Changes in salivary analytes in cows due to the in vitro presence of feed.

BACKGROUND: The effect in a sialochemistry profile of the presence of usually available feed in dairy cows was evaluated by an in vitro experiment. For this purpose, a pooled clean saliva from five healthy dairy cows was incubated five times with a standard feed based on a total mixed ration (F), wheat hay (H), and grass (G). The salivary panel was integrated by biomarkers of stress (cortisol -sCor-, salivary alpha-amylase -sAA-, butyrylcholinesterase -BChE-, total esterase -TEA-, and lipase -Lip-), immunity (adenosine deaminase -ADA-), oxidative status (Trolox equivalent antioxidant capacity -TEAC-, the ferric reducing ability of saliva -FRAS-, the cupric reducing antioxidant capacity -CUPRAC-, uric acid, and advanced oxidation protein products -AOPP-), and enzymes, proteins, and minerals of general metabolism and markers of liver, muscle, and renal damage (aspartate aminotransferase -AST-, alanine aminotransferase -ALP-, γ-glutamyl transferase -gGT-, lactate dehydrogenase -LDH-, creatine kinase -CK-, creatinine, urea, triglycerides, glucose, lactate, total protein, phosphorus, and total calcium). RESULTS: Most of the evaluated analytes showed a coefficient of variations (CV) higher than 15% and/or significant changes compared with the clean saliva when feed was present. Some analytes, such as the oxidative status biomarkers (CV > 80%), AST (CV > 60%), or glucose (CV > 100%), showed significant changes with all the feed types tested. Others showed significant differences only with certain types of feed, such as LDH with F (CV > 60%) or triglycerides with F (CV > 100%) and H (CV > 95%). However, sCor or gGT remained unchanged (CV < 15%, P > 0.05) in all the treatments. CONCLUSIONS: The presence of feed can produce changes in most of the analytes measured in cows' saliva, being of high importance to consider this factor when saliva is used as a sample to avoid errors in the interpretation of the results.

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.

Chemical synthesis and optical, structural, and surface characterization of InP-In2O3 quantum dots.

InP-In2O3 colloidal quantum dots (QDs) synthesized by a single-step chemical method without injection of hot precursors (one-pot) were investigated. Specifically, the effect of the tris(trimethylsilyl)phosphine, P(TMS)3, precursor concentration on the QDs properties was studied to effectively control the size and shape of the samples with a minimum size dispersion. The effect of the P(TMS)3 precursor concentration on the optical, structural, chemical surface, and electronic properties of InP-In2O3 QDs is discussed. The absorption spectra of InP-In2O3 colloids, obtained by both UV-Vis spectrophotometry and photoacoustic spectroscopy, showed a red-shift in the high-energy regime as the concentration of the P(TMS)3 increased. In addition, these results were used to determine the band-gap energy of the InP-In2O3 nanoparticles, which changed between 2.0 and 2.9 eV. This was confirmed by Photoluminescence spectroscopy, where a broad-band emission displayed from 2.0 to 2.9 eV is associated with the excitonic transition of the InP and In2O3 QDs. In2O3 and InP QDs with diameters ranging approximately from 8 to 10 nm and 6 to 9 nm were respectively found by HR-TEM. The formation of the InP and In2O3 phases was confirmed by X-ray Photoelectron Spectroscopy.

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.

Use of acute phase proteins for the clinical assessment and management of canine leishmaniosis: general recommendations.

BACKGROUND: Dogs with canine leishmaniosis (CanL) due to Leishmania infantum can show a wide spectrum of clinical and clinicopathological findings at the time of diagnosis. The aim of this paper is to describe the possible application of acute phase proteins (APPs) for the characterization and management of this disease, based on previously published information on the utility of APPs in CanL and the experience of the authors in using APPs as analytes in the profiling of canine diseases. MAIN BODY: Dogs diagnosed with L. infantum infection by serology, polymerase chain reaction, cytological or histopathological identification, can be divided into three groups based on their clinical condition at physical examination and their APPs concentrations: Group 1: dogs with no clinical signs on physical examination and APPs in reference range; Group 2: dogs with changes in APPs but no clinical signs on physical examination; Group 3: dogs with clinical signs and changes in APPs. This report describes the main characteristics of each group as well as its association with the clinical classification schemes of CanL. CONCLUSION: APPs concentration can be a useful clinical tool to characterize and manage CanL.

Plasma markers of inflammation and hemostatic and endothelial activity in naturally overweight and obese dogs.

BACKGROUND: Obesity is one of the most prevalent health problems in the canine population. While haemostatic parameters and markers of endothelial function have been evaluated in various disease conditions in dogs, there are no studies of these markers in canine obesity. This study was designed to evaluate the effect of naturally gained weight excess and obesity on inflammatory, hemostatic and endothelial biomarkers in dogs. A total of 37 overweight and obese dogs were compared with 28 normal weight dogs. RESULTS: Overweight and obese dogs had significantly elevated concentrations of serum interleukin-6 (IL-6) and C-reactive protein (hsCRP). Number of platelets, activity of factor X and factor VII were significantly higher, while activated partial thromboplastine time (aPTT) and soluble plasminogen activator receptor (suPAR) were significantly decreased. Statistical analysis of high mobility group box - 1 protein (HMGB-1), soluble intercellular adhesive molecule -1 (sICAM-1) and plasminogen activator inhibitor type 1 (PAI-1) concentrations did not show significant differences between the total overweight and obese group and the normal weight group of dogs. CONCLUSIONS: Analytical changes in the dogs in our study reflects that weight excess in dogs can be associated with a chronic low degree of inflammation and a hypercoagulable state, where primary and secondary hemostasis are both affected. However obesity is not associated with impairment of endothelial function in dogs.

Changes in serum proteins after endotoxin administration in healthy and choline-treated calves.

BACKGROUND: This study aimed to investigate the possible serum protein changes after endotoxin administration in healthy and choline-treated calves using proteomics. These results are expected to contribute to the understanding of the pathophysiological mechanisms of endotoxemia and the beneficial effect of choline administration in this clinical situation. METHODS: Healthy-calves (n = 20) were divided into 4 groups: Control, Choline treated (C), Lipopolysaccharide administered (LPS), and LPS + C. Control calves received 0.9 % NaCl injection. Calves in C and LPS + C groups received choline chloride (1 mg/kg/iv). Endotoxin (LPS) was injected (2 µg/kg/iv) to the calves in LPS and LPS + C groups. Serum samples were collected before and after the treatments. Differentially expressed proteins (> 1.5 fold-change relative to controls) were identified by LC-MS/MS. RESULTS: After LPS administration, 14 proteins increased, and 13 proteins decreased within 48 h as compared to controls. In the LPS group, there were significant increases in serum levels of ragulator complex protein (189-fold) and galectin-3-binding protein (10-fold), but transcription factor MafF and corticosteroid binding globulin were down regulated (≥ 5 fold). As compared with the LPS group, in LPS + C group, fibrinogen gamma-B-chain and antithrombin were up-regulated, while hemopexin and histone H4 were down-regulated. Choline treatment attenuated actin alpha cardiac muscle-1 overexpression after LPS. CONCLUSIONS: LPS administration produces changes in serum proteins associated with lipid metabolism, immune and inflammatory response, protein binding/transport, cell adhesion, venous thrombosis, cardiac contractility and blood coagulation. The administration of choline is associated with changes in proteins which can be related with its beneficial effect in this clinical situation.

Stratification of Metal and Sulphate Loads in Acid Mine Drainage Receiving Water Dams - Variables Regionalization by Cluster Analysis.

The Sancho Reservoir (Iberian Pyrite Belt, SW Spain) is nourished by the waters of the river Meca, which is affected by acid mine drainage (AMD) processes from the abandoned Tharsis mine. The aim of the present work is to study the hydrochemical variations in this reservoir, in order to define potential stratification processes in metal load and sulphates. A stratified sampling from the surface, with one meter deep intervals to the bottom of the dam, was performed. The results show a clear stratification of temperature, pH, electric conductivity, dissolved oxygen, metal and sulphate loads associated with depth. There is an increase of metal loads at the bottom of the reservoir, though previous studies only detect iron. The proximity between pH and aluminium suggests that water chemistry is strongly influenced by aluminium precipitation processes. This indicates the buffer effect that aluminium exercises, which precipitates as amorphous or low crystalline phases, introducing hydrogen ions to the system, while alkalinity input tends to raise pH.

Herd- and cow-level risk factors associated with subclinical mastitis in dairy farms from the High Plains of the northern Antioquia, Colombia.

Mastitis is the main disease entity affecting dairy farms in the Colombian High Plains of northern Antioquia, Colombia. However, no previous epidemiologic studies have determined the characteristics that increase the risk of infection in this region, where manual milking is still the prevailing system of milking. A 24-mo longitudinal study was designed to identify the predominant mastitis pathogens and important herd- and cow-level risk factors. Monthly visits were made to 37 commercial dairy farms to collect herd- and cow-level data and milk samples. Herd size varied from 6 to 136 cows (mean 37.0, median 29). Herd-level factors included type of milking system (manual or mechanical) and a range of management practices recommended by the National Mastitis Council (Madison, WI) to prevent mastitis. Individual cow-level risk factors included parity, stage of lactation, breed, udder hygiene, and lameness. A logistic regression analysis was used to investigate associations between herd- and cow-level risk factors with the presence of subclinical mastitis and infection caused by Streptococcus agalactiae at the quarter level. A quarter was considered to have subclinical mastitis if it had a positive California Mastitis Test and was subsequently confirmed to have a somatic cell count of ≥200,000 cells/mL. Any cow with one or more quarters with subclinical mastitis was considered to have subclinical mastitis at the cow level. Using 17,622 cow observations, the mean prevalence of subclinical mastitis at the cow level was 37.2% (95% confidence interval: 31.2, 43.3) for the first month and did not substantially change throughout the study. The predominant microorganisms isolated from quarters meeting the subclinical mastitis definition were contagious pathogens, including Strep. agalactiae (34.4%), Corynebacterium spp. (13.2%), and Staphylococcus aureus (8.0%). Significant variables associated with subclinical mastitis risk at the quarter level included being a purebred Holstein cow, higher parity, and increased months in milk. Variables that were protective for mastitis risk included being a crossbreed cow and adequate premilking udder hygiene. Significant variables associated with Strep. agalactiae infection were higher parity, increased months in milk, and manual milking. Variables that were protective were postmilking teat dipping and adequate cleaning of the udder. The results highlight the importance of hygiene practices in contagious mastitis control in manually milked herds.

Salivary oxytocin changes and effect of the season in sows kept in different farrowing systems: Farrowing crate and farrowing pen with temporary crating.

Alternative farrowing systems that have been developed in recent years could have a positive effect on the welfare of sows during farrowing and lactation. Oxytocin measurements in saliva may provide information about positive animal welfare status. The purpose of this study was to evaluate the changes in salivary oxytocin concentrations in sows during the lactation period in three different farrowing systems and in two different seasons. Crossbred Duroc sows (n = 34, average parity = 3.6 ± 1.80) were housed in conventional farrowing crates (FC) (n = 10) or in farrowing pens with temporary crating (TC), including SWAP (n = 12) and JFL15 (n = 12) in two different seasons: summer and winter. Saliva samples were collected for six days during lactation: days 2, 4, 12, 23, 25 (i.e., 1-day post-weaning) and 26 (i.e., 2-day post-weaning) after farrowing. Moreover, behavioral data from sows was recorded on days 2, 4, 12 and 23 after farrowing, using a 30-s scan sampling method for 3 min per pen to record the behaviors which were assessed by the same observer. The results showed that the salivary oxytocin concentrations were 472.5 pg/mL and 399.4 pg/mL higher in both TC (SWAP and JLF15, respectively) than in the FC in early-lactation period, and these differences were more pronounced in summer and at the end of lactation in winter. In terms of behavior, higher number of mother-young interactions were observed in TC than FC in early- and mid-lactation period. In conclusion, TC is associated to a higher salivary oxytocin concentration that could indicated an increased mother-young interaction, although oxytocin concentration can be influenced by other factors, such as season or day of lactation.

Changes in cortisol, cortisone and 11ß-hydroxysteroid dehydrogenase type II activity in saliva during pregnancy and lactation in sows.

The activity of the enzyme 11ß-hydroxysteroid dehydrogenase type II, which can be estimated by the combined measurement of cortisol and cortisone, is gaining importance as a marker for the assessment of stress in pigs. The aim of this study was to investigate the activity of this enzyme and the salivary concentrations of cortisol and cortisone in pigs during pregnancy, farrowing and lactation and to compare it with other stress-related biomarkers such as CgA, S100A12 and alpha-amylase. Salivary cortisol concentrations and 11ß-hydroxysteroid dehydrogenase isoenzyme type 2 activity decreased after farrowing, while cortisol concentrations increased. Enzyme activity did not show significant correlations with any of the other stress-related biomarkers measured in this study. Overall, the results of this report indicate a different regulation of 11ß-HSD type II activity and of cortisol and cortisone during pregnancy and lactation, which should be considered when evaluating these analytes in saliva during these periods.

Changes in biomarkers of the redox status in whole blood and red blood cell lysates in canine hypothyroidism.

Hypothyroidism is the most commonly diagnosed endocrine disease in dogs. The objective of this study was to evaluate the changes in the redox status in canine hypothyroidism using whole blood (WB) and red blood cell (RBCs) lysates. For this purpose, a panel of five antioxidants and five oxidants biomarkers was measured in WB and RBCs lysates of 30 dogs with hypothyroidism, 26 dogs with non-thyroidal illnesses and 15 healthy dogs. The antioxidants measured were cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of plasma (FRAP), Trolox equivalent antioxidant capacity (TEAC), thiol and paraoxonase type-1 (PON-1). Oxidants measured include the total oxidant status (TOS), peroxide-activity (POX-Act), reactive oxygen-derived metabolites (d-ROMs), advanced oxidation protein products (AOPP) and thiobarbituric acid reactive substances (TBARS). WB showed a significant decrease of the antioxidants CUPRAC, TEAC and thiol, and also an increase in TBARS and a decrease in AOPP in dogs with hypothyroidism compared to healthy dogs. Meanwhile, RBCs lysates showed a significant increase in FRAP and PON-1 in dogs with hypothyroidism. The changes in the redox biomarkers in this study show that WB in canine hypothyroidism had a higher number of changes in biomarkers of the redox status than RBCs lysates, making it a promising sample type for the evaluation of the redox status in this disease. In addition, WB is easier and simpler to process than RBCs lysates and unlike serum, it does not have any hemolysis interference.

Why working with porcine circulating serum amyloid A is a pig of a job.

Serum amyloid A (SAA) is a major acute phase protein in most species, and is widely employed as a health marker. Systemic SAA isoforms (SAA1, and SAA2) are apolipoproteins synthesized by the liver which associate with high density lipoproteins (HDL). Local SAA (SAA3) isoforms are synthesized in other tissues and are present in colostrums, mastitic milk and mammary dry secretions. Of systemic SAA the bulk is monomeric and bound to HDL, and a small proportion is found in serum in a multimeric form with a buried HDL binding site. In most species, systemic SAA could easily be studied by purifying it from serum of diseased individuals by hydrophobic interaction chromatography methods. For years, we were not able to isolate systemic pig SAA using the latter methods, and found that the bulk of pig SAA did not reside in the HDL-rich serum fractions but in the soluble protein fraction mainly as a multimeric protein. Based on these surprising results, we analysed in silico the theoretical properties and predicted the secondary structure of pig SAA by using the published pig primary SAA amino acid sequence. Results of the analysis confirmed that systemic pig SAA had the highest homology with local SAA3 which in other species is the isoform associated with non-hepatic production in tissues such as mammary gland and intestinal epithelium. Furthermore, the primary sequence of the pig SAA N-terminal HDL binding site did differ considerably from SAA1/2. Secondary structure analysis of the predicted alpha-helical structure of this HDL binding site showed a considerable reduction in hydrophobicity compared to SAA1/2. Based on these results, it is argued that systemic acute phase SAA in the pig has the structural properties of locally produced SAA (SAA3). It is proposed that in pig SAA multimers the charged N-terminal sequence is buried, which would explain their different properties. It is concluded that pig systemic SAA is unique compared to other species, which raises questions about the proposed importance of acute phase SAA in HDL metabolism during inflammation in this species.

Effect of estradiol and progesterone on metabolic biomarkers in healthy bitches.

The aim of this study was to evaluate the possible association between hormonal changes that occur during oestrus and biomarkers related with glucose metabolism (glucose and insulin), lipid metabolism (lipidic profile and BChE) and adipokines (adiponectin and ghrelin) in healthy bitches. For this purpose, we measured these analytes in serum of bitches, at two times: before (T1) and after (T2) the LH peak that were established according to progesterone concentrations. Increased levels of total cholesterol (p < 0.01), high density lipoprotein cholesterol (HDL-C) (p < 0.01), low density lipoprotein cholesterol (LDL-C) (p < 0.01), adiponectin (p < 0.01) and ghrelin (p < 0.05) were observed at T2 in comparison with T1. No statistically significant changes were observed in serum glucose, insulin, homoeostasis model assessment for insulin sensitivity (HOMA), triglycerides and BChE. When all data of T1 and T2 were pooled, serum adiponectin showed positive correlation with progesterone (r = 0.353; p = 0.022) and HDL-C (r = 0.307; p = 0.048), and negative with insulin (r = -0.429; p = 0.005), HOMA (r = -0.446; p = 0.003) and BChE (r = -0.522; p < 0.001). Ghrelin showed negative correlation with estradiol (r = -0.701; p = 0.004). BChE was negatively correlated with estradiol (r = -0.441; p = 0.018) and glucose (r = -0.343; p = 0.028), and positively with insulin (r = 0.460; p = 0.003) and HOMA (r = 0.505; p < 0.001). In conclusion, changes in metabolic biomarkers occur in bitches after LH peak, characterized by increased lipids (total cholesterol, HDL cholesterol and LDL cholesterol) without changes in BChE activity, and increased adiponectin and ghrelin concentrations, without significant changes in glucose and insulin.

Validation of assays for measurement of oxidant compounds in saliva of pigs: Thiobarbituric acid reactive substances (TBARS), carbonyl, and reactive oxygen species (ROS).

Thiobarbituric acid reactive substances (TBARS), carbonyls and reactive oxygen species (ROS) are oxidant compounds that can provide useful information on the oxidative status. Pigs can be affected by oxidative stress in different situations including physiological conditions such as lactation and also in different diseases, and the measurement of these three analytes in saliva could be potentially useful as biomarkers of the redox status in this species. Assays for the measurement of TBARS and carbonyls by spectrophotometry and ROS by luminol-based chemiluminescence in pigs' saliva were analytically validated and were applied in saliva of pigs after an in vitro incubation with different doses of ascorbic acid (AA). All the assays showed a satisfactory analytical precision and accuracy. The 240 h-incubation of saliva samples with 60 mM of AA induced to an increased TBARS and carbonyls production. TBARS, carbonyls and ROS can be estimated in saliva of pigs by the assays validated in this report. In addition, these assays can detect changes in the concentration of these analytes associated to incubation of saliva samples with AA.

Diagnostic performance of glycated haemoglobin (HbA1c) for diabetes mellitus in dogs.

The objective of this study was to determine the diagnostic performance of glycated haemoglobin (HbA1c) for canine diabetes mellitus (DM) and compare it with that of serum fructosamine. Aliquots of blood samples collected for diagnostic purposes from adult dogs were used. HbA1c was measured using a previously validated capillary electrophoresis assay. The dogs were allocated into four groups: (1) DM; (2) hyperadrenocorticism (HAC); (3) long-term corticosteroid therapy (CST); and (4) various chronic diseases (VD). In total, 88 dogs were included as follows: DM (n = 11), HAC (n = 10), CST (n = 14), and VD (n = 53). Fructosamine was measured in all four groups as follows: DM (n = 6), HAC (n = 7), CST (n = 9), and VD (n = 42). Median (range) serum glucose concentration was higher (P < 0.001) in the DM group (22.8 mmol/L; range, 15.6-29.3 mmol/L) compared to HAC (5.9 mmol/L; range, 4.2-6.8 mmol/L), CST (5.6 mmol/L; range, 4.3-23.3 mmol/L), and VD (5.5 mmol/L; range, 4.1-9.4 mmol/L) groups. Mean (± standard deviation) HbA1c was higher (P < 0.001) in the DM group (6.3% ± 1.5%) compared to HAC (1.9% ± 0.5%), CST (1.7% ± 0.5%), and VD (1.9% ± 0.5%) groups. All diabetic dogs and none of the other dogs had HbA1c levels above the cut-off value for DM (3.3%), indicating an accuracy of 100% in diagnosing DM. Significant differences (P < 0.01) were observed in median fructosamine between the DM group (389 µmol/L; range, 348-865 µmol/L) and the HAC (306 µmol/L; range, 167-348 µmol/L) and the VD (316 µmol/L; range, 189-500 µmol/L) groups. Fructosamine had an accuracy of 84.4% for the diagnosis of DM. When used for the diagnosis of canine DM, HbA1c measured with this specific assay had excellent diagnostic accuracy and was superior to serum fructosamine.

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.

Using integrated wildlife monitoring to prevent future pandemics through one health approach.

In the One Health context, Integrated Wildlife Monitoring (IWM) merges wildlife health monitoring (WHM) and host community monitoring to early detect emerging infections, record changes in disease dynamics, and assess the impact of interventions in complex multi-host and multi-pathogen networks. This study reports the deployment and results obtained from a nationwide IWM pilot test in eleven sites representing the habitat diversity of mainland Spain. In each study site, camera-trap networks and sampling of indicator species for antibody and biomarker analysis were used to generate information. The results allowed identifying differences in biodiversity and host community characteristics among the study sites, with a range of 8 to 19 relevant host species per point. The Eurasian wild boar (Sus scrofa) was the most connected and central species of the host communities, becoming a key target indicator species for IWM. A negative relationship between biodiversity and disease risk was detected, with a lower number and prevalence of circulating pathogens in the sites with more species in the community and larger network size. However, this overall trend was modified by specific host-community and environmental factors, such as the relative index of wild boar - red deer interactions or the proximity to urban habitats, suggesting that human-driven imbalances may favour pathogen circulation. The effort of incorporating wildlife population monitoring into the currently applied WHM programs to achieve effective IWM was also evaluated, allowing to identify population monitoring as the most time-consuming component, which should be improved in the future. This first nationwide application of IWM allowed to detect drivers and hotspots for disease transmission risk among wildlife, domestic animals, and humans, as well as identifying key target indicator species for monitoring. Moreover, anthropogenic effects such as artificially high wildlife densities and urbanisation were identified as risk factors for disease prevalence and interspecific transmission.

Long gamma-ray bursts and core-collapse supernovae have different environments.

When massive stars exhaust their fuel, they collapse and often produce the extraordinarily bright explosions known as core-collapse supernovae. On occasion, this stellar collapse also powers an even more brilliant relativistic explosion known as a long-duration gamma-ray burst. One would then expect that these long gamma-ray bursts and core-collapse supernovae should be found in similar galactic environments. Here we show that this expectation is wrong. We find that the gamma-ray bursts are far more concentrated in the very brightest regions of their host galaxies than are the core-collapse supernovae. Furthermore, the host galaxies of the long gamma-ray bursts are significantly fainter and more irregular than the hosts of the core-collapse supernovae. Together these results suggest that long-duration gamma-ray bursts are associated with the most extremely massive stars and may be restricted to galaxies of limited chemical evolution. Our results directly imply that long gamma-ray bursts are relatively rare in galaxies such as our own Milky Way.

Validation of two ELISA assays for total ghrelin measurement in dogs.

The aim of this study was to validate two commercially available ELISA assays for total ghrelin measurement in dogs: one canine-specific and one originally designed for measuring human ghrelin. The two assays showed intra-assay coefficient of variations (CVs) lower than 10%, while the inter-assay CVs exceeded the 15% limit. Sample dilutions resulted in linear regression equations with correlation coefficients close to 1. In order to compare methods and verify ability of the ghrelin assays to differentiate between low and high levels, ghrelin concentrations were measured in plasma samples obtained before and at different times after glucose administration in five Beagle dogs. A statistically significant changes in ghrelin after glucose administration was recorded only with assay B. In conclusion, the human ELISA validated in this study showed a good intra-assay precision, accuracy, and when applied to the glucose injection study, was better in distinguishing high and low canine ghrelin levels than the canine ELISA assay.