Analytical characterization of trace elements (zinc, copper, cadmium, lead and selenium) in saliva of pigs under common pathological conditions in the field: a pilot study.

BACKGROUND: This study is focused on the measurement of trace elements (zinc, copper, cadmium, lead and selenium) in the saliva of pigs in order to study their levels on different porcine pathological conditions in the field. The experiment involved 15 pigs without clinical signs of disease and 42 diseased pigs (suffering from lameness, rectal prolapse, fatigue or growth rate retardation). Individual saliva samples were collected, allowing the pigs to chew a sponge each for trace element quantifications through atomic absorption spectrometry (AAS). Since this is the first report on the measurements of trace elements in porcine saliva, a routine analytical validation study was performed for the quantification of all the studied elements. Moreover, the acute phase proteins C-reactive protein (CRP) and haptoblobin (Hp), the total antioxidant capacity (TAC) and adenosine deaminase (ADA) were quantified in the saliva samples for the animal's health status assessment. RESULTS: Modifications in the levels of acute phase proteins or ADA were only recorded in animals with lameness and rectal prolapse and those with fatigue respectively. Moreover, TAC level changes were observed in pigs with growth-rate retardation. However, alterations in the levels of two or more trace elements were reported for all the different groups of diseased pigs with evident variations within pathologies. CONCLUSIONS: The salivary quantification of trace elements could be considered as a complementary tool to acute phase proteins, TAC and ADA determinations for disease detection and differentiation in the pig and should be explored in greater depth.

Gender influence on the salivary protein profile of finishing pigs.

A study on gender differences in the normal range of biomarkers in porcine saliva samples has the scope for further attention. In the present study, the salivary protein profiles of age-matched healthy male and female finishing pigs were compared. The levels of salivary adenosine deaminase (ADA) activity, haptoglobin (Hp) and C-reactive protein (CRP) have been quantified in 32 male and 32 female pigs to ensure the presence of gender effect on the median levels of salivary biomarkers. Moreover, the total salivary protein content was quantified and compared. The overall salivary protein distribution was compared with SDS-PAGE in 14 male and 14 female pigs and the possible gender influence in the salivary protein profile was analysed by 2DE in 6 male and 6 female pigs. Statistically significant differences were observed in the median values of Hp, CRP, and ADA between male and female pigs (p<0.005). Although the total salivary protein content was not different between the sexes, the salivary protein distribution and profile showed specific gender differences in three proteins of the lipocalin family: the odorant-binding protein, salivary lipocalin and lipocalin 1. These proteins have been related to animal immune status and should be further explored as possible porcine salivary biomarkers. SIGNIFICANCE: The biological relevance of the reported research is based on the possible gender influence on the discovery of salivary biomarkers in porcine production. As differences have been reported in the salivary protein distribution in male pigs in comparison to that of female pigs, the normal-range values, according to gender, of the newly discovered biomarkers should be explored and defined prior to its application in the porcine production system. A hormonal sexual influence is highly hypothesized.

Easy and non-invasive disease detection in pigs by adenosine deaminase activity determinations in saliva.

The quantification of adenosine deaminase (ADA) in porcine saliva samples has been analyzed for its use as a marker of disease. First, an analytical validation of the enzymatic assay used for ADA measurements was performed. Afterwards, saliva samples were collected from 50 healthy animals and 64 animals with different symptoms of disease, which were divided into local inflammation, gastrointestinal disorder, respiratory disorder and growth retardation. To optimize ADA measurements, total ADA (tADA), specific ADA (sADA) and ADA isoforms 1 and 2 activities were calculated. Moreover, to preliminarily estimate the diagnostic value of tADA activity measurements for disease detection, receiver operating characteristic (ROC) analyses was performed and compared to the results obtained for salivary acute phase proteins, haptoglobin (Hp) and C-reactive protein (CRP). The salivary levels of tADA activity were significantly elevated in animals with local inflammation, gastrointestinal disorder and respiratory disorder. The calculation of the different ADA activities did not provide additional information to tADA activity quantification for disease detection. The diagnostic value of tADA activity was superior to those observed for Hp and CRP measurements in the present study. It might be concluded that ADA analysis in saliva could be used as a simple, rapid, economic and non-invasive diagnostic tool in porcine production in field conditions.

Cortisol, adrenocorticotropic hormone, serotonin, adrenaline and noradrenaline serum concentrations in relation to disease and stress in the horse.

No detailed comparative data are available on the hormonal parameters of horses suffering from a number of diseases. The aim of our study was to measure concentrations of cortisol, adrenocorticotropic hormone (ACTH), serotonin, adrenaline and noradrenaline in horses with various diseases and following surgery, to assess the response of the HPA axis and adrenal medulla. Blood samples were obtained from six groups of horses comprising a total of 119 animals as follows: laminitis, acute abdominal syndrome (AAS), castration surgery, acute diseases, chronic diseases and healthy controls. Serum hormonal concentrations were determined for each group for comparison. Statistically significant differences between all groups and controls were found for cortisol, ACTH (except for castration), serotonin and adrenaline concentrations but only in horses with laminitis and AAS for noradrenaline. No statistically significant differences were found between males and females. The largest changes in the pituitary-adrenal axis activity occurred mainly in acute diseases, laminitis and in the AAS group.

Effects of different variables on whole blood cholinesterase analysis in dogs.

The influence of several variables such as sample and reagent storage, anticoagulants, reaction temperature, pH, and substrate concentration on whole blood cholinesterase determination was studied. Storage of nondiluted whole blood samples at room temperature or under refrigeration (4 C) was adequate for short-term storage (3 days to 2 weeks). However, freezing would be more appropriate for long-term storage (> or = 1 month), and successive thawing and freezing did not produce any loss of cholinesterase activity. All reagents (2,2'-dithiodipyridine as chromophore and acetylthiocholine and butyrylthiocholine as substrates) were stable for 3 months when frozen. Heparin and ethylenediaminetetraacetic acid were the most suitable anticoagulants for whole blood acetylcholinesterase and butyrylcholinesterase determination, because citrate yielded lower acetylcholinesterase values and fluoride inhibited butyrylcholinesterase. Increases in reaction temperature and pH yielded higher cholinesterase values but also increased nonenzymatic substrate hydrolysis. Higher cholinesterase and nonenzymatic substrate hydrolysis values were obtained as higher substrate concentrations were used.