Basics for the potential use of saliva to evaluate stress, inflammation, immune system, and redox homeostasis in pigs.

The use of saliva as a biological sample has many advantages, being especially relevant in pigs where the blood collection is highly stressful and painful, both for the animal and the staff in charge of the sampling. Currently one of the main uses of saliva is for diagnosis and detection of infectious diseases, but the saliva can also be used to measure biomarkers that can provide information of stress, inflammation, immune response and redox homeostasis. This review will be focused on the analytes that can be used for such evaluations. Emphasis will be given in providing data of practical use about their physiological basis, how they can be measured, and their interpretation. In addition, some general rules regarding sampling and saliva storage are provided and the concept of sialochemistry will be addressed. There is still a need for more data and knowledge for most of these biomarkers to optimize their use, application, and interpretation. However, this review provides updated data to illustrate that besides the detection of pathogens in saliva, additional interesting applicative information regarding pigs´ welfare and health can be obtained from this fluid. Information that can potentially be applied to other animal species as well as to humans.

Correction to: Inflammatory markers before and after farrowing in healthy sows and in sows affected with postpartum dysgalactia syndrome.

The original article [1] contains an error whereby the caption in Figure 8 is incorrect; the correct caption can be seen ahead alongside its respective image.

Uterine artery Doppler and sFlt-1/PlGF ratio: prognostic value in early-onset pre-eclampsia.

OBJECTIVE: To evaluate the performance of the mean uterine artery pulsatility index (UtA-PI) and the automated measurement of the soluble fms-like tyrosine kinase-1 (sFlt-1)/placental growth factor (PlGF) ratio for the prognostic assessment of both maternal and perinatal outcomes, and the time-to-delivery interval in early-onset (≤ 34 + 0 weeks) pre-eclampsia (PE) cases with attempted expectant management. METHODS: Fifty-one singleton pregnancies with early-onset PE were enrolled in the study. Mean UtA-PI and sFlt/PlGF ratio were measured at diagnosis. The association of each marker and their combinations with adverse maternal and perinatal outcomes was assessed by univariable comparisons and multivariable logistic regression analysis and time-to-delivery interval by survival analysis. RESULTS: Twenty-six (51%) had adverse maternal outcome and 14 (27%) had adverse perinatal outcome. At the time of onset of PE, only gestational age was significantly related to maternal complications. Gestational age at onset, mean UtA-PI and sFlt-1/PlGF ratio were significantly associated with perinatal complications, their combination reaching a sensitivity of 64% with 95% specificity, and an area under the receiver-operating characteristics curve of 0.89 (95% CI, 0.79-0.99). Regarding the time until delivery, 92% (12/13) of cases with sFlt-1/PlGF ratio > 655 and 39% (15/38) of cases with a ratio ≤ 655 delivered within the first 48 h, 8% (1/13) and 19% (7/38), respectively, delivered between 48 h and 7 days and 0% (0/13) and 42% (16/38), respectively, delivered after 7 days. CONCLUSION: Mean UtA-PI and sFlt-1/PlGF ratio in combination with gestational age are useful for the prognostic assessment of perinatal complications at the time of diagnosis of early-onset PE, but this combination has limited value for the prediction of maternal complications. Moreover, sFlt-1/PlGF ratio > 655 is closely related to the need to deliver within 48 h. [[ArtCopyrightmsg]].

Teaching cardiopulmonary resuscitation using virtual reality: A randomized study.

BACKGROUND: The main functions of healthcare professionals include training and health education. In this sense, we must be able to incorporate new technologies and serious game to the teaching cardiopulmonary resuscitation. METHODS: a multicenter, comparative and cross-sectional study was carried out to assess the learning of resuscitation of a group that was trained with the use of serious gaming with virtual reality, as compared to a control group trained with conventional classroom teaching. RESULTS: the mean quality obtained in chest compressions for the virtual reality group was 86.1 % (SD 9.3), and 74.8 % (SD 9.5) for the control group [mean difference 11.3 % (95 % CI 6.6-16.0), p < 0.001]. Salivary Alpha-Amylase was 218.882 (SD 177.621) IU/L for the virtual reality group and 155.190 (SD 116.746) IU/L for the control group [mean difference 63.691 (95 % CI 122.998-4.385), p = 0.037]. CONCLUSION: using virtual reality and serious games can improve the quality parameters of chest compressions as compared to traditional training.

Prediction of coarctation of the aorta in the second half of pregnancy.

OBJECTIVE: To determine which combination of cardiac parameters provides the best prediction of postnatal coarctation of the aorta (CoAo) in fetuses with cardiac asymmetry. METHODS: We selected all cases of disproportion of the ventricles and great vessels prenatally diagnosed between 2003 and 2010 at the Hospital Universitario '12 de Octubre', Madrid, Spain. Only appropriate-for-gestational age liveborn fetuses with isolated cardiac asymmetry and with complete postnatal follow-up were included in the study. Eighty-five cases were retrieved and analyzed. Logistic regression analysis was used to select the best predictors of CoAo. Optimal cut-offs for these parameters were identified and the corresponding likelihood ratios used to calculate the post-test probability of CoAo in each fetus. RESULTS: CoAo was confirmed in 41/85 neonates (48%). The parameters selected by logistic regression and their cut-off values were: gestational age at diagnosis ≤ 28 weeks, Z-score of diameter of the ascending aorta ≤ -1.5, pulmonary valve/aortic valve diameters ratio ≥ 1.6 and Z-score of the aortic isthmus diameter in the three vessels and trachea view ≤ -2. We divided the study group into two subgroups: Group A, in whom the diagnosis was made at ≤ 28 weeks' gestation (80% CoAo (32/40)); and Group B, in whom the diagnosis was made at > 28 weeks (20% CoAo (9/45)). The mean post-test probabilities of CoAo were higher in fetuses with CoAo than in normal fetuses in both subgroups (Group A, 82 vs 55%; P = 0.002 and Group B, 51 vs 20%; P < 0.001). In addition, a rate of growth of the aortic valve of ≤ 0.24 mm/week provided 80% sensitivity and 100% specificity for predicting CoAo in Group A. CONCLUSIONS: We have derived a multiparametric scoring system, combining size-based cardiac parameters and gestational age at diagnosis, which may improve the accuracy of fetal echocardiography for the stratification of the risk of CoAo. The objectivity and simplicity of its components may allow its implementation in fetal cardiology units.

Measurement of haptoglobin in saliva of cows: Validation of an assay and a pilot study of its potential application.

In recent years, the use of saliva as a matrix for the measurement of biomarkers of health and welfare is gaining importance due to its non-invasive collection. Haptoglobin (Hp) is an acute-phase protein involved in the inflammatory response and changes in its concentration can provide information about the health status of the animals. This study aimed to develop and validate an assay based on luminescent amplification (AlphaLISA technology) for the measurement of Hp in bovine saliva and to study the possible changes in different inflammatory situations such as peripartum period and lameness. The assay proved to be accurate, reliable, and sensitive for the measurement of Hp in cow saliva (coefficient of variation (CV) 7.57%; coefficient of determination (R2) 0.992; recovery test 105.15%; lower limit of quantification (LLQ) 7.9 ng/ml). Significant differences were observed between Hp levels in saliva of cows before (13 days before) and after (7 and 20 days after) calving and at the moment of calving (p < 0.0001), and between lame and healthy cows (p < 0.008). In conclusion, this assay can detect Hp in a precise, sensitive, and accurate way in saliva of cows. Future studies with a larger population and different disease conditions should be conducted to determine the potential of Hp as an inflammatory biomarker in cow saliva.

Predictive value of sequential models of uterine artery Doppler in pregnancies at high risk for pre-eclampsia.

OBJECTIVE: To evaluate the performance of models described previously for the prediction of pre-eclampsia (PE), based on the sequential evaluation of uterine artery resistance at 11-13 weeks and 19-22 weeks, in a high-risk population. METHODS: This was a prospective study in 135 women with singleton pregnancies and at least one of the following high-risk conditions: PE and/or intrauterine growth restriction in a previous pregnancy, chronic hypertension, pregestational diabetes, renal disease, body mass index > 30 kg/m(2) , autoimmune disease (systemic lupus erythematosus, antiphospholipid syndrome or rheumatoid arthritis) and thrombophilia. Mean uterine artery pulsatility index (mUtA-PI) at 11-13 and at 19-22 weeks' gestation was measured and analyzed according to quantitative and semi-quantitative models, to predict late PE (resulting in delivery ≥ 34 weeks) and early PE (delivery < 34 weeks). RESULTS: Late PE developed in 21 (15.6%) pregnancies and early PE in six (4.4%). Using mUtA-PI, the detection rates of late and early PE for a false-positive rate of 10% were 14.3% and 33.3%, respectively, at 11-13 weeks, and 19.0% and 66.7%, respectively, at 19-22 weeks. Using a semi-quantitative approach, the group of pregnant women with mUtA-PI ≥ 90(th) percentile at both 11-13 and 19-22 weeks had a greater risk for early PE (odds ratio, 21.4 (95% CI, 2.5-184.7)) compared with the group with mUtA-PI < 90(th) percentile at both periods. Using a quantitative approach, there was relative worsening in the mUtA-PI (multiples of the median) from the first to the second trimester in all cases of early PE. CONCLUSION: The application of semi-quantitative and especially quantitative models to evaluate sequential changes in uterine artery Doppler findings between the first and second trimesters could be of additional value in assessing high-risk women regarding their true risk of developing early PE.

Tetralogy of Fallot: prediction of outcome in the mid-second trimester of pregnancy.

OBJECTIVE: To evaluate the usefulness of fetal echocardiography in the mid-second trimester in predicting postnatal outcome of tetralogy of Fallot (ToF), focusing on the need for early intervention (EI) and surgery type: pulmonary valve-sparing surgery (PVSS) versus placement of transannular patch (TAP). METHODS: Assessment of cardiac morphological and functional parameters in 23 live-born fetuses with isolated ToF was performed at 19 to 22 and 34 to 38 weeks. Comparisons were made between outcome groups (EI vs non-EI and PVSS vs TAP). EI was considered as requirement either of palliative procedure or corrective surgery before three months. RESULTS: Overall survival was 96%. EI was required in 32% of cases and TAP in 50%. At 19 to 22 weeks, a pulmonary valve peak systolic velocity (PVPSV) ≥87.5 cm/s predicted EI with 100% sensitivity and 93.3% specificity (p < 0.01). At 34 to 38 weeks, the size of the pulmonary valve, pulmonary valve/aortic valve and main pulmonary artery/ascending aorta were significantly different, but the PVPSV again yielded the best performance: all cases undergoing EI and/or TAP were selected using cut-off of ≥144.5 cm/s. CONCLUSION: The postnatal outcome of fetuses with ToF may be established using PVPSV from the mid-second trimester. This may be useful in providing the most appropriate perinatal management and accurate parental counselling.

Changes in saliva analytes during pregnancy, farrowing and lactation in sows: A sialochemistry approach.

Salivary biomarkers were studied in 17 healthy Large White sows from early gestation to the end of lactation. Saliva samples were obtained at 34 ± 3 days from insemination (G30), 24 ± 4 days before farrowing (G90), within the first 24 h after farrowing (L1) and at the end of a lactation period of 21 days (L21). The measurements in saliva included stress-related biomarkers (cortisol, chromogranin A, α-amylase, butyrylcholinesterase [BChE] and lipase [Lip]), inflammatory biomarkers (adenosine deaminase isoenzymes 1 [ADA1] and 2 [ADA2], and haptoglobin [Hp]) and oxidative stress biomarkers (cupric reducing antioxidant capacity, trolox equivalent antioxidant capacity, ferric reducing ability, uric acid, advanced oxidation protein products [AOPP] and hydrogen peroxide [H2O2]), as well as routine biochemistry analytes (aspartate aminotransferase [AST], alkaline phosphatase [ALP], γ-glutamine transferase [GGT], lactate dehydrogenase [LDH], creatine kinase [CK], urea, creatinine, triglycerides, lactate, calcium and phosphorus). The main changes were observed at farrowing, with increases in biomarkers of stress (cortisol and BChE), inflammation (ADA isoenzymes and Hp) and oxidative stress (AOPP and H2O2), as well as muscle and hepatic enzymes (CK, AST, ALP, GGT and LDH). Lactate and triglycerides increased at the end of gestation and remained at high concentrations until the end of lactation. Lip was higher in gestation than at lactation. Thus, changes in biomarkers of stress, immune function, oxidative stress, hepatic and muscle integrity, and energy mobilization occur in sow saliva during pregnancy, farrowing and lactation. These changes, caused by physiological conditions, should be taken into consideration when these biomarkers are used for the evaluation of sow health and welfare.

Effect of reduction and alkylation treatment in three different assays used for the measurement of oxytocin in saliva of pigs.

Oxytocin is a hormone that is increasingly being used for welfare evaluation in animals. Although several types of samples have been used for oxytocin measurement, saliva can be a suitable option for pigs producing less stress than blood sampling. In this study, 3 different methods for oxytocin measurements, 2 based on alphaLISA technology (one with a monoclonal and other with a polyclonal antibody) and one commercially available kit, were compared in saliva of pigs. These methods were used in saliva samples obtained from female pigs at 3 different days during gestation and lactation, with and without a reduction/alkylation (R/A), which is a procedure for breaking the links between oxytocin and proteins of the sample. The assays showed a different behavior after the R/A procedure, with no significant changes in the oxytocin results in case of the alphaLISA monoclonal method, a significant decrease with the alphaLISA polyclonal method, and a significant increase with the commercial kit. Although all assays showed a similar tendency in detecting the changes in oxytocin during gestation and lactation, they showed changes of different magnitude and statistical signification. This report indicates that different assays can measure different forms of oxytocin present in saliva and can have a different behavior after R/A of the sample and when are used to measure oxytocin in gestation and lactation.

Changes in saliva proteins in cows with mastitis: A proteomic approach.

This study aimed to evaluate the possible saliva proteome changes in cows with mastitis using a Tandem Mass Tags (TMT) proteomics approach. For this purpose, the salivary proteomes from healthy cows and cows with mastitis were analysed, and their serum proteomes were also studied for comparative purposes. A total of eight saliva and serum paired samples for each group were used for the proteomic study, and eight additional samples for each group were analysed in the analytical and overlap performance studies. In saliva samples, 2192 proteins were identified, being sixty-three differentially modulated in mastitis. In serum, 1299 proteins were identified, being twenty-nine differentially modulated in mastitis. Gamma glutamyl transferase (γGT) in saliva and serum amyloid A (SAA) were validated by commercially available automated assays. In conclusion, there are changes in protein expression and metabolic pathways in saliva and serum proteomes of cows with mastitis, showing different response patterns but complementary information.

Salivary D-dimer in pigs: Validation of an automated assay and changes after acute stress.

D-dimer is a peptide found in serum and is derived from the degradation of blood clots. Even though it has been analysed in human saliva, D-dimer has not been previously evaluated in the saliva of any veterinary species, and its source and role remain unknown. The objectives of this research were firstly, to validate the use of an automated method for the measurement of D-dimer in porcine saliva, and secondly, to evaluate whether D-dimer concentration changes in pig saliva after an acute stress stimulus. For this purpose, a complete analytical validation of a commercially-available immunoturbidimetric assay was carried out. In addition, an experimental acute stress model was induced in 11 pigs based on a technique involving restraint by nose-snare immobilisation for 1 min. Saliva samples were subsequently collected at different times and D-dimer, salivary alpha-amylase (sAA) and cortisol were assessed in order to evaluate changes in its concentrations after the stress induction. The D-dimer automated assay showed adequate reproducibility and sensitivity, with coefficients of variation below 10% and a limit of quantification of 0.167 µg/mL fibrinogen equivalent units (FEU). It also showed a high accuracy, determined by linearity under dilution and recovery tests. In the stress model, a significant increase (P < 0.05) in salivary D-dimer 15 min after the stress stimulus and a positive correlation between D-dimer and sAA (r = 0.51; P < 0.001) were observed. These results indicate that D-dimer can be measured in porcine saliva with an automated method and suggest that its concentration can be influenced by stressful conditions.

Oxytocin in saliva of pigs: an assay for its measurement and changes after farrowing.

Oxytocin is a hormone of interest in reproduction, but also in the field of psychology and behavior, being considered as a biomarker of positive emotions. Saliva can be a noninvasive way to measure oxytocin, which is very useful in species such as the pig where blood collection can produce a high degree of stress. In this study, a new assay for oxytocin measurement was developed, analytically validated, and used to measure possible changes in oxytocin in saliva of female pigs at different days after farrowing. The assay showed an adequate accuracy and precision and does not need a previous extraction step. In addition, oxytocin concentrations were significantly higher at day 1 of lactation than at day 9 after farrowing, but levels increased at day 20 again. This assay can contribute to a wider use of oxytocin measurements in pigs as it is a noninvasive sampling procedure that minimizes stress.

SuperCam Calibration Targets: Design and Development.

SuperCam is a highly integrated remote-sensing instrumental suite for NASA's Mars 2020 mission. It consists of a co-aligned combination of Laser-Induced Breakdown Spectroscopy (LIBS), Time-Resolved Raman and Luminescence (TRR/L), Visible and Infrared Spectroscopy (VISIR), together with sound recording (MIC) and high-magnification imaging techniques (RMI). They provide information on the mineralogy, geochemistry and mineral context around the Perseverance Rover. The calibration of this complex suite is a major challenge. Not only does each technique require its own standards or references, their combination also introduces new requirements to obtain optimal scientific output. Elemental composition, molecular vibrational features, fluorescence, morphology and texture provide a full picture of the sample with spectral information that needs to be co-aligned, correlated, and individually calibrated. The resulting hardware includes different kinds of targets, each one covering different needs of the instrument. Standards for imaging calibration, geological samples for mineral identification and chemometric calculations or spectral references to calibrate and evaluate the health of the instrument, are all included in the SuperCam Calibration Target (SCCT). The system also includes a specifically designed assembly in which the samples are mounted. This hardware allows the targets to survive the harsh environmental conditions of the launch, cruise, landing and operation on Mars during the whole mission. Here we summarize the design, development, integration, verification and functional testing of the SCCT. This work includes some key results obtained to verify the scientific outcome of the SuperCam system.

Role of uterine artery Doppler in interpreting low PAPP-A values in first-trimester screening for Down syndrome in pregnancies at high risk of impaired placentation.

OBJECTIVES: Low maternal serum levels of pregnancy-associated plasma protein-A (PAPP-A) are associated with both increased risk of aneuploidies and impaired trophoblastic invasion, while high uterine artery (UtA) resistance is associated with impaired trophoblastic invasion but not with an increased risk of aneuploidies. The aim of this study was to determine whether high UtA resistance plays a role in explaining low PAPP-A levels in the absence of aneuploidies. METHODS: This was a prospective study of 116 singleton pregnancies at high risk for impaired placentation (having at least one major risk factor: prior history of pre-eclampsia, pregestational diabetes mellitus, chronic hypertension, chronic kidney disease, body mass index >30, autoimmune disorder, thrombophilia or recurrent pregnancy loss), booked for routine assessment of risk for aneuploidies by means of the first-trimester combined screening test (nuchal translucency thickness (NT) + PAPP-A + beta-human chorionic gonadotropin (beta-hCG)). Measurement of NT and the mean UtA pulsatility index (PI) were carried out at the 11 to 13 + 6-week scan. All values were calculated in multiples of the median (MoM) adjusted for gestational age. A cut-off risk of 1/270 at time of sampling was adopted to differentiate high- from low-risk groups for trisomy 21. RESULTS: There were 108 patients deemed to be at low risk for trisomy 21 and eight at high risk. None had chromosomal defects, giving a false-positive rate for trisomy 21 of 6.9%. The greatest differences between patients at low risk and those at high risk for trisomy 21 were found in their PAPP-A (0.98 vs. 0.38 MoM, P < 0.01) and beta-hCG (1.09 vs. 1.77 MoM, P = 0.04) values. Greater NT thickness (1.02 vs. 0.90 MoM) and higher mean UtA-PI (1.05 vs. 0.96 MoM) were recorded in the high-risk group, although the differences did not reach statistical significance (P = 0.19 and 0.40, respectively). After log-transformation there were no significant correlations between mean UtA-PI and NT and between mean UtA-PI and beta-hCG. There was a significant negative linear correlation between mean UtA-PI and PAPP-A (r = -0.331; P < 0.01). After adjusting the PAPP-A values by UtA-PI, the false-positive rate for trisomy 21 decreased to 2.6%. CONCLUSION: Mean UtA-PI at the 11 to 13 + 6-week scan may be an effect-modifier variable for PAPP-A that should be taken into account in the first-trimester combined screening for aneuploidies, at least in pregnancies at high risk for impaired placentation.

Prenatal diagnosis of right aortic arch: associated findings, pregnancy outcome, and clinical significance of vascular rings.

OBJECTIVES: To analyze the characteristics and clinical implications of right aortic arch (RAA) detected in fetal life. METHODS: Retrospective study of all cases of RAA diagnosed prenatally in high-risk patients who underwent fetal echocardiography between 2000 and 2007. RESULTS: There were 48 RAA; 18 had vascular ring (Group 1), including 15 RAA with aberrant left subclavian artery and 3 double aortic arch, and 30 had not vascular ring (Group 2), all RAA with mirror-image branching. The prenatal diagnosis was confirmed in 94%. In Group 1 most fetuses had normal heart (89%), and none had 22q11 deletion. There were 16 live births and all infants but one are asymptomatic (mean follow-up of 31 months). In Group 2 almost all fetuses had congenital heart defects (CHDs) (97%), and five were chromosomally abnormal (17%), including four 22q11 deleted. The 1-year survival rate was 69%, which was significantly higher in Group 1 (89% vs 57%, p < 0.05). CONCLUSIONS: RAA can be accurately diagnosed by fetal echocardiography. The outlook for these patients largely depends on the presence of associated defects, mainly CHDs, and their severity. If isolated, aortic arch anomalies are asymptomatic vascular variants in most cases. This information is important to assist parental counseling in pregnancy.

Glucose, fructosamine, and insulin measurements in saliva of dogs: variations after an experimental glucose administration.

The aim of this study was to evaluate if glucose, fructosamine, and insulin levels can be measured in saliva of dogs and assess the changes in these compounds after an experimental glucose administration. Automated spectrophotometric assays for glucose and fructosamine and an ELISA assay for insulin measurements were validated in saliva of dogs, by evaluating precision, accuracy, and limits of detection. In addition, an intravenous glucose bolus was administrated to 10 beagles and fasting serum and saliva samples were obtained immediately before and 5, 10, 20, 30, and 45 min after glucose infusion. The results of the between-run imprecision gave mean CVs of 6.16, 9.40, and 3.10% for glucose, fructosamine, and insulin, respectively. Linearity under dilution showed coefficient of correlation of 0.999, 0.994, and 0.990 for glucose, fructosamine, and insulin, respectively. The LDs were 0.04 mg/dL, 4.08 µmol/L, and 0.02 µg/mL for glucose, fructosamine, and insulin, respectively. The glucose administration caused an increase in serum and salivary levels of glucose with a peak in salivary levels at 30 min and of insulin with a peak in salivary levels at 45 min, while fructosamine did not change. No correlations between serum and salivary concentrations were found for any compound. It is concluded that glucose, fructosamine, and insulin can be measured in saliva of dogs, and an experimental administration of glucose in this species can lead to increases in glucose and insulin in saliva.

Salivary alpha-amylase activity and concentration in horses with acute abdominal disease: Association with outcome.

BACKGROUND: Salivary biomarkers could be useful to objectively evaluate critical illness and prognosis for survival in horses with acute abdominal disease. OBJECTIVES: To compare salivary alpha-amylase (sAA) activity and concentration in healthy horses and horses with acute abdominal disease, and evaluate the association between sAA activity and concentration with disease severity and outcome. STUDY DESIGN: A prospective cohort. METHODS: sAA activity, measured using a colorimetric commercial kit, and concentration, measured using a Time-resolved immunofluorometric assay, in 25 healthy horses and in 33 horses with acute abdominal disease was compared using an ANOVA. Associations between survival to discharge and sAA activity and concentration and other clinical parameters were examined using univariable logistic regression and Spearman correlation. RESULTS: sAA activity and concentration were different between healthy (median = 4.3 [2.6-11.2] IU/L and 58.4 [53.4-80.6] ng/mL, respectively) and diseased (median = 29.8 [14.2-168.9] IU/L and 388.3 [189.1-675.8] ng/mL, respectively) (P<0.001). The sAA activity was higher in non-survivors (median = 479.0 [78.7-2064.0] IU/L, n = 8) compared to survivors (median = 19.3 [12.1-103.7] IU/L, n = 25, P<0.001) and sAA activity and concentration correlated (P<0.001) moderately with HR (r = 0.66 and r = 0.61, respectively). sAA activity correlated weakly with salivary cortisol (r = 0.45, P<0.001) and systemic inflammatory response syndrome score (r = 0.43, P<0.05), while activity and concentration correlated (P<0.001) moderately with plasma lactate concentration (r = 0.57 and r = 0.60, respectively). The sAA activity was significantly (P = 0.01) associated with increased risk of nonsurvival. MAIN LIMITATIONS: Pain scores were not recorded. The sample population was small. CONCLUSIONS: The sAA activity, but not concentration, shows potential as a biomarker of prognosis for survival in horses with acute abdominal disease. The summary is available in Spanish - see Supporting Information.

Evaluation of new biomarkers of stress in saliva of sheep.

Some routine handling procedures can produce stress in farm animals, and an adequate control of these stressors is important to avoid the negative effects on animal health and production. The measurement of biomarkers in saliva can be a suitable tool for the evaluation and control of stress. In this report, lipase, butyrylcholinesterase (BChE), total esterase (TEA) and adenosine deaminase (ADA) activities in the saliva of sheep were evaluated as biomarkers of stress. For this purpose, they were measured after inducing stress by facing a dog (experiment 1) and shearing (experiment 2), and comparing them to other stress salivary biomarkers such as α-amylase (sAA) and cortisol, as well as heart rate (HR). Each analyte was measured at the basal time, and during and just after the end of the stressful stimulus, and at various times for the first hour after the period of stress induction. Values were compared with those obtained from a control group. Lipase was the only analyte that showed significant changes between the stress and the control group in both experiments. Although TEA and ADA increased after stress, no significant differences were seen compared with the control group. Lipase was correlated highly with sAA and HR, in experiment 1; and correlated moderately with cortisol and HR in experiment 2. Lipase showed the greatest percentage increase after the stressful stimuli and less overlap with the control group in the two experiments. From the results of this study it can be concluded that lipase, TEA, BChE and ADA are enzymes present in the saliva of sheep and that they can be measured by using simple and fast colorimetric methods. Further studies should be undertaken with regard to the possible application of lipase as a biomarker of stress in sheep.

Evaluation of adenosine deaminase in saliva and serum, and salivary α-amylase, in canine pyometra at diagnosis and after ovariohysterectomy.

An assay for adenosine deaminase (ADA) was validated in serum and saliva in dogs. Changes in ADA and salivary α-amylase activities were analysed in 26 bitches diagnosed with pyometra and compared with activities in 19 healthy bitches. All animals were classified according to the American Society of Anaesthesiologists (ASA) scoring for physical status. In the validation study, the ADA assay had an imprecision<12% and determination coefficients>0.90 in linearity under dilution experiments, with recoveries of 99.2-114.4%. On the day of presentation, salivary ADA activity was significantly higher in dogs with pyometra than in healthy dogs (median values 7.1IU/L vs. 0.8IU/L, respectively; P<0.01). ADA had a moderate positive correlation with leucocyte and band neutrophil counts, haptoglobin, salivary α-amylase and ASA score, and a low positive correlation with C-reactive protein. There were no significant differences in salivary α-amylase activity between dogs with pyometra and healthy dogs (57.3IU/L vs. 27.4IU/L, respectively). Salivary α-amylase had a low correlation with ASA grade, and leucocyte and band neutrophil counts. In 7/26 bitches with pyometra that were sampled 3 and 10days after ovariohysterectomy, there were no significant changes in α-amylase or ADA activities. These results indicate that ADA activity is increased in the saliva of bitches with pyometra, probably related to systemic inflammation.