Bacterial infection within a joint, septic arthritis, is a serious condition in horses that can lead to long-term joint disease if the infection is not resolved quickly. Equine septic arthritis is diagnosed primarily based on clinical signs and synovial fluid cytology. Septic synovial fluid is characterized by significant elevations in total protein (TP) and total nucleated cell count (TNCC). However, in some cases it can be difficult to distinguish between septic arthritis and non-septic joint inflammation (synovitis) based on clinical signs and synovial fluid cytology alone. A rapid assay to help confirm septic arthritis would be advantageous. A new assay to quantify the major equine acute phase protein, serum amyloid A (SAA) may fulfill this need. Serum amyloid A increases in the body in response to injury, infection, and inflammation and shows promise as a useful tool in confirming a diagnosis of sepsis, as inflammation causes mild increases in SAA and infection causes marked elevations. In our study, serial serum and synovial fluid samples were collected from horses with experimental models of synovitis and septic arthritis, synovial fluid cytology was performed, and serum and synovial fluid SAA were quantified. Synovial fluid TNCC and TP concentrations increased significantly following induction of both models. Serum and synovial fluid SAA concentrations remained normal in synovitis horses and increased significantly in septic arthritis horses. Any elevation in serum or synovial fluid SAA above normal values may be supportive of synovial sepsis since synovial inflammation alone did not result in SAA elevations in our model. / Master of Science

The analysis of acute phase protein serum amyloid A (SAA) has recently been brought into clinical use in veterinary medicine. Some of the difficulties with incorporating the SAA method in clinical practice have been the expensive and rather large equipment required for the method. Due to these difficulties only larger clinics can afford to use the SAA analysis. The company Equinostic has recently developed a smaller instrument that costs one-tenth of a larger instrument. The instrument is named EVA1 and has so far only been used to analyze SAA in horses. The aim of this study was to investigate if the EVA1 instrument could be used to analyze SAA in cats. This study included 24 serum samples from cat, which were first analyzed twice on the EVA1 instrument and then sent to the Strömsholm Referral Animal Hospital in Sweden where they reanalyzed the samples using a validated reference method. Both instruments are based on an immunoturbidimetric assay. The correlation between the two instruments was good (r=0.97) but the EVA1 instrument showed constantly lower results than the reference method. The difference between the duplicates when analyzed on the EVA1 instrument was larger than expected. The conclusion is that EVA1 could be used to analyze SAA in cats. However, before it could be used clinically in veterinary practice an extended study is recommended.

Die Bestimmung der Plasmaviskositaet und des Haptoglobin beim Rind erweisen sich als sinnvolle Ergaenzungen in der Labordiagnostik. Ihre Bestimmung als Suchtest zur Fruehdiagnostik von praxisrelevanten Rinderkrankheiten ist zu empfehlen.

Die Bestimmung der Plasmaviskositaet und des Haptoglobin beim Rind erweisen sich als sinnvolle Ergaenzungen in der Labordiagnostik. Ihre Bestimmung als Suchtest zur Fruehdiagnostik von praxisrelevanten Rinderkrankheiten ist zu empfehlen.

C-reactive protein (CRP) is synthesized in hepatocytes. The serum concentration of CRP increases dramatically during the acute phase response. In human hepatoma Hep3B cells, maximal CRP expression occurs in cells treated with the combination of IL-6 and IL-1β. IL-6 induces transcription of the CRP gene and IL-1β synergistically enhances the effects of IL-6. We investigated the role of IL-6-activated transcription factor STAT3, also known as STAT3α, in inducing CRP expression since we identified four consensus STAT3-binding sites centered at positions - 72, - 108, - 134 and - 164 on the CRP promoter. It has been shown previously that STAT3 binds to the site at - 108 and induces CRP expression. We found that STAT3 also bound to the other three sites, and several STAT3-containing complexes were formed at each site, suggesting the presence of STAT3 isoforms and additional transcription factors in the complexes. Mutation of the STAT3 sites at - 108, - 134 or - 164 resulted in decreased CRP expression in response to IL-6 and IL-1β treatment, although the synergy between IL-6 and IL-1β was not affected by the mutations. The STAT3 site at - 72 could not be investigated employing mutagenesis. We also found that IL-6 activated two isoforms of STAT3 in Hep3B cells: STAT3α which contains both a DNA-binding domain and a transactivation domain and STAT3β which contains only the DNA-binding domain. Taken together, these findings raise the possibility that IL-6 not only induces CRP expression but also regulates the induction of CRP expression by activating STAT3 isoforms and by utilizing all four STAT3 sites.

Orientador: Helio Langoni / Resumo: A mastite bovina é responsável por grandes perdas econômicas na bovinocultura de leite, além de aspectos de saúde pública que devem também ser considerados. Animais com mastite clínica ou subclínica apresentam diversas alterações reprodutivas. No primeiro artigo da tese, os resultados obtidos demonstraram que a mastite causada por patógenos maiores (20,1%) ou por gram-negativos (15,4%) levaram a queda na taxa de prenhez na primeira inseminação comparado com o grupo controle (32,6%). Foi observado maior taxa de perda gestacional para os grupos de patógenos maiores (22,2%) e gram-negativos (30,1%) comparados com o controle (12,8%). Foi observado diferença entre todos os grupos para o número de dias em aberto. O grupo controle apresentou o menor intervalo (126,5 dias) seguido pelo grupo patógenos menores (162,0 dias) e pelo grupo de patógenos maiores (175,1 dias). Comportamento similar observado para patógenos gram-positivo (172,7 dias) e patógenos gram-negativos (191,1 dias). Portanto, é importante controlar a mastite em rebanhos leiteiros e, para tanto, o diagnóstico é uma etapa importante. Diferentes proteínas de fase aguda foram descritas como bons biomarcadores para o diagnóstico da mastite. No segundo artigo da tese, foi observado que diferentes patógenos podem alterar a liberação de proteínas de fase aguda no leite. Bactérias que causam inflamações mais graves (E. coli e Klebsiella pneumoniae) levaram maior concentração das APP. Patógenos que causam infecções crônica (Str... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Bovine mastitis is responsible for major economic losses in dairy cattle farming, as well as public health aspects that should also be considered. Animals with clinical or subclinical mastitis present several reproductive alterations. In the first article of this thesis, the results showed that mastitis caused by major pathogens (20.1%) or gram-negative (15.4%) led to a decrease in pregnancy rate in the first insemination compared to the control group (32.6%). Higher gestational loss rates were observed for the major (22.2%) and gram-negative (30.1%) pathogen groups compared with the control (12.8%). The difference was observed between all groups for the number of days open. The control group had the shortest interval (126.5 days) followed by the minor pathogens group (162.0 days) and the major pathogens group (175.1 days). Similar behavior observed for gram-positive pathogens (172.7 days) and gram-negative pathogens (191.1 days). Therefore, it is important to control mastitis in dairy herds and, so, diagnosis is a critical step. Different acute phase proteins have been described as good biomarkers for the diagnosis of mastitis. In the second article of this thesis, it was observed that different pathogens may alter the release of acute phase proteins in milk. Bacteria that cause more severe inflammation (E. coli and K. pneumoniae) led to a higher concentration of APP. Pathogens causing chronic infections (Environmental Streptococcus, S. aureus, Mycoplasma spp.) led to interm... (Complete abstract click electronic access below) / Doutor

Os objetivos deste estudo foram: determinar a dinâmica de concentrações séricas de proteínas de fase aguda em cadelas gestantes e aspectos ultra-sonográficos da gestação, além de estabelecer uma possível correlação entre níveis séricos das mesmas proteínas, idade gestacional e aspectos ultra-sonográficos da gestação. Examinou-se 09 cadelas gestantes da referida raça com um aparelho portátil da marca GE®, modelo Logic &alpha; 100 MP, equipado com um transdutor convexo 5,0 MHz e outro linear de 7,5 MHz, de 2 a 3 vezes por semana a partir do 18&ordm; dia de gestação até o parto com a finalidade de mensurar as seguintes estruturas no concepto: DBP, DC e EP. Nos mesmos dias, coletou-se soro para dosagem de Hp, GA&alpha;1 e CRP por meio de kits comerciais (Phase Haptoglobin Assay ®, ,Canine &alpha;1 Acid Glycoprotein measurement kit ®; Phase Canine C - Reactive Protein Assay ®, Tridelta Development Limited, respectivamente). As cadelas do grupo NG tiveram o soro coletado a partir do proestro ao fim do diestro. Foi possível dividir o período gestacional em 9 semanas, cada uma delas, da terceira à nona semana de gestação, foi associada a um determinado conjunto de características ultra-sonográficas no concepto. As mensurações ultra-sonográficas foram altamente correlacionadas à idade gestacional e resultou em um modelo ; Y = 22,886 + 1,26 DBP + 0,107 DC, onde Y = idade gestacional (p < 0001 e R2 = 0,99). As concentrações séricas de Hp, GA&alpha;1 e CRP, no grupo G, foram significativamente maiores que no grupo NG a partir da 4&ordf; semana de gestação. Essas variáveis não foram tão bem correlacionadas com a idade gestacional, quanto às mensurações ultra-sonográficas no concepto, contudo, pode-se afirmar que as concentrações séricas máximas de Hp, GA&alpha;1 e CRP (11,11 ± 1,7 mg/ml; 449,44 ± 146,91 &micro;g/ml e 58,30 ± 14,69 µg/ml) foram atingidas na 5&ordf semana de gestação. Foi possível ilustrar a dinâmica das proteínas e fase aguda (Hp, GA&alpha;1 e CRP) e das mensurações ultra-sonográficas (DBP, DC e EP), possibilitando a visualização do comportamento dessas variáveis, uma em relação às outras, durante a gestação de cadelas da raça boxer. / The aims of this study were: to set up the dynamics of acute phase proteins serum concentrations in pregnant bitches and ultrasonographis features of conceptus and make a correlation among acute phase proteins serum levels, ultrasonographic features of conceptus and gestational age. Serial ultrasonographic examinations were performed on 09 boxer bitches, 2 or 3 times per week from 18º day of gestation until parturition. The used device is a portable GE®, Logic &alpha; 100 MP, equipped with a sectorial transducer 5,0 MHz and another linear one of 7,5 MHz. Embryonic and fetal parameters had been measured: DBP, DC e EP. Serum samples were collected in the same day and acute phase proteins (Hp, GA&alpha;1 e CRP) were measured by commercial kits (Phase Haptoglobin Assay ®, Canine &alpha;1 Acid Glycoprotein measurement kit ®; Phase Canine C - Reactive Protein Assay ®, Tridelta Development Limited). Serum samples of non pregnant bitches were collected from the beginnig of proestrous to the end of diestrous. The gestational period was divided into 9 weeks, each one of them (from the 3rd to the 9th week of gestation) was associated to a range of ultrasonographic features of the conceptus. Ultrasonographic mensurations (DBP, DC e EP) were linearly correlated to gestational age. The R2 values were > 0,9 in all cases, whitch indicates that more than 90% of the variability of gestational age was explained by any one of these measurements. Those mensurations were put in a multivariated model and resulted in a equation: Y = 22,886 + 1,26 DBP + 0,107 DC (p < 0,0001 e R2 = 0,99) that is able to estimate the gestational age. Hp, GA&alpha;1 e CRP seric concentrations were statisticaly more elevated in pregnant than in non pregnant bitches, from the 4th week of gestation until parturition. The acute phase proteins didn´t presented goog correlation to the gestational age like the ultrasonographic mensurations did, however it is possible to affirm that their concentrations reached maximum values (Hp = 11,11 ± 1,7 mg/ml, GA&alpha;1 = 449,44 ± 146,91 µg/ml e CRP = 58,30 ± 14,69 µg/ml) on the 5th week. The dynamic of the acute phase proteins (Hp, GA&alpha;1 e CRP) and ultrasonographic mensurations (DBP, DC e EP) was performed in order to visualise the profile of all mean measurements, one regarding to the other, during gestation in boxer bitches.

A manheimiose pneumônica bovina (MPB) gera significativa lesividade quando predomina no Complexo Doença Respiratória Bovina. Visto isso, considera-se importante o estudo do diagnóstico precoce para melhor estadiamento e tratamento da doença. As concentrações plasmáticas e séricas das proteínas de fase aguda (PFA) têm sido adotadas na medicina veterinária como marcadores precoces de doenças inflamatórias e algumas vezes, relacionadas ao bem-estar animal. O presente estudo objetivou revisar o painel inflamatório da MPB; investigar a contribuição que o emprego da flunixina meglumina no tratamento de bovinos manifestando MPB; e investigar marcadores inflamatórios para o diagnóstico precoce da doença e evolução do tratamento, empregando referências de bem estar animal. Para isto foram utilizados 12 bezerros da raça holandesa com seis meses de idade, hígidos e divididos em dois grupos experimentais, o primeiro tratado com antimicrobiano e o segundo com antimicrobiano e anti-inflamatório. Os animais foram submetidos à inoculação intratraqueal de M. haemolytica e tiveram suas amostras de lavado broncoalveolar e de sangue colhidas em quatro momentos, onde foram realizados exame clínico, coleta de sangue e lavado broncoalveolar para realização de hemograma, determinação das proteínas de fase aguda, análise de cortisol e cortisona. A análise estatística foi realizada utilizando programa SAS 9.3 para Windows. Os dados foram testados quanto à normalidade pelo teste de Shapiko Wilk, considerando significância de 5%. Foi efetuada a análise de variância simples com comparação entre médias por meio do teste de LSD, sendo considerado significante p&lt;0.05. O resultados mostraram que a MPB é uma doença de grande importância no cenário mundial; não foram encontradas diferenças estatísticas entre os marcadores precoces entre os grupos com ou sem anti-inflamatório, porém o uso concomitante com o antimicrobiano promoveu incremento do bem estar. / The bovine pneumonic mannheimiosis (MPB) generates significant lesivity when it predominates in the Complex Bovine Respiratory Disease. Thus being important the study of the previous diagnosis for better understanding and treatment of the disease. As plasma and serum concentrations of acute phase proteins (PFA) have been adopted in veterinary medicine as early markers of inflammatory diseases and sometimes related to animal welfare. The aim of the study was review the inflammatory panel of MPB; to investigate a contribution that the use of flunixin meglumina without treatment of bovines manifesting MPB; and to investigate inflammatory markers for the previous diagnosis of the disease and treatment evolution, using references of animal welfare. Twelve six-month-old, healthy and divided into two experimental groups, the first treated with antimicrobial and the second with antimicrobial and anti-inflammatory were used in this study. The animals were submitted to intratracheal inoculation of M. haemolytica and had their bronchoalveolar lavage and blood samples collected at four moments, where clinical examination, blood collection and bronchoalveolar lavage were carried out to perform hemogram, determination of the acute phase proteins, analysis of cortisol and cortisone. Statistical analysis was performed using SAS 9.3 software for Windows. The data were tested for normality by the Shapiko Wilk test, considering a significance of 5%. The analysis of simple variance was performed with a comparison of means by means of the LSD test, being considered significant p &lt;0.05. The results showed that MPB is a disease of great importance in the world scenario; no statistical differences were found between the early markers between the groups with or without anti-inflammatory, but the concomitant use with the antimicrobial promoted an increase in animal welfare.

Orientador: José Jurandir Fagliari / Banca: Rafael Resende Faleiros / Banca: Delphim da Graça Macoris / Banca: José Dantas Ribeiro / Banca: Daniela Gomes da Silva / Resumo: Este estudo teve como objetivo geral comparar os efeitos da laparotomia exploratória e do uso da fenilbutazona em bovinos e bubalinos. Para isso foram avaliados o perfil bioquímico, o proteinograma e o hemograma de 28 animais dos quais 14 eram búfalas e 14 vacas, todas fêmeas não lactantes, distribuídas em quatro grupos - VT: vacas tratadas com fenilbutazona; VC: vacas controle; BT: búfalas tratadas com fenilbutazona e BC: búfalas controle. Os animais dos grupos VT e BT receberam dose única de 10 mg/kg de fenilbutazona, por via intravenosa, uma hora antes da cirurgia. Foram coletadas amostras de sangue antes da laparotomia (0h) e 1(1h), 6(6h), 12(12h), 24(24h), 48(48h), 72(72h), 96(96h) e 120 horas (120h) e 15(15d) e 30(30d) dias após a intervenção. Os valores de creatinina foram mais elevados nas búfalas, e não sofram alteração devido à cirurgia, ou ao tratamento. O tratamento com fenilbutazona aumentou a concentração sérica de HDL em vacas e búfalas. Dentre as proteínas de fase aguda, a concentração de ceruloplasmina se elevou em todos os grupos de 48 horas a 120 horas após a cirurgia. A concentração sérica de haptoglobina se elevou mais nos grupos das búfalas do que nas vacas. A hemopexina foi verificada apenas na espécie bubalina e apenas até o 5º dia. O tratamento com fenilbutazona não alterou a contagem de hemácias nem o volume globular. Todas as vacas e búfalas apresentaram neutrofilia 12 horas após a laparotomia. O teor de fósforo aumentou no grupo controle das búfalas (BC) e no grupo tratado (BT), às 72h e 96h, respectivamente. O magnésio se apresentou, no início do experimento (0h) e ao final (30d), significativamente maior nos grupos das búfalas (BT e BC). A laparotomia e ou o tratamento com fenilbutazona não interferiram nas concentrações de sódio, potássio e cálcio ionizado / Abstract: This study aims to compare the effect of exploratory laparotomy and treated with phenylbutazone in bovine and buffaloes. To pursue this purpose, biochemical profile, proteinogram and blood count cell tests were performed in 28 animals, of which 14 were buffaloes and 14 cows, all of then non-nursing, distributed in four groups - VT: cows treated with phenylbutazone; VC: control cows; BT: buffaloes treated with phenylbutazone and BC: control buffaloes. Animals from VT and BT received a single administration of 10 mg/kg of phenylbutazone, intravenously one hour before surgery. Blood samples were taken before laparotomy (0h) and 1(1h), 6(6h), 12(12h), 24(24h), 48(48h), 72(72h), 96(96h) e 120 horas (120h) e 15(15d) e 30(30d) days after surgery. Creatinine values were higher in buffaloes, and there were no changes, despite surgery or treatment. HDL showed different values in both species, due to surgical procedure. The use of phenylbutazone increased serum HDL in cows and buffaloes. Among acute phase proteins, serum haptoglobin showed higher values in buffaloes than in cows. Hemopexina was foundetected only in buffaloes and until the 5th day. Treatment with phenylbutazone did not change RBCs or globular volume values. Cows and buffaloes showed neutrophilia 12 hours after laparotomy. Phosphorus content increased in control buffaloes (BC) and in the treated buffaloes (BT) at 72h and 96h, respectively. Magnesium presented higher itself at the beginning of the experiment (0h) and end (30d) in buffaloes groups (BT and BC). Laparotomy or treatment with phenylbutazone did not changed sodium, potassium and calcium ionized values / Doutor

Orientador: José Jurandir Fagliari / Banca: Daniela Gomes da Silva / Banca: Luiz Paulo Martins Filho / Resumo: Foi avaliada a importância de proteínas de fase aguda (PFA) como biomarcadores de enfermidades em ruminantes domésticos. Para tal se determinou o proteinograma sérico e leucograma de 15 bovinos sadios e 15 doentes, 15 ovinos sadios e 15 doentes, e 15 caprinos sadios e 15 doentes. Todos foram submetidos a colheitas diárias de sangue durante sete dias, enquanto permaneciam internados no Hospital Veterinário da FCAV-UNESP, Câmpus de Jaboticabal. Em bovinos a haptoglobina teve destaque como biomarcador precoce e confiável de foco inflamatório/infeccioso. Em ovinos ceruloplasmina, haptoglobina, 1-glicoproteína ácida mostraram moderada atividade frente às enfermidades avaliadas. Caprinos acometidos por ectima contagioso não apresentaram boa responsividade das PFAs. No entanto, ceruloplasmina, haptoglobina, 1-glicoproteína ácida destacaram-se como biomarcadores de foco inflamatório/infeccioso em caprinos acometidos por mastite e artrite secundária à infecção pelo vírus da artrite encefalite caprina. Transferrina, embora tenha se mostrado biologicamente como uma PFA negativa em todos os grupos estudados, apenas apresentou comportamento estatisticamente diferenciado em bovinos acometidos por mastite. Por fim, o proteinograma sérico obtido por meio da técnica eletroforética SDS-PAGE, mostrou-se mais útil como exame complementar ao diagnóstico das enfermidades avaliadas que o leucograma / Abstract: The importance of acute phase proteins (APP) as biomarkers of disease was evaluated in domestic ruminants. It was determined the serum proteinogram and leucocyte count of 15 healthy and 15 clinically ill cattle; 15 healthy and 15 clinically ill sheep and 15 healthy and 15 clinically ill goats. All animals were submitted to daily blood sampling during 7 days, while kept interned in the Veterinarian Hospital from FCAV-UNESP, Jaboticabal campus. In cattle, haptoglobin was a very reliable inflamatory/infeccious early biomarker. In sheep ceruloplasmin, haptogobin and 1-acid glycoprotein showed moderate activity to the diseases studied. Goats with contagious ectima did not show a good response in APP serum concentrations. However, ceruloplasmin, haptoglobin and 1-acid glycoprotein showed to be reliable soon biomarkers in goats with mastitis and arthritis due to caprine arthritis encephalitis. Alhtough transferrin biologically behaved as a negative APP in all diseases and species studied, it only showed a statistically significant behaviour as such in cows with mastitis. Moreover, the assessment of APP profile through the electrophoretic SDS-PAGE technique was more useful as a complementary tool to the establishment of diagnosis than the leucocyte count / Mestre