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Ticks have saliva rich in immunoregulatory molecules that interfere with the host's physiology in order to feed. This study aimed to evaluate the concentration of acute phase proteins and circulating oxidative stress in response to infestation by Amblyomma sculptum and Dermacentor nitens in two breed horses, Mangalarga Marchador and Breton Postier, to define resistance or susceptibility to ticks. Among the oxidative stress markers, we observed lower malondialdehyde and nitric oxide in horses with tick infestation, consequently not altering the antioxidant enzymes. Breton Postier with tick infestation showed a reduction in the ferric reducing ability of plasma (FRAP), which may be due to lower feeding of the host due to the stress caused by the infestation or even to sequestration of components induced by the tick during blood feeding. The alpha-1-antitrypsin, an acute phase protein, showed an increase in Mangalarga Marchador with tick infestation; curiously it is related to a protective action against tissue damage, pathogens and parasites. We could assume that Mangalarga Marchador showed a better response to ticks when compared to Breton Postier. However, it is still early to define the resistance or susceptibility to ticks, as we did not observe significant changes in most of the analyzed variables. Further studies are needed to understand the compounds and mechanisms of action of the tick saliva in the acute phase proteins and the possible relationships of oxidative stress in the host and the tick during blood feeding.

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BACKGROUND: Electrophoresis can be used to aid in the diagnosis of different diseases in avian species. Capillary zone electrophoresis (CZE) is an automated method that is proposed to be superior to the dye methods used in agarose gel electrophoresis (AGE). However, reference intervals (RIs) for CZE in avian species and comparison studies between electrophoretic methods are lacking. OBJECTIVES: The goals of the current study were to compare AGE and CZE methods and determine reference intervals for CZE using plasma from bald eagles (Haliaeetus leucocephalus). METHODS: Heparinized plasma samples from 44 bald eagles (mean age 18.7 years) under managed care were examined by AGE and CZE. Method comparison analyses were completed, as well as the generation of preliminary RIs using the CZE method and ASVCP guidelines. RESULTS: Passing-Bablok regression and Bland-Altman plots demonstrate that these methods are not equivalent. All fractions were significantly correlated between the methods except for alpha 1 globulin. Inter-assay and intra-assay CVs for CZE were lower or comparable to AGE and ranged from 2.4% to 15.4%, and 0.8% to 8.3%, respectively. CZE resolved more fractions than AGE with two fractions observed in the beta and gamma region vs one for AGE in each region. CONCLUSIONS: CZE provided improved resolution and reproducibility for the quantitation of protein fractions in the bald eagle. Although most fraction results correlated with AGE, these methods were judged as not equivalent, necessitating method-specific Rls. Reference intervals generated from a limited number of mostly aged individuals under managed care should be considered preliminary; additional studies will aid in the production of more robust intervals.

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Introduction: Horses submitted to carbohydrate overload can develop laminitis due to changes in cecal pH and microbiota, followed by an increase in transmural absorption of luminal content, including bacterial toxins. In response to acute injury there is hepatic overproduction of several proteins known as acute phase proteins (APP). Few studies have evaluated protein fractionation to characterize the inflammatory response in acute laminitis. The aim of this study was to test the viability of an experimental model to induce acute laminitis, using a single carbohydrate overload, and the influence of a buffering solution on the development of the disease; also, study the kinetics of APP during acute laminitis, as well as the correlation between these proteins and clinical signs associated to this syndrome. Methods: Ten healthy horses were divided in a factorial and randomized way into four groups (n = 5): control group (CG), starch group (SG), buffer group (BG), and starch C buffer group (SBG). They were evaluated at seven times (T0h, T4h, T8h, T12h, T24h, T48h, and T72h), which included clinical evaluation and blood sample collection. Total serum protein and albumin concentrations were determined by colorimetry and the other APP by polyacrylamide gel electrophoresis containing sodium dodecyl sulfate and commercial ELISA kits. Data were analyzed by two-way ANOVA, followed by Tukey's test (p < 0.05). The correlation between clinical signs and APP were verified using the Pearson's correlation coefficient. Results and discussion: 40% of the animals from SG and 60% from SBG developed clinical laminitis. A single administration of buffer solution was not able to prevent clinical signs of laminitis. There was no difference between groups on total serum protein, albumin, serum amyloid A and C-reactive protein concentrations (p > 0.05). Transferrin, considered a negative APP, showed a positive response pattern in SG and SBG. Ceruloplasmin had a positive correlation with Obel grade, heart rate on animals from SGB and number of steps on horses submitted to starch overload (SG and SBG). Ceruloplasmin, α-1-antitrypsin and haptoglobin concentrations increased in SBG, suggesting an inflammatory response in animals of this group. Changes in clinical parameters were also more evident in the SBG, corroborating the protein fractionation findings.

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The acute phase response (APR), coordinated by a complex network of components of the immune and neuroendocrine systems, plays a key role in early immune defense. This response can be elicited by a wide variety of pathogens at different intensities (frequencies and doses), hence experimental immune challenges with antigen gradients makes it possible to evaluate sickness progression with a better representation of what occurs in natural systems. However, how infection intensity could shape the APR magnitude in wild species is still poorly understood. Here, the immune response was activated in the subterranean rodent Ctenomys talarum with a gradient of lipopolysaccharide (LPS) doses (0.5, 1, 1.5, and 2 mg/kg of body mass). Changes in body temperature, body mass, and energetic costs were evaluated over time. We also assessed cortisol levels, white blood cells counts and neutrophil: lymphocyte ratios, before and after injection. Results indicated that during the APR, C. talarum shows a hyperthermic response, which is maintained for 6 h, with slight differences among antigen doses in the pattern of thermal response and body mass change. A maximum increase in body temperature of 0.83°C to 1.63°C was observed during the first hour, associated with a metabolic cost that ranged from 1.25 to 1.41 ml O2 /gh. Although no clear effects of treatment were detected on leukocyte abundance, we found increments in neutrophil: lymphocyte ratios and gradual increases in cortisol levels corresponding to the intensity of simulated infection, which may indicate redistribution of immune cells and enhancement of immune function. An evident sickness syndrome was observed even at the lowest LPS dose that was characterized by an increase in body temperature, energy expenditure, and N: L ratio, as well as a dose-dependent increase in cortisol levels. Although in nature, other constraints and challenges could affect the magnitude and costs of immune responses, C. talarum mounts an effective APR with a low increase in their daily energy expenditure, regardless of LPS dose.

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Horse transport is a common practice and is usually associated as a cause of stress in animals, with consequences for their well-being. There are several of evidence that stress can increase an acute phase response. The aim of this study was to verify whether the road transport of horses over distances of 50 and 300 kilometers induces changes in the values of acute phase proteins. To do this, the serum SDS-PAGE was performed and the bands obtained were identified by mass spectrometry (MALDI-TOF). The blood samples were collected in tubes without anticoagulant to obtain the serum, and the evaluations occurred before the road transportation (T0), immediately after the journey (T1), six hours later (T2), and 24 hours (T3), 48 hours (T4), 72 hours (T5), 96 hours (T6), 120 hours (T7) and 144 hours (T8) after the end of the trip. All analyzes were performed using the Minitab 17 statistical package, and significance was considered when P<0.05. The APPs found through SDS-PAGE and properly identified were α2-macroglobulin, ceruloplasmin, transferrin, albumin, α1-antitrypsin, haptoglobin, apolipoprotein alpha 1, and α1-acid glycoprotein. No differences were observed in the concentration values between 50 and 300 km or between the moments after each route. The distances covered with the horses were not challenging enough to provoke an acute phase response reflected in changes in APPs.

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Sturgeons are chondrostean fish critically endangered due to anthropogenic loss and degradation of natural habitat and overfishing for meat and caviar production. Consequently, sturgeon aquaculture has extensively developed lately, being Russian sturgeon (Acipenser gueldenstaedtii) the second most important species reared for caviar production. However, Russian sturgeon aquaculture in subtropical countries, such as Uruguay, confronts difficulties because fish have to endure excessive summertime warm temperatures, which weaken their innate defences facilitating opportunistic infections. To address this problem, we look for identifying putative acute phase proteins (APPs), which might be robust serum biomarkers of both infection and chronic thermal stress, applied to monitoring Russian sturgeon health status in farms. We focused on the C-Reactive Protein/Serum Amyloid P (CRP/SAP) pentraxin since the pentraxin family includes well-known APPs, better characterised in mammals than fish. We identified A.gueldenstaedtii CRP/SAP (AgCRP/SAP), as a member of the universal CRP/SAP pentraxin sub-family, and studied AgCRP/SAP involvement in sturgeon response to bacterial challenge and chronic thermal stress, in comparison with A. gueldenstaedtii Serum Amyloid A (AgSAA), a previously described positive APP. Results showed that AgCRP/SAP is a constitutive serum component that remained constant upon Aeromonas hydrophila challenge and chronic thermal stress. Contrastingly, serum AgSAA was subjected to regulation by bacterial and thermal stress challenges, showing a 50-fold increase and 3-fold decline in serum levels, respectively. Overall, results highlight the potential value of AgSAA, but not of AgCRP/SAP, as a biomarker of bacterial infection and the need to continue searching for robust chronic thermal stress biomarkers in sturgeons.

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BACKGROUND: This study aimed to evaluate the inflammatory response of miniature horses subjected to open and half-closed orchiectomy by physical examination, blood cell count, peritoneal fluid evaluation, total plasma protein, fibrinogen, and serum amyloid A (SAA) concentrations. METHODS: Thirteen male healthy miniature horses were divided into two groups, according to the surgical approach: half-closed technique (HCT) and open technique (OT). The HCT group was subjected to ligation of the spermatic cord followed by its sharp incision, and closure of the vaginal tunic, and the OT group was only submitted to cord ligation. Prior to, and at 1, 2, 3 and 5 days after the surgery, a general and specific physical examination, blood cell counts, total plasma protein, peritoneal fluid evaluation, fibrinogen, and SAA concentrations were performed. RESULTS: Higher postoperative perilesional oedema, rectal temperature, and fibrinogen were observed in the HCT group. Groups did not differ as to SAA concentrations. The evaluated local and systemic inflammatory profile demonstrated that, as expected, surgery resulted in inflammation in both groups. CONCLUSIONS: The group subjected to the HCT showed a more intense and lasting inflammatory response. However, despite the different postoperative inflammatory profiles, both groups presented a favourable outcome and recovery.

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The objective of this study was to evaluate the serum proteinogram, identifying and quantifying the acute-phase proteins (APPs) of horses used in show jumping activity with obstacles of a meter in height. As it is an equestrian sport that involves high intensity and excessive impact, the possibility of injury is relevant. The serum of 10 horses was evaluated in a competition for beginners. The material was collected at rest (T0), immediately after exercise (T1), 30 minutes after the effort (T2), 1 hour after the effort (T3), and 24 hours after the effort. Acute-phase proteins were separated by polyacrylamide gel electrophoresis and their concentrations determined by computerized densitometry. Protein identification was performed using mass spectrometry. The data were evaluated using analysis of variance for repeated measures, considering the level of significance of P < .05. Eight APPs were identified: α2-macroglobulin (α2-macro), ceruloplasmin (Cp), transferrin (Trf), albumin (Alb), α1-antitrypsin (α1-atp), haptoglobin (Hp), acid glycoprotein (AGP), and apolipoprotein A1 (Apo A1). There was a difference in Cp, AGP, and Apo A1 between moments. As the other proteins were not influenced by exercise, they were established as a valuable resource in the monitoring of inflammatory processes and an important complementary element in controlling the impact of training on the animals, thus guaranteeing their welfare.

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The acute phase response is a response to injury and depends on the severity of the trauma. Heparin is routinely used for postsurgical treatment of horses to prevent abdominal adhesions; however, its effect on inflammation is unknown. This study aimed to assess systemic inflammatory response of horses subjected to small colon enterotomy and to evaluate heparin effects on postsurgical inflammation. Ten adult horses were subjected to small colon enterotomy and were assigned to a control or a treatment group. Both groups received prophylactic antibiotics and flunixin, and the treatment group received 150 IU/kg heparin subcutaneously after surgery and every 12 hours for five days. WBC counts, peritoneal fluid evaluation, determination of serum and peritoneal haptoglobin (Hp), and serum amyloid A (SAA) were performed before, 12 hours, and 1, 2, 4, 6, 10, and 14 days after enterotomy. Forty-eight hours after surgery, a significant increase in serum Hp was observed in the control group, and SAA concentrations increased significantly in the both groups between 24 hours, 48 hours, and 4 days after surgery. The SAA and serum Hp concentrations produced no significant differences between the groups. Peritoneal Hp increased significantly in the control group 4 days after surgery and was significantly higher in the control group than in the treated group 14 days after surgery. Serum Hp and SAA identified the acute phase response changes faster, however, were not able to identify differences between groups. Peritoneal Hp concentrations identified inflammatory differences between the groups 14 days after surgery; the difference suggests that heparin may act decreasing inflammation.

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Obesity in human and veterinary medicine is one of the most complex challenges within emerging diseases in the context of health. The problem of obesity in horses results in severe comorbidities; therefore, acute-phase proteins should be investigated for fluctuations increasing or decreasing by at least 25% in response to inflammation; these are candidates for future biomarkers and might provide new perspectives on early diagnosis and prognosis. Serum samples were analyzed in nine healthy animals (C) and nine obese animals (O). The proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the concentrations determined by computerized densitometry; matrix assisted laser desorption ionization time of flight mass spectrometry/TOF mass spectrometry was performed to identify the proteins, and the results obtained were compared to the Equidae and Metazoa taxon protein database deposited in UNIPROT using the MASCOT application. Three proteins presented a difference between the groups; ceruloplasmin (Cp), α1-antitrypsin (α1-antitryp), and haptoglobin (Hp). The behavior of the Cp and Hp proteins was compatible with the available literature for obesity in other species. The α1-antitryp protein was positively correlated with leptin, demonstrating the need for further investigations. The initial study of these proteins was important due to the lack of information available on the influence of obesity on inflammatory biomarkers in horses in Brazil; therefore, we sought to verify a possible association between overweight and changes in the studied variables.

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