Comparison of the Effect of Two Different Handling Conditions at Slaughter in Saliva Analytes in Pigs.

In this report, different handling conditions at slaughterhouse were studied to assess changes in salivary biomarkers. For this purpose, finishing pigs were divided into two groups, one in which handling was improved to minimize stress (Group A, n = 24, transported and stabled at the slaughterhouse at low density without mixing with unfamiliar animals throughout the whole process) and another one in which animals had a more stressful handling process (Group B, n = 24, transported and stabled at high density with unfamiliar animals). Saliva samples were taken the day before transport to the slaughterhouse at 8:00 a.m. (B0) and 12:00 a.m. (B4), and the day of slaughter just after unloading animals at the slaughterhouse at approximately 8:00 a.m. (S0) and after 4 h of lairage at approximately 12:00 a.m. (S4). Group B showed significantly higher cortisol, total esterase activity, oxytocin, adenosine deaminase and haptoglobin levels than the Group A at both S0 and S4 sampling times, and higher levels of calprotectin and creatine kinase at S4 sampling time. This report indicates that differences in the way in which the pigs are handled at the slaughterhouse can lead to changes in salivary biomarkers and opens the possibility of the use of biomarker at slaughter to monitor handling conditions.

Saliva Sampling Material Matters: Effects on the Results of Saliva Analysis in Pigs.

The use of saliva as a biological sample from pigs is of high practical interest because blood collection from pigs is difficult and stressful. In this study, the influence of two different materials, a cotton roll and a polypropylene sponge, in porcine saliva collection was evaluated. For this purpose, the effect of the material used for sampling was evaluated in a panel of 13 analytes, including those related to stress (cortisol and oxytocin), inflammation and immunity (adenosine deaminase, haptoglobin and myeloperoxidase), redox homeostasis (the cupric reducing ability of saliva, the ferric reducing activity of saliva, and the Trolox equivalent antioxidant capacity), and sepsis (procalcitonin), as well as other routine analytes related to metabolism and different tissues and organs, such as lactate dehydrogenase, creatine kinase, urea, and total protein concentration. The polypropylene sponge provided a higher sample volume than the cotton roll. Although the results of some salivary analytes were equivalent for both materials, other analytes, such as creatine kinase, haptoglobin and total proteins, showed significant differences depending on the material used for saliva collection. Therefore, the type of material used for salivary collection in pigs should be considered when interpreting the results of analyses of the salivary analytes.

Gaining knowledge about biomarkers of the immune system and inflammation in the saliva of pigs: The case of myeloperoxidase, S100A12, and ITIH4.

An assay for the measurement of myeloperoxidase (Mpx) in porcine saliva was developed and validated, and factors influencing Mpx and another two biomarkers of inflammation and immune system, the protein S100A12 and the inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), were studied. The spectrophotometric method for Mpx measurement validated in this assay showed an adequate analytical performance including precision and accuracy. When a group of twenty healthy pigs was sampled every 4 h from 8 a.m. until 8 p.m., Mpx and S100A12 showed significant increases at 4 p.m., whereas ITIH4 concentration showed a significant decrease at 12 a.m. Increases were also seen in salivary Mpx, S100A12, and ITIH4 levels 24 h after the intramuscular administration of Escherichia coli lipopolysaccharide in five pigs; whereas in a non-septic inflammation after the subcutaneous administration of turpentine oil to five pigs changes were seen in S100A12 at 3 h and in ITIH4 at 48 h. When a stressful situation consisting of the transportation and stay of 4 h to a slaughterhouse of 24 pigs was performed, all analytes were increased after 4 h of lairage in the slaughterhouse compared with the values that were obtained the day before at the same time of the day. Mpx can be measured in the saliva of pigs with the automated assay described in this report. Mpx, S100A12, and ITIH4 salivary levels can change depending on the hour of the day in which the sample is taken, and increases can be produced due to sepsis, non-septic inflammation and stress.

Changes in S100A8/A9 and S100A12 and Their Comparison with Other Analytes in the Saliva of Pigs with Diarrhea Due to E. coli.

The family of calgranulins includes S100A8 (calgranulin A), S100A9 (calgranulin B), which can appear as a heterodimer known as S100A8/A9 or calprotectin, and S100A12 (calgranulin C). These proteins are related to different inflammatory conditions, immune-mediated diseases, and sepsis and are considered biomarkers of potential interest. This study aims to evaluate if S100A8/A9 and A12 could change in pigs with diarrhea due to E. coli and to compare the changes of S100A8/A9 and A12 with other analytes in order to explore the possible causes or mechanisms involved. For this purpose, a panel integrated by analytes related to inflammation (haptoglobin, inter-alpha trypsin inhibitor 4 (ITIH4), and total protein); immune system (adenosine deaminase, ADA); stress (alpha-amylase); tissue damage (lactate and lactate dehydrogenase (LDH)); sepsis (aldolase) and redox status (ferric-reducing ability of saliva (FRAS) and advanced oxidation protein products (AOPP)) was evaluated. S100A8/A9 and A12 and the other analytes measured in this study showed increases in the saliva of pigs with diarrhea due to E. coli. S100A8/A9 and/or A12 showed a significant correlation of different magnitude with some of the other analytes evaluated. Further studies should be conducted to gain knowledge about the possible practical applications as biomarkers of the measurements of S100A8/A9 and A12 in the saliva of pigs.

Oxytocin measurements in saliva: an analytical perspective.

Oxytocin has traditionally been known for its physiological effects on muscle contraction associated with birth and lactation, but in the last years is widely used as a biomarker of "positive experiences" in psychology and behavior. Different types of samples have been used for oxytocin measurements with saliva samples having the particular advantage of an easy and non-stressful collection. However, the low concentration of oxytocin in saliva can represent a limitation for its use. For this reason, sensitive assays and even a previous sample treatment in some cases are required for saliva oxytocin quantification. In addition, the lack of standardized and generally agreed-upon approach to peripheral oxytocin measurement leads to large discrepancies between different laboratories, that use different sample treatment protocols and different assays. The main objectives of this review are to describe the current status of the use of saliva for oxytocin measurement, provide details of the different sample processing techniques that can be applied and inform about the analytical techniques and assays available in different animal species, and also in humans for comparative purposes. It is expected that this information can contribute to an increase in the knowledge about the measurements of oxytocin in saliva and to its wider use in the future.

Different Types of Glucocorticoids to Evaluate Stress and Welfare in Animals and Humans: General Concepts and Examples of Combined Use.

The main glucocorticoids involved in the stress response are cortisol and cortisone in most mammals and corticosterone in birds and rodents. Therefore, these analytes are currently the biomarkers more frequently used to evaluate the physiological response to a stressful situation. In addition, "total glucocorticoids", which refers to the quantification of various glucocorticoids by immunoassays showing cross-reactivity with different types of glucocorticoids or related metabolites, can be measured. In this review, we describe the characteristics of the main glucocorticoids used to assess stress, as well as the main techniques and samples used for their quantification. In addition, we analyse the studies where at least two of the main glucocorticoids were measured in combination. Overall, this review points out the different behaviours of the main glucocorticoids, depending on the animal species and stressful stimuli, and shows the potential advantages that the measurement of at least two different glucocorticoid types can have for evaluating welfare.

Evaluation of the Effect of Sampling Time on Biomarkers of Stress, Immune System, Redox Status and Other Biochemistry Analytes in Saliva of Finishing Pigs.

This study aims to evaluate the possible variations due to the sampling time in the day in 26 analytes of pigs' saliva, related to stress, the immune system, redox status and other biomarkers related to metabolism and selected tissues and organs, in order to know the possible effects of the hour of the day in their interpretation. These analytes were measured in saliva obtained from a population of 40 clinically healthy pigs from 8 a.m. to 8 p.m., every 4 h in the same day. In our experimental conditions, daily variations were observed in cortisol, salivary α-amylase, total esterase activity, butyrylcholinesterase, lipase, adenosine deaminase isoenzyme 1, uric acid, superoxide dismutase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatine kinase, lactate dehydrogenase, lactate and triglycerides. These changes appeared in both sexes, except for adenosine deaminase isoenzyme 1 and superoxide dismutase which only showed differences in females. In conclusion, this report indicates that, in the experimental conditions of this trial, the time of the day and sex can influence the values obtained in various salivary analytes in pigs. These variations should thus be taken into consideration for an adequate interpretation of these analytes when used for the evaluation of health and welfare in this species.

Changes in a Comprehensive Profile of Saliva Analytes in Fattening Pigs during a Complete Productive Cycle: A Longitudinal Study.

A comprehensive panel of 29 salivary analytes was measured in fattening pigs to evaluate its possible changes along their productive cycle. The identification of those changes would allow a better interpretation of the results according to the productive phase of the animal. Saliva samples were obtained from 49 Large-White pigs (24 females, 25 males) in suckling phase, at the beginning and the end of the nursery phase, and at the beginning and the end of the growing phase. Several analytes changed according to the phase of the productive cycle, with most of the analytes showing higher values at lactation and at the beginning of nursery. Additionally, differences were seen due to sex. When possible relations between performance parameters and analytes were evaluated, significant positive but weak relationships were found between weight at birth and salivary γ-glutamyl transferase, and between back-fat thickness and salivary lactate dehydrogenase. In conclusion, differences in the values of salivary analytes can be found in fattening pigs depending on the productive phase and sex of the animals.

Pharmacokinetics of Tildipirosin in Plasma, Milk, and Somatic Cells Following Intravenous, Intramuscular, and Subcutaneous Administration in Dairy Goats.

Tildipirosin is a macrolide currently authorized for treating respiratory diseases in cattle and swine. The disposition kinetics of tildipirosin in plasma, milk, and somatic cells were investigated in dairy goats. Tildipirosin was administered at a single dose of 2 mg/kg by intravenous (IV) and 4 mg/kg by intramuscular (IM) and subcutaneous (SC) routes. Concentrations of tildipirosin were determined by an HPLC method with UV detection. Pharmacokinetic parameters were estimated by non-compartmental analysis. Muscle damage, cardiotoxicity, and inflammation were evaluated. After IV administration, the apparent volume of distribution in the steady state was 7.2 L/kg and clearance 0.64 L/h/kg. Plasma and milk half-lives were 6.2 and 58.3 h, respectively, indicating nine times longer persistence of tildipirosin in milk than in plasma. Moreover, if somatic cells are considered, persistence and exposure measured by the area under concentration-time curve (AUC) significantly exceeded those obtained in plasma. Similarly, longer half-lives in whole milk and somatic cells compared to plasma were observed after IM and SC administration. No adverse effects were observed. In brief, tildipirosin should be reserved for cases where other suitable antibiotics have been unsuccessful, discarding milk production of treated animals for at least 45 days or treating goats at the dry-off period.

Somatic Support with Veno-venous ECMO in a Pregnant Woman with Brain Death: A Case Report.

Supporting homeostasis in a pregnant woman with brain death to achieve fetal viability is called somatic support. We present a case of young pregnant woman at 21 weeks' gestation who developed acute respiratory distress syndrome secondary to influenza A H2N3 infection requiring veno-venous extracorporeal membrane oxygenation (VV ECMO) support for refractory hypoxemia. The clinical course was complicated by intracranial hemorrhage and subsequent brain death. After multidisciplinary team discussion with her family, consensus was reached to continue somatic support with VV ECMO to enable fetal development to attain extrauterine viability. The challenging clinical, ethical, and legal concerns are discussed.

Experience with veno-venous ecmo in patients with adult respiratory distress syndrome secondary to viral pneumonia. Case series / Experiencia con ecmo veno-venoso en pacientes con síndrome de dificultad respiratoria del adulto secundario a neumonía viral. Serie de casos

Abstract Introduction The use of extracorporeal membrane oxygenation (ECMO) has increased exponentially in recent years and has shown to be effective in treating adult respiratory distress syndrome (ARDS) secondary to HiNi-related pneumonia. However, evidence remains controversial. This study describes a case series of ECMO in ARDS secondary to viral pneumonia. Methods A search was conducted in the ECMO database of Fundación Cardiovascular de Colombia for the 20132017 period. A case series report was written of patients diagnosed with ARDS secondary to confirmed or suspected viral pneumonia. Results Nineteen patients with ECMO support and ARDS due to viral pneumonia were included in the study. The survival rate upon discharge was 11 patients (58%) and weaning from ECMO support was successful in 13 patients (68%). Hemorrhagic complications were the most frequent: gastrointestinal bleeding, 10 patients (53%); intracranial bleeding, 2 (10%); alveolar hemorrhage, 2 (10%);' hemothorax requiring thoracostomy with chest tube drainage, 2 (10%); cannulation site bleeding, 9 patients (47%); and surgical site bleeding in 3 patients (25%) who required tracheostomy. Other complications were: pneumothorax, 1 patient (5%); sepsis, 6 patients (32%); and growth of microorganisms in bronchial lavage, 6 patients (32%). Conclusions This study supports the use of veno-venous ECMO to achieve a higher survival rate than expected in patients with severe ARDS and refractory hypoxemia secondary to viral pneumonia. Early initiation of the therapy should improve overall results.

Resumen Introducción El uso de la oxigenación por membrana extracorpórea (ECMO) ha tenido un incremento exponencial en los últimos años y ha demostrado ser efectivo en el manejo del síndrome de dificultad respiratoria del adulto (SDRA) secundario a neumonía por H1N1, si bien la evidencia sigue siendo controvertida. En este estudio describimos una serie de casos de ECMO por SDRA secundario a neumonía viral. Métodos Se realizó una búsqueda en la base de datos de ECMO de la Fundación Cardiovascular desde el año 20132017. Reportamos una serie de casos donde se incluyeron pacientes diagnosticados con SDRA secundario a neumonía viral sospechosa o confirmada. Resultados Se incluyeron en el estudio 19 pacientes con soporte de ECMO y SDRA por neumonía viral. La sobrevida al alta fue 11 pacientes (58%) y el destete del ECMO fue exitoso en 13 pacientes (68%). Las complicaciones hemorrágicas presentadas fueron: sangrado digestivo, 10 pacientes (53%), sangrado cerebral, 2 (10%), hemorragia alveolar, 2 (10%), hemotórax con requerimiento de toracostomía a drenaje cerrado, 2 (10%), sangrado activo por sitio de canulación, 9 pacientes (53%), y 3 pacientes traqueostomizados (25%) que sangraron por el sitio quirúrgico. Otras complicaciones presentadas fueron: neumotórax, 1 paciente (5%), septicemia, 6 (32%) y crecimiento de microorganismos en lavados bronquiales 6 (32%). Conclusion El presente estudio permite indicar que el uso de la ECMO VV viabiliza una sobrevida mayor a la esperada en pacientes con SDRA severo e hipoxemia refractaria secundario a neumonía viral. Su inicio tempranamente debe mejorar los resultados globales.

A Comprehensive Study on the Occurrence of Mycotoxins and Their Producing Fungi during the Maize Production Cycle in Spain.

Mycotoxin contamination is one of the main problems affecting corn production, due to its significant risk to human and animal health. The Fusarium and Aspergillus species are the main producers of mycotoxins in maize, infecting both pre-harvest and during storage. In this work, we evaluated the presence of mycotoxins and their producing species along maize production cycles in three different stages (anthesis, harvest, and storage) during three consecutive seasons (2016-2018). Fungal occurrences were studied using species-specific PCR protocols, whereas mycotoxin levels were determined by LC-MS/MS. Fumonisin-producing Fusarium species (F. verticillioides and F. proliferatum), as well as the aflatoxin producer Aspergillus flavus, were the most predominant species at all stages; although, during some seasons, the presence of F. graminearum and A. niger aggregate species were also identified. Contrastingly, fumonisins were the only mycotoxins detected and levels were always under legal regulations. The results presented here demonstrate that even when fungal contamination occurs at the early stages of the maize production cycle, the application of good agricultural and storage practices might be crucial to ensure mycotoxin-free grains.

SCHeMA EU Project Summer School Report (Bilbao June 16 - 17, 2016).

This conference report describes the training activities that took place in the frame of the Integrated in Situ Chemical MApping probe (SCHeMA) summer school organized from the 14th to the 16th of June 2016 in Bilbao (Spain).

Effects of foliar sprays containing calcium, magnesium and titanium on plum (Prunus domestica L.) fruit quality.

An experiment was performed in which Ti(4+)-ascorbate was sprayed onto plum trees in several combinations with other commercial compounds containing Ca2+ and Mg2+ to study the effects on the commercial quality of fruits, with special focus on improving their resistance against postharvest handling damage. All the treatments containing titanium increased the tree performance (branch elongation, flowering and fruit setting intensities) and fruit size. At harvest fruits from the Ti-treated trees showed improved resistance to compression and penetration, as well as a decrease in weight-loss during postharvest storage. A similar response was obtained for the external colour, though all the treatments seemed to delay somewhat the apparent ripening status. Nevertheless, the fruits from Ti-treated trees showed a better behaviour in the evolution of the colour parameters during storage than did the control fruits. Titanium application significantly increased the calcium, iron, copper and zinc concentrations in peel and flesh. This improvement in the calcium absorption is explained as a consequence of the beneficial effect of titanium on the absorption, translocation and assimilation processes.