Collection and short-term preservation of semen from free-ranging eastern grey kangaroos (Macropus giganteus: Macropodidae)

Objectives To evaluate electro-ejaculation of free-range eastern grey kangaroos in the field and assess the efficacy of four diluents to preserve sperm motility over a 48-h period at 5°C.
Procedure and design Under gaseous anaesthesia, 25 free-range kangaroos were electro-ejaculated and characteristics of the ejaculate noted. Spermatozoa obtained from eight ejaculates were diluted in phosphate buffered saline containing various combinations of egg yolk and glucose and refrigerated at 5°C for 48 h.
Results Spermatozoa were recovered from 24 of 28 ejaculates. Mean (± SEM) semen volume (mL) and pH were 25.0 ± 1.9 and 7.1 ± 0.1 respectively. The forward motility (%), rate of movement of sperm (0 to 5) and sperm concentration (x 10⁶/mL) were 77.4 ± 1.5, 3.8 ± 0.9 and 31.2 ± 7.3 respectively. There was no significant difference between the four diluents in their ability to maintain forward motility of spermatozoa over 48 h. However, rate of movement over the same period was significantly (P < 0.01) improved when sperm were diluted in phosphate buffered saline containing 10% egg yolk.
Conclusions Electro-ejaculation is a safe and reliable method for collecting semen from free-ranging eastern grey kangaroos. Preliminary attempts at short-term preservation showed that the motility of kangaroo spermatozoa could be adequately stored for 24 h and that the addition of egg yolk to the semen diluent was beneficial for improving the rate of sperm movement.     

Rapid detection and identification of viral and bacterial fish pathogens using a DNA array-based multiplex assay

Fish diseases can be caused by a variety of diverse organisms, including bacteria, fungi, viruses and protozoa, and pose a universal threat to the ornamental fish industry and aquaculture. The lack of rapid, accurate and reliable means by which fish pathogens can be detected and identified has been one of the main limitations in fish pathogen diagnosis and fish disease management and has consequently stimulated the search for alternative diagnostic techniques. Here, we describe a method based on multiplex and broad-range PCR amplification combined with DNA array hybridization for the simultaneous detection and identification of all cyprinid herpesviruses (CyHV-1, CyHV-2 and CyHV-3) and some of the most important fish pathogenic Flavobacterium species, including F. branchiophilum, F. columnare and F. psychrophilum. For virus identification, the DNA polymerase and helicase genes were targeted. For bacterial identification, the ribosomal RNA gene was used. The developed methodology permitted 100% specificity for the identification of the target species. Detection sensitivity was equivalent to 10 viral genomes or less than a picogram of bacterial DNA. The utility and power of the array for sensitive pathogen detection and identification in complex samples such as infected tissue is demonstrated in this study.     

The net energy value of balanced diets for growing pigsValeur enérgétique nette des rations équilibrées pour les porcs en croissanceDer nettoenergiegehalt von ausgewogenen rationen für wachsende schweine

Experiments with 80 growing pigs of Danish Landrace were performed to study factors influencing the net energy value of balanced diets and to develop an energetic feed evaluation system for pigs.Although balanced with regard to essential nutrients, the cereal based diets varied widely in chemical composition and accounted for 90% of the variation in the chemical composition of the diets used in practice. The pigs were distributed on dietary treatments on a within-litter basis taking sex and weight into account. The experimental period lasted from 20–90 kg live weight. The daily intake of the different diets was regulated in such a way that the daily gain in the different treatments was almost identical through the entire experimental period.Six digestibility and balance experiments were performed with each pig; at approximately 90 kg live weight the pigs were killed, dissected, ground, mixed and then chemically analysed. The diet composition had a highly significant influence on digestibility, metabolizability and efficiency of utilization of metabolizable energy. Energy concentration (ME/kg DM) accounted for 90% of the variation in the net energy value of metabolizable energy. This result was adopted as a basis for the official energy evaluation in Denmark in 1976, and the net energy content of diets and feedstuffs is calculated from the equation Net energy ($\text{MJ}\text{kg DM}\text{) = ?1.88 + 0.75 ×}\text{ME}\text{(}\text{MJ}\text{kg DM}\text{)}$The net energy value of 1 kg common barley, i.e., 7.72 MJ is defined as a feed unit for pigs. The investigations are continued to elucidate, in greater detail, the influence of the individual nutrients, sites of absorption, etc.     

Determination of donor-type chimerism using a semi-quantitative PCR-based method in a canine model for bone marrow transplantation

Dogs are used in preclinical transplantation models to study methods of allogeneic bone marrow transplantation (BMT). The evaluation of chimerism is of major significance for the investigation of graft-vs.-host (GvH) and host-vs.-graft (HvG) reactions. To detect and quantitate male donor cells after a sex-mismatched (male to female) allogeneic BMT, we established a semi-quantitative polymerase chain reaction (PCR) assay. Based on the canine Y-chromosome sex-determining region (Sry) sequence, we designed primer specific for the detection of male DNA and optimised PCR conditions and cycle numbers. Artificial mixtures of male and female leukocytes were used to analyse the sensitivity of the assay. To validate our established method, we determined the percentage of chimerism in three transplanted female dogs. Under optimised conditions, the established PCR assay specifically detected male cells down to 0.01%, which corresponds to 0.1ng of transplanted male DNA. The percentage of chimerism could be quantitated either by agarose gel analysis or Southern blot analysis. Using our assay, we could confirm the percentage of chimerism in blood samples of three transplanted female canines, previously determined by karyotype analysis as 0, 100 and 100%, respectively. The established semi-quantitative PCR assay offers a quick, simple, accurate and sensitive way of evaluating and quantitating the percentage of chimerism in a sex-mismatched canine BMT model.     

Dorsolateral approach for arthrocentesis of the centrodistal joint in horses

OBJECTIVE: To develop a dorsolateral approach to the centrodistal (distal intertarsal) joint in horses and compare its success rate with that of the traditional medial approach in that joint. SAMPLE POPULATION: 25 cadaveric equine hind limbs, ultrasonographic images, and radiographic views of the tarsal region of 5 and 59 healthy horses, respectively, and 22 horses with a clinical indication for centrodistal joint centesis. PROCEDURES: The dorsolateral approach was established anatomically (3 cadaveric limbs), ultrasonographically (5 horses), and radiographically (59 horses). Centrodistal joint arthrocentesis was performed in 22 cadaveric hind limbs and 22 horses; the number of needle repositionings required for procedure completion via the medial (in vitro) and the dorsolateral approach (in vitro and in vivo) was determined. RESULTS: For the dorsolateral approach to the centrodistal joint, the injection site was 2 to 3 mm lateral to the long digital extensor tendon and 6 to 8 mm proximal to a line drawn perpendicular to the axis of the third metatarsal bone through the proximal end of the fourth metatarsal bone. The needle was directed plantaromedially (angle of approx 70 degrees from the sagittal plane). The number of needle repositionings required to complete centrodistal joint centesis via the dorsolateral and medial approaches was not significantly different. CONCLUSION AND CLINICAL RELEVANCE: In a clinical setting, the dorsolateral approach to the centrodistal joint in horses appears to have some advantages over the traditional medial approach. The success rate of arthrocentesis was similar via either approach, and palpation of the anatomic landmarks was easy.