of 59 cases. Microvessel density (MVD) and endothelial area (EA) were determined by immunolabelling using an antibody specific for von Willebrand factor (vWF). Mean MVD (expressed as the number of microvessels per mm(2)) was 129 ± 14 in melanocytomas, 191 ± 16 in cutaneous melanomas and 208 ± 16 in oral melanomas. Mean EA (expressed as the percentage of the total area) was 1.5 ± 0.14 in melanocytomas, 2.6 ± 0.2 in cutaneous melanomas and 2.4 ± 0.3 in oral melanomas. The differences in MVD and EA between melanocytomas and melanomas were significant (P = 0.001 and P = 0.003, respectively). MVD and EA were significantly correlated between cutaneous and oral MTs (r = 0.54; P <0.001 and r = 0.63; P <0.001, respectively). MVD and EA were not related to survival in cutaneous and oral MTs. In conclusion, tumour vascularization was higher in melanomas than in melanocytomas, but it seemed to have no prognostic significance in these tumours.

Equine arteritis virus gP5 protein induces apoptosis in cultured insect cells

Equine Arteritis Virus (EAV) has been shown to induce apoptosis in vitro but the induction of this mechanism has not been previously associated with any viral gene product. In this work, we found a cytotoxicity effect of the EAV gP5 protein on baculovirus-insect cells and a low yield of protein recovery. Besides, different morphological features by electron transmission microscopy, DNA fragmentation in agarose gel, TUNEL analysis and caspase 3 activity were found. All these findings indicate that the EAV gP5 protein induces apoptosis in insect cells.

Constrictura rectal en cerdos necropsiados en una granja de ciclo completo en confinamiento. Consideraciones sobre su prevalencia, hallazgos anatomopatológicos y etiopatogenia

Se estudiaron durante dos periodos (anos 1996 y 1998-99), la prevalencia, los hallazgos anatomopatologicos y las probables causas de la constrictura rectal en cerdos de una granja intensiva en confinamiento. En el primer periodo se realizaron 660 necropsias de las categorias recria II, Desarrollo y engorde, siendo de 7.5% la mediana porcentual mensual de la constrictura rectal como causa primaria de muerte. Se observaron valores superiores al 10% en los meses de verano para las tres categorias de cerdos, en septiembre para la categoria recria II y en diciembre para la categoria Desarrollo. En el segundo periodo se realizaron 879 necropsias de las categorias Desarrollo y engorde. El porcentaje de constrictura rectal fue de 10, 3% y el de prolapso rectal 7.7%. La edad media fue de 164.2±6. L y 115.8±4.2 dias, respectivamente. Los cerdos presentaron una marcada distension abdominal y pobre desarrollo. A la necropsia se remarco la distension del recto, colon espiroideo y ciego, generalmente asociado a la presencia de exudado serofibrinoso, tanto en el peritoneo como en el pericardio. La lesion se caracterizo por una estrechez de recto ubicada entre 2.5 y 5 cm en craneal del ano, variando su longitud entre 3 y 8 cm. Los estudios bacteriologicos fueron negativos para patogenos primarios del cerdo. Se realizaron 122 analisis de aflatoxinas y zearalenona y 36 de deoxinivalenol de las materias primas del alimento mediante una prueba de ELISA por competicion. Si bien en el estudio realizado no se consignaron valores altos de zearalenona en correspondencia con los meses de alta prevalencia de constrictura rectal, con excepcion del mes de marzo, lo constante, a lo largo del ano, fue su consumo, asociado o no, con dioxinivalenol. Se desconoce lo que ocurre en condiciones de campo, por lo que no se puede excluir que dicha micotoxina participe en la patogenesis de la constrictura rectal, si la consideramos como una secuela del prolapso rectal.

Double immunoenzymatic labelling of intermediate filaments in bovine urinary bladder tumours

Double immunoenzymatic labelling made possible the simultaneous staining of two antigens with a mixture of polyclonal and monoclonal commercial antibodies. Immunocharacterization of intermediate filament proteins was found to be an accurate indicator of histogenesis in urinary bladder tumours of cattle.

Citrus reticulata peel oil inhibits non-small cell lung cancer cell proliferation in culture and implanted in nude mice

Non-small cell lung cancer (NSCLC) accounts for most cases of lung cancer. The peel oil of mandarin Citrus reticulata Blanco cv. Dancy (MPO) is a natural source of essential oils and carotenoids. Volatile and non-volatile lipid compounds were characterized by chromatographic methods. We demonstrate that MPO causes a dose-dependent growth inhibition of NSCLC model cells (A549) in culture and tumour growth in vivo of the same cells implanted in nude mice fed with MPO-supplemented diets. MPO induced cell cycle arrest mainly at the G0/G1 phase and reduced the amount of membrane-bound Ras protein along with apoptosis induction. No toxicological effect was found in liver parameters analysed in treated mice and histopathological analyses of their organs did not show any morphological changes. In conclusion, the data suggest that MPO possesses significant antitumor activity without causing systemic toxicity, proposing it as a dietary supplement that may be helpful in cancer prevention.

First report of cerebellar abiotrophy in an Arabian foal from Argentina

Evidence of cerebellar abiotrophy (CA) was found in a six-month-old Arabian filly with signs of incoordination, head tremor, wobbling, loss of balance and falling over, consistent with a cerebellar lesion. Normal hematology profile blood test and cerebrospinal fluid analysis excluded infectious encephalitis, and serological testing for Sarcocystis neurona was negative. The filly was euthanized. Postmortem X-ray radiography of the cervical cephalic region identified not abnormalities, discounting spinal trauma. The histopathological analysis of serial transverse cerebellar sections by electron microscopy revealed morphological characteristics of apoptotic cells with pyknotic nuclei and degenerate mitochondria, cytoplasmic condensation and areas with absence of Purkinje cells, matching with CA histopathological characteristics. The indirect DNA test for CA was positive in the filly, and DNA test confirmed the CA carrier state in the parents and the recessive inheritance of the disease. To our knowledge this is the first report of a CA case in Argentina.

A SURVEY OF PARASITE LESIONS IN WILD RED DEER (CERVUS ELAPHUS) FROM ARGENTINA

Abstract In Argentina there is little information about diseases that affect exotic ungulates and the health risks that they pose to native wildlife, livestock, and humans. The aim of this study was to evaluate the health status of red deer (Cervus elaphus) in the Nahuel Huapi National Park and surrounding areas in Patagonia, Argentina. During three consecutive hunting seasons, necropsies were performed on 101 red deer, and tissues were examined histologically. The most common lesions were those associated with hepatic and pulmonary parasites. Fasciola hepatica was observed in 15 red deer and was associated with cholangiohepatitis (8%) and/or cholangitis (10%). Dictyocaulus sp. (likely Dictyocaulus eckerti) was associated with interstitial pneumonia (5%), bronchitis (5%), pulmonary emphysema (13%), and bronchus-associated lymphoid tissue hyperplasia (13%). Other findings included Sarcocystis spp. cysts in the myocardium (89%) associated with interstitial, focal, lymphoplasmacytic myocarditis (8%); periportal lymphoplasmacytic hepatitis (8%); hepatic centrilobular necrosis (6%); lymphoplasmacytic interstitial nephritis (25%); and follicular hyperplasia in mediastinal, prescapular, and prefemoral lymph nodes (86%). Our report of lesions caused by endemic parasites of livestock in free-ranging exotic red deer in Patagonia sets the foundation for a health-monitoring and -surveillance system of wildlife in this region, which is essential for the sustainable management of threatened Argentinean native fauna.

Extranodal follicular lymphoma in the lung of a free-ranging red deer (Cervus elaphus)

A hunted free-ranging female red deer (Cervus elaphus) from a region near the Nahuel Huapi National Park, Northern Patagonia, Argentina, had a focally extensive peribronchial lymphoid proliferative lesion in the lung characterized by formation of multiple follicles, with prominent germinal centers lacking mantle zone cells and antigen-related polarity. On examination of immunohistochemically stained tissues, a predominance of B cells (cluster of differentiation [CD]20 positive) with only a few scattered T cells (CD3 positive) were present. The histologic and immunohistochemical characteristics are consistent with follicular lymphoma, which is frequently seen in human beings and less frequently in domestic animals.