Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Novel markers of inflammatory response and hepatic dysfunction in canine leishmaniasis

Dogs are the main host of Leishmania infantum, and the clinical presentation may range from asymptomatic to systemic manifestations. The immune mechanisms in infected, but clinically healthy dogs, prevails Th1 response mediated by cytokines. In this sense, adenosine deaminase (ADA) and butyrylcholinesterase (BChE) are considered as key enzymes in several physiological processes, including the modulation of inflammatory process. Considering the variable immune response against Leishmania and the known participation of ADA and BChE, the aim of this study was to assess the relation between these two enzymes with the inflammatory response as well as hepatic function in dogs naturally infected with L. infantum. For this purpose, the activity of ADA and BChE was assessed in sera of 24 dogs naturally infected with L. infantum, plus 17 healthy dogs. The naturally infected dogs had clinical signs compatible with leishmaniasis and sera activities of ADA (P<0.01) and BChE (P<0.05) decreased, when compared to the healthy group. The reduction of ADA activity probably represented an effect on inflammatory response, especially due to the decreased hydrolysis of extracellular adenosine, might in order to protect against tissue damage and, also, setting a down-regulation on pro-inflammatory cytokines. BChE enzyme had no effect on modulating the immune response in leishmaniasis, but it decreased, a fact may related to deficiency of synthesis in the liver. Therefore, ADA and BChE activities reduced probably in order to protect against extra tissue damage and due failure in synthesis, respectively.

The shadows of a ghost: a survey of canine leishmaniasis in Presidente Prudente and its spatial dispersion in the western region of São Paulo state, an emerging focus of visceral leishmaniasis in Brazil

Visceral leishmaniasis is an emerging zoonosis and its geographic distribution is restricted to tropical and temperate regions. Most of the individuals infected in Latin America are in Brazil. Despite the control measures that have been adopted, the disease is spreading throughout new regions of the country. Domestic dogs are involved in the transmission cycle and are considered to be the main epidemiologic reservoir of Leishmania infantum (syn. L. chagasi). Our aim was to determine the prevalence of canine leishmaniasis (CL) and Ehrlichiosis infection in Presidente Prudente as well as the spatial dispersion of the disease in the western region of São Paulo state. Dogs underwent clinical examination and symptoms related to CL were recorded. Anti- Leishmania antibodies were detected using ELISA, rK39-immunocromatographic tests (DPP), and an indirect fluorescent antibody test (IFAT). Anti-E. canis antibodies were detected by IFAT. A follow-up was conducted in dogs that were positive in the ELISA at the baseline study. Data on the spatial distribution of L. longipalpis and CL in São Paulo state were obtained from Brazilian public health agencies. Serum samples from 4547 dogs were analyzed. The seroprevalence of CL was 11.2 % by ELISA and 4.5 % by IFAT. In the follow-up, seroprevalence was 32.9 % by ELISA, 15.3 % by IFAT, 11.8 % by DPP test, and 66.5 % for E. canis. There was a significant positive association between Leishmania and E. canis infection (P < 0.0001). In the follow-up, clinical examinations revealed symptoms compatible with CL in 33.5 % of the dogs. L. longipalpis was found in 24 and CL in 15 counties of the Presidente Prudente mesoregion. The dispersion route followed the west frontier of São Paulo state toward Paraná state. Low CL and high ehrlichiosis prevalence rates were found in Presidente Prudente city. This emerging focus of CL is moving through the western region of São Paulo state toward the border of Paraná state. Integrated actions to fight the vector, parasites, infected dogs, and humans are needed to monitor the disease and implement strategies for epidemiologic control.

Immunohistochemical expression of B and T-lymphocytes and TGF-beta in experimentally transplanted canine venereal tumor

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological, immunohistochemical and molecular study of renal lesions in canine visceral leishmaniasis (CVL)

Canine visceral leishmaniasis (CVL) is an anthropozoonosis characterized by a clinically chronic progressive disease. Non lymphoid organs are also affected, especially the kidneys. Dogs with leishmaniasis usually die with renal failure despite treatment. Haematoxylin-eosin (HE) staining in kidney tissue sections has low sensitivity for parasite identification. Immunohistochemistry (IHC) and polymerase chain reaction (PCR) are efficient methods for Leishmania sp. antigen and DNA detection in cases of low parasite burden. The present study aims to identify renal lesions of CVL and correlate them with microscopic findings determined by histochemistry, IHC and PCR. Both IHC and PCR provided similar positivity for amastigote identification, 3/20 animals (15%), thus increasing detection of the parasite in renal tissues when compared with histopathologic examination. The lesion most commonly observed with visceral leishmaniasis-positive canine kidney tissue was membranoproliferative glomerulonephritis, followed by interstitial nephritis without correlation to the number of amastigotes.

Leishmania sp. amastigotes identification in canine transmissible venereal tumor

Leishmaniasis is a vector-borne disease with Leishmania chagasi being the etiological agent of canine visceral leishmaniasis in South America. Canine venereal tumor is a transplantable round cell tumor of histiocytic origin which is mostly observed in sexually active male and female intact dogs. It has been shown that Leishmania amastigotes have higher tropism for the canine male genital tract tissues and venereal leishmaniasis transmission has been documented in dogs but, to date, a canine venereal tumor-dependent transmission route has not been fully demonstrated. In this report, a 10-year-old, mixed breed, intact female dog presented a vaginal venereal transmissible tumor but no other clinical abnormalities otherwise. Unexpectedly, tumor tissue imprint smears examination revealed Leishmania sp. amastigotes within infiltrating macrophages. In addition to the cytological direct identification, the protozoan was confirmed within the neoplastic tissue by means of immunohistochemistry and polymerase chain reaction. This report illustrates an asymptomatic Leishmania sp. infection that may have started on or from the canine venereal tumor tissue, the latter option further supporting previous evidence of such an alternative vector-independent route of transmission for canine visceral leishmaniasis in areas where these diseases coexist.

Healing and expression of growth factors (TGF-β and PDGF) in canine radial ostectomy gap containing platelet-rich plasma

Objectives: To evaluate bone healing in an experimental radial ostectomy in dogs treated with autologous platelet-rich plasma (PRP), through histological, densitometric, radiographic studies, as well as expression of growth factors in the ostectomy gap. Methods: Twenty-one dogs were randomly divided into either a control or a PRP group. All underwent unilateral ostectomy of the radius to generate a gap of 2.0 mm, that was stabilized with external skeletal fixation. The ostectomy gap was either filled with PRP or left empty as a control. The radiographic and densitometric studies were performed after surgery, then at intervals until 60 days during the post-surgery period. Histological and immunohistochemical evaluations were performed at seven or 60 days post-surgery. Analyses were performed using a statistical analysis system, and the level of significance was set at p <0.05. Results: The median radiographic healing score in the PRP group increased significantly between day 0 and day 60. Furthermore, at 60 days, the median healing score and the proportion of healed ostectomies in the control group (score 1; 1/6 healed) and the PRP group (score 5; 4/5 healed) were significantly different. There were differences between groups in radiographic and densitometric values at days 45 and 60. The histological evaluation showed advanced bone healing at 60 days in the PRP group and signs of delayed union in the control group. Clinical relevance: Platelet-rich plasma can be used as an adjuvant therapy because it may promote better bone healing of a radial ostectomy treated with external skeletal fixation in dogs.

Tumour necrosis factor-alpha-induced protein 8 (TNFAIP8) expression associated with cell survival and death in cancer cell lines infected with canine distemper virus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detection of the tumour suppressor gene TP53 and expression of p53, Bcl-2 and p63 proteins in canine transmissible venereal tumour

Canine transmissible venereal tumour (CTVT) is a neoplasm transmitted among healthy dogs by direct contact with injured skin and/or mucous tissue. This study aimed to identify the TP53 gene, messenger RNA (mRNA) as well as the expression of p53, Bcl-2 and p63 proteins in histological sections of 13 CTVT samples at different stages of evolution. The in situ hybridization (ISH) and in situ reverse transcriptase polymerase chain reaction (RT-PCR) assays were used, which showed the DNA homologous to TP53 and its respective mRNA in 92.3% of the samples. We detected p53, p63 and Bcl-2 proteins in most of the cell samples in different grades of intensity. In addition, 46% of the samples were in the progressive and 54% in the regression phase. This is the first description of these proteins and a detailed study of their role in CTVT cells needs to be addressed in or to verify how these cells undergo apoptosis.

Immunohistochemical detection of metalloproteinase-9 (MMP-9), anti-oxidant like 1 protein (AOP-1) and synaptosomal-associated protein (SNAP-25) in the cerebella of dogs naturally infected with spontaneous canine distemper

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of Protein A in Detection of Canine Distemper Virus through immunosensor construction

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ex vivo torsional properties of the Targon Vet Nail System in canine femurs: comparison with the 2.4 mm LC-DCP Plate

Pós-graduação em Cirurgia Veterinária - FCAV

Discrimination of production environments of specialty coffees by means of stable isotopes and discriminant model

The south of Minas Gerais, Brazil stands out among various regions through its capacity for production of specialty coffees. Its potential, manifested through being one of the most award-winning Brazilian regions in recent years, has been recognized by the Cup of Excellence (COE). With the evident relationship between product quality and the environment in mind, the need arises for scientific studies to provide a foundation for discrimination of product origin, creating new methods for combating possible fraud. The aim of this study was to evaluate the use of carbon and nitrogen isotopes in discrimination of production environments of specialty coffees from the Serra da Mantiqueira of Minas Gerais by means of the discriminant model. Coffee samples were composed of ripe yellow and red fruits collected manually at altitudes below 1,000 m, from 1,000 to 1,200 m and above 1,200 m. The yellow and red fruits were subjected to dry processing and wet processing, with five replications. A total of 119 samples were used for discrimination of specialty coffee production environments by means of stable isotopes and statistical modeling. The model generated had an accuracy rate of 89% in discrimination of environments and was composed of the isotope variables of δ15N, δ13C, %C, %N, δD, δ18O (meteoric water) and sensory analysis scores. In addition, for the first time, discrimination of environments on a local geographic scale, within a single municipality, was proposed and successfully concluded. This shows that isotope analysis is an effective method in verifying geographic origin for specialty coffees.