Application of sperm cryopreservation in selective breeding of the Pacific oyster,Crassostrea gigas (Thunberg)

The robustness of Pacific oyster, Crassostrea gigas (Thunberg), sperm cryopreservation in the context of selective breeding based on family lines was investigated. Irrespective of egg density, high fertilization success was achieved with cryopreserved sperm when sperm:egg ratios of 1000:1 to 10 000:1 were used. Variation among replicate runs on the same oyster batches was minimal, indicating that cryopreservation and larval rearing procedures were repeatable. Twenty independent single male–female crosses were made to assess the utility of cryopreserved sperm in selective breeding. The fertility of unfrozen sperm was generally a poor predictor of cryopreserved sperm fertility. Based on D‐larval yields, 17 of the 20 crosses were likely to yield adequate spat for selective breeding (>10⁵ D‐larvae from 1 million eggs), two were marginal (5 × 10⁴ D‐larvae) and one was inadequate (4 × 10³ D‐larvae). An alternative fertilization strategy to improve D‐yield from a given number of sperm was then tested. Fertilizing 10 million eggs at a sperm:egg ratio of 200:1 increased the total D‐yield when compared with fertilizing 1 million eggs at a sperm:egg ratio of 2000:1 for the same male–female pair. We conclude that, despite wide variation in fertility, cryopreserved sperm is useful for family production.     

A method for inducing muscle relaxation in the abalone, Haliotis iris

Four chemical and two non-chemical methods for induction of relaxation in the abalone, Haliotis iris, were tested for prospective use in pearl seeding. Requirements were for rapid onset of relaxation, full extension and softening of the foot and epipodium muscle for at least 10 min, and recovery with minimal stress and no mortality. Nembutal at 1 ml/l was most successful, producing relaxed and soft muscle with good accessibility to operation sites, and no mortality. Benzocaine and MS-222 made animals release from container walls within 30–60 min, but the animal's muscle was hard and contracted. Propylene phenoxytol caused muscle contraction and mortality at 2.5 ml/l. The effects of the chemical relaxants were similar in warm (21–22.5°C) and cool (15–16°C) water. Elevation of the water temperature from 18°C to 23°C (without chemicals) caused some softening of the muscle, but was much less effective than Nembutal. Air exposure for up to 4 h was ineffective. This work provides a simple method to relax live abalone for pearl nucleation or other manipulations.     

The effect of level of nutrition on the development of resistance to Haemonchus contortus in sheep

Merino wether weaners, 5-months old at the start of the experiment and fed lucerne-based pellets at 600 g per head per day, developed resistance to Haemonchus contortus given as a continuous or "trickle" infection at the rate of 500 larvae 3 times per week over a period of 17 weeks. By contrast, the development of resistance was impaired in weaners fed pellets at 400 g per head per day. Low food intake and protein-energy deprivation, therefore, have a major effect in impairing the development and expression of host-protective immunity against haemonchosis. The point at which low food intake causes outbreaks of haemonchosis in sheep at pasture is critical in determining procedures for prevention and control of the disease.     

A single screw technique compared to a two screw and wire technique as a temporary transphyseal bridge for correction of fetlock varus deformities

A single screw technique is described as a temporary transphyseal bridge for the treatment of fetlock varus angular limb deformity in foals. This has been compared to tension band wiring with regards to rate of correction, cosmetic result and incidence of complications. The single screw was found to be an effective technique resulting in more rapid improvement of deviations in foals aged up to 5 months. Complications were minimal following both techniques and the final cosmetic result was better following single screw surgery.     

Milk yield of primiparous beef cows from three calving systems and varied weaning ages

Primiparous beef cows produced in 3 calving systems were used in a 2-yr study with a completely random design to measure milk yield throughout a 190-d lactation (2002, n = 20; 2003, n = 24 per calving system). Calving occurred in late winter (average calving date = February 4 +/- 2 d), early spring (average calving date = March 30 +/- 2 d), and late spring (average calving date = May 26 +/- 1 d). Additionally, cows used in this study had been weaned at varied ages as calves, creating 6 dam treatments. Dam age at weaning was 140 (late spring), 190 (late winter, early spring, late spring), or 240 (late winter, early spring) d of age. Milk production was measured by using the weigh-suckle-weigh technique at an average of 20, 38, 55, 88, 125, 163, and 190 d in milk. Milk yield for the 190-d lactation period was calculated as area under the curve by trapezoidal summation. Data were analyzed with a model containing treatment, year, and their interaction. Orthogonal contrasts were used to separate effects when treatment was significant (P < 0.10). Total milk yield did not differ (P = 0.42) between cows in the late winter and early spring systems, but cows in the late spring system tended to differ (P = 0.09) from the average of the other 2 systems. Cows in the late spring calving system had increased milk yield in 2002 and lesser milk yield in 2003 compared with the other calving systems (treatment x year interaction, P < 0.001). Cows born in late spring that had been weaned at 140 d of age produced more (P = 0.05) total milk than those weaned at 190 d of age. Peak milk yield was affected (P < 0.001) by treatment and showed a treatment x year interaction (P = 0.006). Day of peak lactation differed among treatments (P = 0.002), with cows in the late winter system peaking later (P = 0.007) than early spring cows, and late spring cows peaking earlier (P = 0.004) than the average of late winter and early spring cows. The average date of peak lactation was May 4 for the late winter system, May 31 for the early spring system, and July 19 for the late spring system. Calf ADG differed (P < 0.001) for the late spring system compared with the average of the late winter and early spring systems, but the relationship interacted with year (P < 0.001). Cow BW and BW change differed among treatments (P < 0.004), with much of the difference associated with the amount of milk produced or the timing of peak lactation. Season of calving affects milk yield of primiparous cows grazing Northern Great Plains rangelands and ADG of their calves.     

Insulin-like growth factor binding proteins in granulosa and thecal cells from bovine ovarian follicles at different stages of development

Because IGFBP inhibit IGF-stimulated cellular proliferation and differentiation, it is hypothesized that variations among IGFBP in individual follicles might contribute to the regulation of recruitment, selection, dominance, and turnover of ovarian follicles. Sources of IGFBP in fluid of bovine follicles are not well established; thus, objectives of this study were to determine levels of IGFBP binding activities and messenger RNA (mRNA) in granulosa and theca interna cells at different stages of follicular development (small [< 6 mm], medium [6 to < 8 mm], and large [> or = 8 mm]) and to characterize associations of these levels measured in the cells with levels of IGFBP and steroids in follicular fluid. Thecal and granulosa cells from large healthy follicles contained two- to twentyfold less (P < 0.05) IGFBP-2, -3, and -5 than cells from small, medium, and large atretic follicles. Thecal cells from small, medium, and large atretic follicles contained more (P < 0.05) IGFBP-3 and -4 than granulosa cells from these follicles, whereas granulosa cells from these follicles contained more IGFBP-2 activity than thecal cells. Differences in IGF binding activity were paralleled by differences in levels of mRNA for the respective IGFBP. Developmental differences in IGFBP activity in follicular fluid were positively associated with activity in granulosa and/or thecal cells, with the exception of IGFBP-4, which was low in fluid from large healthy follicles but markedly increased (mRNA and binding activity) in granulosa cells from these follicles. It is concluded that developmental changes in follicular fluid IGFBP-2 and -5 binding activities seem to be controlled in part by alterations in synthesis of these IGFBP by granulosa and thecal cells, whereas diminished IGFBP-4 in fluid from large healthy follicles occurs concomitantly with increased levels of IGFBP-4 mRNA and activity in granulosa cells, implicating posttranslational regulation by specific proteases.     

Effect of immunosuppressive doses of cyclosporine on pancreatic beta cell function in pigs

OBJECTIVE: To evaluate whether immunosuppressive doses of cyclosporine (CsA) have an adverse effect on the liver, kidney, and pancreatic beta cells of pigs. ANIMALS: 8 juvenile 8-week-old Landrace X Large White crossbred pigs. PROCEDURE: CsA (100 to 140 mg/kg) was administered orally to euglycemic pigs to reach whole blood trough concentrations of approximately 1500 ng/mL. To determine pancreatic beta cell function, plasma C-peptide and insulin concentrations were measured in response to i.v. administration of glucose, glucagon, arginine, and oral administration of glucose. Effects on liver and kidney were determined by monitoring serum measurements of liver function and serum creatinine concentrations, respectively. RESULTS: Plasma concentrations of C-peptide were significantly lower in euglycemic CsA-treated pigs, compared with control pigs, following i.v. administration of glucose, glucagon, arginine, and oral administration of glucose. Furthermore, the glucose clearance rate was decreased in euglycemic CsA-treated pigs, compared with control pigs. Serum creatinine concentrations and 4 of 7 serum measurements of liver function were not adversely affected by CsA administration. Serum concentrations of bilirubin and albumin were significantly increased, and serum alanine aminotransferase activity was significantly decreased in CsA-treated pigs, compared with control pigs. Histologic evaluation of liver and kidney sections revealed no pathologic findings in CsA-treated or control pigs. CONCLUSIONS AND CLINICAL RELEVANCE: In our study, immunosuppressive doses of CsA caused an impairment of porcine pancreatic beta cell function, but did not have toxic effects on the kidney. However, on the basis of changes in serum bilirubin and albumin concentrations and alanine aminotransferase activity, subclinical toxic effects on the liver did occur when immunosuppressive doses of CsA were administered.     

Comparison of a new sheep dipping technique with saturation-type methods

Three methods of dipping sheep for ectoparasite control, plunge dipping, shower dipping and two models of a James “Mini-shower”^(R) (Mk I, Mk II) were compared for efficiency. Usingdiazinon and fenthion-etliyl, at rates recommended by the manufacturers, both plunge and shower dipping gave 6 to 8 weeks protection from flystrike and at least 10 week sprotection from re-infestation by lice. Insecticides applied by both models of “Mini-shower” gave significantly less protection from flystrike (2–3 weeks) than either plunge or shower dipping. Four weeks' protection from louse re-infestation was achieved using insec-ticides applied by the Mk I “Mini-shower”.     

Identity and pathogenicity of Rhizoctonia spp. associated with bare patch disease of cereals at a field site in Western Australia

Roots of seedlings of wheat and barley affected by bare patch disease at a field site in Western Australia were assessed for root damage and plated to isolate fungi. The patches were variable in shape and size and had the most severely affected plants in the centre. Of the 165 isolates ofRhizoctonia spp. obtained, 90% were multinucleate and 10% binucleate, the former being predominant in the plants at the centre of the patch. The relative frequency of binucleate isolates increased with proximity to the periphery. The increase in activity of avirulent binucleate isolates towards the periphery of the patch may be related to the sharp and abrupt edging of the patch. A variety of other species of fungi such asFusarium spp.,Mortierella spp.,Bipolaris sorokiniana, Pythium sp. andTrichoderma sp. were encountered within the patches. The multinucleate isolates belonging to anastomosis groups (Ag) 2–1, 2–2 and 8 (Thanatephorus cucumeris) were most pathogenic to wheat. The binucleate isolates of Ag C, D, E, and K (Ceratobasidium sp.) were less pathogenic. It is suggested that the bare patch disease is caused by a complex of root rot fungi composed of one or more anastomosis groups ofRhizoctonia spp. and other associated fungi.Samenvatting Van kiemplanten van tarwe en gerst, afkomstig van een met kale-plekkenziekte besmet perceel in West Australië werd de mate van wortelbeschadiging bepaald en werden schimmels uit de wortels geïsoleerd. De plekken waren verschillend van vorm en afmeting; de zwaarst aangetaste planten werden in het centrum ervan aangetroffen. Van de 165 verkregen isolaten vanRhizoctonia spp. was 90% meerkernig en 10% tweekernig. De meerkernige overheersten in de centra van de plekken. Relatief gezien nam het aantal tweekernige isolaten toe naarmate de herkomst dichter bij de periferie van de plekken was. De scherpe begrenzing van de ziekte aan de randen van de plekken zou in verband kunnen staan met het toenemen van de activiteit van de avirulente tweekernige isolaten in de nabijheid van de periferie van de plekken. Een aantal andere schimmels, zoalsFusarium spp.,Mortierella spp.,Bipolaris sorokiniana, Pythium sp. enTrichoderma sp. werd eveneens in de plekken aangetroffen. De meerkernige isolaten die tot de anastomosegroepen Ag 2–1, 2–2 en 8 (Thanatephorus cucumeris) behoren, waren voor tarwe het meest pathogeen. De tweekernige isolaten van de anastomosegroepen Ag C, D, E en K (Ceratobasidium sp.) waren minder pathogeen. Gesuggereerd wordt, dat de kale-plekkenziekte veroorzaakt wordt door een complex van verschillende wortelschimmels, die behoren tot een of meer anastomosegroepen vanR. solani en andere daarmee geassocieerde schimmels.     

The response of monocyte derived dendritic cells following exposure to a nematode larval carbohydrate antigen

The glycolipid CarLA (carbohydrate larval antigen) is present on the epicuticle of the infective-stage larvae of gastrointestinal nematode parasites infecting livestock. The molecule is lost from the surface of the larvae in the few days post-ingestion by a host animal, and the resulting anti-CarLA antibody response has been demonstrated to be protective in vivo. Both the anti-CarLA response, and anti-parasite immunity in general, are slow to develop, and several months of natural exposure to ingested larvae is required. The current study was designed to provide information on how the anti-CarLA response develops, and focuses on the initial recognition of the molecule by human monocyte derived dendritic cells (mdDC) in vitro. Immunofluorescence and flow cytometry demonstrated that mdDC recognise and internalise both the purified and the native form of CarLA, in the case of the latter once it is shed from the larval surface. However, the recognition of CarLA did not result in classical maturation of DC, while there was only transient or minor up-regulation of CD86, CD83, HLA-DR and CD40. Exposure of mdDC to purified CarLA resulted in the increased production of the pro-inflammatory cytokines IL-6 and to a lesser extent of IL-8 and TNF-α, and a reduced production of the anti-inflammatory cytokine IL-1RA. CarLA therefore has little ability to mature and functionally alter monocyte derived dendritic cell function.