Ciliary rootlet coiled coil protein (CROCC), the structural component that originates from the basal body at the proximal end of the ciliary rootlet, plays a crucial role in maintaining the cellular integrity of ciliated cells. In the current study, we cloned Xenopus CROCC and performed the expression analysis. The amino acid sequence of Xenopus laevis was related to those of Drosophila, cow, goat, horse, chicken, mouse and human. Reverse transcription polymerase chain reaction analysis revealed that CROCC mRNA encoding a coiled coil protein was present maternally, as well as throughout early development. In situ hybridization indicated that CROCC mRNA occurred in the animal pole of embryo during gastrulation and subsequently in the presumptive neuroectoderm at the end of gastrulation. At tailbud stages, CROCC mRNA expression was localized in the anterior roof plate of the developing brain, pharyngeal epithelium connected to gills, esophagus, olfactory placode, intestine and nephrostomes of the pronephric kidney. Our study suggests that CROCC may be responsible for control of the development of various ciliated organs.

Bovine parainfluenza virus type 5 (bPIV5) was isolated from cattle with downer cow syndrome in 2012, and included both respiratory and neurotropic pathogens from a variety of animals. In the current study, we conducted serosurveillance using sera obtained from seven Korean farms and optimized a reverse transcription-polymerase chain reaction (RT-PCR) assay to detect bPIV5. The overall seropositive rate for Korean cattle was 21.4% (163/760). A farm located near the city of Milyang in Gyeoungnam province had a markedly elevated seropositive rate for bPIV5 compared to that of the other six farms. The regional seropositive rates were 4.2% (8/192) for Haman, 19.5% (18/ 55) for Hwasung, 73.9% (65/88) for Milyang, 26.0% (50/192) for Namwon, 1.0% (1/96) for Uljin, 13.5% (13/96) for Yeongju, and 32.7% (8/41) for Yongin. The sensitivity and specificity of three RT-PCR primer sets used to amplify the conserved fusion gene of bPIV5 were also evaluated. An RT-PCR assay using the bPIVFR3 primer set was 10- fold more sensitive than the assays using the two other primer sets and did not result in non-specific amplification. These results demonstrated that the bPIFR3 primer set can be used to detect bPIV5.

4 , 4 - diaminodiphenyl sulfone (dapsone) is a sulfone drug that has antibacterial effects on a variety of bacteria, especially Mycobacterium leprae ; thus, it has been used to treat leprosy. Previous studies demonstrated that dapsone inhibits integrin-mediated adherence of neutrophils and production of prostaglandin E2 by polymorphonuclear leukocytes. Hence, dapsone may act in immune cells and regulate cell-mediated inflammation processes. However, its antiinflammatory effects remain unclear. The present study demonstrated that dapsone modulates the production of inflammation-related cytokines in immune cells. We employed the spleen cells of mice, which are major immune cells, and lipopolysaccharide (LPS) as a causative agent of inflammation for experiments. Dapsone induced a proportional change in splenocyte subsets and the apoptosis of spleen cells. Interestingly, dapsone decreased the production of tumor necrosis factor-alpha and interleukin (IL)-10, but not IL-6, in LPS-treated spleen cells. In other assays, we measured the dapsone-induced production of nitric oxide (NO) and the expression of activation markers of spleen cells. Dapsone decreased NO production in LPS-treated spleen cells. Taken together, our results demonstrate that dapsone has antiinflammatory effects in immune cells and provide new insight into the potential uses of this agent.

Cardiopulmonary depression of long-term constant rate infusion (CRI) administration of multiple analgesic drugs is important, especially in critically ill dogs. Therefore, this study was conducted to evaluate the effects of lidocaine, ketamine or combined lidocaine-ketamine combination CRI treatment on vital signs and left ventricular (LV) function in healthy dogs. Six adult Beagle dogs were administered either ketamine (initial loading dose of 0.5 mg/ kg followed by 10 ¥ìg/kg/min CRI), lidocaine (initial loading dose of 2 mg/kg followed by 0.025 mg/kg/min CRI), or combined lidocaine-ketamine intravenously. Arterial blood pressure (BP), heart rate (HR), respiratory rate (RR), body temperature (BT) and echocardiographic LV dimensions were measured before administration of medications, immediately after administration of drugs, and then every 10 min for 2 h. There were no significant changes in HR, RR, BT and BP after the administration of either lidocaine CRI, ketamine CRI, or combined lidocaine and ketamine CRI. There were also no significant changes in LV dimensions and stroke volume. The results revealed that treatment with either lidocaine, ketamine or combined lidocaine-ketamine may not cause cardiopulmonary suppression in healthy dogs.

OBJECTIVE To measure incidence and estimate temporal and spatial dynamics of porcine reproductive and respiratory syndrome virus (PRRSV) infection in US sow herds. ANIMALS 371 sow herds in the United States from 14 production companies. PROCEDURES The exponentially weighted moving average was used to monitor incident PRRSV infections for onset of an epidemic. The spatial scan statistic was used to identify areas at significantly high risk of PRRS epidemics. A χ2 test was used to estimate whether there were significant differences in the quarterly and annual PRRS incidence among time periods, and a bivariable logistic regression model was used to estimate whether PRRSV infection during a given year increased the odds of that herd being infected in the following year. RESULTS During the 4-year period of this study, 29% (91/319; 2009 to 2010), 33% (106/325; 2010 to 2011), 38% (135/355; 2011 to 2012), and 32% (117/371; 2012 to 2013) of the herds reported new infections. Weekly incidence was low during spring and summer and high during fall and winter. The exponentially weighted moving average signaled the onset of a PRRSV epidemic during the middle 2 weeks of October each year. Disease incidence was spatially clustered. Infection in the previous year increased the odds of infection in 2010 to 2011 and 2011 to 2012. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated a striking repeatability in annual PRRSV temporal and spatial patterns across 4 years of data among herds from 14 production companies, which suggested that efforts to control PRRSV at a regional level should continue to be supported.

The objective of this study was to determine the effect of additional human chorionic gonadotrophin (hCG) on the ovarian response of gilts previously treated with 200 IU hCG combined with 400 IU equine chorionic gonadotrophin (eCG) (eCG/hCG). Seventy-one prepuberal gilts (105 ± 7.5 kg) were assigned to groups: i) eCG/hCG (hCG-0; n = 25); ii) eCG/hCG followed by 100 IU of hCG at 24 h (hCG-100; n = 24); iii) eCG/hCG followed by 200 IU hCG at 24 h (hCG-200; n = 10); and iv) controls (CON; n = 12). Ovulation response was assessed by ovarian dissection or real-time ultrasonography. Additional hCG did not significantly improve numbers of gilts ovulating. Numbers of corpora lutea increased with hCG, and was higher in hCG-200 (P < 0.01). Compared to hCG-0, the frequency of cysts in gilts was higher in hCG-100 (P < 0.05) and further increased in hCG-200 (P < 0.01). The number of cysts per gilt was dose-dependently increased by additional hCG. We conclude that supplemental hCG will increase the number of corpora lutea but will be associated with follicular cyst development in a dose dependent manner.

OBJECTIVE To assess changes in biochemical and biophysical properties of canine RBCs during cold (1° to 6°C) storage in a licensed RBC additive solution (the RBC preservation solution designated AS-1) supplemented with ascorbic acid. SAMPLE Blood samples from 7 neutered male Greyhounds; all dogs had negative results when tested for dog erythrocyte antigen 1.1. PROCEDURES Blood was collected into citrate-phosphate-dextrose and stored in AS-1. Stored RBCs were supplemented with 7.1mM ascorbic acid or with saline (0.9% NaCl) solution (control samples). Several biochemical and biophysical properties of RBCs were measured, including percentage hemolysis, oxygen-hemoglobin equilibrium, and the kinetic rate constants for O2 dissociation, carbon monoxide association, and nitric oxide dioxygenation. RESULTS Greyhound RBCs stored in AS-1 supplemented with ascorbic acid did not have significantly decreased hemolysis, compared with results for the control samples, during the storage period. CONCLUSIONS AND CLINICAL RELEVANCE In this study, ascorbic acid did not reduce hemolysis during storage. Several changes in stored canine RBCs were identified as part of the hypothermic storage lesion.

Mannheimia haemolytica is an important cause of pneumonia in feedlot cattle. Nuclear factor erythroid-2-related factor 2 (Nrf2) is a redox-sensitive transcription factor responsible for the induction of antioxidant enzymes, such as heme oxygenase 1 (HO-1), within the lung. The expression of Nrf2 and HO-1 was immunohistochemically evaluated in 4 calves 24 h after experimental infection with M. haemolytica. Calves receiving normal saline served as controls. In the infected lungs, cytoplasmic Nrf2 expression was high in macrophages and bronchioles and low in alveolar epithelium, whereas nuclear expression was high in endothelial cells, macrophages, and bronchioles and lowest in alveolar epithelium. Normal lung samples displayed only faint Nrf2 cytoplasmic staining within bronchiolar epithelium. Expression of HO-1 was detected within the cytoplasm of macrophages and bronchiolar epithelial cells in all infected lung samples, whereas normal lungs displayed only weak cytoplasmic staining in bronchiolar epithelial cells. These findings suggest that bronchiolar epithelial cells and macrophages up-regulate Nrf2 expression early in the course of infection, which results in increased expression of HO-1 within these cells.

Pododermatitis is a disease of concern for mink breeders in Canada and worldwide, as it causes discomfort and lowers the breeding rates on farms affected by the disease. Unfortunately, the etiology and pathogenesis of pododermatitis are still unknown. In this study, we compared Staphylococcus spp. and Streptococcus canis isolates from healthy mink with isolates from animals with pododermatitis on 2 farms in Ontario. Almost all hemolytic Staphylococcus spp. isolated were shown to be Staphylococcus delphini Group A by 16S ribosomal ribonucleic acid (rRNA) sequence analysis and polymerase chain reaction (PCR). Pulsed-field gel electrophoresis (PFGE) did not reveal any S. delphini or S. canis clonal lineages specifically associated with pododermatitis, which suggests that these bacteria do not act as primary pathogens, but does not dismiss their potential roles as opportunistic pathogens. While S. delphini and S. canis were the most prevalent bacterial pathogens in mink pododermatitis, they were also present in samples from healthy mink. Arcanobacterium phocae is occasionally isolated from pododermatitis cases, but is difficult to recover with conventional culture methods due to its slow growth. A quantitative real-time PCR was developed for the detection of A. phocae and was tested on 138 samples of footpad tissues from 14 farms. The bacterium was detected only in pododermatitis-endemic farms in Canada and was at higher concentrations in tissues from infected footpads than in healthy tissues. This finding suggests that A. phocae is involved in the pathogenesis of pododermatitis.

Indirect enzyme-linked immunosorbent assays (ELISAs) are frequently run as endpoint ELISAs (e-ELISAs). However, kinetic ELISAs (k-ELISAs) have certain advantages over e-ELISAs. The objective of this study was to understand the relationship between e-ELISA and k-ELISA results. Specifically, to determine whether it was possible to run both k-ELISA and e-ELISA on the same plate and establish an appropriate time interval for k-ELISA measurements. A normalization method for k-ELISA slopes (slope ratio) is proposed. Using an indirect e-ELISA test measuring antibodies against Ostertagia ostertagi in milk from dairy cattle, we found that running a k-ELISA had no effect on optical density ratio results of an e-ELISA on the same plate, and that agreement was very strong at 10, 15, and 28 min, allowing for a reduction in the total processing time for ELISA tests.