Total Esterase Research Articles

Saliva as a Potential Source of Biomarkers in Cows with Metritis: A Pilot Study.

Metritis affects 5-20% of cows after parturition, negatively impacting animal welfare and the profitability of dairy farms, increasing culling rates and costs, and decreasing productivity and reproduction rates. This study compared the results of a comprehensive biochemical panel consisting of 25 salivary and 31 serum analytes between healthy cows (n = 16) and cows with metritis (n = 12). Descriptive parameters such as depression, rectal temperature, body condition score (BCS), heart rate, respiratory rate, mucous color, ruminal motility, vaginal discharge, milk production, and complete hematology analyses were also assessed for comparative purposes. The biochemistry analytes comprised five analytes related to stress, five to inflammation, five to oxidative status, and nineteen to general metabolism. The two-way ANOVA analysis revealed that, in saliva, eight biomarkers (lipase, adenosine deaminase (ADA), haptoglobin (Hp), total proteins, g-glutamyl transferase (gGT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and creatine kinase (CK)) were significant higher in cows with metritis. In serum, eight biomarkers (ADA, Hp, serum amyloid A (SAA), fibrinogen, ferritin, AOPPs/albumin ratio, non-esterified fatty acids (NEFAs), and bilirubin) were significantly higher in cows with metritis, whereas six (total esterase (TEA), albumin, urea, lactate, phosphorus, and calcium) were lower. Of the total number of 23 biomarkers that were measured in both saliva and serum, significant positive correlations between the two biofluids were found for six of them (Hp, FRAP, CUPRAC, AOPPs, urea, and phosphorus). Urea showed an R = 0.7, and the correlations of the other analytes were weak (R < 0.4). In conclusion, cows with metritis exhibited differences in biomarkers of stress, inflammation, cellular immune system, and general metabolism in both salivary and serum biochemistry profiles. These changes were of different magnitudes in the two biofluids. In addition, with the exception of ADA and Hp, the analytes that showed changes in the saliva and serum profiles of cows affected by metritis were different. Overall, this report opens a new window for the use of saliva as potential source of biomarkers in cows with metritis.

Non-invasive methods for analysing pig welfare biomarkers.

Atpresent, the assessment ofpig welfare quality has gained significant importance, prompting theexploration ofnovel biomarkers for this purpose. Traditionally, these biomarkers have been monitored inthe blood; however, blood sampling isconsidered aninvasive procedure. Currently, non-invasive methods for collecting samples are emerging asviable alternatives for assessing these biomarkers. This article aims topresent the current knowledge regarding the use ofnon-invasive methods for analysing pig welfare biomarkers, specifically focusing onthe saliva, hair, faeces, and urine asmatrices todetermine these biomarkers. The saliva analysis encompasses various biomarkers, such ascortisol, alpha-amylase, chromogranin A, the total esterase, oxytocin, acute phase proteins, adenosine deaminase, immunoglobulins and parameters ofredox homeostasis. Cortisol, aspecific biomarker, can bedetermined inthe hair, urine and faeces, while urine samples allow for the analysis ofcatecholamines asnon-invasive markers ofpig welfare.

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.

Mulberry branch fiber improved lipid metabolism and egg yolk fatty acid composition of laying hens via the enterohepatic axis

BackgroundThe utilization of mulberry branch fiber (MF), the largest by-product of the sericulture industry, is an important issue. Supplementation with MF as a dietary fiber for poultry may serve as a useful application. However, little is known about the effects of MF on liver lipid metabolism and egg yolk fatty acid composition of laying hens and their underlying mechanisms. In this study, we performed a multi-omics investigation to explore the variations in liver lipid metabolism, egg yolk fatty acid composition, gut microbiota, and the associations among them induced by dietary MF in laying hens.ResultsDietary MF had no harmful effects on the laying performance or egg quality in laying hens. The enzyme activities associated with lipid metabolism in the liver were altered by the addition of 5% MF, resulting in reduced liver fat accumulation. Furthermore, dietary 5% MF induced the variation in the fatty acid profiles of egg yolk, and increased the polyunsaturated fatty acid (PUFA) content. We observed a significant reduction in the diversity of both gut bacteria and changes in their compositions after the addition of MF. Dietary MF significantly increased the abundance of genes involved in fatty acid biodegradation, and short-chain fatty acids biosynthesis in the gut microbiota of laying hens. The significant correlations were observed between the liver lipid metabolism enzyme activities of hepatic lipase, lipoprotein lipase, and total esterase with gut microbiota, including negative correlations with gut microbiota diversity, and multiple correlations with gut bacteria and viruses. Moreover, various correlations between the contents of PUFAs and monounsaturated fatty acids in egg yolk with the gut microbiota were obtained. Based on partial-least-squares path modeling integrated with the multi-omics datasets, we deduced the direct effects of liver enzyme activities and gut bacterial compositions on liver fat content and the roles of liver enzyme activities and gut bacterial diversity on egg yolk fatty acid composition.ConclusionsThe results indicate that dietary MF is beneficial to laying hens as it reduces the liver fat and improves egg yolk fatty acid composition through the enterohepatic axis.CByDGCz_6LcoMs-GQqtpHGVideo Graphical

Effects of repeated intramuscular injections on sow behaviour reactions and stress-related saliva biomarkers – A pilot study

Medicating large production animals, such as sows, individually can be challenging, especially when repeated treatments are needed. Intramuscular (i.m.) injections have been shown to be aversive, and an increased reaction could be expected over consecutive days of injections. The aim of this pilot study was to assess if daily repeated i.m. injections cause sows to react differently with time and if injection stress is reflected in increased stress- and pain-related saliva biomarkers. Additionally, the aim was to assess if oxytocin was related to the behaviour reaction of sows during injections. In total 37 sows were allocated randomly into four treatment groups: 1. Sham-injected (SHAM: 9 sows), 2. Control (CON: 11 sows), 3. Penicillin (PEN: 9 sows) and 4. Tetracyclin (TET: 8 sows). The reaction of the sows to injections over three days was subjectively assessed by two observers. Further, SHAM and TET sows were saliva sampled for stress-related biomarkers before and after injections. Treatment (p < 0.001) influenced the medication reaction. Pairwise comparisons showed that the reaction to SHAM treatment was less compared to CON (p = 0.002) and PEN treatments (p < 0.001), while reactions to SHAM did not differ from TET (p > 0.1). Reaction to PEN was higher than that to TET (p = 0.01), while PEN and CON did not differ from each other (p > 0.1). Day tended to affect medication reaction (p = 0.08). The reaction to all other treatments decreased numerically from day 1 to day 3, while this was not the case for the TET treatment. Changes in biomarkers were subtle. Oxytocin tended to increase after TET injection on day 3 (p = 0.05). For the SHAM treatment, some of the biomarkers (chromogranin A, alpha-amylase activity and total esterase activity) showed significant or numerically decreasing trends in POST vs PRE-treatment samples on both days. The oxytocin level before injections was higher in sows that did not react to the medication at all than in sows that showed any kind of reaction (p = 0.008). In conclusion, sows appeared to habituate to the handling and injection, depending on the level of discomfort caused by the injectable substance. Further, the results suggest a potential role of oxytocin in decreasing aversive reactions in sows to human handling.

Decreased retronasal olfaction and taste perception in obesity are related to saliva biochemical and microbiota composition

Understanding the individual factors that modulate flavor perception is a central issue for the development of personalized diets strategies to fight obesity. This study aimed to investigate differences in flavor perception between adults with normal weight and those with obesity, as well as some potential biological factors related to these differences. To do that, liking and flavor perception intensity were measured against retronasal olfactory (pineapple, butter, tropical and chocolate) and taste attributes (sweetness, umami and bitter) in 77 individuals grouped as normalweight or obese, according to their body mass index (BMI). Unstimulated saliva was collected from all participants and characterized in terms of salivary flow, total protein content, total antioxidant capacity, total esterase activity and bacterial composition through 16S rDNA amplicon sequencing. The results showed that participants displayed differences in flavor perception according to their BMI group. Thus, the group with obesity showed significant lower liking and intensity scores for low calorie related food aroma (pineapple and tropical), lower taste intensity scores for sweet and umami, and a higher acceptability for umami than the group with normal weight. Significant differences between BMI groups were observed for salivary biochemical variables and specific bacterial taxa, some of which were significantly correlated to flavor intensity. This work suggests for the first time the existence of an oral-brain axis that might contribute to the development or perpetuation of obesity, which opens new and interesting avenues of research.

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

Simple SummarySaliva is a sample with a high potential in pigs since it is usually easy to obtain and its collection from animals causes less stress than blood sampling. However, the possible effects of daily variations in many salivary biomarkers are still unknown in this species. In our report, the possible variations depending on the sampling time in the day in a panel of 26 salivary biomarkers related to stress, immune system, redox status and other physiological functions in the saliva of pigs were evaluated. In our experimental conditions, daily variations were observed in cortisol, α-amylase, total esterase, butyrylcholinesterase, lipase, adenosine deaminase isoenzyme 1, uric acid, superoxide dismutase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatine kinase, lactate dehydrogenase, triglycerides and lactate. In some analytes, these differences appeared in both sexes, whereas others only showed differences in one sex. These variations should be taken into consideration for an appropriate interpretation of these analytes in the saliva of healthy 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.

Bacterial symbionts influence host susceptibility to fenitrothion and imidacloprid in the obligate hematophagous bed bug, Cimex hemipterus

The use of insecticides remains important in managing pest insects. Over the years, many insects manifested physiological and behavioral modifications resulting in reduced efficacy of insecticides targeted against them. Emerging evidence suggests that bacterial symbionts could modulate susceptibility of host insects against insecticides. Here, we explore the influence of host microbiota in affecting the susceptibility of insect host against different insecticides in the blood-sucking bed bug, Cimex hemipterus. Rifampicin antibiotic treatment resulted in increased susceptibility to fenitrothion and imidacloprid, but not against deltamethrin. Meanwhile, the host fitness parameters measured in the present study were not significantly affected by rifampicin treatment, suggesting the role of bacterial symbionts influencing susceptibility against the insecticides. 16S metagenomics sequencing revealed a drastic shift in the composition of several bacterial taxa following rifampicin treatment. The highly abundant Alphaproteobacteria (Wolbachia > 90%) and Gammaproteobacteria (Yersinia > 6%) in control bed bugs were significantly suppressed and replaced by Actinobacteria, Bacilli, and Betaproteobacteria in the rifampicin treated F1 bed bugs, suggesting possibilities of Wolbachia mediating insecticide susceptibility in C. hemipterus. However, no significant changes in the total esterase, GST, and P450 activities were observed following rifampicin treatment, indicating yet unknown bacterial mechanisms explaining the observed phenomena. Re-inoculation of microbial content from control individuals regained the tolerance of rifampicin treated bed bugs to imidacloprid and fenitrothion. This study provides a foundation for a symbiont-mediated mechanism in influencing insecticide susceptibility that was previously unknown to bed bugs.

Changes in Saliva Analytes in Dairy Cows during Peripartum: A Pilot Study.

Simple SummaryThe use of saliva as a biological fluid to assess welfare and health status is gaining interest nowadays since it can be collected by non-invasive methods without specialized staff. The possibility of measuring analytes in saliva by techniques adapted to automated analyzers is cost-effective, reliable, and replicable. These analytes can provide information useful for the evaluation of welfare and health in dairy cows. In this pilot study, a total of 26 salivary analytes were measured in healthy dairy cows along their peripartum period to assess possible changes and associations with their inflammatory, energy, and milk yield status. Salivary analytes related to stress (cortisol, salivary alpha-amylase, butyrylcholinesterase, and total esterase), immunity (adenosine deaminase), oxidative status (Trolox equivalent antioxidant capacity and the advanced oxidation protein products), and general metabolism (creatine kinase, γ-glutamyl transferase, urea, triglycerides, glucose, and lactate) had significant changes throughout this period. This study indicated that the saliva of dairy cows shows changes in its composition during the transition period and potentially can be a source of biomarkers for monitoring health and welfare.This pilot study aimed to study the possible changes in a profile of 26 salivary analytes in thirteen healthy dairy cows along their peripartum period. Analytes associated with the stress (salivary cortisol, salivary alpha-amylase, butyrylcholinesterase, and total esterase), inflammation (adenosine deaminase), oxidative status (total antioxidant capacity and the advanced oxidation protein products), and general metabolism (creatine kinase, γ-glutamyl transferase, urea, triglycerides, glucose, and lactate) varied along the sampling times. A positive correlation between the white blood cells counts, and the lipase, Trolox equivalent antioxidant capacity, advanced oxidation protein products, and lactate levels in saliva were observed at the delivery. A linear association between selected salivary analytes at different sampling times and the milk yield after calving was observed. In conclusion, in our experimental conditions, it was observed that the peripartum period in dairy cows can induce changes in salivary analytes. Some of them were associated with inflammatory status and the capacity of milk production after calving.

Toxicity and genetic analysis of Scaptotrigona bipunctata Lepeletier, 1836 contaminated with the pyrethroid cypermethrin

Stingless bees are important pollinators for the native forest of tropical and subtropical regions, predominantly in Latin America. This group contains more than 300 species, many of them native from Brazil. Their colonies present various types of structures, formats and materials. Scaptotrigona bipunctata (Lepeletier, 1836) is a species of stingless bee that builds large colonies within tree trunks. They are ecologically important as pollinators, maintaining the ecological balance. However, studies indicate that the number of bees has been decreasing drastically over the years due to habitat destruction and intensive use of agrochemicals. High doses of insecticides can lead to the death of bees, but low concentrations may promote behavioral changes that affect the colonies and its services to ecosystem and agricultural crops. Around 40% of all insecticides applied in Brazil are toxic to bees. Cypermethrin insecticide belongs to the pyrethroid group and it is widely used in several crops, such as cotton, potato, coffee, maize and tomato. This study aimed to investigate the mortality rate, alterations in total protein and esterase expression in S. bipunctataafter contamination by ingestion and contact with cypermethrin. At the end of the bioassays, there was found a low mortality rate, however, sublethal effects were expressive. Esterases showed different expression patterns after contamination, both by ingestion and contact and total protein analysis presented changes in expression, as well.

Resistance status and detoxification enzyme activity in ten populations of Panonychus citri (Acari: Tetranychidae) from Turkey

Acaricide resistance monitoring in phytophagous mites is crucial for effective pest management. Citrus growers in Cukurova region, a region that comprised more than 75% of total citrus production in Turkey, complained that acaricides have become inefficient in controlling outbreaks of the citrus red mite, Panonychus citri McGregor (Acari: Tetranychidae). To assess whether such control failures were caused by acaricide resistance, we bio-assayed P. citri populations collected from nine commercial orchards and a susceptible laboratory strain. Based on the mode of action (MoA) of acaricides, protonymphs were bio-assayed with etoxazole, spirodiclofen and spirotetramat, whereas abamectin and pyridaben were used for adults. Additionally, we determined kinetic esterase and glutathione S-transferase (GST) enzyme activities using a micro plate reader. Based on LC50 values, the resistance ratios of the orchard populations compared to the laboratory population varied from 2.81 to 34.82-fold, 1.22–18.35-fold, 1.23–40.43-fold, 1.76–27.50-fold and 2.24–75.06-fold for abamectin, etoxazole, spirodiclofen, spirotetramat and pyridaben, respectively. Total esterase and GST activities of the populations were in the ranges of 15.86–43.28 and 2.07–5.29 mOD/min/mg protein, respectively. The highest enzyme activity levels were detected in the Yakapinar population which was among one of the most resistant populations to all tested acaricides. The activities of detoxification enzymes may play an important role in the metabolic breakdown of the acaricides in resistant P. citri populations. Rotation of acaricides with different modes of action is fundamental to prevent potential resistance problems.

Changes in Saliva Analytes Associated with Lameness in Cows: A Pilot Study.

Simple SummarySaliva may contain useful biomarkers which provide information about animal welfare using convenient and non-invasive sampling methods. In addition, the development of automated techniques of measuring analytes in saliva provides advantages from the technical point of view since they are cost-effective, reliable, and replicable. In this study, 21 salivary analytes measured by automated assays were tested as potential biomarkers of lameness, one of the most prevalent diseases in dairy cows producing significant economic losses. As a result, total esterase (TEA) showed increases in saliva in a group of 11 cows with lameness, decreasing when the lameness was solved after a specific treatment consisting of a hoof trimming and a medical treatment. In addition, TEA activity correlated with the severity of the lameness. Further studies using a larger population of cows with different causes of lameness and severity should be performed to determine the potential of TEA as a biomarker of lameness in cows.The possible changes in a panel of 21 salivary analytes on a population of cows with lameness before and after treating lameness by hoof trimming were analyzed. Then, the analytes that showed significant changes were studied in a larger population of cows with lameness and compared with healthy cows For this purpose, two groups of cows were made by a specialized veterinarian. One consisted of healthy cows with no external signs of diseases and no hematological or biochemical abnormalities, and showing no signs of lameness according to the numerical rating system of severity (NRS, 5-point scale); and the other composed of cows showing only lameness with a NRS of 3.1 ± 0.87 and a lesion scoring system (LSS, 4-point scale) of 3.3 ± 0.89. Both groups did not differ in parity (p = 0.140), days in milk (DIM) (p = 0.780), and body condition score (BCS) (p = 0.074). Initially, 21 biochemical analytes were determined in the saliva of six cows with lameness at the diagnosis time (T0) and twenty days after hoof trimming that successfully solved the lameness (TF). This exploratory study only showed significantly higher values in lipase (Lip) and total esterase (TEA) at T0 compared to TF (p < 0.001 and p = 0.034, respectively). When both analytes were measured in the additional five lame cows and the results of all the animals of the lame group (n = 11) were compared with the healthy group (n = 11), only TEA showed higher activities in the group of lame cows than healthy cows (p = 0.004). TEA was positively correlated with both NRS and LSS (r = 0.43, p = 0.004 and r = 0.35, p = 0.003). In conclusion, this study showed that cows with lameness in our experimental conditions had higher TEA values than healthy cows, and these values decreased after treatment. This is a pilot study, and further studies using a larger population of cows with lameness due to different causes and severity should be performed to determine the potential of TEA as a biomarker of lameness in cows.

Changes in oxytocin concentrations in saliva of pigs after a transport and during lairage at slaughterhouse

Oxytocin is associated with reproductive physiology but also with welfare and positive emotions. In this study, oxytocin was measured in saliva samples of 45 pigs that were collected before being transported to the slaughterhouse, at the time of arrival and 4 h after arrival to the slaughterhouse. Two previously validated assays, one that measures free oxytocin and other that measures oxytocin linked to proteins, were used. In addition, cortisol, salivary alpha-amylase (sAA), total esterase activity (TEA), butyrylcholinesterase (BChE) and lactate dehydrogenase (LDH), which are biomarkers associated with stress and pain in pigs, were measured. The results showed a decrease in free and protein-linked oxytocin concentrations at 4 h after transport compared with the time before transport, while cortisol, sAA, TEA, BChE and LDH showed an increase at 4 h after transport compared with the time before transport. Based on these results it can be concluded that the transport and lairage at slaughterhouse in the conditions of this study produce a decrease in oxytocin in the saliva of pigs that could indicate a reduced emotional well-being.

Changes in Salivary Analytes of Horses Due to Circadian Rhythm and Season: A Pilot Study.

Simple SummaryThe use of salivary biomarkers is gaining interest in the veterinary field, since saliva is usually easy to obtain and its collection from animals causes less stress than blood sampling. However, our knowledge of the possible factors related to daily and seasonal variations in salivary biomarkers is still in its infancy. In our study, the possible circadian or circannual variations in a panel of salivary biomarkers in horse saliva were evaluated. The results showed that daily and/or seasonal variations can be observed in cortisol, salivary alpha-amylase, total esterase, butyrylcholinesterase, adenosine deaminase, and creatine kinase. Therefore, these factors should be considered for the interpretation of these analytes when measured in horse saliva.This study aims to evaluate the circadian and circannual variations in a panel of analytes in horse saliva that have been previously described as biomarkers related to stress and disease, in order to interpret them correctly when they are measured in this species. This panel of analytes integrated cortisol, salivary alpha-amylase (sAA), lipase (Lip), total esterase (TEA), butyrylcholinesterase (BChE), adenosine deaminase (ADA), γ-glutamyl transferase (gGT), creatine kinase (CK), urea, total bilirubin, total protein (TP), and phosphorus. These analytes were measured in saliva obtained from a population of five clinically healthy mares from 06:30 to 20:30, every 2 h over two consecutive days in two different photoperiod seasons, winter and spring. The temperature and relative humidity did not change between the two consecutive days sampled in each sampled season, and no thermal discomfort was observed. Changes throughout the course of the day were observed for cortisol, sAA, TEA, BChE, ADA, and CK. However, a circadian pattern was only observed for cortisol, TEA, BChE, ADA, and CK. Moreover, the values obtained for sAA, Lip, and BChE were significantly different between seasons, with different daily rhythms for cortisol, TEA, BChE, and ADA depending on the season. In conclusion, this pilot study indicates that the time of the day and the season influence salivary analytes in horses, showing a rhythmic pattern for cortisol, TEA, BChE, ADA, and CK. These factors should thus be taken into consideration for the interpretation of analytes in horse saliva.

Efficacy of essential oil from Clausena anisata and its impact on biochemical changes of Sitophilus oryzae.

Essential oil (EO), from Clausena anisata, was evaluated for their efficacy (fumigant and repellent activities) against Sitophilus oryzae. Result revealed that C. anisata showed 82% fumigant toxicity (insect mortality) at 160μL/L concentration after 24h observation period. C. anisata exhibited LC50 value of 17.84μL/L concentration for fumigant toxicity. The insect movement behaviour (repellent) was observed in two different concentrations of 10 and 50μL for a period of 3h. Maximum repellent activity (- 0.25) was observed at 50μL/L concentration against S. oryzae after 3-h observation. At 3, 6 and 12μL/L concentrations of C. anisata oil significantly reduced the protein content and total esterase activity of S. oryzae.

Effect of food contamination and collection material in the measurement of biomarkers in saliva of horses

This study aims to evaluate the effect of the presence of food and the material used in a panel of biomarkers in saliva of horses.For the food effect study, clean saliva was incubated with a known amount of food consisting of oats, hay or grass. Significant changes were observed when saliva was incubated with oats for total protein (P = .050) and phosphorus (P = .008), with grass for total protein (P = .037), salivary alpha-amylase (sAA, P = .018), total esterase (TEA, P = .018), butyrilcholinesterase (BChE, P = .037), adenosine deaminase (ADA, P = .037), and total bilirubin (P = .018), and with hay for sAA (P = .018), phosphorus (P = .037), γ-glutamyl transferase (gGT, P = .004), and creatine kinase (CK, P = .016). For the material-based collection study, saliva using a sponge and a cotton role at the same time were collected and compared. Lower values were obtained in clean saliva collected with cotton role compared to sponge for sAA (P = .030), TEA (P = .034), BChE (P = .003), gGT (P = .002) and cortisol (P < .001)In conclusion, the presence of food and the material used for its collection, can influence the results obtained when analytes are measured in saliva of horses.

Changes in Saliva Analytes Correlate with Horses' Behavioural Reactions to An Acute Stressor: A Pilot Study.

Simple SummaryEmotionality is an individual characteristic defined as the propensity to respond to stress-inducing stimuli such as fear-inducing objects or social separation. Evaluation of emotionality in horses is important as it may impact their learning performance. Although emotionality is usually assessed by measuring behavioural patterns, biomarkers could provide additional information about stress response, especially with respect to its temporal dynamics. In this study, behavioural responses were measured as well as a panel of salivary biomarkers related to stress, including salivary alpha-amylase, lipase, total esterase, butyrylcholinesterase, adenosine deaminase, and cortisol, in riding horses after acute experimental stress (the sudden opening of an umbrella). We found significant changes in most of the salivary biomarkers evaluated after the induced stress, where increases in butyrylcholinesterase were more closely related to behavioural responses to acute stress and low salivary alpha-amylase values were more closely related to quietness behaviours. Therefore, this preliminary research provides information about the relationship between behaviour patterns and biomarkers of stress in saliva in horses, and opens the possibility of wider use of selected biomarkers in saliva, such as butyrylcholinesterase or alpha-amylase, for the evaluation of acute stress in horses.Acute stress induces an array of behavioural reactions in horses that vary between individuals. Attempts to relate behavioural patterns and physiological responses have not always given clear-cut results. Here, we measured the changes in a panel of salivary components: salivary alpha-amylase (sAA), lipase, total esterase (TEA), butyrylcholinesterase (BChE), adenosine deaminase (ADA), and cortisol, and their potential link with horses’ behaviours after acute stress. Saliva samples were collected in nine riding horses subjected to a test consisting of opening an umbrella. Saliva sampling was obtained at a basal time point in the stall (T1), in the test indoor arena (T2), at a time of stress (T3), and 30 min (T4) and 60 min (T5) later. The horses’ behaviour was recorded at T3 for 1 min. sAA, lipase, TEA, and BChE showed significant changes along time, increasing at T3 for BChE, and decreasing at T4 for sAA and BChE. Butyrylcholinesterase appeared to be the most reliable predictor of behavioural responses, as it correlated with the index of emotionality, of laterality, and the occurrence of alarm signals, while sAA decreased when horses expressed quieter behaviours. These first results bring promising lines for novel, more precise physiological markers of acute stress in horses that can bridge the gap between behaviour and physiology.

Determination of LC values to some insecticides and amount of total esterase in Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations in cotton in Söke (Aydın, Turkey)

Amaç:Bu çalışma, bazı insektisitler inAydın ili Söke ilçesi pamuk alanlarından toplanan dört farklı Pamuk beyazsineği Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) popülasyonu için öldürücü konsantrasyon (LC) değerlerinin belirlenmesi amacıyla gerçekleştirilmiştir.Materyal ve Yöntem:Yaprak daldırma yöntemi ile yörede en çok kullanılan insektisitler seçilerek yapılan denemelerde, zararlının nimf dönemlerinde pyriproxyfen ve buprofezine, ergin dönemlerinde acetamiprid ve bifenthrin için LC değerleri belirlenmiştir. Ayrıca biyokimyasal çalışmalar da yürütülerek söz konusu aktif maddelerin toksisite değer farklılıklarının enzimlerle ilişkisinin belirlenmesine çalışılmıştır.Bulgular:Dört farklı beyazsinek populasyonundan elde edilen LC50 değerleri sırasıyla, acetamiprid için; 37.51, 28.22, 52.12, 75,61 ppm; bifenthrin için 2.72, 4.39, 1.08, 1.89 ppm, buprofezin için 30.95, 29.05, 46.46, 25.95 ppm ve pyriproxyfen için 38.85, 76.06, 108.91, 34.15 ppm olmuştur.Sonuç:Ayrıca, biyokimyasal analizlerde söz konusu dört populasyonda sadece toplam esteraz değerleri tespit edilmiş ve enzim seviyelerinde önemli bir fark görülmemiştir.

Application of a score for evaluation of pain, distress and discomfort in pigs with lameness and prolapses: correlation with saliva biomarkers and severity of the disease

A score system was used to evaluate pain, distress and discomfort in healthy pigs and pigs with two different diseases: lameness and rectal prolapse. In addition, correlations between the results of this score and a panel of salivary biomarkers and severity of disease were studied. This panel included biomarkers of stress (cortisol, salivary alpha-amylase (sAA), total esterase activity (TEA), butyrylcholinesterase (BChE) and lipase (Lip)), immunity (adenosine deaminase isozymes 1 (ADA1) and 2 (ADA2)) and oxidative status (uric acid (UA), Trolox equivalent antioxidant capacity (TEAC), cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of saliva (FRAS), advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2)). Based on their score, diseased animals were subdivided in those without and with evident pain. Lame pigs and prolapsed pigs with pain showed higher salivary levels of cortisol, sAA, TEA, BChE, ADA1 and ADA2 compared with the healthy pigs. In addition, the prolapsed pigs with pain showed higher levels of FRAS, AOPP and H2O2 compared with the healthy animals. Salivary cortisol, TEA, BChE, ADA isozymes 1 and 2, FRAS and AOPP correlated with the pain score. This five-point pain score system can be easily applied to lame and prolapsed pigs, and salivary biomarkers could be used as an additional tool for pain assessment in those pigs.

Physiological and ecological responses and changes of Phaeodactylum tricornutum under long-term stress of naphthalene

Naphthalene, as a common polycyclic aromatic hydrocarbon (PAHs), attracts broad attention due to its toxicity to human and marine organisms. This study investigate the effects of naphthalene on growth, maximum photochemical efficiency (Fv/Fm), total esterase activity and reactive oxygen species production of Phaeodactylum tricornutum. The study explored how algal physiological characteristics change under naphthalene’s long-term exposure and analyzed the correlation between photosynthetic efficiency with growth, total esterase activity and reactive oxygen species production, respectively. The study shows that low-concentration naphthalene promotes slightly or doesn’t affect algal growth, total esterase activity and reactive oxygen species while it promotes Fv/Fm significantly. On the other hand, high-concentration naphthalene inhibits algal growth, total esterase activity and Fv/Fm significantly, while it stimulates reactive oxygen species production and the stimulation increases with time. The study also shows that growth and Fv/Fm of P. tricornutum are correlative and the correlation is related with naphthalene concentration. Fv/Fm of P. tricornutum is significantly positively-related with total esterase activity, while is significantly negatively-related with reactive oxygen species.