Estudo da infecção por Besnoitia besnoiti : aplicação da imunohistoquímica na avaliação da resposta celular inflamatória peri-quística

Tese de mestrado em Doenças Infecciosas Emergentes, apresentada à Faculdade de Medicina da Universidade de Lisboa, 2009A besnoitiose bovina é uma doença infecciosa causada pelo Apicomplexa Besnoitia besnoiti sendo reemergente no sul da Europa. O presente trabalho visa o estudo morfológico da resposta celular inflamatória peri-quística que ocorre na pele de bovinos com besnoitiose crónica recorrendo, para isso, ao exame histológico de biópsias de pele de bovinos naturalmente infectados. Para além da classificação morfológica do tipo de resposta inflamatória, avaliou-se o número de quistos presentes na pele, por fragmento de biópsia, bem como, de forma qualitativa e quantitativa, as diferentes células envolvidas.Foram estudadas 52 biópsias de pele contendo quistos de B. besnoiti, todos exibindo inflamação granulomatosa em seu redor, assumindo diversos tipos de organização celular. Assim, 34 amostras apresentaram inflamação nodular, 15 inflamação nodular discreta e, apenas três, inflamação difusa intensa. Na maioria dos casos analisados (28 em 52), o número de quistos por fragmento de biópsia foi superior a dez, o que é manifestamente indicador de elevada carga parasitária na pele. Não foram encontradas diferenças estatisticamente significativas entre as contagens celulares nas diferentes respostas inflamatórias, o que nos permitiu concluir pela existência de um padrão celular de resposta comum, independente da sua organização morfológica, constituído maioritariamente por linfócitos e plasmócitos, eosinófilos, macrófagos e por último, neutrófilos. A avaliação específica das sub-populações celulares que compõem os infiltrados celulares peri-quísticos foi efectuada com recurso a técnicas de imunohistoquímica. A imunomarcação realizou-se com sucesso para os linfócitos T, os linfócitos B, a sub-população de linfócitos T γδ e para os macrófagos. Não foi obtida marcação para as sub-populações de linfócitos CD4+ e CD8+, apesar de várias tentativas efectuadas. O infiltrado inflamatório que rodeia o quisto é maioritarBovine besnoitiosis is an re-emergent infectious disease in the south of Europe, caused by the protozoa Besnoitia besnoiti. The present study aims to characterize the peri-cystic inflammatory response in the skin of chronically infected bovine using skin biopsies. Morphological classification of the inflammatory response was accomplished as well as identification and counting of type of inflammatory cells involved.The study involved 52 skin biopsies with cysts in variable numbers, which showed granulomatous inflammatory response around the cysts, classified as follows: 34 nodular, 15 discretely nodular and only 3 were classified as intensely diffuse. For the majority of cases (28 in 52), the number of cysts per fragment of skin biopsy was higher than ten, which is certainly an indicator of a heavy parasitic skin infection.No statistical differences were found for the cell counting parameters regarding the different types of inflammatory response, showing that there is a common reaction pattern, regardless of the cellular organisation around the cysts. Cell types involved were, by decreasing order, lymphocytes, eosinophils, macrophages and finally, the neutrophils.Immunohistochemistry techniques were applied for characterization of cell sub-populations in peri-cystic infiltrates. Those techniques were successful for T lymphocytes, B lymphocytes, T γδ lymphocytes and also for macrophages. Unfortunately and in spite of several attempts, we did not succeed in the identification of sub-populations of CD4+ and CD8+ lymphocytes. The inflammatory infiltrate around the cyst was mainly composed by T lymphocytes, followed by eosinophils and macrophages. The B lymphocytes appeared peripherally in small groups with plasmocytoid differentiation. The T γδ sub-population of lymphocytes occurred isolated and dispersed all over the inflammatory infiltrate.The present study's conclusions add to the knowledge of the cutaneous lesions in the chronic phase of bovine besnoitiosi

Optimization of the effect of pineapple by-products enhanced in bromelain by hydrostatic pressure on the texture and overall quality of silverside beef cut

Dehydrated pineapple by-products enriched in bromelain using a hydrostatic pressure treatment (225 MPa, 8.5 min) were added in marinades to improve beef properties. The steaks from the silverside cut (2 ± 0.5 cm thickness and weight 270 ± 50 g), characterized as harder and cheaper, were immersed in marinades that were added to dehydrated and pressurized pineapple by-products that corresponded to a bromelain concentration of 0–20 mg tyrosine, 100 g−1 meat, and 0–24 h time, according to the central composite factorial design matrix. Samples were characterized in terms of marination yield, pH, color, and histology. Subsequently, samples were cooked in a water-bath (80 °C, 15 min), stabilized (4 °C, 24 h), and measured for cooking loss, pH, color, hardness, and histology. Marinades (12–24 h) and bromelain concentration (10–20 mg tyrosine.100 g−1 meat) reduced pH and hardness, increased marination yield, and resulted in a lighter color. Although refrigeration was not an optimal temperature for bromelain activity, meat hardness decreased (41%). Thus, the use of pineapple by-products in brine allowed for the valorization of lower commercial value steak cuts.The first author acknowledges the financial support from Fundação para a Ciência e a Tecnologia (FCT),Portugal, through a doctoral fellowship (SFRH/BD/109124/2015). This work was supported by the national fundingof FCT, under the scope of the strategic funding to the research units LEAF, Linking Landscape, Environment,Agriculture and Food, School of Agriculture, University of Lisbon, 1349-017 Lisbon (UIDP/04129/2020), CIISA,Interdisciplinary Centre of Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon,1300-477, Lisbon (UIDB/00276/2020) FMV and PDR2020-1.0.1-FEADER-031359, funded by the European RegionalDevelopment Fund (ERDF)), QOPNA (UID/QUI/00062/2019), LAQV-REQUIMTE, Department of Chemistry,University of Aveiro, 3810-193 Aveiro (UIDB/50006/2020) and CEB, Centre of Biological Engineering, Departmentof Biological Engineering, University of Minho, 4710–057 Braga (UIDP/04469/2019) through national funds andwhere applicable co-financed by the FEDER, within the PT2020 Partnership Agreement.info:eu-repo/semantics/publishedVersio

Surgical Excision of an Adrenal Neuroblastoma in a Dog

A 11-month-old, intact male, Rhodesian Ridgeback was presented to the Veterinary Teaching Hospital with signs of inappetence, lethargy, and abdominal pain for 3 days. A large and well-defined abdominal retroperitoneal mass, related with the left kidney, at the expected location of the adrenal gland, was revealed by radiography, ultrasound, and computed tomography. The mass extended caudally to the iliac artery bifurcation, compressing the aorta, caudal vena cava, and both kidneys. Cytology findings were compatible with a malignant round cell tumor. The most probable diagnosis was neuroblastoma. Following a comprehensive discussion with the owners about a treatment plan, surgical excision was performed. Because a wide excision would compromise major vessels, excision was performed after careful dissection of the aorta and vena cava. The left kidney was removed because the proximal ureter could not be separated from the tumor. The animal recovered successfully. Diagnosis was confirmed by histopathology and immunohistochemistry, but the owners decided not to pursue any further treatment. Clinical signs of abdominal pain recurred within 1 month following surgery. Therefore, the animal was euthanized upon the owners' request. This report describes the diagnosis, surgical treatment, and follow-up of a dog with an abdominal peripheral neuroblastoma

The significance of cephalopod beaks as a research tool: An update

The use of cephalopod beaks in ecological and population dynamics studies has allowed major advances of our knowledge on the role of cephalopods in marine ecosystems in the last 60 years. Since the 1960's, with the pioneering research by Malcolm Clarke and colleagues, cephalopod beaks (also named jaws or mandibles) have been described to species level and their measurements have been shown to be related to cephalopod body size and mass, which permitted important information to be obtained on numerous biological and ecological aspects of cephalopods in marine ecosystems. In the last decade, a range of new techniques has been applied to cephalopod beaks, permitting new kinds of insight into cephalopod biology and ecology. The workshop on cephalopod beaks of the Cephalopod International Advisory Council Conference (Sesimbra, Portugal) in 2022 aimed to review the most recent scientific developments in this field and to identify future challenges, particularly in relation to taxonomy, age, growth, chemical composition (i.e., DNA, proteomics, stable isotopes, trace elements) and physical (i.e., structural) analyses. In terms of taxonomy, new techniques (e.g., 3D geometric morphometrics) for identifying cephalopods from their beaks are being developed with promising results, although the need for experts and reference collections of cephalopod beaks will continue. The use of beak microstructure for age and growth studies has been validated. Stable isotope analyses on beaks have proven to be an excellent technique to get valuable information on the ecology of cephalopods (namely habitat and trophic position). Trace element analyses is also possible using beaks, where concentrations are significantly lower than in other tissues (e.g., muscle, digestive gland, gills). Extracting DNA from beaks was only possible in one study so far. Protein analyses can also be made using cephalopod beaks. Future challenges in research using cephalopod beaks are also discussed.info:eu-repo/semantics/publishedVersio

Cross Species Analysis and Comparison of Tumors in Dogs and Cats, by Age, Sex, Topography and Main Morphologies. Data from Vet-OncoNet

The animal cancer burden is essential for the translational value of companion animals in comparative oncology. The present work aims to describe, analyze, and compare frequencies and associations of tumors in dogs and cats based on the Animal Cancer Registry created by Vet-OncoNet. With 9079 registries, regarding 2019 and 2020, 81% (n = 7355) belonged to dogs. In comparison, cats have a general one-year right advance in the mean age of cancer diagnosis compared to dogs. The multivariate topography group analysis shows a distinct pattern between the two species: dogs have higher odds of cancer in the genito-urinary system, spleen, soft tissue tumors and skin, while cats show higher odds for tumors in the eyes, digestive organs, nasal cavity, lymph nodes, bones and mammary glands. Regarding morphologies, dogs are overrepresented in mast cell tumors (MCT), melanomas, and hemangiosarcomas. While cats are overrepresented in fibrosarcomas, lymphomas (T and B-cell), in malignant mammary tumors, and squamous cell carcinoma (SCC). Females have greater odds only in the mammary gland, with males having greater odds in six of twelve topographies. This study is the first outcome of continuous animal cancer registration studies in Portugal

The significance of cephalopod beaks as a research tool: An update

The use of cephalopod beaks in ecological and population dynamics studies has allowed major advances of our knowledge on the role of cephalopods in marine ecosystems in the last 60 years. Since the 1960’s, with the pioneering research by Malcolm Clarke and colleagues, cephalopod beaks (also named jaws or mandibles) have been described to species level and their measurements have been shown to be related to cephalopod body size and mass, which permitted important information to be obtained on numerous biological and ecological aspects of cephalopods in marine ecosystems. In the last decade, a range of new techniques has been applied to cephalopod beaks, permitting new kinds of insight into cephalopod biology and ecology. The workshop on cephalopod beaks of the Cephalopod International Advisory Council Conference (Sesimbra, Portugal) in 2022 aimed to review the most recent scientific developments in this field and to identify future challenges, particularly in relation to taxonomy, age, growth, chemical composition (i.e., DNA, proteomics, stable isotopes, trace elements) and physical (i.e., structural) analyses. In terms of taxonomy, new techniques (e.g., 3D geometric morphometrics) for identifying cephalopods from their beaks are being developed with promising results, although the need for experts and reference collections of cephalopod beaks will continue. The use of beak microstructure for age and growth studies has been validated. Stable isotope analyses on beaks have proven to be an excellent technique to get valuable information on the ecology of cephalopods (namely habitat and trophic position). Trace element analyses is also possible using beaks, where concentrations are significantly lower than in other tissues (e.g., muscle, digestive gland, gills). Extracting DNA from beaks was only possible in one study so far. Protein analyses can also be made using cephalopod beaks. Future challenges in research using cephalopod beaks are also discussed

Osteomyelitis caused by Aspergillus terreus complex in a dog: a case report

Abstract Background In dogs, the most frequently reported mycosis associated with Aspergillus spp. are respiratory infections. Systemic aspergillosis is uncommon, with reported cases been associated with several Aspergillus species. Aspergillus terreus species complex are ubiquitous organisms, unfrequently associated with local or systemic disease in animals and humans, and treatment of osteomyelitis caused by this species is usually unfavorable. Case presentation This case report describes the case of a 5-year-old dog, referred to the Veterinary Hospital of the Faculty of Veterinary Medicine of the University of Lisbon, Portugal, with a history of lameness of the right thoracic limb. Radiographs and CT scan revealed two different lesions on right humerus and radio, which were biopsied. The samples collected were submitted to cytological and histopathological evaluation and bacterial and mycological culture. Environmental samples, including of the surgery room and of the biopsy needle were also evaluated for the presence of fungi. Regarding biopsy samples, bacterial culture was negative, but mycological analysis originated a pure culture of a fungal species later identified as Aspergillus terreus by Sanger sequencing. Results were compatible with histopathologic examination, which revealed periosteal reaction and invasion of hyphae elements. Also, mycological analysis of both environmental samples evaluated were negative. The virulence profile of the fungal isolate was phenotypically characterized using specific media, allowing to reveal its ability to produce several enzymes involved in its pathogenicity, namely lipase, hemolysin and DNAse, corresponding to a Virulence Index (V. Index.) of 0.43. The patient was submitted to itraconazole therapy for 8 weeks. After 3 weeks, the patient showed significant clinical improvement, and after 6 weeks no radiographic signs were observed. Conclusions Antifungal therapy with itraconazole can contribute to the remission of canine infections promoted by Aspergillus terreus complex with a relevant V. Index