Porcine salivary analysis by 2-dimensional gel electrophoresis in 3 models of acute stress: A pilot study

The purpose of this research was to study changes in the salivary proteome of healthy pigs in stressful situations to identify any potential new salivary biomarker of stress. Three groups of animals were subjected to 3 stress models: snaring restraint followed by simulated sampling of vena cava blood; brief transport by road; and restriction of movement in a digestibility cage. Saliva was obtained from each animal before and 15 and 30 min after the induction of stress. The samples from the animals that showed the greatest increase in salivary cortisol concentration were pooled and run on 2-dimensional gels. Coomassie Brilliant Blue R-250 was used for spot detection and mass spectrometry for spot identification. Statistical analyses showed that 2 proteins had significant differences in expression before and after the induction of stress. These proteins were identified as odorant-binding protein and fragments of albumin. Further studies will be necessary to confirm the value of using these proteins as salivary biomarkers of stress in pigs.     

Epidemiological Analysis of Leptospira spp. Infection in Equids from the Brejo Paraibano Microregion of Brazil

Leptospirosis is a zoonotic disease that occurs in humans and several animal species, including cattle, pigs, and horses, and it has a worldwide distribution. In equids, leptospirosis manifests through recurrent uveitis and abortion and other reproductive disorders. The objective of the present study was to investigate the epidemiological situation of Leptospira spp. infection in equids from the Brejo Paraibano microregion in the northeast of Brazil, that is, to assess the prevalence of and identify the major risk factors for infection. For this purpose, blood samples were taken from 257 equids (204 horses, 46 mules, and 7 donkeys), and a microscopic agglutination test (MAT) was used as serological analysis, with 24 serovars of Leptospira spp. as antigens. Antibodies were observed in 16.2% of horses, 13.0% of mules, and 28.6% of donkeys. Animals between the ages of 2.5 and 11 years had a risk of infection 0.4 times lower than those younger than 2.5 years old (odds ratio, 0.4; 95% confidence interval, 0.18-0.90). Further studies are needed to identify potential wild reservoirs and the main routes of infections.     

Subependymal Reaction Secondary to Choroid Plexus Papilloma in a Horse

Choroid plexus papilloma (CPP) is a rare, benign, intracranial neoplasm. The present report described a choroid plexus papilloma of the fourth ventricle with unusual proliferative reaction of ependymal/subependymal cells in a stallion. A single, soft neoformation, red-gray in color, with an irregular surface, resembling a cauliflower, expanding from the fourth ventricle to the cerebellar peduncles was observed. Microscopically, the neoformation was composed of a single layer of simple cuboidal to columnar epithelium with a well-vascularized papillary-arboriform pattern. Multifocal subependymal aggregates forming rosettes were observed near the mesencephalic aqueduct (MA). Glial-fibrillary-acidic-protein (GFAP) was markedly positive only in the neuropil adjacent to the proliferative areas of the MA. Ependymal/subependymal reactions secondary to CPP have never been described in equine or human neuropathology. The capabilities of ependymal cells to regenerate and eventually proliferate are controversial and unclear and, thus, require additional investigation to understand the adaptive mechanisms of this rare pathogenic condition.     

Breed Differences in Natriuretic Peptides in Healthy Dogs

Background

Measurement of plasma concentration of natriuretic peptides (NPs) is suggested to be of value in diagnosis of cardiac disease in dogs, but many factors other than cardiac status may influence their concentrations. Dog breed potentially is 1 such factor.

Objective

To investigate breed variation in plasma concentrations of pro‐atrial natriuretic peptide 31‐67 (proANP 31‐67) and N‐terminal B‐type natriuretic peptide (NT‐proBNP) in healthy dogs.

Animals

535 healthy, privately owned dogs of 9 breeds were examined at 5 centers as part of the European Union (EU) LUPA project.

Methods

Absence of cardiovascular disease or other clinically relevant organ‐related or systemic disease was ensured by thorough clinical investigation. Plasma concentrations of proANP 31‐67 and NT‐proBNP were measured by commercially available ELISA assays.

Results

Overall significant breed differences were found in proANP 31‐67 (P < .0001) and NT‐proBNP (P < .0001) concentrations. Pair‐wise comparisons between breeds differed in approximately 50% of comparisons for proANP 31‐67 as well as NT‐proBNP concentrations, both when including all centers and within each center. Interquartile range was large for many breeds, especially for NT‐proBNP. Among included breeds, Labrador Retrievers and Newfoundlands had highest median NT‐proBNP concentrations with concentrations 3 times as high as those of Dachshunds. German Shepherds and Cavalier King Charles Spaniels had the highest median proANP 31‐67 concentrations, twice the median concentration in Doberman Pinschers.

Conclusions and Clinical Importance

Considerable interbreed variation in plasma NP concentrations was found in healthy dogs. Intrabreed variation was large in several breeds, especially for NT‐proBNP. Additional studies are needed to establish breed‐specific reference ranges.     

Neutrophil Gelatinase–Associated Lipocalin in Dogs with Naturally Occurring Renal Diseases

Background

Neutrophil gelatinase–associated lipocalin (NGAL) is released from renal tubular cells after injury and serves in humans as a real‐time indicator of active kidney damage, including acute kidney injury (AKI) and chronic kidney disease (CKD). However, NGAL concentrations in dogs with naturally occurring AKI or CKD rarely have been explored in detail.

Hypothesis/Objectives

The goal of this study was to evaluate whether NGAL can serve as a useful biomarker in dogs with naturally occurring renal disease.

Animals

Client‐owned dogs with renal disease (57) and control dogs without any disease (12) were examined.

Methods

Serum NGAL (sNGAL) and urine NGAL (uNGAL) concentrations were measured in each animal by a newly developed ELISA system. Demographic, hematologic, and serum biochemical data were recorded. Survival attributable to AKI and CKD was evaluated at 30 days and 90 days, respectively.

Results

Serum and urine NGAL concentrations in azotemic dogs were significantly higher than in nonazotemic dogs and were highly correlated with serum creatinine concentration (P < .05). Among CKD dogs, death was associated with significantly higher sNGAL and uNGAL concentrations compared with survivors. Receiver‐operating characteristic curve (ROC) analysis showed that sNGAL was better than serum creatinine concentration when predicting clinical outcomes for CKD dogs (P < .05). The best cutoff point for sNGAL was 50.6 ng/mL, which gave a sensitivity and a specificity of 76.9 and 100%, respectively. Furthermore, dogs that had higher concentrations of sNGAL survived for a significantly shorter time.

Conclusion

sNGAL is a useful prognostic marker when evaluating dogs with CKD.     

Turnover of Se in adequately fed chickens using Se‐75 as a tracer

Inorganic selenium (Se) in the form of selenite is applied to livestock to avoid Se deficiency. Selenite is, however, an artificial Se source in diets of unsupplemented chickens. It is therefore hypothesized that organic Se sources, such as Se‐enriched yeast and wheat, could be a more suitable Se supply in animal nutrition, although information on the transition of Se from organic Se sources in fast‐growing chickens is scarce. In this work, chickens were fed a low Se diet (0.27 ± 0.01 mg Se/kg, Se‐enriched yeast) until 20 days of age, after which the Se concentration was increased to maximum concentration allowed by the poultry industry in Europe (0.5 p.p.m. Se). At the same time, a daily contribution of carrier‐free ⁷⁵Se tracer from labelled wheat was administered from day 20 to 27. The chickens showed S and Se homeostasis, as the concentration of S and Se in liver, blood or kidney remained about constant, and steady state of S and Se in the other organs was reached 1 day after the diet shift. The uptake of ⁷⁵Se was readily seen in all organs. After 1 week, the depuration of the ⁷⁵Se tracer was followed, and biological half‐lives and retention in individual organs were determined. The shortest biological half‐lives were observed in major metabolic organs, the liver, kidney and pancreas with half‐lives close to 4 days. There was a significant (p < 0.05) uptake in lung, brain and muscle that reached steady state when the administration of ⁷⁵Se was terminated. The half‐life of ⁷⁵Se in heart was 9 days and 7 days in blood. The longest half‐lives were observed in muscle (12 days), brain and lungs (13 days). All half‐lives were shorter than in Se deplete animals.     

Effect of feeding dried distillers' grains with solubles on milk yield and milk composition of cows in mid‐lactation and digestibility in sheep

We evaluated the effect of three sources of dried distillers' grains with solubles (DDGS) in diets of mid‐lactating dairy cows on milk production and milk composition and on digestibility in sheep. DDGS from wheat, corn and barley (DDGS₁), wheat and corn (DDGS₂) and wheat (DDGS₃) were studied and compared with a rapeseed meal (RSM). RSM and DDGS were characterized through in situ crude protein (CP) degradability. Nutrient digestibility was determined in sheep. Twenty‐four multiparous cows were used in a 4 × 4 Latin square design with 28‐day periods. Treatments included total mixed rations containing as primary protein sources RSM (control), DDGS₁ (D1), DDGS₂ (D2) or DDGS₃ (D3). RSM contained less rapidly degradable CP (fraction a), more potentially degradable CP (fraction b) and more rumen undegradable CP (UDP) than the three DDGS. In vivo digestibility of RSM organic matter was similar to DDGS. Calculated net energy for lactation (NE_L) was lower for RSM (7.4 MJ/kg DM) than for DDGS, which averaged 7.7 MJ/kg DM. Cows' dry matter intake did not differ between diets (21.7 kg/day). Cows fed D1 yielded more milk than those fed D3 (31.7 vs. 30.4 kg/day); no differences were found between control and DDGS diets (31.3 vs. 31.1 kg/day). Energy‐corrected milk was similar among diets (31.2 kg/day). Diets affected neither milk fat concentration (4.0%) nor milk fat yield (1.24 kg/day). Milk protein yield of control (1.12 kg/day) was significantly higher than D3 (1.06 kg/day) but not different form D1 and D2 (1.08 kg/day each). Feeding DDGS significantly increased milk lactose concentration (4.91%) in relation to control (4.81%). DDGS can be a suitable feed in relation to RSM and can be fed up to 4 kg dry matter per day in rations of dairy cows in mid‐lactation. However, high variation of protein and energy values of DDGS should be considered when included in diets of dairy cows.