Circulating oxidative stress and acute phase protein levels in horses infested with ticks.

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.

Comparison of agarose gel and capillary zone electrophoresis methods with preliminary reference interval generation using capillary zone electrophoresis in bald eagles (Haliaeetus leucocephalus).

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.

Acute phase proteins levels in horses, after a single carbohydrate overload, associated with cecal alkalinization.

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.

Varying intensity of simulated infection partially affects the magnitude of the acute-phase immune response in the subterranean rodent Ctenomys talarum.

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.

Influence of Short and Medium Distance Road Transport on the Acute Phase Proteins in Horses.

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.

Different response of Acipenser gueldenstaedtii CRP/SAP and SAA to bacterial challenge and chronic thermal stress sheds light on the innate immune system of sturgeons.

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.

Inflammatory response of miniature horses subjected to open and half-closed orchiectomy techniques.

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.

Profile of Acute-Phase Proteins of Horses Submitted to Low-Level Show Jumping Classes.

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.

Inflammatory Response of Healthy Horses Subjected to Small Colon Enterotomy and Treated or Not With Heparin.

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.

Influence of Obesity on Serum Concentrations of Acute-Phase Proteins in Horses.

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.

Acute-Phase Proteins of Healthy Horses and Horses Naturally Affected by Colic Syndrome.

The purpose of this study was to evaluate and compare the serum proteinogram, identifying and quantifying the acute-phase proteins (APPs) of healthy horses and those naturally affected by colic syndrome. Blood samples were collected from 9 healthy horses and 17 horses with clinical (G1) or surgical (G2) colic at the time of arrival at the veterinary hospital (M0) and 24 (M1), 48 (M2), and 72 (M3) hours after the initiation of treatment. The APPs were separated by polyacrylamide gel electrophoresis (SDS-PAGE), and their concentrations were determined by computerized densitometry. Protein identification was performed using matrix assisted laser desorption/ionization-time of flight mass spectrometry. Nine APPs were identified: α2-macroglobulin, ceruloplasmin, transferrin, albumin, α1-antitrypsin, haptoglobin, α1-acid glycoprotein, apolipoprotein A1, and serum amyloid A. Ceruloplasmin (Cp) presented an elevation at M0 in G2 in relation to G1 and the control group (CG). Serum amyloid A (SAA) presented an elevation with a difference at M3 in G1 and G2, compared with CG. In the comparison between the time points within each group (G1 and G2), there was an increase in SAA in G2 with a difference at M1, M2, and M3. Cp and SAA were suitable for evaluating the acute-phase response in horses with colic. Cp was an indicator of colic with surgical resolution. SAA presented favorable behavior for its use as a biomarker to establish the prognosis in horses with colic. The serial measurement of these proteins was useful in differentiating the clinical or surgical treatment and prognosis of horses with this disorder.

Response to lipopolysaccharide in Octodon degus pups produces age-related sickness behavior but does not have effects in juveniles.

During vertebrate development, the immune function is inefficient and is mainly controlled by innate defense. While there have been detailed studies of various aspects of innate immune function, the effects of this function in the growth of vertebrates is still not well known. Similarly, there is little information regarding how early endotoxin exposure would affect juvenile phenotypes, specifically in a non-model mammal like a precocial rodent. We evaluated the response to an antigen and its cost in offspring of the rodent Octodon degus. We inoculated pups at 4 different ages (8, 15, 22 and 30 days after birth) with an antigen to determine the ontogeny and costs of the response to an endotoxin. We assessed changes in body mass, body temperature, sickness behavior and the levels of a key mediator of the inflammatory response, the cytokine interleukin-1ß. We also determined the effects of early endotoxin exposure on the resting metabolic rate of juvenile animals (i.e. 90 days after birth). The cytokine levels, body mass and body temperature were unaffected by time of inoculation and treatment. However, pups subjected to inoculation at 22 days after birth with the antigen showed reduced locomotion. Juvenile resting metabolic rate was not affected by early endotoxin exposure. These results suggest that the magnitude of O. degus responses would not change with age. We discuss whether the lack of effect of the response on body mass or body condition is caused by environmental variables or by the precocial characteristics of O. degus.

Vitamin C Status in Hospitalized Adults According to Subjective Global Assessment Method.

BACKGROUND: The subjective global assessment (SGA) is a powerful tool for nutrition status assessment. Our aim was to compare vitamin C serum levels among patients classified as A, B, or C in the SGA. METHODS: One hundred-and-fifty adults in the wards of the University Hospital participated in this study. Besides SGA, all cases were submitted to anthropometry, bioelectrical impedance analysis (BIA), and 24-hour dietary recall. Laboratory data included blood hemoglobin, serum ferritin, serum albumin, transferrin, C-reactive protein (CRP), and vitamin C. Acute phase response was defined by serum CRP > 0.5 mg/dL; low serum vitamin C was defined by serum levels < 0.4 mg/dL. Analysis of variance and χ2 tests were used to compare groups; P < 0.05 was considered significant. RESULTS: Patients were diagnosed as SGA A (n = 76), B (n = 38), or C (n = 36) and showed different anthropometry and BIA. The same occurred, respectively, with vitamin C (median; range, in mg/day) intake (55.0; 4.7-140.6 vs 34.0; 10.3-244.2 vs 15.8; 2.3-124.0) and high (%) CRP (88.3 vs 65.8 vs 48.7) and low (%) vitamin C serum levels (21.1 vs 34.2 vs 63.9). CONCLUSION: Patients with worst nutrition parameters (SGA C) showed lower ascorbic acid serum levels than those classified as SGA A or B. These results are in accordance with reduced vitamin C intake and the presence of acute phase response.

Food restriction, but not seasonality, modulates the acute phase response of a Neotropical bat.

Season and food intake are known to affect immune response of vertebrates yet their effects on metabolic rate have been rarely explored. We tested the effect of season and acute food restriction and their interaction on the energetic cost of immune response activation of a tropical vertebrate, the Seba's short-tailed fruit bat (Carollia perspicillata). We specifically stimulated the acute phase response (APR) with bacterial lipopolysaccharide (LPS) to measure metabolic changes along with changes in body temperature (Tb), body mass (Mb), white blood cell counts and the Neutrophil/Lymphocyte ratio (N/L). We found no effect of season on the different factors associated to the activation of the APR. In contrast to our expectations, unfed bats reached similar Tb increments and RMR peak values and had higher RMR scope values and higher caloric costs than fed bats after LPS injection. However, food deprivation led to delayed metabolic response indicated by longer time required to reach peak RMR values in unfed bats. Both food-deprived and fed bats did not present leukocytosis after APR activation and their WBC counts were similar, but unfed bats had a significant increase of N/L. APR activation represented a small fraction of the bat daily energy requirements which might explain why unfed bats were not limited to mount a metabolic response. Our study adds to recent evidence showing that activating the innate immune system is not an energetically expensive process for plant-eating bats.

The energetic cost of mounting an immune response for Pallas's long-tongued bat (Glossophaga soricina).

The acute phase response (APR) is the first line of defense of the vertebrate immune system against pathogens. Mounting an immune response is believed to be energetically costly but direct measures of metabolic rate during immune challenges contradict this assumption. The energetic cost of APR for birds is higher than for rodents, suggesting that this response is less expensive for mammals. However, the particularly large increase in metabolic rate after APR activation for a piscivorous bat (Myotis vivesi) suggests that immune response might be unusually costly for bats. Here we quantified the energetic cost and body mass change associated with APR for the nectarivorous Pallas's long-tongued bat (Glossophaga soricina). Activation of the APR resulted in a short-term decrease in body mass and an increase in resting metabolic rate (RMR) with a total energy cost of only 2% of the total energy expenditure estimated for G. soricina. This increase in RMR was far from the large increase measured for piscivorous bats; rather, it was similar to the highest values reported for birds. Overall, our results suggest that the costs of APR for bats may vary interspecifically. Measurement of the energy cost of vertebrate immune response is limited to a few species and further work is warranted to evaluate its significance for an animal's energy budget.

Acute phase proteins in serum and cerebrospinal fluid in healthy cattle: possible use for assessment of neurological diseases / Proteínas de fase aguda no soro e no líquido cefalorraquidiano de bovinos hígidos: possíveis usos na avaliação de doenças neurológicas

Use of acute-phase proteins (APPs) for assessment of health and disease in animals has increased greatly within the last decade. The objective was to determine the normal concentration of APPs in the serum and cerebrospinal fluid (CSF) of healthy cattle by polyacrylamide gel electrophoresis. Fifty crossbred animals (350±70kg of BW and 18±1.2 months of age), 25 heifers and 25 steers were used. CSF samples were collected from atlanto-occipital (AO) site and blood samples were obtained from the jugular vein. CSF and serum protein electrophoresis were performed by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-seven proteins with molecular weights ranging from 7 and 37kDa were identified in CSF of all animals. These eight were nominally identified with immunoglobulin A and G, celuloplasmin, transferrin, albumin, α1-antitripsin, acidic glycoprotein, and haptoglobin. All protein fractions in CSF did not differ between heifers and steers. In sera, 34 proteins with molecular weights between 7 and 244kDa were identified in heifers and steers. Similar proteins were nominally identified in the sera, but only the CSF presented α1-antitripsin. The serum values of acidic glycoprotein and immunoglobulin G were significantly higher in steers compared with heifers. In conclusion, measurement of CSF acute phase protein concentrations can be useful in diagnosing and monitoring the progression of bovine neurological diseases, perhaps even to guide therapeutic procedures. The CSF electrophoretic profile of healthy cattle does not change depending on gender.(AU)

O uso de proteínas de fase aguda (PFAs) para a avaliação da saúde e da doença em animais de produção tem aumentado consideravelmente na última década. O objetivo deste estudo foi determinar a concentração normal de PAFs no soro e no líquido cefalorraquidiano (LCR) de bovinos sadios por meio da eletroforese em gel de poliacrilamida. Foram avaliados cinquenta animais mestiços (350±70kg de PV e 18±1,2 meses de idade), 25 novilhas e 25 novilhos. As amostras de LCR foram colhidas no espaço atlanto-occipital (AO) e as amostras de sangue obtidas da veia jugular. As PAFs do soro e do LCR foram determinadas através da eletroforese em gel poliacrilamida. Trinta e sete proteínas com pesos moleculares que variaram entre 7 e 37kDa foram identificadas no LCR de todos os animais, independente do sexo. Estas oito proteínas foram nominalmente identificadas como imunoglobulina A e G, ceruloplasmina, transferrina, albumina, α1-antitripsina, glicoproteína ácida, e haptoglobina. As frações de proteínas presentes no LCR não diferiram entre novilhas e novilhos. No soro de machos e fêmeas, 34 proteínas com pesos moleculares entre 7 e 244 kDa foram identificadas. As proteínas do soro foram similarmente identificadas, entretanto a α1-antitripsina foi identificada somente no LCR. Os valores séricos de glicoproteína ácida e imunoglobulina G foram significativamente mais elevados nas novilhas em comparação aos novilhos. Em conclusão, a determinação das concentrações de proteínas de fase aguda presentes do LCR pode ser útil no diagnóstico e monitoramento da progressão de doenças neurológicas bovinas, talvez possa ainda direcionar procedimentos terapêuticos. O perfil eletroforético do LCR de bovinos hígidos não se altera em função do sexo.(AU)

Acute phase proteins in serum and cerebrospinal fluid in healthy cattle: possible use for assessment of neurological diseases / Proteínas de fase aguda no soro e no líquido cefalorraquidiano de bovinos hígidos: possíveis usos na avaliação de doenças neurológicas

Use of acute-phase proteins (APPs) for assessment of health and disease in animals has increased greatly within the last decade. The objective was to determine the normal concentration of APPs in the serum and cerebrospinal fluid (CSF) of healthy cattle by polyacrylamide gel electrophoresis. Fifty crossbred animals (350±70kg of BW and 18±1.2 months of age), 25 heifers and 25 steers were used. CSF samples were collected from atlanto-occipital (AO) site and blood samples were obtained from the jugular vein. CSF and serum protein electrophoresis were performed by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-seven proteins with molecular weights ranging from 7 and 37kDa were identified in CSF of all animals. These eight were nominally identified with immunoglobulin A and G, celuloplasmin, transferrin, albumin, α1-antitripsin, acidic glycoprotein, and haptoglobin. All protein fractions in CSF did not differ between heifers and steers. In sera, 34 proteins with molecular weights between 7 and 244kDa were identified in heifers and steers. Similar proteins were nominally identified in the sera, but only the CSF presented α1-antitripsin. The serum values of acidic glycoprotein and immunoglobulin G were significantly higher in steers compared with heifers. In conclusion, measurement of CSF acute phase protein concentrations can be useful in diagnosing and monitoring the progression of bovine neurological diseases, perhaps even to guide therapeutic procedures. The CSF electrophoretic profile of healthy cattle does not change depending on gender.(AU)

O uso de proteínas de fase aguda (PFAs) para a avaliação da saúde e da doença em animais de produção tem aumentado consideravelmente na última década. O objetivo deste estudo foi determinar a concentração normal de PAFs no soro e no líquido cefalorraquidiano (LCR) de bovinos sadios por meio da eletroforese em gel de poliacrilamida. Foram avaliados cinquenta animais mestiços (350±70kg de PV e 18±1,2 meses de idade), 25 novilhas e 25 novilhos. As amostras de LCR foram colhidas no espaço atlanto-occipital (AO) e as amostras de sangue obtidas da veia jugular. As PAFs do soro e do LCR foram determinadas através da eletroforese em gel poliacrilamida. Trinta e sete proteínas com pesos moleculares que variaram entre 7 e 37kDa foram identificadas no LCR de todos os animais, independente do sexo. Estas oito proteínas foram nominalmente identificadas como imunoglobulina A e G, ceruloplasmina, transferrina, albumina, α1-antitripsina, glicoproteína ácida, e haptoglobina. As frações de proteínas presentes no LCR não diferiram entre novilhas e novilhos. No soro de machos e fêmeas, 34 proteínas com pesos moleculares entre 7 e 244 kDa foram identificadas. As proteínas do soro foram similarmente identificadas, entretanto a α1-antitripsina foi identificada somente no LCR. Os valores séricos de glicoproteína ácida e imunoglobulina G foram significativamente mais elevados nas novilhas em comparação aos novilhos. Em conclusão, a determinação das concentrações de proteínas de fase aguda presentes do LCR pode ser útil no diagnóstico e monitoramento da progressão de doenças neurológicas bovinas, talvez possa ainda direcionar procedimentos terapêuticos. O perfil eletroforético do LCR de bovinos hígidos não se altera em função do sexo.(AU)

Cinética da proteína total, fibrinogênio e ceruloplasmina em cordeiros Santa Inês durante os primeiros cinco meses de vida / Kinetics of total protein, fibrinogen, and ceruloplasmin in Santa Inês lambs during the first five months of life

Objetivou-se estudar a cinética da proteína total, fibrinogênio e ceruloplasmina durante os primeiros cinco meses de vida, em cordeiros saudáveis da raça Santa Inês, no município de São Gonçalo dos Campos, Bahia, Brasil. Amostras de sangue foram colhidas de 22 animais, ao longo de onze momentos: logo após o parto (T0), 12 (T1), 24 (T2), 48 horas (T3), sete (T4), 15 (T5), 30 (T6), 60 (T7), 90 (T8), 120 (T9) e 150 dias de vida (T10). A proteína total e o fibrinogênio plasmáticos foram analisados por meio de refratômetro clínico e pela técnica de desnaturação pelo calor, respectivamente, enquanto que a determinação da ceruloplasmina sérica se baseou em sua atividade oxidásica. Para análise estatística utilizou-se o programa SPSS versão 18; os dados com distribuição não paramétrica foram submetidos ao teste de Friedman para avaliar o efeito do tempo, enquanto que as comparações múltiplas pelo teste de Wilcoxon permitiram a identificação das diferenças entre os momentos, adotando-se nível de significância de 5% (p<0,05). A proteína total apresentou o menor valor no T0 diferindo estatisticamente dos demais tempos, com pico às 12 horas (T1), porém estabilizando-se até o final do experimento. O fibrinogênio não apresentou diferença estatística entre os tempos. De T1 (12h) a T3 (48h) constatou-se baixos valores de ceruloplasmina, muito embora às 24 horas (T2) tenha diferido estatisticamente (p<0,05), em relação ao T0. A partir do sétimo dia (T4) a concentração desta proteína aumentou significativamente, atingindo pico nos tempos T8 (90 dias) e T9 (120 dias). Foi possível estabelecer a cinética das proteínas estudadas, identificar os principais momentos com alterações e sugerir os fatores associados com as mudanças observadas.(AU)

The objective was to study the kinetics of the total protein, fibrinogen, and ceruloplasmin during the first five months of life in healthy lambs Santa Inês, in São Gonçalo dos Campos, Bahia, Brazil. Blood samples were collected from 22 animals, over eleven times: immediately after birth (T0), 12 (T1), 24 (T2), 48 hours (T3), seven (T4), 15 (T5), 30 (T6), 60 (T7), 90 (T8) 120 (T9), and 150 days of age (T10). The total protein and plasma fibrinogen were analyzed by means of a clinical refractometer and the heat denaturation technique, respectively, while the determination of serum ceruloplasmin was based on its oxidase activity. For statistical analysis, the SPSS version 18 program was used; the non-parametric data were submitted to the Friedman test to evaluate the effect of time, whereas the multiple comparisons by the Wilcoxon test allowed the identification of the differences between the moments, adopting a significance level of 5% (P < 0.05). The total protein presented the lowest value at T0 differing statistically from the other times, with a peak at 12 hours (T1), but stabilizing until the end of the experiment. Fibrinogen is not able to differentiate between the times. Between T1 (12h) and T3 (48h), low values of ceruloplasmin were observed, although at 24 hours (T2) it differed statistically (p <0.05) in relation to T0. On the seventh day (T4) the concentration of this protein increased significantly, reaching a peak at T8 (90 days) and T9 (120 days). It was possible to establish the kinetics of the proteins studied, to identify the main moments with alterations and to suggest the factors associated with the observed changes.(AU)

Cinética da proteína total, fibrinogênio e ceruloplasmina em cordeiros Santa Inês durante os primeiros cinco meses de vida / Kinetics of total protein, fibrinogen, and ceruloplasmin in Santa Inês lambs during the first five months of life

Objetivou-se estudar a cinética da proteína total, fibrinogênio e ceruloplasmina durante os primeiros cinco meses de vida, em cordeiros saudáveis da raça Santa Inês, no município de São Gonçalo dos Campos, Bahia, Brasil. Amostras de sangue foram colhidas de 22 animais, ao longo de onze momentos: logo após o parto (T0), 12 (T1), 24 (T2), 48 horas (T3), sete (T4), 15 (T5), 30 (T6), 60 (T7), 90 (T8), 120 (T9) e 150 dias de vida (T10). A proteína total e o fibrinogênio plasmáticos foram analisados por meio de refratômetro clínico e pela técnica de desnaturação pelo calor, respectivamente, enquanto que a determinação da ceruloplasmina sérica se baseou em sua atividade oxidásica. Para análise estatística utilizou-se o programa SPSS versão 18; os dados com distribuição não paramétrica foram submetidos ao teste de Friedman para avaliar o efeito do tempo, enquanto que as comparações múltiplas pelo teste de Wilcoxon permitiram a identificação das diferenças entre os momentos, adotando-se nível de significância de 5% (p<0,05). A proteína total apresentou o menor valor no T0 diferindo estatisticamente dos demais tempos, com pico às 12 horas (T1), porém estabilizando-se até o final do experimento. O fibrinogênio não apresentou diferença estatística entre os tempos. De T1 (12h) a T3 (48h) constatou-se baixos valores de ceruloplasmina, muito embora às 24 horas (T2) tenha diferido estatisticamente (p<0,05), em relação ao T0. A partir do sétimo dia (T4) a concentração desta proteína aumentou significativamente, atingindo pico nos tempos T8 (90 dias) e T9 (120 dias). Foi possível estabelecer a cinética das proteínas estudadas, identificar os principais momentos com alterações e sugerir os fatores associados com as mudanças observadas.

The objective was to study the kinetics of the total protein, fibrinogen, and ceruloplasmin during the first five months of life in healthy lambs Santa Inês, in São Gonçalo dos Campos, Bahia, Brazil. Blood samples were collected from 22 animals, over eleven times: immediately after birth (T0), 12 (T1), 24 (T2), 48 hours (T3), seven (T4), 15 (T5), 30 (T6), 60 (T7), 90 (T8) 120 (T9), and 150 days of age (T10). The total protein and plasma fibrinogen were analyzed by means of a clinical refractometer and the heat denaturation technique, respectively, while the determination of serum ceruloplasmin was based on its oxidase activity. For statistical analysis, the SPSS version 18 program was used; the non-parametric data were submitted to the Friedman test to evaluate the effect of time, whereas the multiple comparisons by the Wilcoxon test allowed the identification of the differences between the moments, adopting a significance level of 5% (P < 0.05). The total protein presented the lowest value at T0 differing statistically from the other times, with a peak at 12 hours (T1), but stabilizing until the end of the experiment. Fibrinogen is not able to differentiate between the times. Between T1 (12h) and T3 (48h), low values of ceruloplasmin were observed, although at 24 hours (T2) it differed statistically (p <0.05) in relation to T0. On the seventh day (T4) the concentration of this protein increased significantly, reaching a peak at T8 (90 days) and T9 (120 days). It was possible to establish the kinetics of the proteins studied, to identify the main moments with alterations and to suggest the factors associated with the observed changes.

Acute phase proteins in serum and cerebrospinal fluid in healthy cattle: possible use for assessment of neurological diseases

ABSTRACT: Use of acute-phase proteins (APPs) for assessment of health and disease in animals has increased greatly within the last decade. The objective was to determine the normal concentration of APPs in the serum and cerebrospinal fluid (CSF) of healthy cattle by polyacrylamide gel electrophoresis. Fifty crossbred animals (350±70kg of BW and 18±1.2 months of age), 25 heifers and 25 steers were used. CSF samples were collected from atlanto-occipital (AO) site and blood samples were obtained from the jugular vein. CSF and serum protein electrophoresis were performed by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-seven proteins with molecular weights ranging from 7 and 37kDa were identified in CSF of all animals. These eight were nominally identified with immunoglobulin A and G, celuloplasmin, transferrin, albumin, 1-antitripsin, acidic glycoprotein, and haptoglobin. All protein fractions in CSF did not differ between heifers and steers. In sera, 34 proteins with molecular weights between 7 and 244kDa were identified in heifers and steers. Similar proteins were nominally identified in the sera, but only the CSF presented 1-antitripsin. The serum values of acidic glycoprotein and immunoglobulin G were significantly higher in steers compared with heifers. In conclusion, measurement of CSF acute phase protein concentrations can be useful in diagnosing and monitoring the progression of bovine neurological diseases, perhaps even to guide therapeutic procedures. The CSF electrophoretic profile of healthy cattle does not change depending on gender.

RESUMO: O uso de proteínas de fase aguda (PFAs) para a avaliação da saúde e da doença em animais de produção tem aumentado consideravelmente na última década. O objetivo deste estudo foi determinar a concentração normal de PAFs no soro e no líquido cefalorraquidiano (LCR) de bovinos sadios por meio da eletroforese em gel de poliacrilamida. Foram avaliados cinquenta animais mestiços (350±70kg de PV e 18±1,2 meses de idade), 25 novilhas e 25 novilhos. As amostras de LCR foram colhidas no espaço atlanto-occipital (AO) e as amostras de sangue obtidas da veia jugular. As PAFs do soro e do LCR foram determinadas através da eletroforese em gel poliacrilamida. Trinta e sete proteínas com pesos moleculares que variaram entre 7 e 37kDa foram identificadas no LCR de todos os animais, independente do sexo. Estas oito proteínas foram nominalmente identificadas como imunoglobulina A e G, ceruloplasmina, transferrina, albumina, 1-antitripsina, glicoproteína ácida, e haptoglobina. As frações de proteínas presentes no LCR não diferiram entre novilhas e novilhos. No soro de machos e fêmeas, 34 proteínas com pesos moleculares entre 7 e 244 kDa foram identificadas. As proteínas do soro foram similarmente identificadas, entretanto a 1-antitripsina foi identificada somente no LCR. Os valores séricos de glicoproteína ácida e imunoglobulina G foram significativamente mais elevados nas novilhas em comparação aos novilhos. Em conclusão, a determinação das concentrações de proteínas de fase aguda presentes do LCR pode ser útil no diagnóstico e monitoramento da progressão de doenças neurológicas bovinas, talvez possa ainda direcionar procedimentos terapêuticos. O perfil eletroforético do LCR de bovinos hígidos não se altera em função do sexo.