Sheep naturally infected with Anaplasma ovis - evaluation of cardiac and inflammatory biomarkers

Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

Early maturation of corpus luteum in rabbits - effect of sildenafil citrate on luteolytic capacity in the early luteal period

Background: Prostaglandin F2 alpha (PGF2 α) binds to the specific receptor (PTGFR) on the corpus luteum (CL) in mammals, inducing regression of the CL structure (luteolysis) and initiating a new cycle. While PGF2 α is effective only on mature CL, the immature CL structure (early luteal phase) resists PGF2 α. In this study, sildenafil citrate, which is used to increase blood flow in the genital organs for treating specific pregnancy issues in women, was administered during the early luteal phase in a rabbit model to test the hypothesis of enhancing blood flow to the CL, thereby promoting earlier maturation and enabling a response to PGF2 α. Materials, Methods & Results: The study was conducted in 2 sub-studies: clinical and molecular. A large number of rabbits were initially included in the sub-studies to ensure a sufficient number of pseudo-pregnant rabbits. Ovulation in rabbits was induced with buserelin acetate and was considered as day 0 of the study. The sub-studies were continued with rabbits whose pseudo-pregnancies were confirmed according to progesterone (P4 ) results. As a result, the studies were continued with a total of 41 pseudo-pregnant New Zealand female rabbits, 21 of which were included in the clinical sub-study and 20 in the molecular sub-study. In both sub-studies, on day 3 of the luteal period, rabbits in the treatment group received 5 mg/kg sildenafil citrate and all rabbits received a single dose of exogenous PGF2 α on day 4 to induce luteolysis. In the clinical sub-study, echotexture and intraovarian blood flow changes in the ovaries were determined by ultrasonography (USG) examination. In the molecular sub-study, the expression changes of Hypoxia Inducible Factor 1 Alpha (HIF1A) and Vascular Endothelial Growth Factor (VEGF) related to angiogenesis, Steroidogenic Acute Regulatory Protein (StAR) related to P4 metabolism, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) related to prostaglandin (PG) mechanism and 15-Hydroxyprostaglandin Dehydrogenase (HPGD) genes at mRNA level were determined using Real Time Polymerase Chain Reaction (RT-PCR) in CL tissues obtained with ovariohysterectomy (OVH) at 1 and 12 h after PGF2 α injection. In addition, blood samples were collected for determine P4 levels from all rabbits. In the clinical sub-study; there was no difference between the groups in mean gray values (MGV), whereas there was a significant decrease in both pulsatile index (PI) and resistance index (RI) values at 40 min after PGF2 α injection (P < 0.05). In the molecular sub-study, it was determined that sildenafil citrate had no significant effect (P > 0.05) on the expression levels 1 and 12 h after PGF2 α injection. According to the results of the molecular sub-study, no significant effect of sildenafil citrate on the mRNA expression levels in the investigated genes was detected (P > 0.05). However, within each group, differences were found according to OVH time after PGF2 α injection. It was observed that PTGS2 and HPGD mRNA expressions decreased at the 12th h compared to the 1st h, while HIF1A expression increased (P < 0.05). Discussion: According to the results obtained from clinical and molecular sub-studies, it was determined that a single dose of sildenafil citrate (5 mg/kg) applied on the 3rd day of the luteal period did not contribute to the maturation process of the CL, did not increase blood flow, and was insufficient to break the resistance of the CL against PGF2 α applied on the 4th day of the luteal period. However, a significant decrease in the PI value at the 40th min after PGF2 α injection suggests that sildenafil citrate has a supportive effect, and that this decrease is also seen in the RI value, suggesting that its effect is insufficient against the vasoconstrictive effect of PGF2 α.

Acute phase protein and vitamin D concentration in dogs with multicentric lymphoma / Concentração de proteínas de fase aguda e vitamina D em cães com linfoma multicêntrico

This study aimed to evaluate the serum concentration of vitamin D (25-Hydroxyvitamin D) and acute phase proteins (APPs; alpha-1 acid glycoprotein, haptoglobin, transferrin, ceruloplasmin, albumin, IgA, IgG and alpha-1 - antitrypsin) as potential biomarkers for prognostic and therapy response in dogs with multicentric lymphoma submitted to the CHOP (Cyclophosphamide, Doxorubicin, Vincristine and Prednisone) chemotherapy protocol. Thirteen dogs with multicentric lymphoma classified as high grade by cytology were included in the treatment group (GL), while ten healthy dogs were included in the control group (GC). Serum was collected in the weeks T0, T5 and T10 of CHOP chemotherapy protocol, for the GL group, and in a single collection, for the GC group. All the collected samples were evaluated for the APPs and vitamin D concentrations through electrophoresis and chemiluminescence methods, respectively. Diagnostic and staging tests were performed for all the dogs in the GL group, and included cytopathology, histopathology and immunohistochemistry of the affected lymph node. Of these dogs, 9 achieved a complete response and 4 a partial response to the treatment. Data analysis was performed with the R software. The results demonstrated that serum concentrations of IgA, haptoglobin and α1-acid glycoprotein were significantly different between the groups and also between the different chemotherapy times analyzed (p<0.05), indicating that these proteins can be considered as sensitive biomarkers for lymphoma in dogs. Furthermore, the α1-acid glycoprotein showed prognostic value for the disease, with 63% specificity. However, vitamin D concentration was not correlated with prognosis of the dogs with lymphoma.

Objetivou-se caracterizar a concentração sérica da vitamina D e das PFAs (Proteínas de Fase Aguda) (alfa-1 glicoproteína ácida, haptoglobina, transferrina, ceruloplasmina, albumina, IgA, IgG e alfa-1 ­ antitripsina) em cães com linfoma multicêntrico, submetidos ao tratamento quimioterápico com protocolo CHOP (Ciclofosfamida, Doxorrubicina, Vincristina e Prednisona), determinando o valor prognóstico desses marcadores para a doença. Foram avaliadas as concentrações séricas das PFAs, através do método da eletroforese e as concentrações da vitamina D, através da quimioluminescência em dois grupos experimentais, um grupo de 13 cães com linfoma multicêntrico classificados como alto grau pela citologia (GL) durante as semanas T0, T5 e T10 do tratamento com protocolo quimioterápico antineoplásico e em um grupo de 10 animais saudáveis para compor o grupo controle (GC), em coleta única. Para isso, foi realizado o diagnóstico, estadiamento e avaliação de resposta terapêutica dos 13 pacientes com linfoma multicêntrico através de técnicas de citopatologia, histopatologia, imuno-histoquímica do linfonodo periférico acometido. Foi observado que 9 pacientes tiveram resposta completa e 4 pacientes tiveram resposta parcial ao tratamento. Os dados foram analisados através do software R. Os resultados indicam que as diferenças entre as variáveis IgA, haptoglobina e α1-glicoproteína ácida foram significativas entre os grupos, e entre os diferentes momentos da quimioterapia (p< 0,05), indicando que podem ser marcadores sensíveis ao linfoma em cães. A α1-glicoproteína ácida apresentou valor prognóstico para o linfoma, com 63% de especificidade. Porém a vitamina D não apresentou valor prognóstico para o linfoma multicêntrico em cães.

Can intramammary infections change immunoglobulins and acute phase proteins of colostrum and transition milk in dairy goats? / A infecção intramamária pode alterar imunoglobulinas e proteínas de fase aguda do colostro e do leite de transição de cabras leiteiras?

Proteiongram analysis is useful for the early diagnosis of intramammary infections during the period of colostrogenesis. This study aimed to evaluate the profile of total proteins, immunoglobulins, lactoferrin, and gamma-glutamyl transferase (GGT) in the colostrum of dairy goats with intramammary infections. Animals were divided in groups: GI (n=12) of goats without mammary gland infections, and GII (n=8) of goats with mammary gland infections. Intramammary infections were diagnosed using microbiological isolations and somatic cell counts Cs). Total protein was evaluated in the samples using SDS-PAGE shortly after parturition, and 24 and 48 hours after that event. Non-aureus Staphylococcus (NAS) were detected all isolates. At 48 h, GII had high IgG levels and a SCC of 1660.25 x 103/mL. Levels of total protein were high in this group at 24 and 48 h. Albumin levels were high in goats with mastitis at 24 h. Overall, the IgG, lactoferrin, and albumin levels differed between animals with and without intramammary infections at M0. GGT activity was not influenced by the intramammary infection. The results of this study reinforce the importance of the proteinogram as an auxiliary tool in the diagnosis of mastitis in dairy goats.

O proteinograma do colostro/leite pode ser útil para diagnóstico precoce de infecções intramamárias, assim como para a avaliação da intensidade da resposta inflamatória. Este estudo teve como objetivo avaliar o perfil de proteínas, imunoglobulinas, lactoferrina e gamaglutamiltransferase no colostro de cabras leiteiras portadoras s de infecções intramamárias. Os animais foram distribuídos em dois grupos, GI (n 12), composto por cabras sem isolamento microbiológico das glândulas mamárias, e GII (n 8), composto por cabras com resultados positivos na cultura do leite de pelo menos uma das glândulas mamárias. O diagnóstico de infecção intramamária foi realizado logo após o parto, por meio de isolamento microbiológico e de contagem de células somáticas (CCS). Os níveis de proteína total, imunoglobulina A (IgA), imunoglobulina (G) (IgG), lactoferrina, albumina e atividade da gamaglutamiltransferase (GGT) foram avaliados em amostras de colostro/leite de transição, usando-se SDS-PAGE logo após o parto bem como às 24 e às 48 horas após esse evento. Staphylococcus não aureus (NAS) foram encontrados em todos os isolamentos. A concentração de IgG de 186 foi superior no GII apenas às 48h, ao mesmo tempo em que o CCS foi de 1660,25 x 10/mL. No entanto, a proteína total foi maior neste grupo às 24 e às 48h. A albumina foi maior nas cabras com mastite às 24h, e a lactoferrina no momento 0h (M0) e às 48h. Em geral, os valores de IgG, lactoferrina e albumina diferiram entre os animais com e sem infecções intramamárias no M0. A atividade da gamaglutamiltransferase não foi influenciada pela infecção intramamária. Os resultados deste estudo reforçam a importância do proteinograma como uma ferramenta auxiliar no diagnóstico da mastite em cabras leiteiras.

Effects of 48-hour feed deprivation on acute-phase response in horses / Efeitos da privação alimentar de 48 horas na resposta de fase aguda em cavalos

The objective of this study was to evaluate the effect of feed restriction on acute-phase response in horses. Twenty horses were deprived of food for 48 h and others 12 animals (control) had free access to water and hay. They were closely monitored and examined, and blood samples were taken at the beginning (0) of the study and 6, 12, 18, 24, 30, 36, 42 and 48 hours afterward. Data were submitted to two-way analysis of variance with repeated measures and statistical significance was P ≤ 0.05. The horses tolerated feed restriction without serious clinical complications. Feed restriction induced an increase in the acute-phase response by elevating serum concentrations of α2-macroglobulin (24-38 h), ceruloplasmin (36-48 h), α1-antitrypsin (30-48 h), α1-acid glycoprotein (42-48 h) and haptoglobin (42-48 h). Nutrient deprivation raised the levels of circulating cortisol, which acts on the innate immune system, which then induces the acute-phase response. In conclusion, food restriction is a physical stressor for horses, capable of inducing an acute-phase protein reaction, characterized by increased production of α2-macroglobulin, ceruloplasmin, α1- antitrypsin, α1-acid glycoprotein and haptoglobin.

O objetivo deste estudo foi avaliar o efeito da restrição alimentar na resposta de fase aguda em equinos. Vinte cavalos foram submetidos à restrição alimentar por 48 h enquanto outros 12 animais (controle) tiveram livre acesso à água e alimento. Os animais foram monitorados, examinados e amostras de sangue foram coletadas no início (0) do estudo e com 6, 12, 18, 24, 30, 36, 42 e 48 horas de restrição alimentar. Os dados foram submetidos à análise de variância bidirecional com medidas repetidas e a significância estatística foi P ≤ 0,05. Os cavalos toleraram a restrição alimentar sem complicações clínicas relevantes. A restrição alimentar induziu uma resposta de fase aguda caracterizada pela elevação das concentrações séricas de α2-macroglobulina (24-38 h), ceruloplasmina (36-48 h), α1-antitripsina (30-48 h), α1-glicoproteína ácida (42-48 h) e haptoglobina (42-48 h). A privação de nutrientes eleva os níveis de cortisol circulante, que atua no sistema imunológico inato o qual, então induz a resposta de fase aguda. Em conclusão, a restrição alimentar é um fator estressor físico para equinos, capaz de induzir uma reação proteica de fase aguda, caracterizada pelo aumento na produção de α2-macroglobulina, ceruloplasmina, α1-antitripsina, α1-glicoproteína ácida e haptoglobina.

Effects of flunixin meglumine, meloxicam, and firocoxib on the acute-phase proteins in horses following standing castration / Efeitos do flunixin meglumine, meloxicam e firocoxib na concentração de proteínas da fase aguda após em equinos após castração

Excessive infection and inflammation are the most common complications associated with castration. The objective of this study was to compare the efficacy of flunixin meglumine (FM), meloxicam (MX), or firocoxib (FX) for inflammation control after castration in horses using acute-phase proteins (APP) as markers of inflammation. Thirty healthy, unbroken, mixed-breed horses (body weight 358.62±45.57kg and age 4.99±2.63 years) were randomly (n=10 animals/group) allocated to receive one of three different post-castration anti-inflammatory medicines: Group 1 (FM 1.1mg/kg bwt, IV, s.i.d for 5 days); Group 2 (MX 0.6mg/kg bwt, IV, s.i.d for 5 days); and Group 3 (FX 0.1mg/kg bwt, IV, s.i.d for 5 days). All horses were castrated in standing position, using the open technique. Serum and peritoneal APP concentrations were measured by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and determined before castration (0), and 3, 5, 24, 48, 72, 120 and 168 hours after castration. The results were submitted to analysis of variance using the SAS statistical program, and means were compared by the Student-Newman-Keuls test (p<0.05). Three animals from the MX group developed hyperthermia (with rectal temperatures of 39.8, 39.3 and 38.9°C on day 4, 5 and 6, respectively) and showed local clinical signs of inflammation (inguinal and excessive scrotal edema) and reluctance to walk, as well as a rigid gait of the hind limbs. The same complications were observed in one FX horse. No complications were observed among the FM animals. The castration resulted in significant changes in serum and peritoneal values of total proteins, ceruloplasmin (Cp), transferrin (Tf), albumin (Alb), haptoglobin (Hp) and α1-acid glycoprotein (Gp) in animals of all experimental groups. However, the animals of the MX and FX groups presented more intense acute phase response compared to the animals of the FM group. Changes in the APP were associated with the surgical trauma of castration, but the differences between groups were associated with the ability of the nonsteroidal anti-inflammatory drug to control the inflammation. In conclusion, and based on the findings of acute phase proteins, flunixin is more efficient to control the magnitude of inflammation following castration as compared to meloxicam and firocoxib.(AU)

Infecção e inflamação excessivas são as complicações mais comuns associadas à castração. O objetivo deste estudo foi comparar a eficácia do flunixin meglumine (FM), meloxicam (MX) ou firocoxib (FX) no controle da inflamação após a castração em cavalos usando proteínas da fase aguda (APP) como marcadores de inflamação. Trinta equinos saudáveis (358,62±45,57kg; 4,99±2,63 anos) foram em função dos anti-inflamatórios utilizados após as castrações aleatoriamente (n= 10 animais/grupo) alocados em três diferentes grupos: Grupo 1 (FM 1,1mg/kg de peso, IV, sid por 5 dias); Grupo 2 (MX 0,6mg/kg de peso, IV, s.i.d por 5 dias); e Grupo 3 (FX 0,1mg/kg de peso, IV, s.i.d por 5 dias). Todos os cavalos foram castrados em posição quadrupedal, utilizando a técnica aberta. As concentrações de APP sérica e peritoneal foram separadas por eletroforese em gel de poliacrilamida (PAGE) com dodecil-sulfato de sódio (SDS) e determinadas no momento 0 (antes da castração) e com 3, 5, 24, 48, 72, 120 e 168 horas após a castração. Os resultados foram submetidos à análise de variância pelo programa estatístico SAS e as médias foram comparadas pelo teste de Student-Newman-Keuls (p<0,05). Três animais do grupo MX desenvolveram hipertermia (com temperatura retal de 39,8, 39,3 e 38,9° C nos dias 4, 5 e 6, respectivamente) e mostraram sinais clínicos locais de inflamação (edema inguinal e escrotal excessivo) e relutância em andar, bem como marcha rígida dos membros posteriores. As mesmas complicações foram observadas em um cavalo do FX. Não foram observadas complicações entre os animais do FM. Independente do grupo, a castração resultou em alterações significativas nos valores séricos e peritoneais de proteínas totais, ceruloplasmina (Cp), transferrina (Tf), albumina (Alb), haptoglobina (Hp) e glicoproteína ácida α1 (Gp). No entanto, os animais dos grupos MX e FX apresentaram resposta de fase aguda mais intensa quando comparados aos animais do FM. Alterações na resposta de fase aguda deveram-se ao trauma cirúrgico da castração, mas as diferenças entre os grupos foram associadas à capacidade do anti-inflamatório em controlar a inflamação. Em conclusão, baseado da resposta de fase aguda, o flunixin em comparação com o meloxicam e o firocoxib é mais eficiente no controle da inflamação após a castração em equinos.(AU)

Effects of flunixin meglumine, meloxicam, and firocoxib on the acute-phase proteins in horses following standing castration / Efeitos do flunixin meglumine, meloxicam e firocoxib na concentração de proteínas da fase aguda após em equinos após castração

Excessive infection and inflammation are the most common complications associated with castration. The objective of this study was to compare the efficacy of flunixin meglumine (FM), meloxicam (MX), or firocoxib (FX) for inflammation control after castration in horses using acute-phase proteins (APP) as markers of inflammation. Thirty healthy, unbroken, mixed-breed horses (body weight 358.62±45.57kg and age 4.99±2.63 years) were randomly (n=10 animals/group) allocated to receive one of three different post-castration anti-inflammatory medicines: Group 1 (FM 1.1mg/kg bwt, IV, s.i.d for 5 days); Group 2 (MX 0.6mg/kg bwt, IV, s.i.d for 5 days); and Group 3 (FX 0.1mg/kg bwt, IV, s.i.d for 5 days). All horses were castrated in standing position, using the open technique. Serum and peritoneal APP concentrations were measured by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and determined before castration (0), and 3, 5, 24, 48, 72, 120 and 168 hours after castration. The results were submitted to analysis of variance using the SAS statistical program, and means were compared by the Student-Newman-Keuls test (p<0.05). Three animals from the MX group developed hyperthermia (with rectal temperatures of 39.8, 39.3 and 38.9°C on day 4, 5 and 6, respectively) and showed local clinical signs of inflammation (inguinal and excessive scrotal edema) and reluctance to walk, as well as a rigid gait of the hind limbs. The same complications were observed in one FX horse. No complications were observed among the FM animals. The castration resulted in significant changes in serum and peritoneal values of total proteins, ceruloplasmin (Cp), transferrin (Tf), albumin (Alb), haptoglobin (Hp) and α1-acid glycoprotein (Gp) in animals of all experimental groups. However, the animals of the MX and FX groups presented more intense acute phase response compared to the animals of the FM group. Changes in the APP were associated with the surgical trauma of castration, but the differences between groups were associated with the ability of the nonsteroidal anti-inflammatory drug to control the inflammation. In conclusion, and based on the findings of acute phase proteins, flunixin is more efficient to control the magnitude of inflammation following castration as compared to meloxicam and firocoxib.(AU)

Infecção e inflamação excessivas são as complicações mais comuns associadas à castração. O objetivo deste estudo foi comparar a eficácia do flunixin meglumine (FM), meloxicam (MX) ou firocoxib (FX) no controle da inflamação após a castração em cavalos usando proteínas da fase aguda (APP) como marcadores de inflamação. Trinta equinos saudáveis (358,62±45,57kg; 4,99±2,63 anos) foram em função dos anti-inflamatórios utilizados após as castrações aleatoriamente (n= 10 animais/grupo) alocados em três diferentes grupos: Grupo 1 (FM 1,1mg/kg de peso, IV, sid por 5 dias); Grupo 2 (MX 0,6mg/kg de peso, IV, s.i.d por 5 dias); e Grupo 3 (FX 0,1mg/kg de peso, IV, s.i.d por 5 dias). Todos os cavalos foram castrados em posição quadrupedal, utilizando a técnica aberta. As concentrações de APP sérica e peritoneal foram separadas por eletroforese em gel de poliacrilamida (PAGE) com dodecil-sulfato de sódio (SDS) e determinadas no momento 0 (antes da castração) e com 3, 5, 24, 48, 72, 120 e 168 horas após a castração. Os resultados foram submetidos à análise de variância pelo programa estatístico SAS e as médias foram comparadas pelo teste de Student-Newman-Keuls (p<0,05). Três animais do grupo MX desenvolveram hipertermia (com temperatura retal de 39,8, 39,3 e 38,9° C nos dias 4, 5 e 6, respectivamente) e mostraram sinais clínicos locais de inflamação (edema inguinal e escrotal excessivo) e relutância em andar, bem como marcha rígida dos membros posteriores. As mesmas complicações foram observadas em um cavalo do FX. Não foram observadas complicações entre os animais do FM. Independente do grupo, a castração resultou em alterações significativas nos valores séricos e peritoneais de proteínas totais, ceruloplasmina (Cp), transferrina (Tf), albumina (Alb), haptoglobina (Hp) e glicoproteína ácida α1 (Gp). No entanto, os animais dos grupos MX e FX apresentaram resposta de fase aguda mais intensa quando comparados aos animais do FM. Alterações na resposta de fase aguda deveram-se ao trauma cirúrgico da castração, mas as diferenças entre os grupos foram associadas à capacidade do anti-inflamatório em controlar a inflamação. Em conclusão, baseado da resposta de fase aguda, o flunixin em comparação com o meloxicam e o firocoxib é mais eficiente no controle da inflamação após a castração em equinos.(AU)

Haptoglobin and its association with naturally occurring diseases in Holstein heifer calves / [Haptoglobina e sua associação com doenças ocorridas naturalmente em bezerras da raça Holandesa]

The present study evaluated the use of haptoglobin (Hp) as an indicator of health and performance in 166 Holstein heifer calves reared in an intensive production system. Calves were evaluated at D6-9; D10-13; D20-23; D35-38 and D65-68, corresponding to the days of life. The absence or presence of diseases was evaluated by physical examination and classification of scores. The performance parameters evaluated were body weight, height at withers and hind width. Hp was measured by spectrophotometric technique. The highest prevalence of diarrhea (59.4%; 98/165) was observed in D10-13, bovine respiratory disease (BRD) was on D35-38 (25.8%; 42/163), and umbilical inflammations in D6-D9 (7.8%; 13/166). Highest values of Hp were observed in animals with diarrhea (P=0.02), and umbilical inflammation (P=0.057), in comparison with the group of healthy calves. A significant negative correlation was observed between Hp and performance index. This protein presented an important relation with diarrhea and performance of the calves, opening perspectives on its utilization as a biomarker of diseases.(AU)

O presente estudo avaliou o uso da haptoglobina (Hp) como indicadora de sanidade e desempenho em 166 bezerras Holandesas criadas em um sistema de produção intensivo. As bezerras foram avaliadas nos momentos D6-9; D10-13; D20-23; D35-38 e D65-68, sendo estes correspondentes aos dias de vida. A ausência ou a presença de doenças foi avaliada por meio do exame físico e da classificação por escores. Os parâmetros de desempenho avaliados foram peso corporal, altura de cernelha e largura de garupa. A Hp foi mensurada por técnica espectrofotométrica. A maior prevalência de diarreia (59,4%; 98/165) foi observada em D10-13, doença respiratória bovina (DRB) ocorreu em D35-38 (25,8%; 42/163) e inflamações umbilicais em D6-D9 (7,8%; 13/166). O valor de Hp foi maior nos animais que apresentaram diarreia (P=0,02) e inflamações umbilicais (P=0,057), em comparação ao grupo de bezerras saudáveis. Houve correlação negativa significativa entre a Hp e os índices de desempenho. Essa proteína apresentou uma importante relação com a diarreia e com o desempenho das bezerras, abrindo perspectivas sobre a sua utilização como biomarcadora de doenças.(AU)

Correlation of serum acute-phase proteins with bone repair in rabbits (Oryctolagus cuniculus) submitted to partial radio ostectomy / Correlação do proteinograma sérico com a reparação óssea em coelhos (Oryctolagus cuniculus) submetidos à ostectomia parcial do rádio

Bone trauma triggers an acute inflammatory response, which can alter the serum concentration of acute-phase proteins (PFA). We aimed to evaluate the effectiveness of PFA measurement as a laboratory biomarker for inflammation related to bone regeneration. A partial ostectomy of the right radius was performed on 22 young male rabbits. The complete blood count, albumin, ceruloplasmin, haptoglobin, C-reactive protein, and transferrin levels were recorded and correlated with the radiographic evolution of bone healing. Statistically the best correlations with the inflammatory process and radiological findings were reported with haptoglobin and transferrin compared to the other measured PFAs. Haptoglobin and transferrin showed a maximum peak 24 and 36 hours after surgery, respectively. Transferrin displayed signs of decrease in the first 6 hours after surgery, in advance of the leukocyte response. The measurement of acute-phase proteins proved to be viable, considering the conditions in which the present study was carried out. Among all variables studied, haptoglobin and transferrin showed best correlation with the inflammatory process of bone healing. Additional studies are needed to determine the sensitivity of acute-phase proteins as predictors of complications in the treatment of fractures.(AU)

Os traumas ósseos desencadeiam resposta inflamatória aguda e consequentemente alteram a concentração sérica das proteínas de fase aguda (PFA), podendo essas representarem um parâmetro para avaliação da evolução do processo inflamatório relacionado à cicatrização óssea, assim como complicações. Objetivou-se avaliar a inter-relação da mensuração de PFAs com a repação óssea em coelhos submetidos à ostectomia parcial do rádio. Foram utilizados 22 coelhos, machos, jovens (160 a 180 dias) e peso médio (3,8 ± 0,3). Para acompanhamento da evolução do processo inflamatório foram realizados hemograma completo e mensuração de albumina, ceruloplasmina, haptoglobina, proteína C reativa e transferrina, além do acompanhamento radiológico. Foi observado a melhor correlação da haptoglobina e da transferrina junto ao processo inflamatório e achados radiológicos frente às demais PFAs mensuradas. A haptoglobina apresentou pico máximo 24 horas do pós-operatório e a transferrina após 36 horas, entretanto, essa última já mostrou indícios de diminuição nas primeiras 6 horas do pós-cirúrgico, antecipadamente à resposta leucocitária. Nas condições em que o presente estudo foi realizado, a mensuração das proteínas de fase aguda mostrou-se factível. Das proteínas estudadas, a haptoglobina e transferrina foram as que mostraram maior correlação com o processo inflamatório da cicatrização óssea. Estudos adicionais são necessários para determinar a sensibilidade das proteínas de fase aguda como previsores de complicações do tratamento das fraturas.(AU)

Leukocytes and albumin in canine leishmaniasis

Canine Leishmaniasis (CanL) is a multisystemic and chronic inflammatory disease characterized by nonspecific clinical manifestations. In CanL, inflammatory cells and chemical mediators released in response to the parasite play a role in disease development and progression. Alterations on hematological parameters have been documented in CanL. These changes can also be assessed in relation to systemic inflammation caused by this disease. The circulating leukocyte counting, such as neutrophils, as well as the albumin level, are considered direct indicators of an inflammatory host environment. Several studies point to the use of biomarkers on the assistance in diagnosis and prognosis of several canine pathologies. The present study investigated the Neutrophils to Lymphocyte Ratio (NLR), Albumin to Globulin Ratio (AGR), and Neutrophils to Albumin Ratio (NAR) on systemic inflammatory response induced by Canine Leishmaniasis (CanL). For this purpose, adult dogs with confirmed diagnosis to CanL were divided into symptomatic (SD, n = 33) and asymptomatic (AD, n = 20) dogs for L. infantum and control dogs (CD, n = 20). Routine hematological and biochemical parameters were determined in blood samples using a veterinary automatic hematology and biochemical analyzers. Asymptomatic dogs (AD) had a higher number of white blood cells and neutrophils (16.48 ± 4.93; 13.41 ± 3.60, respectively) in relation to symptomatic dogs (SD) (13.54 ± 5.13; 10.42 ± 3.69, respectively) (P = 0.015 and P < 0.0001, respectively). Neutrophils to Lymphocyte Ratio (NLR) was higher in dogs with leishmaniasis (9.45 ± 3.76) than in healthy dogs (3.39 ± 1.19) (P < 0.0001). Serum total proteins (STP) and globulins increased in CanL, while albumin and AGR decreased in CanL, when compared to CD and references values to canine species. Neutrophils to Albumin Ratio (NAR) was higher in AD and SD (5.02 ± 1.14; 4.79 ± 1.07, respectively) when compared to CD (2.36 ± 0.55) (P < 0.0001). As reported in scientific researches, dogs with Leishmaniasis present alterations in circulating cell counts. Based on these data, we decided to expand this information using the NLR as a parameter in an attempt to better clarify the changes in these cells in CanL. We observed that NLR was increased on CanL in relation to healthy dogs, which could be a consequence of relative neutrophilia rather than lymphopenia. Neutrophils to Lymphocyte Ratio (NLR) is a biomarker that conveys information about inflammatory conditions. An elevated NLR can reflect an upregulated innate immune response, since neutrophils are effector cells of innate immunity and are involved in several acute and chronic inflammatory processes. Albumin is an acute phase protein that is considered an immune-inflammatory biomarker, which can be found reduced systemically in progressive inflammatory response. Serum total proteins (STP) and globulins were increased in CanL. These data are already well documented in CanL, which serum globulins are mainly associated with the increase of acute phase proteins, cytokines, and increase of specific antibodies to Leishmania infantum. Our results showed neutrophilia with hypoalbuminemia in CanL. So, in an attempt to assess the relationship of these two available markers, we used NAR calculation in order to evaluate the changes induced by CanL. In this study NAR was higher in CanL when compared to control dogs. Thus, our data indicate that NLR and NAR could be used as biomarkers in veterinary medical clinics in order to assess inflammatory profile in CanL, mainly in asymptomatic dogs. These parameters obtained from routine blood tests might be useful as cost-effective, easily accessible and helpful markers in order to distinguish the inflammatory response intensity in CanL.(AU)

Perfil clínico e laboratorial de cadelas sororeativas para erliquiose tratadas em um Hospital Veterinário Universitário em Niterói, Estado do Rio de Janeiro, Brasil / Clinical and laboratory profile of dogs seroreactive to ehrlichiosis treated at the Veterinary Medical Teaching Hospital in Niterói, State of Rio de Janeiro, Brazil

Ehrlichiosis is a tick-borne disease highly prevalent in Brazil, and is relevant in canine clinical practice due to its high morbidity and mortality. Its clinical signs are nonspecific and its phases are acute, lasting 2 to 4 weeks; subclinical, i.e., asymptomatic; and chronic, resembling an autoimmune disease. The purpose of this study was to identify the occurrence of reactivity to Ehrlichia canis of bitches treated at the Veterinary Medical Teaching Hospital of the Universidade Federal Fluminense (UFF) - Niterói, RJ, Brazil, based on serological examination by iELISA, and to compare the hematological, biochemical, urinary protein-creatinine and urinary density profiles of reactive and non-reactive animals. This study involved solely bitches, regardless of breed, starting at 1 year of age. One hundred and thirty bitches, 1 to 16 year-old (mean age 7.02 ± 4.00), weighing 1.5 to 50 kg (mean weight 12.12 ± 10.65) were subjected to clinical examination and abdominal ultrasound. Complete blood count, biochemical measurements, urinalysis and serology for E. canis were also performed. The serum was used in the iELISA to identify immunoglobulin G (IgG), using a canine Ehrlichia Imunotest® diagnostic kit (Imunodot®, Jaboticabal, SP, Brazil) according to the manufacturer's instructions. Sixty animals (46.20%) were reactive to E. canis. According to their owners, only 5 (8.3%) of the 60 seroreactive animals had a history of tick-borne disease. The most common profile was that of mixed breed animals living with their owners, older than 7 years, who had not been treated preventatively with specific drugs against ectoparasites. Laboratory tests showed significant differences between groups in terms of total protein (TP), and calcium and urinary protein-creatinine ratio (UPC). TP and UPC were elevated in the non-reactive group, while the only significant change in the reactive group was mild hypocalcemia. In this study, 30% (18/60) of the bitches were seroreactive to E. canis and had hypocalcemia. Of these, 50% (9/18) had a UPC above 0.5. Furthermore, 66.7% (12/18) of this group with hypocalcemia also showed urine density (UD) of less than 1024. Among these 18 bitches, 5 had both alterations, i.e., UPC > 0.5 and UD < 1024. In this study, a high prevalence of bitches seroreactive to Ehrlichia canis was observed, despite the absence of clinical and/or laboratory signs indicative of the disease. In the investigation of IgG class antibodies, it is not possible to determine the exact time of infection, and titers may remain high for a period of more than 11 months, even after treatment and elimination of the bacterium. The fact that most seroreactive bitches showed no symptoms compatible with the disease either before or during the study suggests that they were in the subclinical phase of ehrlichiosis. The main reason for calcium metabolism disorders is a phosphorus imbalance, a condition that occurs in kidney diseases. Isosthenuria reflects the kidney's inability to concentrate urine. This finding may be one of the first clinical manifestations of chronic kidney disease (CKD), especially in dogs. On the other hand, the UPC ratio may increase with the progression of CKD. The presence of hypocalcemia, isosthenuria and increased UPC associated with seroreactivity suggests that infection by E. canis may be associated with the onset of CKD. Veterinarians should keep in mind the complexity of the pathophysiology of ehrlichiosis to ensure the disease is not underdiagnosed in any of its phases, thereby ensuring the correct treatment is provided. Such awareness is expected to reduce the chronicity of the disease and underlying sequelae among dogs.(AU)

Hematologic response and serum and peritoneal fluid proteinogram of cattle affected by intestinal diseases and traumatic reticuloperitonitis / Resposta hematológica e proteinograma sérico e do líquido peritoneal de bovinos acometidos com enfermidades intestinais e reticuloperitonite traumática

This study aimed to evaluate the hematologic response and the serum and peritoneal fluid (PF) proteinogram of cattle affected by digestive diseases. Twenty-seven animals were distributed in two groups: GI (intestinal diseases) and GII (traumatic reticuloperitonitis, TRP). The animals were previously submitted to a physical exam. Subsequently, blood samples were collected to perform the complete blood count, determine the plasma protein and fibrinogen, and obtain the serum for proteinogram in polyacrylamide gel (SDS-PAGE). Simultaneously, PF was collected to perform physical and chemical evaluation and the electrophoretic profile (SDS-PAGE). ANOVA at the 5% probability level was used to compare the groups. The animals showed signs of apathy, dehydration, and gastrointestinal hypomotility in both groups. However, GI animals showed more significant clinical changes. The blood count of both groups (P > 0.05) showed leukocytosis due to neutrophilia and a regenerative left shift with hyperfibrinogenemia. The proteinogram of both body fluids allowed the identification of proteins albumin (ALB), transferrin (TRF), ceruloplasmin, haptoglobin, α1-acid glycoprotein (α1-AGP), MW 23000 Da, α1-antitrypsin, IgA, and IgG. The [PT] PF/[PT] blood serum ratio of each of the identified proteins increased, showing statistical differences between groups (P < 0.05) regarding PT, ALB, TRF, α1-AGP, and IgG values, with GI animals showing the highest ratio. Intestinal diseases and TRP triggered a systemic and local response characterized by clinical, hematological, and serum and PF proteinogram alterations. The proteins α1-GPA, haptoglobin, and TRF measured in PF were good inflammation biomarkers and useful as an auxiliary tool for the diagnosis and prognosis of digestive diseases in cattle.(AU)

O objetivo deste estudo foi avaliar a resposta hematológica e o proteinograma sérico e do líquido peritoneal (LP) de bovinos acometidos com doenças digestivas. Foram avaliados 27 bovinos distribuídos em dois grupos, GI (enfermidades intestinais) e GII (reticuloperitonite traumática-RPT). Os animais foram previamente submetidos ao exame físico. Posteriormente foram colhidas amostras de sangue para realização do hemograma, determinação plasmática da proteína e do fibrinogênio e obtenção do soro para realização do proteinograma em gel de poliacrilamida (SDS-PAGE). Simultaneamente foi colhido o LP para avaliação física e química, assim como a realização do perfil eletroforético (SDS-PAGE). Empregou-se a análise de variância ao nível de 5% de probabilidade visando à comparação entre os grupos. Em ambos os grupos os animais demonstraram sinais de apatia, desidratação e hipomotilidade gastrointestinal, no entanto, os animais do GI apresentaram alterações clínicas mais expressivas. No hemograma observou-se em ambos os grupos (P > 0,05) leucocitose por neutrofilia e desvio à esquerda regenerativo com hiperfibrinogenia. O proteinograma de ambos os fluidos corpóreos permitiu a identificação das proteínas albumina (ALB), transferrina (TRF), ceruloplasmina, haptoglobina, α-1 glicoproteína ácida (α1-GPA), PM 23.000 Da, α-1 anti-tripsina, IgA e IgG. Os valores da relação [PT] LP / [PT] soro sanguíneo de cada uma das proteínas identificadas demonstrou elevação dos mesmos, bem como diferença estatística entre grupos (P < 0,05) nos valores da PT, ALB, TRF, α1-GPA e IgG, nos quais a relação foi mais elevada nos animais do GI. As enfermidades intestinais e a RPT desencadearam resposta sistêmica e local caracterizada pelas alterações clínicas, hematológicas, e do proteinograma sérico e do LP. A α1-GPA, a haptoglobina e a TRF, mensuradas no LP se mostraram bons biomarcadores de inflamação, sendo úteis como recurso auxiliar de diagnóstico e prognóstico das doenças digestivas dos bovinos.(AU)

Acute phase protein response and clinical application in Veterinary Medicine / Resposta das proteínas de fase aguda e sua aplicação clínica em Medicina Veterinária

Acute phase proteins (APP) are the group of blood proteins that the synthesis happens in a quick and intense manner in response to external or internal challenges, such as infection, inflammation, surgical trauma or stress. These proteins have been used as biomarkers of inflammation, infection and trauma in human medicine, but have been relatively under-utilised in the context of veterinary medicine due to the unknown diagnostic possibilities. The purpose of this paper is to update the knowledge about APPs in veterinary medicine, reviewing results from different studies suggesting possible clinical applicability of APP measurement as a valuable tool for veterinary diagnostic and prognostic.

As proteínas da fase aguda (PFA) são um grupo de proteínas presentes no sangue, as quais a síntese ocorre de forma rápida e intensamente em resposta a desafios externos ou internos, como infecção, inflamação, trauma cirúrgico ou estresse. Essas proteínas têm sido usadas como biomarcadores de inflamação, infecção e trauma na medicina humana, mas tem sido relativamente subutilizadas no contexto da medicina veterinária devido às possibilidades de diagnóstico desconhecidas. O objetivo deste artigo é atualizar o conhecimento sobre PFAs em medicina veterinária, revisando resultados de diferentes estudos que sugerem uma possível aplicabilidade clínica da mensuração de PFAs como uma valiosa ferramenta para o diagnóstico e prognóstico veterinário.

Acute phase protein response and clinical application in Veterinary Medicine / Resposta das proteínas de fase aguda e sua aplicação clínica em Medicina Veterinária

Acute phase proteins (APP) are the group of blood proteins that the synthesis happens in a quick and intense manner in response to external or internal challenges, such as infection, inflammation, surgical trauma or stress. These proteins have been used as biomarkers of inflammation, infection and trauma in human medicine, but have been relatively under-utilised in the context of veterinary medicine due to the unknown diagnostic possibilities. The purpose of this paper is to update the knowledge about APPs in veterinary medicine, reviewing results from different studies suggesting possible clinical applicability of APP measurement as a valuable tool for veterinary diagnostic and prognostic.(AU)

As proteínas da fase aguda (PFA) são um grupo de proteínas presentes no sangue, as quais a síntese ocorre de forma rápida e intensamente em resposta a desafios externos ou internos, como infecção, inflamação, trauma cirúrgico ou estresse. Essas proteínas têm sido usadas como biomarcadores de inflamação, infecção e trauma na medicina humana, mas tem sido relativamente subutilizadas no contexto da medicina veterinária devido às possibilidades de diagnóstico desconhecidas. O objetivo deste artigo é atualizar o conhecimento sobre PFAs em medicina veterinária, revisando resultados de diferentes estudos que sugerem uma possível aplicabilidade clínica da mensuração de PFAs como uma valiosa ferramenta para o diagnóstico e prognóstico veterinário.(AU)

Acute-phase protein concentration in bronchoalveolar lavage fluid from healthy horses / Proteínas de fase aguda do lavado broncoalveolar de equinos hígidos

Bronchoalveolar lavage fluid (BALF) was analyzed to obtain information on leakage of acute-phase proteins from the blood into the respiratory lumen and about local synthesis. Ceruloplasmin, transferrin, albumin, α1-antitripsin, immunoglobulin G heavy, immunoglobulin G light, immunoglobulin A, haptoglobin, acidic glycoprotein, and P23 were measured in BALF from 30 horses without inflammatory disease by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In serum, the same proteins were identified except for α1-antitrypsin. In conclusion, this study demonstrated that polyacrylamide gel electrophoresis (SDS-PAGE) can be used for the determination of acute-phase proteins in BALF samples from horses. In healthy horses, the values are very low, but they can be compared with reference values to assist in the diagnosis of animals with respiratory diseases.(AU)

O líquido obtido através da lavagem broncoalveolar (LBA) foi analisado para obter informações sobre as proteínas da fase aguda. Ceruloplasmina, transferrina, albumina, α1-antitripsina, imunoglobulina G pesada, imunoglobulina G leve, imunoglobulina A, haptoglobina, glicoproteína ácida e P23 foram medidas nos LBA de 30 cavalos sem doença inflamatória por eletroforese em gel de poliacrilamida com dodecilsulfato de sódio (SDS-PAGE). No soro, as mesmas proteínas foram identificadas, exceto a α1-antitripsina. Em conclusão, este estudo demonstra que a eletroforese em gel de poliacrilamida (SDS-PAGE) pode ser usada para a determinação de proteínas de fase aguda em amostras de LBA em cavalos. Em cavalos saudáveis, os valores são muito baixos, no entanto, podem ser comparados e auxiliar no diagnóstico de animais com doenças respiratórias.(AU)

Acute-phase protein concentration in bronchoalveolar lavage fluid from healthy horses / Proteínas de fase aguda do lavado broncoalveolar de equinos hígidos

Bronchoalveolar lavage fluid (BALF) was analyzed to obtain information on leakage of acute-phase proteins from the blood into the respiratory lumen and about local synthesis. Ceruloplasmin, transferrin, albumin, α1-antitripsin, immunoglobulin G heavy, immunoglobulin G light, immunoglobulin A, haptoglobin, acidic glycoprotein, and P23 were measured in BALF from 30 horses without inflammatory disease by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In serum, the same proteins were identified except for α1-antitrypsin. In conclusion, this study demonstrated that polyacrylamide gel electrophoresis (SDS-PAGE) can be used for the determination of acute-phase proteins in BALF samples from horses. In healthy horses, the values are very low, but they can be compared with reference values to assist in the diagnosis of animals with respiratory diseases.(AU)

O líquido obtido através da lavagem broncoalveolar (LBA) foi analisado para obter informações sobre as proteínas da fase aguda. Ceruloplasmina, transferrina, albumina, α1-antitripsina, imunoglobulina G pesada, imunoglobulina G leve, imunoglobulina A, haptoglobina, glicoproteína ácida e P23 foram medidas nos LBA de 30 cavalos sem doença inflamatória por eletroforese em gel de poliacrilamida com dodecilsulfato de sódio (SDS-PAGE). No soro, as mesmas proteínas foram identificadas, exceto a α1-antitripsina. Em conclusão, este estudo demonstra que a eletroforese em gel de poliacrilamida (SDS-PAGE) pode ser usada para a determinação de proteínas de fase aguda em amostras de LBA em cavalos. Em cavalos saudáveis, os valores são muito baixos, no entanto, podem ser comparados e auxiliar no diagnóstico de animais com doenças respiratórias.(AU)

Etiology, multidrug resistance, and acute-phase proteins biomarkers as in equine septic arthritis / Etiologia, resistência antimicrobiana e proteínas de fase aguda como biomarcadores na artrite séptica equina

Etiology, multiple antibiotic resistance index, and acute-phase protein profile in the serum and synovial fluid of 44 horses presenting septic arthritis were investigated. Young animals up to 24 months of age not yet submitted to taming were sampled. Synovial samples from the horses were submitted to culture and bacterial isolates subjected to 18 antimicrobials to investigate the in vitro multidrug resistance pattern. Also, hematological aspects, fibrinogen, and serum levels of some acute-phase proteins (total protein, albumin, globulin, amyloid A, and C-reactive protein) were assessed. To statistical analysis, microbiological isolation were divided into four groups: G1 = animals with negative isolation, G2 = isolation of Gram-negative bacteria, G3 = isolation of Gram-positive bacteria), and G4 = isolation of fungi. Microbial isolation was obtained from 77.2% (32/44) of the synovial samples. Escherichia coli (7/44 = 15.9%), Staphylococcus aureus (5/44 = 11.4%), Streptococcus equi subsp. equi (3/44 = 6.8%), Aspergillus niger (3/44 = 6.8%), Pseudomonas aeruginosa (2/44 = 4.5%), Streptococcus equi subsp. zooepidemicus (1/44 = 2.3%), Trueperella pyogenes (1/44 = 2.3%), and Rhodococcus equi (1/44 = 2.3%) were prevalent bacterial agents. Aspergillus niger is reported by the first time as a primary agent of septic arthritis in horses. Gentamicin (84.9%), marbofloxacin (79.3%) and ceftriaxone (72.4%) were the most effective antimicrobials, whereas high resistance of the isolates (>50%) was found to erythromycin (75.9%), clarithromycin (75.9%), penicillin (69%), clindamycin (58.6%), and streptomycin (55.2%). Multiple antibiotic resistance index ( 0.3) was identified in 50% (16/32) of the isolates, with highest resistance in Gram-negative bacterial isolates. Serum levels of amyloid A, C-reactive protein, fibrinogen, globulin, and total protein, as well as the number of nucleated cells in the synovial fluid had values above the reference for horses, reinforcing acute-phase proteins as biomarkers in diagnosis of septic arthritis. We highlighted the diversity of microorganisms that may be involved in equine septic arthritis, the high resistance of bacterial isolates to conventional antimicrobials, the high lethality of young horses with septic arthritis, and importance of in vitro antimicrobial susceptibility pattern tests to therapeutic approaches in the disease.(AU)

Foram investigados a etiologia, o índice de resistência múltipla aos antibióticos e o perfil de proteínas de fase aguda, no soro ou líquido sinovial, de 44 equinos com sinais clínicos compatíveis com artrite séptica. Foram amostrados animais até 24 meses de idade não submetidos a doma. As amostras de líquido sinovial dos 44 animais foram submetidas à cultura bacteriológica e fúngica, e os isolados bacterianos submetidos in vitro diante de 18 antimicrobianos visando investigar a multirresistência. Também foram avaliados os aspectos hematológicos e o fibrinogênio sérico, bem como os níveis séricos de certas proteínas de fase aguda (proteína total, albumina, globulina, amilóide sérico A e proteína C reativa). O isolamento microbiano das amostras foi dividido em quatro grupos: G1 = animais com isolamento negativo; G2 = isolamento de bactérias Gram-negativas; G3 = isolamento de bactérias Gram-positivas e G4 = isolamento fúngico. O isolamento microbiano foi obtido em 77,2% (32/44) das amostras de líquido sinovial. Escherichia coli (7/44 = 15,9%), Staphylococcus aureus (5/44 = 11,4%), Streptococcus equi subsp. equi (3/44 = 6,8%), Aspergillus niger (3/44 = 6,8%), Pseudomonas aeruginosa (2/44 = 4,5%), Streptococcus equi subsp. zooepidemicus (1/44 = 2,3%), Trueperella pyogenes (1/44=2,3%) e Rhodococcus equi (1/44 = 2,3%) foram os patógenos mais frequentes. Aspergillus niger é relatado pela primeira vez como agente primário de artrite séptica em cavalos. Gentamicina (84,9%), marbofloxacino (79,3%) e ceftriaxona (72,4%) foram os antimicrobianos mais eficazes, enquanto elevada resistência dos isolados (>50%) foi encontrada para eritromicina (75,9%), claritromicina (75,9%), penicilina (69%), clindamicina (58,6%) e estreptomicina (55,2%). O índice de resistência múltipla aos antibióticos ( 0,3) foi identificado em 50% (16/32) dos isolados, com maior resistência obervada em bactérias Gram-negativas. Os níveis séricos de amilóide A, proteína C reativa, fibrinogênio, globulina e concentração total de proteínas, bem como o número de células nucleadas no líquido sinovial apresentaram valores acima da referência para equinos saudáveis, reforçando o uso destas proteínas de fase aguda como biomarcadores no diagnóstico da doença. O presente estudo enfatiza a diversidade de microrganismos que podem estar envolvidos na artrite séptica equina, a alta resistência dos isolados aos antimicrobianos convencionais, a alta mortalidade de equinos jovens com artrite séptica e a importância de realizar o tratamento da afecção com respaldo em testes de sensibilidade microbiana in vitro.(AU)

ELISA of amyloid A in paired bronchoalveolar lavage fluid and serum samples of healthy horses / Detecção de amilóide A no lavado broncoalveolar de equinos hígidos

Pulmonary disorders are common in horses, and treatment efficiency depends on an adequate diagnosis. Amyloid A is the most sensitive indicator of pathology in horses. The objective of this study was to establish the concentration of amyloid A of bronchoalveolar lavage fluid (BALF) in healthy horses. Health condition of horses was considered normal based on physical examination, complete blood count, biochemical parameters, and BALF cytology. Blood and BALF were collected from thirty adult female horses. Amyloid A concentrations in serum and BALF were measured using commercial ELISA tests. Amyloid A was detected in serum (mean ± SD = 3.71±2.51) and BALF (mean ± SD = 0.000745±0.000785) of all horses. In conclusion, SAA can also be measured in bronchoalveolar fluid, affording early detection of respiratory infections or inflammatory conditions.(AU)

Distúrbios pulmonares são comuns nos cavalos e a eficiência do tratamento depende de um diagnóstico adequado e precoce. A amilóide A é um biomarcador sensível na deteccção de patologias inflamatórias e infecciososa em cavalos. O objetivo deste estudo foi estabelecer a concentração de amilóide A no líquido broncoalveolar (LBA) em cavalos saudáveis. Os cavalos foram considerados saudaveis baseado nos achados de normalidade do exame físico, hemograma, parâmetros bioquímicos e citologia do LBA. Sangue e LBA foram coletados de 30 fêmeas equinas adultas. Os níveis de Amilóide A no soro e no LBA foram mensurados por meio do teste de ELISA. A amilóide A foi detectada no soro (média ± DP = 3,71±2,51) e no LBA (média ± DP = 0,000745±0,000785) de todos os animais. Conclui-se que a amilóide A também pode ser mensurada no LBA, auxiliando no diagnóstico precoce de processos inflamatórios e infecciosos pulmonares.(AU)

ELISA of amyloid A in paired bronchoalveolar lavage fluid and serum samples of healthy horses / Detecção de amilóide A no lavado broncoalveolar de equinos hígidos

Pulmonary disorders are common in horses, and treatment efficiency depends on an adequate diagnosis. Amyloid A is the most sensitive indicator of pathology in horses. The objective of this study was to establish the concentration of amyloid A of bronchoalveolar lavage fluid (BALF) in healthy horses. Health condition of horses was considered normal based on physical examination, complete blood count, biochemical parameters, and BALF cytology. Blood and BALF were collected from thirty adult female horses. Amyloid A concentrations in serum and BALF were measured using commercial ELISA tests. Amyloid A was detected in serum (mean ± SD = 3.71±2.51) and BALF (mean ± SD = 0.000745±0.000785) of all horses. In conclusion, SAA can also be measured in bronchoalveolar fluid, affording early detection of respiratory infections or inflammatory conditions.(AU)

Distúrbios pulmonares são comuns nos cavalos e a eficiência do tratamento depende de um diagnóstico adequado e precoce. A amilóide A é um biomarcador sensível na deteccção de patologias inflamatórias e infecciososa em cavalos. O objetivo deste estudo foi estabelecer a concentração de amilóide A no líquido broncoalveolar (LBA) em cavalos saudáveis. Os cavalos foram considerados saudaveis baseado nos achados de normalidade do exame físico, hemograma, parâmetros bioquímicos e citologia do LBA. Sangue e LBA foram coletados de 30 fêmeas equinas adultas. Os níveis de Amilóide A no soro e no LBA foram mensurados por meio do teste de ELISA. A amilóide A foi detectada no soro (média ± DP = 3,71±2,51) e no LBA (média ± DP = 0,000745±0,000785) de todos os animais. Conclui-se que a amilóide A também pode ser mensurada no LBA, auxiliando no diagnóstico precoce de processos inflamatórios e infecciosos pulmonares.(AU)

Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests / Expressão e análise antigênica da proteína RTP36 recombinante da amostra São Paulo de Ehrlichia canis para testes sorológicos

Ehrlichia canis is the main etiological agent of canine monocytic ehrlichiosis (CME), a globally canine infectious disease. In Brazil, CME is considered to be endemic, and its prevalence can reach 65% in some states. The diagnosis of ehrlichiosis is important for treatment and epidemiological purposes. The E. canis TRP36 (Tandem Repeat Protein) protein elicits the earliest acute-phase antibody response observed during the course of the disease. This study aimed to generate the recombinant TRP36 protein from E. canis São Paulo strain and to evaluate its potential as a tool for the serologic diagnosis of CME. The E. canis São Paulo isolate was cultivated in DH82 lineage cells, and its genomic DNA was obtained. The bacterial DNA fragment encoding the entire ORF of TRP36 was cloned into the pBAD/Thio-TOPO vector and transformed into Escherichia coli DH10B competent cells with the trp36-bearing plasmid for protein expression. To evaluate the protein antigenicity, 16 canine serum samples were previously tested (by PCR and the commercial SNAP®4Dx® serological test). The results were in accordance with the SNAP®4Dx® test. Experiments using this recombinant protein as an antigen, targeting the development of a serologic test based on ELISA methodology, are the next step to produce a reliable, affordable and useful diagnostic tool for CME in Brazil.(AU)

Ehrlichia canis é o principal agente etiológico da erliquiose monocítica canina (EMC), uma doença infecciosa canina globalmente dispersa. No Brasil, a EMC é considerada endêmica, e a infecção pode atingir 65% em cães em alguns estados. O diagnóstico de erliquiose é importante para fins de tratamento e epidemiológicos. A proteína TRP36 de E. canis leva a uma resposta humoral com produção de anticorpos em fase aguda, encontrada durante o curso da doença. O objetivo deste estudo foi obter a proteína TRP36 recombinante da amostra São Paulo de E. canis e avaliar seu potencial como ferramenta para o diagnóstico sorológico da CME. O isolado de E. canis São Paulo foi cultivado em células da linhagem DH82 e o DNA genômico foi obtido. O fragmento de DNA bacteriano que codifica toda a ORF de TRP36 foi clonado no vetor pBAD / Thio-TOPO e transformado em células competentes Escherichia coli DH10B, com o plasmídeo portador de trp36 para expressão de proteínas. Para avaliar a antigenicidade da proteína, 16 amostras de soro canino foram previamente analisadas (por PCR e teste sorológico comercial SNAP®4Dx®). Os resultados estavam de acordo com o teste SNAP®4Dx®. Os experimentos que utilizam essa proteína recombinante como antígeno, visando ao desenvolvimento de um teste sorológico baseado no ELISA, são o próximo passo para produzir um teste de diagnóstico confiável, acessível e útil para o diagnóstico da EMC no Brasil.(AU)