Antigenic relationship of turkey coronavirus isolates from different geographic locations in the United States

The purpose of the present study was to examine the antigenicity of turkey coronavirus (TCV) isolates from various geographic areas with antibodies to different viruses. Seventeen isolates of TCV were recovered from intestinal samples submitted to Animal Disease Diagnostic Laboratory, Purdue University, from turkey farms located in different geographic areas. The prototype TCV Minnesota isolate (TCV-ATCC) was obtained from the American Type Culture Collection. Intestinal sections were prepared from turkey embryos infected with different TCV isolates and reacted with polyclonal or monoclonal antibodies to TCV, infectious bronchitis virus (IBV), bovine coronavirus (BCV), transmissible gastroenteritis virus (TGEV), reovirus, rotavirus, adenovirus, or enterovirus in immunofluorescent antibody staining. All 18 TCV isolates have the same antigenic reactivity pattern with the same panel of antibodies. Positive reactivity was seen with polyclonal antibodies to the TCV Indiana isolate, the TCV Virginia isolate, TCV-ATCC, and the IBV Massachusetts strain as well as monoclonal antibodies to the TCV North Carolina isolate or the membrane protein of IBV. Antibodies to BCV or TGEV were not reactive with any of the TCV isolates. Reactivity of antibodies to unrelated virus, rotavirus, reovirus, adenovirus, or enterovirus with different TCV isolates was all negative, except positive response was seen between enterovirus antibody and a TCV western North Carolina isolate, suggesting coinfection of turkeys with TCV and enterovirus in that particular case. The results indicated that the TCV isolates from these geographic locations in the U.S. shared close antigenicity and were antigenically related to IBV.     

Properties of the immunosuppressive factor induced by murine cytomegalovirus

Murine cytomegalovirus infection of spleen cultures induced the production of a small (less than 10,000 molecular weight) immunosuppressive factor (VISF), which suppressed concanavalin-A mitogenesis in fresh mouse spleen cells, and in fresh human peripheral blood leukocytes. The factor did not affect the growth of two murine T-cell lines or of mouse fibroblasts. A similar factor was also found in the serum of infected mice, at the time of maximum immune suppression. The properties of VISF indicate that the mechanism of MCMV immune suppression is different from that caused by several other viruses which are important in human and veterinary medicine.     

Pseudomonas fluorescens contamination of a feline packed red blood cell unit and studies of canine units

Background: While screening programs have reduced the risk of infectious disease transmission by donors in human and veterinary blood banking, bacterial contamination of blood products has emerged as a major complication in human medicine. Objectives: To describe a Pseudomonas fluorescens (Pf)‐contaminated feline packed RBC (pRBC) unit and experimentally investigate Pf‐contaminated canine pRBCs. Methods: Canine pRBCs were inoculated with Pf‐rich pRBCs from the sentinel feline unit and stored at 4°C or 20°C for 72 hours. Aliquots from the pRBCs were serially evaluated by microscopy, culture, and a eubacterial 16S rRNA real‐time PCR assay. Results: One Pf‐contaminated feline unit turned black after 22 days of storage and was removed from the blood bank; a source was not found, and no other contaminated units were identified. Canine pRBCs spiked with 5 or 25 μL of the sentinel unit became culture‐ and/or 16S PCR‐positive at ≥8 hours at 20°C and 48 hours at 4°C and developed a color change at ≥24 hours. Sensitivity studies indicated that without incubation, inoculation of ≥100 μL Pf‐rich pRBCs was necessary for a positive 16S PCR test result. Conclusions: P. fluorescens grows in stored pRBCs slowly at 4°C and rapidly at 20°C. Screening of blood products for color change, estimating bacterial concentration with microscopy, and 16S PCR testing are simple and fast ways to detect bacteria in stored blood. Aseptic collection, temperature‐controlled storage, and regular visual monitoring of stored units is recommended. Discolored units should not be transfused, but examined for bacterial contamination or other blood product quality problems.     

Sensitivity,specificity, and predictive values of reagent test strip estimations of blood urea nitrogen

Azostix-reagent-tests(R) strips (Ames, Miles, Inc., Diagnostic Division, Elkhart, IN) were used to measure blood urea nitrogen values in blood samples from 125 dogs and cats at the North Carolina State University, College of Veterinary Medicine. Results of the tests were compared with standard serum urea nitrogen results. Sensitivity, specificity, and negative predictive values were high (86.4, 90.3, and 96.5%, respectively). Positive predictive value was low, 65.5% of the dogs and cats with elevated blood urea nitrogen values were correctly classified as abnormal The test performs well when the prevalence of abnormal values is near 50%.     

The effects of cooling and vitrification of embryos from mares treated with equine follicle-stimulating hormone on pregnancy rates after nonsurgical transfer

Equine embryos can remain viable for 12 to 24 hours when cooled and stored at 5°C.¹ Cryopreservation of embryos would allow for long-term preservation of genetic material and more efficient management of embryo recipients. This study compared pregnancy rates after transfer of equine embryos vitrified within 1 hour of collection or cooled for 12 to 19 hours before vitrification. Mares (N = 40) were superovulated using equine follicle-stimulating hormone (eFSH). Embryos were recovered 6.5 days after ovulation or 8 days after human chorionic gonadotropin. Forty morulae or early blastocysts with a grade of 1 to 2 and <300 mm in diameter were randomly assigned to 1 of 2 treatments: Group 1 (n = 20), washed 4 times in a commercial holding medium and then vitrified; Group 2 (n = 20), washed 3 times and then stored in the same holding medium at 5°C to 8°C in a passive cooling device for 12 to 19 hours before being vitrified. To thaw, embryos were warmed by holding the straw in air at room temperature for 10 seconds and then submerged in a water bath (20°C to 22°C) for an additional 10 seconds. The contents of the straw were transferred directly into a recipient that had ovulated 4 to 6 days previously. There were no differences (P > .05) in embryo diameter, grade, or morphology score between treatment groups before vitrification. Pregnancy rates (day 16) were not different (P > .05) between embryos vitrified immediately after collection (15 of 20; 75%) and embryos cooled for 12 to 19 hours before vitrification (13 of 20; 65%). Based on these results, small equine embryos (<300 mm) can be stored at 5°C to 8°C for 12 to 19 hours before vitrification without a significant loss of viability.     

COLOR DOPPLER AND DOPPLER SPECTRAL ANALYSIS IN THE SPINAL CORD OF NORMAL DOGS

Doppler ultrasonography of the spinal cord was performed in 34 normal, anesthetized dogs following hemilaminectomy. This study was part of an investigation to evaluate the efficacy of a 21-aminosteroid compound and high dose methylprednisolone for the treatment of spinal cord trauma. Grey-scale images of the canine spinal cord were similar to those described for the spinal cord of people. Doppler waveforms of intraparenchymal spinal arteries exhibited high end diastolic blood flow velocities, indicating low resistance to flow. Doppler values (mean ± SD) for arteries immediately ventrolateral to the central canal were: Peak Systolic Velocity = 5.78 ± 2.5 cm/sec, Minimum Diastolic Velocity = 3.5 ± 1.62 cm/sec, Mean Velocity = 4.45 ± 1.96 cm/sec, Minimum Diastolic Velocity = 3.5 ± 1.62 cm/sec, Mean Velocity = 4.45 ± 1.96 cm/sec, Systolic/Diastolic ratio = 169 ± 0.19, Pulsatility Index = 0.53 ± 0.09, and Resistance Index = 0.4 ± 0.06.     

ULTRASONOGRAPHY OF PERIPHERAL NERVES DURING WALLERIAN DEGENERATION AND REGENERATION FOLLOWING TRANSECTION

Ultrasonography was performed on sciatic, tibial and/or peroneal nerves and interosseous muscles in 7 dogs using a ultrasound machine with a 7.5 MHz linear array transducer. A tibial nerve was transected near the distal aspect of the bellies of the gastroenemius muscle. Serial neurologic examinations, electromyography, and ultrasonography were performed before and after surgery. Dogs were euthanized at variable intervals and histopathology performed on nerve samples. In sagittal images, normal nerves had hyperechoic walls with multiple internal linear echoes. In transverse images, the nerves were round or oval hyperechoic structures with internal punctate echoes. After transection, the proximal stump was consistently seen whereas the distal stump and nerve were harder to identify. Neuromas were present in all 5 dogs followed beyond 2 days and appeared as hypoechoic bulbous swellings most apparent at 3 weeks after surgery. Only 1 dog developed a neuroma large enough to be considered of potential clinical significance. Four dogs were followed beyond 2 months. Regeneration was evidenced by a steady growth of nerve with an irregular outline (2 dogs) or by a knobby connection between the proximal and distal stumps (1 dog). Regeneration was not detected in 1 dog.