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.     

On predicting the presence of birds in Eucalyptus forest types

We examine the relationship between the number of bird species and environment within 500 000 ha of eucalypt forests in south-eastern New South Wales, Australia. Birds were surveyed at 39 sites within 31 eucalypt communities, which were, in turn, scored by altitude, temperature, rainfall, basal areas of trees and levels of the nutrients nitrogen, phosphorus, potassium and magnesium in the eucalypt foliage.

Ninety bird species were recorded. Numbers per site, averaged over the whole sample period, ranged between 20 and 38 over all surveys, 24 and 42 in ‘summer’ (October to March) when migrant species were present, and 13 and 38 in ‘winter’ (April to September). A forward stepwise Poisson regression model was used to fit bird species richness to the environmental variables. Over all surveys and in ‘summer’, foliar magnesium and tree basal area (including the basal area of dead trees) were significantly (positive) correlated with the number of bird species. In ‘winter’ the correlations were with altitude and temperature (both negative), presumably the result of emigration of species that avoid cold weather, and tree basal area (positive).

The positive association of number of species over all surveys and in ‘summer’ with tree basal area, including dead trees, and foliar magnesium may index the level of forest maturity and productivity.

Forest management directed solely to timber production reduces the basal area and the number of dead trees and may thereby reduce bird species richness permanently.

A methodology is offered, based on the relationship between the physical environment and the more productive eucalypt communities, for determining forest areas containing the most species of birds.     

Proteomic profiling of plasma-derived small extracellular vesicles: a novel tool for understanding the systemic effects of tick burden in cattle

Cattle ticks pose a significant threat to the health and profitability of cattle herds globally. The investigation of factors leading to natural tick resistance in cattle is directed toward targeted breeding strategies that may combat cattle tick infestation on the genetic level. Exosomes (EXs), small extracellular vesicles (EVs) of 50 to 150 nm diameter, are released from all cell types into biofluids such as blood plasma and milk, have been successfully used in diagnostic and prognostic studies in humans, and can provide essential information regarding the overall health state of animals. Mass spectrometry (MS) is a highly sensitive proteomics application that can be used to identify proteins in a complex mixture and is particularly useful for biomarker development. In this proof of principle study, EXs were isolated from the blood plasma of cattle (Bos taurus) with high (HTR) and low tick resistance (LTR) (n = 3/group). Cattle were classified as HTR or LTR using a tick scoring system, and EXs isolated from the cattle blood plasma using an established protocol. EXs were subjected to MS analysis in data-dependent acquisition mode and protein search performed using Protein Pilot against the B. taurus proteome. A total of 490 unique proteins were identified across all samples. Of these, proteins present in all replicates from each group were selected for further analysis (HTR = 121; LTR = 130). Gene ontology analysis was performed using PANTHER GO online software tool. Proteins unique to HTR and LTR cattle were divided by protein class, of which 50% were associated with immunity/defense in the HTR group, whereas this protein class was not detected in EXs from LTR cattle. Similarly, unique proteins in HTR cattle were associated with B-cell activation, immunoglobins, immune response, and cellular iron ion homeostasis. In LTR cattle, unique exosomal proteins were associated with actin filament binding, purine nucleotide binding, plasma membrane protein complex, and carbohydrate derivative binding. This is the first study to demonstrate that MS analysis of EXs derived from the blood plasma of HTR and LTR cattle can be successfully applied to profile the systemic effects of tick burden.     

Effect of walking velocity on ground reaction force variables in the hind limb of clinically normal horses

OBJECTIVE: To measure the effect of subject velocity on hind limb ground reaction force variables at the walk and to use the data to predict the force variables at different walking velocities in horses. ANIMALS: 5 clinically normal horses. PROCEDURE: Kinematic and force data were collected simultaneously. Each horse was led over a force plate at a range of walking velocities. Stance duration and force data were recorded for the right hind limb. To avoid the effect of horse size on the outcome variables, the 8 force variables were standardized to body mass and height at the shoulders. Velocity was standardized to height at the shoulders and expressed as velocity in dimensionless units (VDU). Stance duration was also expressed in dimensionless units (SDU). Simple regression analysis was performed, using stance duration and force variables as dependent variables and VDU as the independent variable. RESULTS: Fifty-six trials were recorded with velocities ranging from 0.24 to 0.45 VDU (0.90 to 1.72 m/s). Simple regression models between measured variables and VDU were significant (R2 > 0.69) for SDU, first peak of vertical force, dip between the 2 vertical force peaks, vertical impulse, and timing of second peak of vertical force. CONCLUSION AND CLINICAL RELEVANCE: Subject velocity affects vertical force components only. In the future, differences between the forces measured in lame horses and the expected forces calculated for the same velocity will be studied to determine whether the equations can be used as diagnostic criteria.     

The hindlimb in walking horses: 1. Kinematics and ground reaction forces

The objective was to study associations between kinematics and ground reaction forces in the hindlimb of walking horses. Video (60 Hz) and force (2000 Hz) data were gathered for 8 strides from each of 5 sound horses during the walk. Sagittal plane kinematics were measured concurrently with the vertical and longitudinal ground reaction forces. The hindlimb showed rapid loading and braking in the initial 10% stride. The stifle, tarsal and coffin joints flexed and the fetlock joint extended during this period of rapid loading. The vertical ground reaction force showed 2 peaks separated by a dip; this pattern was similar to the fetlock joint angle-time graph. Peaks in the longitudinal ground reaction force did not appear to correspond with kinematic events. Total braking impulse was equal to total propulsive impulse over the entire stride. Flexion and extension of the hip were responsible for protraction and retraction of the entire limb. Maximal protraction occurred shortly before the end of swing and maximal retraction occurred during breakover. During the middle part of stance the tarsal joint extended slowly, while the stifle began to flex when the limb was retracted beyond the midstance position at 28% stride. Flexion cycles of the stifle and tarsal joints were well coordinated during the swing phase to raise the distal limb as it was protracted. The results demonstrate a relationship between limb kinematics and vertical limb loading in the hindlimbs of sound horses. Future studies will elucidate the alterations in response to lameness.