OC4Influence of Seminal Plasma Added to Highly Diluted Semen on Sperm Motility of Young Boar Feed with Selenium and Vitamin E

High dilution rates have been documented as detrimental for boar spermatozoa, shortening their lifespan (Centurion et al. 2003, Biol Reprod 69: 640–646). Addition of seminal plasma (SP) to semen extenders, or selenium (Se) and vitamin E (VE) in diet of boars could increase motility of highly diluted spermatozoa (HDS). The aim of this work was to evaluate the effect of seminal plasma on sperm motility of HDS from boars feed with Se and VE. Sixteen 12 month-old boars were designed to one of four dietary treatments: (i) control, Se 0 ppm–VE 0 IU/kg; (ii) Se 0–VE 250; (iii) Se 0.5–VE 250 and (iv) Se 0.5–VE 0. Boars were treated for 8 weeks before semen collection. Sperm rich fractions from each boar were diluted to 5 × 10⁶ sperm/ml in PBS medium and incubated at 37°C with or without 10% SP. The measurements were done at 0, 2 or 5 h. Data were analyzed as a mixed model for a factorial design [2 (Se) × 2 (VE) × 2 (SP) × 3 (h)]. Percentage of sperm motility (PSM) increases significantly (p < 0.001) with addition of Se (81.3 ± 1.52), VE (81.0 ± 1.62) and SP (81.5 ± 1.57) vs control (73.4 ± 1.61). There was significant interaction Se × VE (p < 0.001) and Se × VE × SP (p < 0.05) in PSM. However, PSM was affected significantly by time (0 h 83.4 ± 1.92; 2 h 80.7 ± 1.92 and 5 h 67.9 ± 1.92; p < 0.001). There was significant interaction SP × Time (p < 0.05) in PSM. These results indicate that Se, VE and SP improve seminal viability. Addition of 10% of SP maintains PSM at least during 5 hours.     

Growth performance, carcass composition, quality, and enhancement treatment of fresh pork identified through deoxyribonucleic acid marker-assisted selection for the Rendement Napole gene

Progeny (n = 70) from unrelated, DNA tested, Rendement Napole carrier (RN-/rn+) Hampshire sires, and DNA tested, Rendement Napole normal (rn+/rn+) Yorkshire dams were genotyped for the segregating RN- allele via DNA marker-assisted methodology. Six slaughter groups ensued, with littermates all being represented within the same slaughter group. Boneless pork loins were removed from right carcass sides after a 48-h chill at 2 degrees C. The anterior portions of the loins were not enhanced, whereas the posterior sections were enhanced with a solution containing 0.5% sodium chloride and 0.5% sodium tripolyphosphate to 110% of their initial weight. Carcasses of carrier pigs had less (P < 0.05) 10th rib fat depth and a greater (P < 0.01) percentage carcass lean than carcasses of normal pigs. Postmortem LM pH of carrier pigs was lower (P < 0.002) at 3, 6, 12, and 24 h, and tended to be lower (P = 0.062) at 48 h compared with that of normal animals. Samples of LM from carrier pigs had greater (P < 0.01) glycolytic potential values, drip loss percentages, and a* values, and lower pH values at fabrication than LM from normal pigs. No genotype differences (P > 0.05) were found for LM lactate, L*, or b* values. Nonenhanced semimembranosus samples from carrier pigs exhibited greater (P < 0.05) purge loss percentages and L* values, and lower (P < 0.01) pH values than samples from normal pigs. Enhanced LM samples exhibited greater (P < 0.05) drip and purge loss percentages, greater pH, and lower L* values at fabrication, regardless of Napole status. These findings suggest that the Napole gene has a positive influence on carcass leanness but detrimental effects for lean quality, which were often further compounded when meat was subjected to enhancement treatment.     

A novel mutation of the CLCN1 gene associated with myotonia hereditaria in an Australian cattle dog

BACKGROUND: Heritable myotonia is a genetic muscle disorder characterized by slow relaxation of skeletal muscles. The main clinical signs are skeletal muscle stiffness, especially after vigorous contraction, and muscle hypertrophy. Muscle stiffness may be enhanced by inactivity, and often is relieved by exercise. Myotonia can be inherited in an autosomal dominant or recessive manner (Thomsen- or Becker-type myotonia, respectively). In mice, goats, Miniature Schnauzer dogs, and most affected humans, the disorder is caused by mutations in CLCN1, which encodes the skeletal muscle voltage-gated chloride channel, Cl1C-1. HYPOTHESIS: We hypothesized that an Australian Cattle Dog with generalized muscle stiffness and hypertrophy examined at the Ontario Veterinary College would have a mutation in the CLCN1 gene. ANIMALS: A pure-bred Australian Cattle Dog from Ontario, Canada, was used. METHODS: Based on clinical signs and electromyographic test results, a diagnosis of myotonia hereditaria was made, and a muscle biopsy was collected for genetic analysis. RESULTS: Sequence data obtained from the affected dog confirmed that it was homozygous for a single base insertion in the CLCN1 coding sequence. This mutation would result in a truncated ClC-1 protein being expressed, which, based on molecular evidence from other studies, would result in functionally compromised chloride conduction in the skeletal muscles of the animal. CONCLUSIONS AND CLINICAL IMPORTANCE: To the authors' knowledge, this report describes the Ist case of myotonia in an Australian Cattle Dog and represents the 1st non-Schnauzer canine myotonia to be genetically characterized. In addition, we developed a polymerase chain reaction-based genetic screen to detect heterozygotes with this mutation in the at-large Australian Cattle Dog population.     

Effects of 2 concentrations of sodium citrate on coagulation test results, von Willebrand factor concentration, and platelet function in dogs

BACKGROUND: Blood collection tubes containing 3.2% (0.109 M) sodium citrate, instead of 3.8% (0.129 M) sodium citrate, have recently become available in the United States. These tubes are visually indistinguishable from the traditional 3.8% sodium citrate tubes, except for wording on the label. Consequently, samples for hemostatic evaluation are frequently collected in tubes containing the lower concentration of sodium citrate. HYPOTHESIS: Results of hemostasis assays are different in samples collected in 3.2% versus 3.8% sodium citrate. ANIMALS: Twenty healthy dogs. METHODS: This study aimed at determining whether results of standard coagulation tests, von Willebrand factor concentration (vWF:Ag), and platelet function with the platelet function analyzer PFA-100a were affected by the different concentrations of sodium citrate. Blood samples were collected in tubes containing either 3.2% or 3.8% sodium citrate concentrations and processed routinely for coagulation assays (one-stage prothrombin time [OSPT], activated partial thromboplastin time [aPTT], fibrinogen concentration, and platelet count), vWF:Ag, and platelet function assays with a PFA-100. RESULTS: There was no significant difference between samples collected in 3.2% versus those collected in 3.8% sodium citrate for OSPT, aPTT, fibrinogen concentration, platelet count, or vWF:Ag. The closure times with collagen/adenosine diphosphate were significantly shorter (66 +/- 8.1 versus 74.8 +/- 9.7 seconds; P < .0001) with the 3.2% than with 3.8% sodium citrate concentration, and the hematocrit was significantly higher (47.9 +/- 5.6 versus 46.0 +/- 4.7 seconds; P = .03) in samples collected in 3.2% than in those collected in 3.8% sodium citrate. CONCLUSIONS AND CLINICAL IMPORTANCE: There is no clinically relevant effect of collection of blood into 3.2% or 3.8% sodium citrate.     

Needle tract implantation after fine needle aspiration biopsy (FNAB) of transitional cell carcinoma of the urinary bladder and adenocarcinoma of the lung

This paper reports three clinical cases of needle tract implantation of neoplastic cells on the abdominal and thoracic wall after ultrasound (US) fine needle aspiration biopsy (FNAB). Primary tumors were two transitional cell carcinomas of the urinary bladder (2 dogs) and one pulmonary adenocarcinoma (1 cat). All three masses grew up along the needle tract. To our knowledge, the seeding of pulmonary adenocarcinoma cells after FNAB on the thoracic wall has never been reported in veterinary medicine.     

Short-term biological variation of clinical chemical values in Dumeril's monitors (Varanus dumerili).

Plasma biochemical values are routinely used in the medical management of ill reptiles, and for monitoring the health of clinically normal animals. Laboratory tests, including clinical biochemical values, are subject to biological and analytical variation, the magnitude of which determines the utility of population-based reference ranges for the detection of abnormal results in the individual animal. Nested analysis of variance of repeated measurements allows the variance to be broken into within-individual, between-individual, and analytical variation. When the within-individual variation is large and the interindividual variation is low, a sample may be accurately classified as normal or abnormal based on a population-based reference interval. However, if the intraindividual variation is low and the interindividual variation high, population-based reference intervals are of limited value as the ranges for an individual encompass only a part of the conventional reference interval. Between-lizard, within-lizard, and analytical components of variance were assessed by nested analysis of variance for 16 commonly measured plasma biochemical parameters in eight healthy adult Dumeril's monitors (Varanus dumerili). Albumin, cholesterol, phosphate, calcium, sodium, and total protein demonstrated levels of individuality suggesting that comparison of a single measurement to a conventional population-based reference range may be too insensitive to detect small but significant alterations in the value for that animal. Only for potassium and AST did the index of individuality suggest that the use of reference values may be warranted. Uric acid, globulin, glucose, and amylase fell in a gray zone, where population-based ranges should be used with caution. The critical difference indicates the difference between two consecutive analytical results that may be safely ascribed to natural variation. In the present study critical difference varied from 7 and 11%, respectively, for sodium and chloride to 75 and 125% for uric acid and AST.     

Resistance to macrocyclic lactone anthelmintics and associated risk factors on sheep farms in the lower North Island of New Zealand

AIM: To estimate the prevalence of resistance to macrocyclic lactone (ML) anthelmintics over a 3-year period on sheep farms in the Taihape area, and to determine associations between the presence of ML resistance and farm management practices. METHODS: All farmers with >1,000 lambing ewes served by one veterinary practice in the Taihape area of the central North Island of New Zealand were invited to take part in the study (n=157); respondents were selected on their willingness to participate. Resistance to ML was measured during 2004-2006, using a standard faecal egg count reduction test (FECRT) and a half dose of oral ivermectin (IVM; 0.1 mg/kg; n=84). Additional FECRTs were conducted separately on some farms, using full doses of IVM (n=47), levamisole (LEV; n=49), a benzimidazole (BZ; n=76), and a BZ/LEV combination (n=42). Resistance was defined as <95% faecal nematode egg count reduction (FECR) 7-10 days post-treatment. Larval cultures were performed on pooled faecal samples from all pre- and post-treatment groups for which any positive faecal nematode egg counts (FEC) were recorded. To ascertain whether particular management or drenching strategies were associated with the presence of resistance, a questionnaire comprising 50 questions was developed, and each farmer interviewed individually. RESULTS: Of all farmers invited, 84/157 (54%) participated in this study. Based on undifferentiated FEC, ML resistance to the half-dose IVM was evident on 46/84 (55%) participating farms. The majority of these cases (71%) involved Ostertagia (Teladorsagia) circumcincta. Resistance was also evident to full doses of IVM on 22/47 (47%), BZ on 41/76 (54%), LEV on 18/49 (37%), and BZ/LEV on 2/42 (5%) farms. Positive associations (p<0.05) were evident between the presence of ML resistance and management practices, such as: weaning over half of the lambs onto paddocks not grazed by lambing ewes since June; not always returning lambs to the same paddock after drenching; and the use of visual signs to assess 'worminess'. Measuring drench efficacy in January to March rather than later in the year, and presence of resistance to BZ or LEV were associated with a higher apparent prevalence of ML resistance (p<0.05). CONCLUSIONS: Anthelmintic resistance, and particularly ML resistance, was widespread on sheep farms in the Taihape area, and mostly involved O. circumcincta. The prevalence of resistance was lower on farms on which management practices, particularly those involving lambs, maintained refugia of unselected nematodes. Further investigation is required to develop control strategies which minimise selection for resistant worms, and how these may vary with climate.     

Risk factors for Thoroughbred racehorse fatality in jump starts in Victoria, Australia (1989-2004)

REASONS FOR PERFORMING STUDY: The risk of fatality is greater in jump than in flat racing in Victoria, Australia. This is the first study to identify risk factors specific to jump starts in Victoria. OBJECTIVE: To identify risk factors for fatality of Thoroughbred racehorses in jump starts on all racecourses in Victoria, Australia between 1989 and 2004. METHODS: Fatalities comprised all horses that died during or immediately after a jump (hurdle or steeplechase) race or official jump trial and all horses that were subjected to euthanasia within 24 h of an event in which an injury was sustained. The retrospective study involved 191 case starts and 2324 control starts. Univariable and multivariable backward stepwise logistic regression was used to identify risk factors for fatality at any one start. A multiple level model was used with racecourse included as a random effect. RESULTS: In the final multivariable model, the duration of the racing career of the horse, the number of flat, hurdle and steeple starts accumulated in the 60 days prior to the case or control start, the number of flat and jump starts accumulated over the racing career, if the horse had had a start between 1 and 14 days prior to the case or control start, the type of jump race (hurdle or steeple), the calendar year of the start and the location of the racecourse were associated with fatality. CONCLUSIONS: The findings highlight the need to investigate further the differences between hurdle and steeplechase events and the adverse effect of prolonged prior flat racing careers on the risk of fatality in jump starts. POTENTIAL RELEVANCE: This is the first study to examine risk factors for fatality in jump starts in Victoria. The results should shape the development of interventions to reduce the risk in jump starts in the future.     

Isolation and characterization of bone marrow-derived equine mesenchymal stem cells

OBJECTIVE: To isolate and characterize bone marrow-derived equine mesenchymal stem cells (MSCs) for possible future therapeutic applications in horses. SAMPLE POPULATION: Equine MSCs were isolated from bone marrow aspirates obtained from the sternum of 30 donor horses. PROCEDURES: Cells were cultured in medium (alpha-minimum essential medium) with a fetal calf serum content of 20%. Equine MSC features were analyzed to determine selfrenewing and differentiation capacity. For potential therapeutic applications, the migratory potential of equine MSCs was determined. An adenoviral vector was used to determine the transduction rate of equine MSCs. RESULTS: Equine MSCs can be culture-expanded. Equine MSCs undergo cryopreservation in liquid nitrogen without altering morphologic characteristics. Furthermore, equine MSCs maintain their ability to proliferate and differentiate after thawing. Immunocytochemically, the expression of the stem cell marker CD90 can be detected on equine MSCs. The multilineage differentiation potential of equine MSCs was revealed by their ability to undergo adipogenic, osteogenic, and chondrogenic differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicate that bone marrow-derived stromal cells of horses can be characterized as MSCs. Equine MSCs have a high transduction rate and migratory potential and adapt to scaffold material in culture. As an autologous cell population, equine MSCs can be regarded as a promising cell population for tissue engineering in lesions of the musculoskeletal system in horses.