Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Factors Affecting the Release of Ascospores of Anisogramma anomala

ABSTRACT Relationships between environmental factors and release of ascospores of Anisogramma anomala, the causal agent of eastern filbert blight, were examined in four European hazelnut (Corylus avellana) orchards during a 2-year period. In each orchard, Burkhard volumetric spore traps and automated weather-monitoring equipment were deployed for 12-week periods beginning at budbreak, when hazelnut becomes susceptible to infection. Ascospores of A. anomala were released when stromata on the surface of hazelnut branches were wet from rain but not from dew. Release of ascospores ceased after branch surfaces dried. The duration of free moisture on branch surfaces regulated the initiation and rate of ascospore release, but no significant effects of temperature, relative humidity, wind, or light on ascospore release were apparent. Most (>90%) ascospores were captured during precipitation events that exceeded 20 h in duration, which represented about 10% of the total precipitation events each season. Quantitative relationships between the hourly capture of A. anomala ascospores and hours since the beginning of a precipitation event were developed. With the onset of precipitation, the hourly rate of ascospore capture increased until the fifth hour of rain, remained relatively constant between the fifth and twelfth hours, and then declined gradually. During the 12-week spore-trapping periods, the likelihood and rates of ascospore release associated with precipitation were highest at budbreak and then declined through April and May until early June, when the reserve of ascospores in the perithecia was depleted. Large numbers of ascospores were captured in the volumetric spore traps, indicating that ascospores may be commonly dispersed long distances on air currents as well as locally by splash dispersal within the canopy, as reported previously. The results indicate that monitoring seasonal precipitation patterns may be useful for estimating the quantity and temporal distribution of airborne inoculum during the period that the host is susceptible to infection.     

Effects of two different anesthetic protocols on 64‐MDCT coronary angiography in dogs

Heart rate is a major factor influencing diagnostic image quality in computed tomographic coronary artery angiography (MDCT‐CA), with an ideal heart rate of 60–65 beats/min in humans. The purpose of this prospective study was to compare effects of two different clinically applicable anesthetic protocols on cardiovascular parameters and 64‐MDCT‐CA quality in 10 healthy dogs. Scan protocols and bolus volumes were standardized. Image evaluations were performed in random order by a board‐certified veterinary radiologist who was unaware of anesthetic protocols used. Heart rate during image acquisition did not differ between protocols (P = 1), with 80.6 ± 7.5 bpm for protocol A and 79.2 ± 14.2 bpm for protocol B. Mean blood pressure was significantly higher (P > 0.05) using protocol B (protocol A 62.9 ± 9.1 vs. protocol B 72.4 ± 15.9 mmHg). The R‐R intervals allowing for best depiction of individual coronary artery segments were found in the end diastolic period and varied between the 70% and 95% interval. Diagnostic quality was rated excellent, good, and moderate in the majority of the segments evaluated, with higher scores given for more proximal segments and lower for more distal segments, respectively. Blur was the most commonly observed artifact and mainly affected the distal segments. No significant differences were identified between the two protocols for optimal reconstruction interval, diagnostic quality and measured length individual segments, or proximal diameter of the coronary arteries (P = 1). Findings indicated that, when used with a standardized bolus volume, both of these anesthetic protocols yielded diagnostic quality coronary 64‐MDCT‐CA exams in healthy dogs.     

Development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosis of Johne''s disease in cattle

An absorbed enzyme immunoassay (EIA) test for Johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. Total test time was reduced to less than 2 h with a minimum of manipulations. The test was evaluated in cattle herds from Johne's disease-endemic and Johne's disease-free regions of Australia. Specificity was 99.8%. Calculations of sensitivity were affected by the history of the herd under test. However, the EIA detected in excess of 80% of animals before onset of clinical disease and 65% of faecal shedders were EIA positive on, or before, first detection of Mycobacterium paratuberculosis in their faeces. The test should aid epidemiological studies and be a useful tool in the management and control of Johne's disease.     

Primary epitheliotropic intestinal T-cell lymphoma in a horse.

A 17-year-old Quarterhorse gelding with a clinical diagnosis of protein-losing enteropathy was submitted for necropsy following a 4-5-month duration of weight loss, decreased appetite, and hypoproteinemia. Gross findings included multiple 1-2-cm diameter ulcers on the luminal surfaces of the duodenum and ileum. Histologic examination revealed individual large, round cells infiltrating much of the mucosal epithelium of the duodenum, jejunum, ileum, and colon in addition to multifocal areas of ulceration. Similar round cells infiltrated Brunner's glands and expanded the submucosa beneath the foci of ulceration. Immunohistochemical staining indicated the round cell population was of T-lymphocyte origin. Several features of this equine neoplasm bear similarities to enteropathy-associated T-cell lymphoma in humans.