Silvia Martínez-Subiela

Serum concentrations of acute phase proteins in dogs with leishmaniasis

The concentrations of haptoglobin, C-reactive protein and ceruloplasmin were measured in symptomatic and asymptomatic dogs naturally infected by Leishmania infantum, and in healthy uninfected dogs to determine the potential value of these proteins for the diagnosis and prognosis of leishmaniasis. The concentrations of the acute phase proteins were significantly higher in the dogs with leishmaniasis than in the control dogs, and the concentration of C-reactive protein was significantly higher in the symptomatic dogs than in the asymptomatic dogs. There were no correlations between the acute phase proteins and the gamma globulins, the albumin/globulin ratio or the titre of anti-leishmanial antibodies.

Acute phase protein response in goats

Acute phase proteins (APPs) are important diagnostic indicators of inflammatory disturbances in animals. The objectives of the current study were to validate analytical methods for measuring haptoglobin (Hp), serum amyloid A (SAA), acid soluble glycoprotein (ASG), fibrinogen, and albumin concentrations in goats and to evaluate their response to an inflammatory stimulus in this species. Intra- and interassay coefficients of variation (CVs) were in the range 0.07–9.31% and 1.83–12.68%, respectively, for all APPs and showed good precision. All assays determined APPs in a linear manner at different sample dilutions with high correlation coefficients with the exception of fibrinogen, which was measured by the heat precipitation method. Subcutaneous injection of turpentine oil induced an increase in Hp, SAA, ASG, and fibrinogen serum concentrations and a decrease in albumin concentration.

C-reactive protein quantification in porcine saliva: a minimally invasive test for pig health monitoring.

Study objectives were to investigate whether C-reactive protein (CRP) in pig saliva could be quantified using an adapted, time-resolved immunofluorometry assay (TR-IFMA), and to determine whether the assay could distinguish healthy from diseased animals. The test method had intra- and inter-assay coefficients of variation of 5.75% and 9.73%, respectively, the limit of detection was 0.47ng/mL and the coefficient of determination was 0.98. Analysis of CRP concentrations in paired serum and saliva samples from 50 pigs gave a positive correlation (r=0.702, P<0.01) and the salivary CRP concentration was able to distinguish healthy from diseased animals in 62 samples from pigs with naturally occurring or experimentally-induced inflammation. The results suggest that this minimally invasive, straightforward and sensitive assay may be useful in pig health and welfare monitoring.

Acute phase protein response in experimental canine leishmaniosis

Acute phase proteins (APPs) have been proposed as useful markers for the diagnosis and monitoring of treatment of dogs infected by Leishmania infantum. However, the kinetics and behavior of these proteins in canine leishmaniasis is still unknown. The aim of this study was to monitor the kinetics of APPs in dogs experimentally infected with L. infantum, before, during and after therapy against canine leishmaniasis. Levels of serum haptoglobin, serum amyloid A and C-reactive protein from 6 infected beagles, positive by both PCR and parasite culture, were monitored for 7 months post-infection. The dogs were then treated for 3 months with allopurinol (20 mg mg/kg/day PO), and their response to therapy was followed for 11 additional months. Levels of Immunoglobulins G and M were recorded during these 21 months and compared. Experimental infection with L. infantum amastigotes induced an increase in all APPs studied which was statistically significant 2 months after infection for all proteins. Clinical recovery was accompanied by a significant decrease of all APPs 1 month after the beginning of treatment. However, differences were found between the APPs in both magnitude and duration of serum level elevations. The increase in total IgG and IgM was delayed in comparison to APPs and contrarily to the APPs, these immunoglobulins did not significantly decrease with treatment. In conclusion, the results of this study suggest that APPs could be used as early markers for disease as well as for monitoring the response to treatment in canine leishmaniasis.

Serum acute phase protein concentrations in dogs with hyperadrenocorticism with and without concurrent inflammatory conditions.

BACKGROUND Acute phase proteins (APPs) are promising markers of inflammation in dogs, because they are more sensitive than WBC counts in detecting clinical and subclinical inflammation. Endogenous corticosteroids can mask an acute phase response and make it more difficult to identify underlying inflammatory disease. OBJECTIVE The purpose of this study was to evaluate the acute phase protein response in dogs with spontaneous hyperadrenocorticism (HAC) with and without concurrent inflammatory conditions. METHODS Serum concentrations of C-reactive protein (CRP), haptoglobin (Hp), fibrinogen, and albumin were measured in 44 healthy adult dogs and 39 dogs with HAC; the HAC group was further divided into dogs with and without concurrent infection/inflammation. A fourth group of dogs with severe sepsis and without HAC was compared with the dogs with HAC and severe sepsis. RESULTS Dogs with uncomplicated HAC had significantly higher Hp and fibrinogen concentrations compared with healthy control dogs (P<.001). Dogs with HAC and severe inflammatory disease also had significantly higher CRP and lower albumin concentrations than control dogs and dogs with HAC without concurrent inflammation. Dogs with sepsis but without HAC had significantly higher CRP concentrations than dogs with HAC and sepsis. CONCLUSIONS Dogs with HAC had increases in the moderate APPs (Hp and fibrinogen), and no significant changes in CRP and albumin compared with healthy dogs. Although concurrent HAC appeared to blunt the CRP response in dogs with sepsis, increased serum CRP concentration in dogs with HAC is likely indicative of severe concurrent inflammation.

Use of saliva for haptoglobin and C-reactive protein quantifications in porcine respiratory and reproductive syndrome affected pigs in field conditions

The purpose of this study was to study the use of saliva samples as alternative to serum for acute phase protein (APP) quantifications in pigs at field conditions. To this end, haptoglobin (Hp) and C-reactive protein (CRP) concentrations were examined in 100 animals at different ages. Sixty pigs were from a farm with chronic PRRS virus infection and 40 from a specific pathogen free (SPF) farm. A serological study was performed to obtain an overview of the immune status of animals and to evaluate possible concomitant infections in animals with PRRS infection. The results reported in this study showed that both saliva and serum samples had higher APP concentrations in PRRS pigs aged 8-9, 17-18 and 24-25 weeks in conventional herds than SPF pigs of the same age (p<0.05). In addition, increases in APP were obtained with age independently of the health status of the animals. According to the ROC analyses performed, saliva could be a better specimen than serum to quantify Hp and CRP levels in field conditions and may contribute to a more efficient detection of diseased animals at farm level.

Evaluation of an immunoassay for determination of haptoglobin concentration in various biological specimens from swine

OBJECTIVE To develop and evaluate an immunoassay based on time-resolved immunofluorometry (TR-IFM) for measurement of haptoglobin concentrations in samples of various body fluids of swine. ANIMALS 20 pigs without clinical signs of disease and seronegative for antibodies against major viruses that affect pigs and 30 pigs with clinical signs of disease. PROCEDURES Haptoglobin concentrations were measured in samples of serum, saliva, and meat juice obtained from both groups of pigs to evaluate the ability of TR-IFM to differentiate between healthy and diseased pigs. Performance of TR-IFM was evaluated by means of its calibration curve and detection limit, analytic precision during routine operation, and linearity of results for serial dilutions for the 3 types of samples. In addition, performance of TR-IFM was compared with that of a commercial spectrophotometric assay. RESULTS The TR-IFM assay involved only 1 step, and the results were obtained in 20 minutes, with good analytic sensitivity and reproducibility. The analytic limit of detection was 0.52 ng/mL. Intra-assay and interassay coefficients of variation ranged from 1.13% to 4.81% and 5.97% to 13.57%, respectively. The method yielded linear results for all sample types. Serum haptoglobin concentrations determined by use of TR-IFM and spectrophotometric assays were highly correlated (r = 0.96). Differences between healthy and diseased pigs with respect to median haptoglobin concentrations were significant for all types of samples. CONCLUSIONS AND CLINICAL RELEVANCE The 1-step TR-IFM assay accurately quantified haptoglobin concentrations in serum, saliva, and meat juice samples from swine and may be useful in laboratory and meat inspection settings.

Critical differences of acute phase proteins in canine serum samples.

The critical difference values for acute phase proteins in canine serum samples were established on a week-to-week basis. Blood samples from 11 apparently clinically healthy dogs were collected once weekly for five consecutive weeks. For each protein the total variance of analytical results was divided into intraindividual variance (S(Intra)(2)), interindividual variance (S(Inter)(2)), and analytical variance (S(Analytical)(2)). The critical difference (d(K)) was then calculated as d(k)=22(S(Intra)(2)+S(Analytical)(2)). The critical difference values were 1.95 g/L for haptoglobin, 4.85 mg/L for C-reactive protein, and 0.016 DeltaAbs/min for ceruloplasmin. When used in conjunction with the corresponding reference interval, critical difference values can be an aid in correctly interpreting acute phase protein results, by determining whether observed differences between two consecutive measurements in individual animals are due to natural variation or due to disease therapy or experimental procedures.

Effect of weight loss on inflammatory biomarkers in obese dogs

The objective of this study was to evaluate the effects of weight loss on selected serum inflammatory biomarkers in obese dogs. An experimentally induced bodyweight reduction of approximately 2.5%/week was accompanied by significant decreases in metabolic markers of obesity (lipidic profile, fructosamine, and insulin-like growth factor-1). The concentrations of acute phase proteins and of selected cytokines remained within reference ranges in obese dogs during weight loss, suggesting that significant inflammation was not a major component of this experimental model. However, adiponectin concentrations increased following the period of weight loss suggesting reduced susceptibility of these animals to obesity-related inflammation.

Validation of an Automated Method for Salivary Alpha-Amylase Measurements in Pigs (Sus Scrofa Domesticus) and its Application as a Stress Biomarker

The aim of the current study was to validate an automated spectrophotometric method for salivary alpha-amylase measurement in pigs and evaluate its possible application as a noninvasive stress biomarker. The analytical validation included intra- and interassay precision, linearity under dilution, and limit of detection. In addition, to study the possible use of salivary alpha-amylase as a possible stress marker, 12 crossbred growing pigs of 3–4 months of age were subjected to restraint stress by a nasal snare for at least 1 min, and saliva samples were obtained at different time points. The results of analytical validation indicated that the method was precise and able to measure alpha-amylase in a linear manner. The results obtained in the stress test showed a significant increase in salivary alpha-amylase activity. Although other factors influencing this enzyme activity should be studied, these preliminary results indicate that salivary alpha-amylase could be a reliable biomarker of stress in pigs.