Evaluation of Sperm-Activating Solutions in Atlantic Salmon Salmo salar Fertilization Tests

The use of saline solutions was tested to determine their efficacy as replacements for ovarian fluid as sperm activators and to eliminate variability encountered with the use of Ovarian fluid. We tested fertilization rate of semen from eight males on Atlantic salmon Salmo salar eggs after five sperm-activating solutions and a non-activating saline were substituted for ovarian fluid. We used solutions shown acceptable for use with other salmonid species. The six solutions tested were a non sperm-activating phosphate-buffered saline (PBS, 7.2 g/L NaCl, 1.48 g/L Na₂HPO₄, 0.43 g/L K H₂PO₄), a Tris buffer (6.99 g/L NaCl, 3.63 g/L Tris and 2.42 g/L glycine), a Borax buffer (12.2 g boric acid/L in solution 1, 76 g disodium tetraborate/L combined 100:118, then 1 L combined with 3.7 L water and 18 g NaCl), and three solutions of 9.25 g/L (125 mM) NaCl buffered to pH 6.0, 7.5, and 8.9. The latter five solutions activated sperm immediately on contact, while PBS required additional water to activate sperm. The PBS solution was the least effective (mean percent eyed eggs 37.6%) for egg fertilization. The mean percent eyed eggs for the other five saline solutions (range 78% to 86%) were not significantly different. Sperm from one male provided significantly lower egg fertilization (39.6%) when compared with the other seven males (67.2–87.4% egg fertilization). Percent egg fertilization was not related to number of live sperm cells per egg. Our results show that osmotically-balanced sperm-activation solutions, even those with a pH range from 6.0 to 8.9 provide adequate media for fertilization of Atlantic salmon eggs. Fertilization in a deactivation saline and water reactivation of sperm resulted in low egg fertilization.     

Effects of phenylbutazone and indomethacin on the post-operative course following experimental orthopaedic surgery in dogs

Randomized placebo-controlled crossover studies were carried out in dogs to evaluate how two non-steroidal anti-inflammatory drugs (NSAID) might modulate an acute post-traumatic inflammatory reaction. Two "identical" surgical interventions were performed on the forelimbs of each animal with an interval of 28 days, to enable a paired comparison of the inflammatory signs and the wound/bone healing processes. At one operation 8 dogs received 300 mg phenylbutazone twice daily for 8 days starting on the day before surgery, and at the other operation matching placebo tablets were given. In a similar placebo-controlled trial another group of 8 dogs received 5 mg indomethacin twice daily. With phenylbutazone the post-operative swelling was not significantly reduced compared to placebo, but there was less pain and limping. With indomethacin the swelling was somewhat reduced, but there was no consistent difference to placebo in the pain and limping assessments. None of the drugs appeared to distinctly effect the wound or fracture healing, as evaluated by clinical inspection, comparison of radiographs and comparison of bone sections from the sites of surgery. It proved difficult to select an appropriate dosage of indomethacin due to its high potential to induce GI ulceration and bleeding in dogs. In this experimental surgical model with an acute inflammation, neither phenylbutazone nor indomethacin showed impressive anti-inflammatory or analgesic properties. In the same model paracetamol has proved to significantly and more efficiently, reduce both swelling and pain without any noticeable adverse effects, and appears to be a better alternative than the two presently tested NSAID.     

Factors affecting the hatchability of eggs from broiler breeders

1. Four experiments were carried out on eggs from broiler breeding flocks between 26 and 60 weeks of age. The effects of storage and incubation conditions on hatchability were tested.

2. Collecting eggs hourly rather than five hours after lay slightly reduced hatchability (P<0.10). Pre‐storage fumigation of almost un‐contaminated eggs had no effect on hatchability even after storage for 8 d. Storing eggs in unsealed polythene bags did not affect hatchability of eggs stored for 5 or 8 d.

3. Eggs stored for 2 d hatched better when held at 18 °C than at 15 °C (P<0.05). Eggs stored for 8 d at 15 °C hatched better than eggs stored for 8 d at 18 °C (P< 0.01). Best hatchability was in eggs stored in unsealed polythene bags at a room temperature of 15 °C. When older eggs were allowed 30 to 40 min more in the setter for each day of storage, the decline in hatchability was 0.5 to 0.6 percentage units per day in storage as compared with a decline of 1.2 percentage units per day when eggs of different storage times, up to 8 d, were set simultaneously.

4. Those eggs which showed a weight loss during incubation of near average for their relative humidity (RH) treatment tended to hatch better than others except under conditions of very low RH (0.36), when best hatchability was associated with lower than average weight loss.

5. In eggs from a young flock (28 to 44 weeks of age) hatchability of fertile eggs was depressed by 1 percentage unit with an increase in RH of 0.17, and by 1 percentage unit with each decrease of 0.06 in RH from a control RH of 0.53. In eggs from the same flock between 48 and 60 weeks of age hatchability was depressed by 1 percentage unit with each 0.037 increase in RH from 0.44 to 0.70.

6. Eggs from a young flock (34–49 weeks) hatched significantly better when maintained at 0.82 rather than at 0.66 (P<0.05) or 0.95 (P<0.10) RH during the hatching period from 19 to 21 d of incubation. Eggs from an older flock (51–61 weeks) hatched better at 0.82 and at 0.‐92 than at 0.72 RH during the same period, but the differences were not significant.     

Equine ocular anatomy and ophthalmic examination.

This article is intended to provide the practitioner with a succinct but complete source regarding equine orbital and ocular anatomy,instrumentation available for ophthalmic examination, a methodical examination technique, sedation and regional nerve blocks, and diagnostic procedures involving the eye. Such knowledge of orbital and ocular anatomy is essential to allow recognition of normal,normal variations, or an abnormality of the equine eye and orbit.     

Diagnosis and treatment of a cerebral infarct in a chimpanzee (Pan troglodytes).

A 29-yr-old chimpanzee (Pan troglodytes) presented after an acute onset of right facial and forearm paresis that progressed to paralysis within 24 hr, with subsequent development of right leg paresis. Magnetic resonance imaging (MRI) of the head revealed an abnormal region of increased signal intensity in the left frontal, parietal, and temporal cerebral hemispheres, corresponding to the vascular territory of the middle cerebral artery, with resultant compression of the left lateral ventricle. The findings were consistent with a cerebral infarct (stroke). MRI is the most sensitive test for early detection of cerebral changes due to ischemia and was essential in obtaining a diagnosis in this case. The chimpanzee responded well to treatment with long-term anticoagulant aspirin and a short, tapered course of prednisone and regained full gross motor function.     

Characterization of a wild-type strain of Francisella tularensis isolated from a cat.

Francisella tularensis type A is the primary cause of tularemia in animals and humans in North America. The majority of research on F. tularensis has been done with the attenuated live vaccine strain (LVS), which is a type B, but very few wild-type F. tularensis strains have been characterized. A gram-negative coccobacillus that was isolated in pure culture from the lungs of a cat that died after being lost for 5 days was received for identification at the Virginia-Maryland Regional College of Veterinary Medicine Teaching hospital. The isolate (strain TI0902) was not identified (or was misidentified) by commercial identification systems; however, it was identified as F. tularensis subspecies tularensis (type A) by sequencing a portion of the 16S ribosomal RNA gene. Furthermore, repetitive extragenic palindromic sequences-polymerase chain reaction amplified a 4-kb DNA fragment from TI0902 that was characteristic of F. tularensis type A but not type B. The electrophoretic profile of the lipopolysaccharide of strain TI0902 was identical to that of the LVS by Western blotting with antiserum to LVS. The protein-enriched outer membrane of strain TI0902 contained 6-8 proteins, which were similar in molecular size to those from the LVS. Electron microscopy of negatively stained and alcian blue-stained LVS and TI0902 cells showed that both strains were coccobacillary in shape and may be encapsulated. However, after mouse challenge, the TI0902 strain was clearly more virulent than the LVS strain. Results of this study indicate that the genotype and phenotype of wild-type F. tularensis type A strain TI0902 is similar, but not identical, to that of the LVS strain. Further studies will help determine whether pathogenesis and host-pathogen interactions are also similar between the 2 strains.     

IMAGING DIAGNOSIS: MAGNETIC RESONANCE IMAGING OF A CERVICAL WOODEN FOREIGN BODY IN A DOG

This article describes the discovery of a chronic cervical wooden foreign body ventral to the left transverse processes of the cranial cervical spine using magnetic resonance imaging (MRI) in a dog that presented with chronic neck pain and lameness. The dog did not exhibit dysphagia or chronic draining tracts, the most common signs of the presumed cause, that of a penetrating oropharyngeal foreign body. The foreign body itself was represented on MR images as an oval straight-edged core within an inflammatory tissue reaction. The wood was slightly hyperintense on T2- and isointense on T1-weighted images relative to muscle. Surrounding this was a more conspicuous contrast-enhancing reactive tissue rim that was hyperintense on all pulse sequences. Adjacent musculature also exhibited diffuse edema and contrast enhancement that extended around the left cervical vertebral transverse processes and local intervertebral nerve roots. The foreign body was found to be a wooden stick upon surgical removal. MRI is an excellent method for visualizing the inflammatory tissue reactions associated with soft-tissue foreign bodies because of its contrast resolution and depiction of anatomy in multiple imaging planes.     

Endophytic-Host Selectivity of Discula umbrinella on Quercus alba and Quercus rubra Characterized by Infection, Pathogenicity and Mycelial Compatibility

The fungal endophytic–host relationships of Discula umbrinella and two oak species, Quercus alba and Quercus rubra, were characterized on the basis of endophytic infection, pathogenicity, and mycelial compatibility. Isolates of D. umbrinella were cultured from leaves of Q. alba and Q. rubra collected from a hardwood forest located in Patuxent Wildlife Research Center in Laurel, Maryland, USA. Endophytic infection was observed on sterile leaf discs and living 2-month-old seedlings of Q. alba and Q. rubra. Fungal-host reciprocal inoculations revealed the presence of conidiomata on both hosts but conidiomata production was more abundant on Q. alba. Isolates from Q. rubra produced milder disease symptoms on both oak species. Mycelial compatibility studies identified seven different MCG groups. MCG groups 1–3 contained isolates from both oak species whereas MCG groups 4–7 contained host specific isolates. Field studies monitored the seasonal appearance of the sexual fruiting structures, perithecia, as a possible source of new genetic variation that might alter host specificity/pathogenicity of the D. umbrinella isolates on Q. alba and Q. rubra hosts. Only 1–2% of the leaves contained perithecia throughout the sampling period (April–September). Isolates collected from Q. alba differed from those collected from Q. rubra in endophytic infection, pathogenic response, and perithecia production. The results of this study suggest that the endophyte–host relationship is one of host selective preference for Q. alba, but that the endophyte has the ability to maintain the endophytic/pathogenic life cycle on the less preferred host species, Q. rubra.     

A spontaneously occurring malignant ovarian Sertoli cell tumor in a young Sprague Dawley rat

Primary ovarian tumors are generally uncommon in rats used in toxicologic studies. A malignant Sertoli cell tumor was present in the ovary of a 19-week-old female Sprague Dawley rat. Macroscopically, the mass was white and firm, 10 × 13 × 17 mm in size, and located in the right ovary. Histopathologically, the mass was composed of nests of pleomorphic cells, which formed seminiferous-like tubules separated by a thin fibrovascular stroma. The tubules were lined by tumor cells, which had basally located nuclei and abundant eosinophilic and vacuolated cytoplasm. In some areas, the tumor cells were arranged in a retiform growth pattern, mimicking a rete testis/ovarii. Disseminated metastases to the surfaces of the mesentery, spleen and liver were also present. Immunohistochemically, many tumor cells were strongly positive for vimentin, estrogen receptor α and Ki 67. Some tumor cells were positive for pancytokeratin and inhibin α. These findings closely resemble those of an ovarian-derived human malignant Sertoli cell tumor. From our review of the literature, we believe this is the first report of a spontaneous malignant Sertoli cell tumor in the ovary of a young laboratory rat. This case might provide useful historical control information for rat toxicity studies.