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.     

Fruit and vegetable fiber fermentation by gut microflora from canines

The objective of this study was to assess fermentability by canine gut microflora to include shortchain fatty acid (SCFA) production, organic matter (OM) disappearance, and gas production of vegetable and fruit fiber sources compared to fiber standards (psyllium, citrus pectin, and Solka Floc). Fiber sources included apple pomace, carrot pomace, flaxseed, fruit blend (mixture of peach, almond, nectarine, and plum), grape pomace, pea hulls, pistachio, and tomato pomace. Substrates were fermented in vitro for 4, 12, and 24 h with fecal flora obtained from three healthy dogs. Citrus pectin had the highest OM disappearance, SCFA production, and gas production at all times of fermentation; psyllium was intermediate and Solka Floc was lowest. A wide variation in fermentability was noted among the vegetable and fruit fiber sources. Apple pomace, carrot pomace, and flaxseed had the greatest fermentability as assessed by OM disappearance. Pea hulls and tomato pomace had intermediate OM disappearances, and fruit blend, grape pomace, and pistachio were poorly fermented. Carrot pomace produced the largest amounts of gas and SCFA. Apple pomace produced high concentrations of gas but intermediate concentrations of SCFA. Pea hulls and tomato pomace produced intermediate concentrations of gas and SCFA, whereas flaxseed, fruit blend, grape pomace, and pistachio produced low amounts of these fermentation products. For all substrates collectively, OM disappearance was highly correlated with both gas production (r2 = 0.782 and 0.723 for 12- and 24-h values, respectively) and SCFA production (r2 = 0.737 and 0.738 for 12- and 24-h values, respectively). In general, OM disappearance, gas production, and SCFA production were related to the insoluble:soluble fiber ratio in the samples; as the insoluble:soluble ratio decreased (increased soluble fiber), the OM disappearance, gas production, and SCFA production increased.     

Fertility and motility of sperm from Atlantic halibut (Hippoglossus hippoglossus) in relation to dose and timing of gonadotrophin-releasing hormone agonist implant

In broodstocks of Atlantic halibut, Hippoglossus hippoglossus, male and female gamete production often becomes unsynchronised towards the end of the spawning season—milt becomes very viscous and difficult to express while the females are still producing batches of good quality eggs. Gonadotrophin-releasing hormone agonist (GnRHa) has been shown to stimulate spermiation in a number of fish species. Therefore, we conducted two experiments where male halibut were implanted intramuscularly with pellets containing GnRHa. The effect of the pellets was tested at three periods: before, at the height of and at the end of spermiation. In the middle period, GnRHa was tested at two doses (5 and 25 μg/kg bodyweight). Measurements were made of milt hydration, sperm motility and fertilisation rate. Implanted males began spermiation at least 4 weeks before control males. Both doses of GnRHa increased the fluidity of the milt. This effect lasted for at least 20 days in the low dose group and for 40 days in the high dose group. When applied at the end of the season, GnRHa reversed the normal trend for the milt to become more viscous. GnRHa treatments did not affect fertilisation rates obtained with the sperm. However, towards the end of the spawning season, sperm motility was enhanced in males treated with the high dose of GnRHa (25 μg/kg) compared to controls. As described previously, plasma concentrations of the gonadal steroids, 5β-pregnane-3β,17,20β-triol 20-sulphate and 17,20-dihydroxy-4-pregnen-3-one, were significantly enhanced by GnRHa treatment. Concentrations of testosterone on the other hand decreased when spermiating males were treated with GnRHa. Our data suggest that 17,20β-dihydroxy-4-pregnen-3-one or its metabolites are involved in milt hydration, possibly through affecting ion transport.     

Effects of large woody debris removal on physical characteristics of a sand-bed river

1. Removal of large woody debris (LWD) is one of the most widely practised stream alterations, particularly in sand-bed rivers of the south-eastern USA. Selective removal of LWD has been proposed as an alternative to orthodox non-selective clearing in order to conserve ecological resources, but methods for comparing hydraulic and environmental effects of selective and non-selective removal have not been developed. Conservation of stream habitats requires quantification of LWD removal impacts on physical habitat. 2. Physical characteristics of straightened, sand-bed reaches of the South Fork Obion River in western Tennessee, USA that were rich in LWD were compared with those in similar reaches where debris had recently been removed using selective removal guidelines. 3. The mean volume of LWD per unit water volume was 0.0545 in the uncleared reaches, but nearly 60% lower (0.0225) in the cleared reach. 4. A simple technique for predicting hydraulic roughness in channels with varying amounts of LWD was developed. Hydraulic roughness, as measured by the Darcy-Weisbach friction factor, was about 400% greater in uncleared reaches at base flow but declined to a level about 35% greater than for the cleared reaches at higher flows. Predicted friction factors were within 35% of measured friction factors at higher flows. 5. Physical habitat diversity in this channelized sand-bed stream was strongly related to the density of LWD. Flow conditions in the uncleared reaches were more heterogeneous than in the cleared reach, especially at low flow. At low flow, uncleared reaches tended to be shallower, have lower velocities, slightly finer bed material, and more heterogeneous conditions overall. Shannon indices based on depth and velocity were an average of 48% higher in uncleared reaches. 6. Bed sediments underneath and immediately adjacent to LWD formations were finer and contained more organic matter than sediments distant from LWD. However, when all bed samples were considered, organic content was positively correlated with median grain size.     

Infection experiments with Aphanomyces invadans in four species of estuarine fish

Along the eastern seaboard of the US, Atlantic menhaden, Brevoortia tyrannus, develop characteristic ulcerative lesions, a condition termed ulcerative mycosis. These lesions are identical to those seen across Asia in fish affected by epizootic ulcerative syndrome, a condition caused by the fungus-like oomycete Aphanomyces invadans. Young-of-the-year menhaden inhabiting estuarine environments are the primary species affected in the USA and little is known about the factors involved in the initiation of the lesions, or why menhaden are predominantly infected. Atlantic menhaden, hogchoker, Trinectus maculatus, striped killifish, Fundulus majalis, and mummichog, Fundulus heteroclitus, were inoculated with A. invadans (80 zoospores per fish) to explore species differences in infection and lesion development. All four species developed lesions. Killifish developed frank lesions similar to those observed in menhaden but the gross lesions occurred later, approximately 5-10 days after those on menhaden. Hogchoker and mummichog did not develop gross skin ulcers; rather, their lesions appeared as reddened areas under the epidermis. Mummichogs also showed evidence of significant healing with a well-developed granuloma and significant myocyte regeneration. These experiments show that species barriers as well as ecological barriers can explain some of the factors involved in the development of lesions in, and specificity of the water mould for, menhaden.     

Fate of lower Mississippi river habitats associated with river training dikes

• 1 Regions of reduced velocity adjacent to spur dikes along the Lower Mississippi River are valuable aquatic habitats. Similar zones along other large rivers have been converted to terrestrial habitats by sediment deposition.
• 2 Repetitive hydrographic surveys of 26 representative groups of dikes are examined to determine the direction and rates of change.
• 3 Since the dikes were constructed, the aquatic volume and area of associated low-velocity habitats have been reduced by 38% and 17%, respectively. Examination of time series shows that most changes occur shortly after construction, and after initial adjustment, habitat area and volume fluctuate about a condition of dynamic equilibrium.
• 4 Sedimentation rates were most rapid for dike fields constructed on the inside of bends to prevent chute development. Dike fields built to force or maintain thalweg crossings exhibited erosion rather than deposition.

     

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.     

Medium for selective isolation of Cryptococcus neoformans

A medium has been developed that permits the selective recovery of Cryptococcus neoformans from heavily contaminated materials. It employs creatinine as a nitrogen source, diphenyl (C(6)H(5)C(6)H(5)) and chloramphenicol as mold and bacterial inhibitors, and Guizotia abyssinica seed extract as a specific color marker. The medium has proved to be effective in the direct isolation of Cryptococcus neoformans from pigeon nests and from the air.