Estimation of Genetic Variation Among Verticillium Isolates using AFLP Analysis

Amplified fragment length polymorphism (AFLP) analysis was used to study genetic variation among 76 isolates of Verticillium. A dendrogram based on the AFLP data revealed three main groups. One group consisted of 35 European isolates derived from Brassica napus together with five Californian isolates taken from B. oleracea. This group displayed a high degree of genetic similarity and included three isolates earlier classified as Verticillium longisporum, indicating that all isolates in this group probably should be regarded as members of V. longisporum. V. dahliae isolates constituted the second group while the third group contained four V. albo-atrum isolates. In addition to these three groups, a cluster of six V. nigrescens isolates was observed. However, the genetic distances between the isolates of V. nigrescens were much higher than those between members in the other groups and the bootstrap value for the V. nigrescens cluster was subsequently low. Four isolates classified as V. tricorpus were highly diverse and did not cluster together. Analysis of molecular variance revealed that the isolates of V. longisporum were separated into four subgroups, based on geographic origin. The study furthermore shows that AFLP is a suitable method for studying population structure in Verticillium.     

Genetic potential for residual feed intake and diet fed during early- to mid-gestation influences post-natal DNA methylation of imprinted genes in muscle and liver tissues in beef cattle

Discovery of epigenetic modifications associated with feed efficiency or other economically important traits would increase our understanding of the molecular mechanisms underlying these traits. In combination with known genetic markers, this would provide opportunity to improve genomic selection accuracy in cattle breeding programs. It would also allow cattle to be managed to improve favorable gene expression. The objective of this study was to identify variation in DNA methylation between beef cattle of differential pre-natal nutrition and divergent genetic potential for residual feed intake (RFI). Purebred Angus offspring with the genetic potential for either high (HRFI) or low (LRFI) RFI were prenatally exposed to either a restricted maternal diet of 0.5 kg/d average daily gain (ADG) or a moderate maternal diet of 0.7 kg/d ADG from 30 to 150 d of gestation. We performed DNA methylation analysis of differentially methylated regions (DMR) of imprinted genes (Insulin-like growth factor 2 (IGF2) DMR2, IGF2/H19 imprinting control region (ICR) and IGF2 receptor (IGF2R) DMR2) using post-natal samples of longissimus dorsi (LD) muscle taken from male and female calves at birth and weaning, and of LD muscle, semimembranosus (SM) muscle, and liver samples collected from steers at slaughter (17 months of age). Interestingly, for all three DMR investigated in liver, LRFI steers had higher levels of methylation than HRFI steers. In LD muscle, IGF2/H19 ICR methylation differences for heifers at birth were due to pre-natal diet, while for steers at birth they were mostly the result of genetic potential for RFI with LRFI steers again having higher levels of methylation than HRFI steers. While results from repeated measures analysis of DNA methylation in steers grouped by RFI revealed few differences, in steers grouped by diet, we found higher methylation levels of IGF2 DMR2 and IGF2R DMR2 in LD muscle of restricted diet steers at weaning and slaughter than at birth, as well as increased methylation in LD muscle of restricted diet steers compared with moderate diet steers at weaning and/or slaughter. Our results suggest that differential pre-natal nutrition, and divergent genetic potential for RFI, induces tissue- and sex-specific alterations in post-natal IGF2 and IGF2R methylation patterns and that these patterns can vary with age in Angus beef cattle.     

Quantification of reservoir bags as airway pressure-limiting devices in a nonrebreathing system

ObjectiveTo evaluate the influence of reservoir bag types, volumes and previous use on the peak pressures (Pmax) and the times to develop 30 cmH₂O pressure (P30) within a nonrebreathing system with a closed adjustable pressure-limiting (APL) valve.Study designIn vitro study using three-way factorial design with repeated measure on one factor.SubjectsA total of 75 new anesthesia reservoir bags (five types, three volumes, five bags from each type × volume). The bag types were reusable latex (RL), disposable latex (DL) and three disposable neoprene (DN-1, DN-2 and DN-3).MethodsEach bag was tested three times (treatments): new, after prestretching and 1 week later. The bags were attached to a Bain system and anesthesia machine with closed APL valve and patient port with O₂ flow 2 L minute^–1 until Pmax was reached. The Pmax and time to reach P30 values were determined from recorded pressure traces. General linear mixed model analysis was used to examine the effects of bag type, volume and treatment. One-sided 95% upper prediction limits of Pmax were calculated to test the null hypothesis that predicted Pmax of new bags would be ≥ 50 cmH₂O for each factor combination.ResultsRL bags were the least and DN-3 bags were the most compliant. Prestretching increased compliance in all bag types. Smaller bags of RL, DL and DN-1 were less compliant than larger ones. The predicted Pmax values were < 50 cmH₂O only for DN-3 bags after prestretching. The time to reach P30 was critically low when using 0.5 L bags (median 17 seconds).Conclusions and clinical relevanceTo minimize the risk of barotrauma, highly compliant reservoir bags (e.g. DN-3) are recommended and reusable bags should be avoided. Bags should be prestretched before first use, 0.5 L bags should be avoided and fresh gas flow minimized.     

Characterization of protection against systemic infection and disease from experimental bovine viral diarrhea virus type 2 infection by use of a modified-live noncytopathic type 1 vaccine in calves

OBJECTIVE: To evaluate protection resulting from use of a modified-live noncytopathic bovine viral diarrhea virus (BVDV) type 1 vaccine against systemic infection and clinical disease in calves challenged with type 2 BVDV. ANIMALS: 10 calves, 5 to 7 months of age. PROCEDURES: Calves were allocated (n = 5/group) to be nonvaccinated or vaccinated SC on day 0 with BVDV 1 (WRL strain). Calves in both groups were challenged intranasally with BVDV type 2 isolate 890 on day 21. Rectal temperatures and clinical signs of disease were recorded daily, and total and differential WBC and platelet counts were performed. Histologic examinations and immunohistochemical analyses to detect lesions and distribution of viral antigens, respectively, were performed. RESULTS: After challenge exposure to BVDV type 2, nonvaccinated calves developed high rectal temperatures, increased respiratory rates, viremia, leukopenia, lymphopenia, and infection of the thymus. Vaccinated calves did not develop high rectal temperatures or clinical signs of respiratory tract disease. Vaccinated calves appeared to be protected against systemic replication of virus in that they did not develop leukopenia, lymphopenia, viremia, or infection of target organs, and infectious virus was not detected in peripheral blood mononuclear cells or the thymus. CONCLUSIONS AND CLINICAL RELEVANCE: The modified-live BVDV type 1 vaccine protected against systemic infection and disease after experimental challenge exposure with BVDV type 2. The vaccine protected calves against infection and viremia and prevented infection of target lymphoid cells.     

Standing enucleation in the horse: a report of 5 cases

Enucleation was performed in 5 horses under local anesthesia and sedation with the horse standing. Minimal hemorrhage occurred during the surgical procedure, and there were no other reported complications. Standing enucleation is a surgery that is safe to perform in horses.     

Investigation of a possible chain of Streptococcus suis infection in a pig breeding community using PCR-based genotyping

Streptococcus (S.) suis plays an important role in pig breeding causing invasive diseases such as meningitis and polyarthritis. Herd problems with Streptococcus suis are common in many pig farms and are frequently characterised by disease outbreaks. In this context, it is often important to identify chains of infection, e.g. between a farrow-to-wean and a grower farm. In the following case report a possible chain of infection among the different farms of a farrow-to-finish system was investigated. In two grower units herd problems with S. suis were present; the mortality was higher than 5 %. It appeared likely, that the streptococci causing these problems were transmitted from a single farrow-to-wean unit to the two different grower farms. In the respective farms swabs were taken from different healthy animals and, in the case of the grower farms, also from the infected animals. Genotypic profiling of strains by a multiplex-PCR and AFLP typing method revealed that two different S. suis pathotypes were responsible for the herd problems. Both pathotypes could not be detected in the farrow-to-wean farm.Thus, a chain of infection originating from the farrow-to-wean farm appeared unlikely. The multiplex PCR was in this case sufficient to elucidate the described problem.     

High-resolution ultrasonography of xenografted domestic cat ovarian cortex

Transplantation of ovarian tissue has high potential for female gamete conservation. However, optimal timing of oocyte recovery for in vitro maturation and fertilization is still critical. Therefore the aim of the present study was to use high-resolution transcutaneous ultrasonography to monitor follicular development within xenografted ovarian tissue. Ovarian cortex fragments (n=44) from domestic cats were transplanted into athymic nude rats (n=12). Graft development in the animals was assessed weekly by high frequency ultrasound (10-22 MHz) under two different FSH regimes. Blood collection for serum estradiol determination and vaginal smears were performed simultaneously. The xenografts were removed at different time points according to the ultrasound findings. The survival rate of the transplants 4 weeks after surgery was 54.5% and antral follicular growth was observed within 10 grafts from 5 different hosts (8.6 +/- 6.43 follicles per graft). Early follicle antrums could be detected from 0.4 mm onwards. The growth rate of the antral cavity was calculated from weekly measurements (0.56 +/- 0.44 mm per week). Although vaginal cells and estradiol levels followed a cyclic pattern, no correlation was found between follicular diameter, estradiol and keratinized vaginal cells. We recovered 5, 1 and 4 cumulus oocyte complexes from three different individuals during weeks 19, 21, and 23 respectively. Extrusion of a polar body (1 oocyte) and germinal vesicle break down (7 oocytes) indicated progression of maturation after in vitro culture. We conclude that ultrasonography und provided a reliable method to examine xenograft survival and follicular development within the grafts. Furthermore, this technique is suitable for assessment of the efficiency of hormonal treatment and narrowing of the optimal time frame for oocyte retrieval. To our knowledge, this is the first report of the in vivo development of early antral follicles in mammalian species.     

Risk factors influencing death prior to discharge in 302 dogs undergoing unilateral adrenalectomy for treatment of primary adrenal gland tumours

Adrenalectomies for canine adrenal tumours are associated with peri-operative morbidity and mortality. Objectives of this study included assessing the prognostic value of tumour- or surgery-related variables in predicting peri-operative mortality and overall survival in dogs undergoing adrenalectomies for primary adrenal tumours as well as pre-treatment with phenoxybenzamine on survival to discharge with pheochromocytomas specifically. A multi-institutional retrospective cohort study was performed across nine institutions. Electronic medical record searches identified 302 dogs which met the inclusion criteria. Data collected included dog-related, tumour-related, treatment-related, surgery-related, and outcome variables. Univariate and multivariable logistic regression and cox proportional hazards models were used to identify variables associated with death prior to discharge and tumour-related survival. Overall, 87% of dogs survived to discharge with a tumour-related survival time of 3.96 years. Post-operative complications were reported in 25%. Increased surgical time (p = 0.002) and pre-surgical medical treatment other than phenoxybenzamine (p = 0.024) were significantly associated with increased peri-operative mortality while ureteronephrectomy (p = 0.021), post-operative pancreatitis (p = 0.025), and post-operative aspiration pneumonia (p < 0.001) were significantly associated with decreased overall survival. Phenoxybenzamine pretreatment had no effect on peri-operative mortality. Thirty-seven of 45 (82%) dogs with pheochromocytomas not pretreated survived to discharge, and 50 of 59 (85%) dogs with pheochromocytomas pretreated with phenoxybenzamine survived to discharge (p = 0.730). This study provides information on risk factors for death prior to discharge and tumour-related survival that may help guide clinical management and owner expectations. In addition, the study findings challenge the previously reported benefit of phenoxybenzamine for pretreatment of dogs undergoing adrenalectomies for pheochromocytomas.     

Schiff-Sherrington syndrome in a 7-year-old zebra mare (Equus quagga)

A 7-year-old plains zebra (Equus quagga) mare was presented to the Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Germany as an emergency. The owner reported that the mare was unable to stand after a suspected traumatic episode at the home yard. The mare was transported in lateral recumbency in a trailer. Clinical examination revealed tachycardia (up to 102 beats/min), tachypnoea (28 breaths/min), mild fever (38.7°C) and clinical dehydration with elastic skin and dry and red mucous membranes. Neurological examination showed paraplegia with complete loss of nociception of the mid-thoracic region and caudal to it. In contrast, examination of the thoracic limbs revealed marked extensor rigidity and hyperaesthesia resulting in an enhanced acute response to minimal touch of the neck and shoulder region. Lateral radiographs of the thoracic vertebral column led to the diagnosis of a complete fracture of the vertebral body of the 7th thoracic (T) vertebra with suspected traumatic compression of the spinal cord. Post-mortem examination confirmed the diagnosis. Gross pathology revealed a complete comminuted fracture of T7 with marked compression of the spinal cord. This is the first report describing the clinical, radiological and pathological findings of a fracture of a thoracic vertebra leading to a Schiff-Sherrington syndrome in a zebra (Equus quagga).