Transfer of MicroRNAs From Epididymal Epithelium to Equine Spermatozoa

All epididymal regions are lined with multiple epithelial cell types, each with different functions to provide the luminal environment for spermatozoal maturation. Epithelial cells also create apical blebs, which are released from the apical surface via apocrine secretion and disintegrate in the lumen, thereby releasing epididymosomes. Epididymosomes transport proteins to spermatozoa and contain microRNAs. We hypothesized that epididymosomes also transfer miRNA from epididymal epithelium to spermatozoa. Quantitative real-time polymerase chain reaction was used to determine miRNA profiles of epididymal tissue from caput and cauda, epididymal spermatozoa from caput and cauda, and epididymosomes and from caput, proximal corpus, distal corpus, and cauda. Pathway analysis was performed using DIANA tools on the miRNA unique to caudal spermatozoa. We found 66 newly acquired miRNAs in spermatozoa located in the caudal epididymis. Predicted pathways targeted by these miRNAs suggest a role in cell motility and viability and factors in oocyte and embryo maturation and development. These findings suggest that miRNAs are transported to spermatozoa from epididymal epithelium via epididymosomes.     

The Equine Science Society: From the 1st to the 25th Symposium

In 1968, the group currently known as the Equine Science Society hosted its first symposium. At the meeting, there were 27 participants representing about a dozen institutions and 23 research presentations were given. In 2017, the Society is hosting its 25^th Symposium and will have over 200 presentations from individuals representing about 120 institutions. Throughout the history of the Society, the objectives have been to promote quality equine research, establish communication amongst those involved with equine research, teaching, extension, and production, conduct periodic symposia, and cooperate with other organizations having similar interests.     

The use of skin delayed-type hypersensitivity as an adjunct test to diagnose brucellosis in cattle: a review

Brucellosis, caused by bacteria of the genus Brucella, is a contagious disease that causes economic loss to owners of domestic animals due to loss of progeny and milk yield. Because cattle, sheep, goats, and to a lesser extent pigs are considered to be the source of human brucellosis, serological tests have been used to screen domestic animals for antibodies against Brucella. Although the serological tests helped to eradicate brucellosis in many countries, serological tests are not always adequate to detect latent carriers of Brucella. Therefore, the use of the skin delayed-type hypersensitivity (SDTH) test, which is independent of circulating antibodies, might improve the diagnosis of brucellosis. In the literature, however, there are conflicting reports as to the value of the SDTH test for the diagnosis of brucellosis. Some studies consider the test unreliable, whereas others advocate its use because it detects brucellosis earlier than serological tests. The objectives of this study were therefore to assess the characteristics of the SDTH test, to select a Brucella strain that will yield a suitable brucellin for use in the field, and to determine whether the use of serological tests in combination with the SDTH test improves the detection of brucellosis. The results of this study clearly show that the SDTH test detects latent carriers of Brucella and confirms brucellosis in cattle with ambiguous serological test results. Brucellins prepared from smooth or mucoid strains of Brucella are better suited for use in the field than brucellins prepared from rough strains because they detect brucellosis in cattle with acute as well as chronic infection. The SDTH test is highly specific (99.3% specificity), and repeated testing of naive cattle or cattle infected with microorganisms that serologically cross-react with Brucella does not sensitize cattle to subsequent SDTH tests. However, it is possible that some naive cattle may serologically react to the injection of brucellin. The effect of these serological reactions on the sero-diagnosis of brucellosis is limited, because cattle may only now and then react serologically either with the serum agglutination test (SAT) or the complement fixation test (CFT). Nevertheless, cattle infected with microorganisms that serologically cross-react with Brucella may test seropositive for brucellosis 4 to 7 weeks after injection of brucellin, depending on the cross-reacting microorganism. The value of the SDTH test for the diagnosis of brucellosis was demonstrated after an outbreak of brucellosis. When the SDTH test was used in combination with SAT and CFT at diagnostic threshold > or =2 mm or > or =1 mm (increase in skinfold thickness), respectively, 39/44 (88%) or 42/44 (95%) of the infected cattle were detected compared with only 27/44 (61%) when SAT and CFT were used. When cattle in areas of low prevalence or in areas free from brucellosis are tested with the SDTH test an increase > or =2 mm in skinfold thickness should be considered indicative of infection. When the control and eradication of brucellosis is based on test-and-slaughter, an increase of > or =1 mm in skinfold thickness should be considered indicative of infection. Repeated serological testing complemented with the SDTH test in this programme will shorten the quarantine (movement control) period of a suspect herd, limiting the financial loss incurred during outbreaks of the disease. Consequently, since the SDTH test usually does not interfere with the serological diagnosis and can safely be used to establish the infection status of cattle in a suspect herd, it is opportune to consider adding the SDTH test to the procedure currently used to diagnose brucellosis in individual animals.     

Ultrasonography as an adjunct to pregnancy assessments in the mare

Ultrasonographic detection of pregnancy was performed in 952 Thoroughbred mares. Characteristic images were obtained between 12 and 55 days after breeding. Twins in the same uterine horn as well as twins in opposite horns were imaged, and for the first few months of gestation appeared similar to single pregnancies. Growth curves as a function of time were developed, utilizing horizontal diameters of the blastocysts versus breeding dates. Comparisons between horizontal and vertical diameters demonstrated no appreciable differences when observed as a group. Growth curves derived from twins did not differ significantly from those for single pregnancies during the period we observed them. Ultrasonographic evaluations were valuable in substantiating mechanical destruction of one of the twin blastocysts immediately following the maneuver. Uterine artifacts such as endometrial and lymphatic cysts also were visualized and portrayed. Although these structures represented a source of false-positive results, they usually could be properly identified on the basis of their morphometric and growth characteristics.     

Equine Protozoal Myeloencephalitis: A Report of Two Cases From Western Canada

Two cases of nonsuppurative myeloencephalitis are reported which clinically and pathologically resemble equine protozoal myeloencephalitis. Lesions in both horses were associated with Toxoplasma-like organisms visible in microscopic sections. Clinical signs and lesions in one case primarily involved the brain and in the other case principally involved the spinal cord and associated meninges. Positive identification of the organisms was not achieved; however, the etiological agent is unlikely to be a species of Toxoplasma. Recently published studies suggest a species of Sarcocystis is involved.     

Failure of Psyllium Mucilloid to Hasten Evacuation of Sand From the Equine Large Intestine

Objective —To examine the efficacy of psyllium mucilloid in evacuating sand from the equine large intestine.
Animals —12 clinically healthy pony geldings.
Procedure—Twelve ponies were assigned to 2 groups of six each. One group was treated with psyllium and the second was a control group. All ponies had an exploratory celiotomy and 10 g/kg body weight of sand was placed into the cecum. Ponies were fed a grain mixture alone at 1 g/kg (controls), a grain mixture plus psyllium pellets, each at 1 g/kg body weight (3 ponies), or fed a grain mixture and given psyllium powder by nasogastric tube at 1 g/kg body weight divided into two daily doses in 3 L of water (3 ponies). Radiographs were taken on days 1 (3 per group), 5 (all ponies), and 11 (3 per group) to monitor sand transit through the large intestine. Ponies were euthanatized 11 days after surgery. Sand was collected from the contents of the cecum, ventral colon, dorsal colon, and small colon. Dry weight of the recovered sand was compared between the two treatment groups as a percentage of the dry weight of sand placed in the cecum.
Results —No significant differences were detected in the mean percentage of sand recovered between the two treatment groups ( P < .05), with 39.2% recovered in ponies treated with psyllium and 27.4% recovered in control ponies.
Clinical Relevance —Psyllium mucilloid had no apparent effect on sand evacuation from the equine large intestine. When intake of sand is prevented, the equine large intestine can reduce and possibly eliminate its sand burden.     

Addition of Glutathione to an Extender for Frozen Equine Semen

The manipulation of equine semen during cryopreservation reduces sperm viability and fertility because of, among other factors, membrane lipid peroxidation that makes cells highly susceptible to free radicals and reactive oxygen species (ROS). The oxidative effect caused by the generation of ROS can be reduced by the addition of antioxidants to the seminal plasma or to the extenders used for freezing. The current study was performed to test the in vitro effect of exogenous glutathione added in five different concentrations (control, 2.5 mM, 5.0 mM, 7.5 mM, and 10 mM [treatments 1-5, respectively]) to the extender for 12 stallions. Analyzed parameters were sperm motility, viability, and acrosome and plasmatic membrane integrity. Total motility was higher in treatments 1 and 2 (P < .05); viability, progressive motility, and plasmatic membrane integrity were higher in treatment 2 (P < .001). As for acrosome membrane integrity, treatment 3 showed the best results (P < .05). The addition of 2.5 mM glutathione to the freezing extender preserves total motility and increases sperm viability, progressive motility, and plasmatic membrane integrity. Concentrations above 2.5 mM were deleterious to spermatozoa.     

Fifty years of the British Equine Veterinary Association as a facilitator of progress in equine clinical science

The British Equine Veterinary Association (BEVA) was established in 1961 and launched the Equine Veterinary Journal (EVJ) in 1968. This review outlines some of the major advances in equine science and practice that have occurred in that time and the role played by the Journal in facilitating those developments.