Maria de Lurdes Pinto

Prenatal administration of vitamin A alters pulmonary and plasma levels of vascular endothelial growth factor in the developing mouse

Vitamin A and the retinoids play a unique role in mammalian embryonic and foetal development and are essential for both cellular differentiation and the establishment of normal morphogenesis. Vascular endothelial growth factor (VEGF) is a known potent mitogenic factor that plays a key role in lung development and function maintenance. In order to contribute to a better knowledge of the modulating effects of vitamin A in lung development, we investigated the effects of the antenatal administration of vitamin A on VEGF expression in lungs and plasma from foetuses and neonates. Pregnant mice were subjected to subcutaneous administration of vitamin A on the 12th gestational day. The lungs and plasma from foetuses and neonates were collected daily from the 15th gestational day till the day of birth. Our results show that vitamin A modulates VEGF concentrations both in lungs and plasma. Statistically significant differences were observed at gestational days 15 (P = 0.004 for lungs; P < 0.0001 for plasma), 16 (P < 0.0001 for lungs and plasma) and 18 (P < 0.0001 for lungs; P < 0.05 for plasma). Vitamin A tends to increase the expression of this factor in the lung, particularly during the critical period of perinatal adaptation to postnatal life. These effects seem to be spatial and temporally regulated, and point out to the important role of vitamin A during lung development.

A case of pulmonary cryptococcosis in a free-living toad (Bufo bufo).

Pulmonary cryptococcosis was observed in a free-living adult female common toad (Bufo bufo) that was killed by a vehicle. Both lungs had various eosinophilic, monomorphic, and spherical to elliptical organisms identified as Cryptoccocus spp. The yeasts were demonstrated by Grocotts silver method and the periodic acid-Schiff reaction and the capsule was positive for mucin with a mucicarmine stain. The agent was confirmed by immunohistochemistry, using the monoclonal antibody anti-Cryptococcus neoformans, and by a polymerase chain reaction-based method using a C. neoformans-specific primer. This report, to the best of our knowledge, represents the first case of cryptococcosis in a common toad.

Pulmonary adiaspiromycosis in a European hedgehog (Erinaceus europaeus) in Portugal

ADIASPIROMYCOSIS (haplomycosis) is a self-limiting pulmonary mycosis caused by the dimorphic fungi Chrysosporium parvum var crescens and C parvum var parvum (Burek 2001, Carter 2002) that occurs worldwide in small mammals (Turner and others 1999, Travis and others 2002). The disease has been reported in 124 mammal species and subspecies (Acha and Szyfres 1989) of the Carnivora, Rodentia and Insectivora (Burek 2001). The infection is widespread, especially among rodents of the Arvicolidae and Muridae families, but it rarely occurs in the Sciuridae and Cricetidae (Fischer 2001); the Mustelidae appear to be particularly susceptible to the infection (Simpson and GavierWiden 2000, Burek 2001). C parvum has also been found in dogs (Jones and others 1997, Carter 2002) and goats, and several cases of adiaspiromycosis have been reported in human beings since 1964 (Jones and others 1997). Human infection is uncommon (England and Hochholzer 1993, Turner and others 1999, Travis and others 2002), but cases have been reported in the USA, Europe, the former Soviet Union, and Central and South America (Chandler and Watts 1988). Most human infections are attributed to C parvum var crescens (Chandler and Watts 1988, Travis and others 2002), but two reports documented disseminated disease caused by C parvum var parvum in patients with AIDS (Echevarria and others 1993, Turner and others 1999). This short communication describes a case of adiaspiromycosis due to C parvum var crescens in a European hedgehog (Erinaceus europaeus). In May 2001, a young male European hedgehog was found dead in a road near Vila Real in northern Portugal. The animal was collected and submitted for postmortem examination at the Histopathology Laboratory of Tras-os-Montes e Alto Douro University. The animal was in normal body condition, weighing 638 g. Postmortem examination revealed multiple traumatic lesions in the mandible and ribs, and rupture of the abdominal wall with evisceration. The pericardial sac was ruptured. The lungs were detached from their normal anatomical position and free in the thoracic cavity. They presented multiple sites of damage, oedema, congestion and extensive haemorrhagic areas, and various grey-white, nodular, focal lesions, measuring 0·5 to 1·0 mm in diameter, were disseminated on the surfaces of both lungs. The lesions appeared to have no predilection for particular areas or lobes of the lungs. No significant abnormalities were observed in the thymus. The liver was pale and friable, and had multiple ruptures, and the spleen was also damaged. The small intestine was ruptured at four points, and its contents had spilled into the peritoneal cavity. Samples of the organs were fixed in 10 per cent buffered formalin and processed for light microscopy using standard methods. Paraffin sections (2 μm) for histological examination were stained with haematoxylin and eosin and by Grocott’s technique. Microscopically, the lung presented atelectasis, emphysema and numerous granulomatous lesions in the interstitium (Fig 1), each containing one or two spherical adiaconidia surrounded by epithelioid cells, giant multinucleated cells in a palisade arrangement, and fibrous tissue. The diameter of the fungi ranged from 119 to 285 μm. The double contour, refractile wall had a thickness of 38 to 59 μm. The outer third of the trilaminar wall was eosinophilic and refractile, and there was a non-staining layer between the outer and inner walls that appeared black with Grocott’s technique (Fig 2). The interior of the adiaconidia was either empty or contained small eosinophilic globules. The structures were consistent with descriptions of the morphology of the adiaspores of C parvum var crescens (Chandler and Watts 1988, Jones and others 1997, Burek 2001, Travis and others 2002). The hedgehog also had a verminous bronchitis. No adiaspores were observed in the regional lymph nodes. The liver showed lipidosis, and the kidney presented pigmentary nephrosis and interstitial nephritis. No significant lesions were observed in the remaining organs. Adiaspiromycotic infection is believed to occur as a result of the inhalation of dustborne spores (conidia) found in soil, which accumulate in the bronchioles and alveoli (England and Hochholzer 1993, Burek 2001, Travis and others 2002). The genus Chrysosporium consists of ubiquitous fungi, which produce 2 μm to 4 μm conidia in the saprophytic form (Simpson and Gavier-Widen 2000, Travis and others 2002). When inhaled by a mammalian host, the conidia progressively enlarge to form thick-walled spherules, designated as adiaconidia or adiaspores (Acha and Szyfres 1989, Simpson and Gavier-Widen 2000, Fischer 2001). The adiaspores of C parvum var parvum grow to up to 20 to 40 μm (Burek 2001), whereas the C parvum var crescens adiaspores have a diameter ranging from 200 to 700 μm (Chandler and Watts 1988, Simpson and Gavier-Widen 2000, Travis and others 2002). C parvum var crescens has also a broader host range and geographical distribution than C parvum var parvum (Peterson and Sigler 1998, Burek 2001). Adiaconidia seem to be incapable of replication or dissemination in the host (Chandler and Watts 1988, Travis and others 2002), yet massive infections have been observed in small mammals (Burek 2001). The disease is not contagious (Jones and others 1997), thus each adiaspore constitutes a separate infection (Simpson and Gavier-Widen 2000). The infection resembles foreign-body inhalation pneumonia, with a granulomatous reaction due to the inhaled spores (Chandler and Watts 1988, Travis and others 2002). The severity of the disease depends on the number of spores inhaled (Acha and Szyfres 1989, England and Hochholzer 1993) and the immune response of the host (Chandler and Watts 1988, Jones and others 1997). When clinical signs are present, they can be attributed to mechanical displacement of the tissues and the inflammatory reaction caused by the proVeterinary Record (2006) 158, 274-275

Granuloma Coinfection with Mycobacterium bovis, Mycobacterium avium subsp. paratuberculosis, and Corynebacterium pseudotuberculosis in Five Hunted Red deer (Cervus elaphus) in Portugal

Abstract We report granulomatous lymphadenitis in red deer (Cervus elaphus) in Portugal caused by coinfection with Corynebacterium pseudotuberculosis, Mycobacterium bovis, and Mycobacterium avium subsp. paratuberculosis, as demonstrated by molecular methods.

SURVEY OF MYCOBACTERIUM AVIUM SUBSPECIES PARATUBERCULOSIS IN ROAD-KILLED WILD CARNIVORES IN PORTUGAL

Abstract:  A survey to determine the occurrence of Mycobacterium avium subsp. paratuberculosis (MAP) in wild carnivores in Portugal was conducted by testing samples from road-killed animals between 2009 and 2012. Postmortem examinations were performed and tissues were collected from wild carnivores representing four families and six different species, with a total of 74 animals analyzed. Cultures were performed by using Löwenstein-Jensen and Middlebrook 7H11 solid media and acid-fast isolates were identified by polymerase chain reaction (PCR) and mycobactin dependency characteristics. Tissues were also screened for MAP by directly extracting DNA and testing for the MAP-specific sequences. The occurrence of infected animals (an animal had at least one tissue that was positive for culture or direct PCR) was 27.0% (n = 20). MAP was isolated from culture of 25 tissue samples (3.8%) and was detected by direct PCR in 40 (6.0%) samples. Infection was recorded in 5/6 studied species: 7/49 (14.3%) red foxes (Vulpes vulpes), 3/3 (100%) beech martens (Martes foina), 2/4 (50.0%) Eurasian otters (Lutra lutra), 7/15 (46.7%) Egyptian mongooses (Herpestes ichneumon), and 1/1 (100%) European badger (Meles meles). These species represent three different taxonomic families: Canidae (14.3% were positive), Mustelidae (75.0% were positive), and Herpestidae (46.7% were positive). The results of this study confirm the presence of MAP infection in wild carnivores in Portugal.

Disseminated Mycobacterium avium subsp. paratuberculosis infection in two wild Eurasian otters (Lutra lutra L.) from Portugal.

Abstract:  Disseminated Mycobacterium avium subsp. paratuberculosis (MAP) infections were found in two Eurasian otters (Lutra lutra, L. 1758) killed by vehicular trauma in February and March 2010 in Castelo Branco, Portugal. At postmortem examination, the organs showed no significant gross alterations; however, microscopically, both animals had diffuse lymphadenitis with macrophage infiltration and deposition of hyaline material in the center of the lymphoid follicles. Acid-fast organisms were isolated from gastrointestinal tissue samples via bacteriologic culture. These organisms were identified as M. avium subsp. paratuberculosis by IS900 polymerase chain reaction (PCR). Additionally, direct IS900 PCR-positive results were obtained for multiple organs of both animals. This is the first report of MAP infection of otters in Portugal.

Cost-benefit analysis of sheep and goat brucellosis vaccination with Rev.1 in the north of Portugal from 2000 to 2005.

No norte de Portugal, procedeu-se a vacinacao massiva contra a brucelose em pequenos ruminantes entre 2001 e 2004. Neste estudo, efetuou-se uma analise de custo beneficio entre 2000 e 2005 para apurar os beneficios economicos dessa estrategia. Para estimar o custo da zoonose, estudaram-se tres variaveis: a indenizacao paga aos proprietarios dos animais eliminados durante a campanha de erradicacao da brucelose, o custo da vacina Rev. 1 e os custos de internamento das pessoas infectadas por brucelose nas unidades de Servicos de Saude. Durante os anos em estudo, observou-se um aumento de custo de aproximadamente US

Mycobacterium avium Complex in Domestic and Wild Animals

110.000, entre 2000 e 2001, principalmente, devido a indenizacao dos animais eliminados e a aplicacao da vacina e os custos por internamento de pessoas infectadas, seguindo-se uma reducao acentuada entre 2001 e 2004 (de US

Coloração de Ziehl-Neelsen como método rápido de diagnóstico de paratuberculose ovina

1.200.000 para US

A seroepidemiological survey of Mycobacterium avium subsp. paratuberculosis in sheep from North of Portugal

180.000 em 2004), o que perfez US