Pseudorabies virus infection in mink: a host-specific pathogenesis

Pseudorabies virus (PRV) is an alphaherpesvirus that causes a neurological disease in many wild and domestic animals. The neuropathology elicited by PRV is quite consistent regardless of the host with the only exception of mink, in which it is characterized by a vasculopathy rather than by an encephalitis. In this study, we aimed to investigate the underlying pathogenic mechanism(s) of PRV infection in mink by using immunohistochemistry and laser capture microdissection (LCM) on material from naturally and experimentally infected animals. The inflammatory reaction induced by PRV was minimal or absent not only in the nervous system, where we identified a low number of macrophages and a few T lymphocytes, but also in the primary replication site, the oropharyngeal mucosa; however, the number of PRV-infected cells detected by immunohistochemistry was extremely high both in the peripheral mucosa and in the nervous tissue. On the other hand, the vascular pathology included parenchymal hemorrhages of various degrees and, in specific cortical areas of the brain, fibrinoid degeneration of the capillary walls. Detection of viral antigens by immunohistochemistry revealed infection of endothelial cells of capillaries situated both in the oropharyngeal mucosa and in the brain stem; the presence of PRV DNA in vessels was further demonstrated by PCR performed on LCM samples of brain capillaries. These results can be interpreted as supporting the idea that the different pathology of the disease in mink may be the consequence of an increased endotheliotropism of PRV in this species. Infection of the vessel wall may then lead to vascular pathology and impairment in endothelial cell function, resulting in a weak immune response to infection.     

A Rapid Detection Method for the Ryanodine Receptor 1 (C7360G) Mutation in Quarter Horses

Background: Anesthetic-induced malignant hyperthermia has been documented in Quarter Horses and is caused by a single-point mutation in the ryanodine receptor 1 gene at nucleotide C7360G generating a R2454G amino acid substitution. An accurate, faster molecular test that is less prone to contamination would facilitate screening for the mutation in horses intended for breeding, in those undergoing surgical procedures, and in those with clinical signs compatible with malignant hyperthermia.
Objective: To report a rapid and accurate method for the detection of the ryanodine receptor 1 C7360G mutation.
Animals: Eleven diseased, 10 healthy, and 225 randomly selected Quarter Horses.
Methods: This study included horses with the ryanodine receptor 1 C7360G mutation as detected by gene sequencing. Available genomic and complementary DNA extracted from whole blood, hair or skeletal muscle was used for genetic analysis. Real-time polymerase chain reaction (RT-PCR) melting curve analysis was performed by equine specific primers and 2 hybridization probes (sensor and anchor probes) that contain the site of the mutation. Results from this method were blinded and compared with nucleic acid sequencing for validation.
Results: A rapid genotyping assay with fluorescence resonance energy transfer probes and melting curve analysis was accurate (100% agreement, K = 1) for identification of affected horses. The prevalence of the mutation in a random population of Quarter Horses was 1.3%.
Conclusions and Clinical Importance: Malignant hyperthermia in Quarter Horses can be rapidly and accurately detected by RT-PCR melting curve genotyping with hybridization probes.     

Di-tri-octahedral smectite for the prevention of post-operative diarrhea in equids with surgical disease of the large intestine: Results of a randomized clinical trial

The aim of this study was to evaluate the effects of a commercially available di-tri-octahedral (DTO) smectite product on clinical signs and prevalence of post-operative diarrhea in horses with colic associated with disease of the large intestine. Sixty-seven horses with surgical disease of the large intestine were randomly assigned to be treated with DTO smectite (n = 37; 0.5 kg via nasogastric intubation every 24 h for 3 days post-operatively) or a placebo (n = 30). The effect of treatment on fecal scores and clinical and hematological parameters, including heart rate, mucous membrane color, temperature, total white blood cell count, total neutrophil count and total plasma protein values, were determined.Horses treated with DTO smectite had a significant reduction in the prevalence of post-operative diarrhea (10.8%), compared with controls (41.4%). A significant improvement in mucous membrane color was observed 72 h post-operatively in horses receiving treatment, compared with placebo. Administration of DTO smectite to colic patients with disease of the large intestine reduced the occurrence of diarrhea in the early post-operative period.     

Prevalence of gastric ulcers in Thoroughbred broodmares in pasture: a preliminary report

Gastroscopic examinations were performed in 62 Thoroughbred broodmares (33 pregnant, 29 non-pregnant) at one breeding farm to investigate the prevalence of gastric ulceration. Age, pregnancy status, race earnings, last race start, herd size, medical history, number of live foals, breeding years, feed type and number of feedings were recorded, plus coat condition and body condition score were determined. Twenty-one mares were re-evaluated after foaling, and the foaling date, foal weight at birth and placenta weight were recorded.The overall prevalence of gastric ulcers was 70.9%, with a median ulcer score of 3.0 (range: 2–5). Most ulcers were present on the squamous portion of the stomach, while two mares had glandular ulcers. There were no differences in the presence, location and severity of gastric ulcers between pregnant and non-pregnant mares. Furthermore, there were no significant associations between the variables measured and the presence of gastric ulceration. The prevalence of gastric ulceration in this specific population of horses was higher than expected and further investigation is warranted to determine the factors that contributed to this finding.     

ORIGINAL ARTICLE: Effects of herbage ingestion upon ileal digestibility of amino acids in heavy Iberian pigs fed on an acorn‐based diet

We conducted two experiments with heavy Iberian pigs to determine the ileal digestibility of amino acids (AA) in acorns and freshly cut herbage, and the effects of adding fresh herbage upon the supply of ileal digestible AA when pigs were fed on holm‐oak acorns. In Experiment 1, carried out in cannulated pigs of 107 kg bodyweight (BW), daily intake of acorns reached 44.9 g DM/kg^0.75 BW. Arg, His and Thr showed the lowest apparent ileal digestibility (AID) values, whereas Met, the branched‐chain AA and Phe had the highest coefficients. The AID of total EAA was 0.716 but only 0.222 for NEAA. Most of the digestive and absorptive processes of acorn protein occurred before the hindgut. Acorn provides (per kg DM) 2.27 g apparent ileal digestible Lys and 22.7 g apparent total digestible AA. Standardized ileal digestibility (SID) values for EAA, NEAA and total AA were 0.924 ± 0.020, 0.784 ± 0.041 and 0.860 ± 0.029. In Experiment 2 fresh herbage was given to six cannulated Iberian pigs of 140 kg either as a single feed (13.7 g DM/kg^0.75 BW) or as a supplement to acorns (28.4 g DM/kg^0.75 BW). When only freshly cut forage was offered the AID of the EAA, NEAA and total AA was close to 0.65 and supplied (per kg DM ingested) 5.61 g AID Lys and 91.7 g digestible AA. Standardized ileal values were 0.744 ± 0.023, 0.912 ± 0.038 and 0.831 ± 0.030 respectively. The addition of fresh forage to the acorns led to a significant decrease in AID of AA in acorn due to digesta transfer to the hindgut: His (p < 0.01), Met (p < 0.001), Phe (p = 0.092), Thr (p < 0.05) and Val (p < 0.05), but Arg, Lys and the branched‐chain AA remained unaffected. The main contribution of herbage to AA nutrition of the grazing Iberian pig relies mainly on increasing the supply of digestible AA for pig tissues.     

Utilization of milk amino acids by the suckling Iberian piglet

Sixteen pure-bred Iberian (IB) sows were used in two trials to determine the efficiency of utilization of milk protein and amino acid (AA) for growth in suckling piglets. It was hypothesized that there may be one or more strongly limiting essential AA (EAA) responsible for the slow rate of growth of the IB piglet. This AA will show the highest fractional retention. Daily milk yield and composition were determined weekly over a 34-day lactation period. Within each litter, one piglet at birth and four piglets on d 35 of life were slaughtered. The protein content of the IB sow milk was similar to that reported for conventional breeds. However, branched-chain AA, Thr, Pro, Asp and Ala were in concentrations somewhat below the range of literature values and Arg and Met, substantially above it. Milk intake per piglet tended to be greater in Trial 2 (832 vs. 893 g/day respectively; p = 0.066). However, the IB piglets grew at 168 ± 3.3 g/day, irrespective of the trial. The whole-body protein of piglets at weaning and the protein deposited in their body during the lactating period showed very close AA pattern. Among EAA, His and Arg show the highest fractional retentions (g AA retained/g AA ingested) in whole-body tissues (1.019 ± 0.025 and 0.913 ± 0.017 respectively) and also the highest body to milk ratios (1.50 and 1.41 respectively). Gly and Ala presented, among non essential AA, the highest efficiencies of utilization for tissue deposition (1.803 ± 0.057 and 1.375 ± 0.026 respectively) and body to milk ratios (2.75 and 2.12 respectively). These results suggest that the low efficiency of utilization of milk protein and the low rate of gain of the IB suckling piglet can be explained by a marked shortage in His supply, in addition to the suboptimal milk provision of Arg, Gly and Ala.