CJVR - April 2021, Vol. 85, No. 2



Classification of porcine reproductive and respiratory syndrome virus in Ontario using Bayesian phylogenetics and assessment of temporal trends

Dylan John Melmer, Robert Friendship, Terri L. O’Sullivan, Amy L. Greer, Dinko Novosel, Davor Ojkic, Zvonimir Poljak (page 83)

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important swine viruses globally, including in Ontario, Canada. Understanding the evolution and relation of the various PRRSV genotypes in Ontario can provide insight into the epidemiology of the virus. The objectives of this study were to i) describe the variability of PRRSV genotypes in Ontario swine herds, and ii) evaluate possible groupings based on PRRSV genomic data. Virus open reading frame 5 (ORF-5) sequences collected from 2010 to 2018 were obtained from the Animal Health Laboratory, University of Guelph and Bayesian phylogenetic models were created from these. The PRRSV population of Ontario was then categorized into 10 distinct clades. Model comparisons indicated that the model with a constant population assumption fit the data best, which suggests that the net change in the PRRS virus variation of the entire population over the last decade was low. Nonetheless, viruses grouped into individual clades showed temporal clustering during distinct time intervals of the entire study period (P < 0.01).

A comparative efficacy test of 1 versus 2 doses of CIRCOQ PCV2 subunit vaccine against naturally occurring PCV2-type d in piglets with high maternally derived antibodies (MDAs) on a Vietnamese swine farm

Duy Tien Do , Khanh Doan Vinh Tran, Anh Tuyet Quach , David Lee, Frank CJ Chang, Carol PY Wu, Toan Nguyen Tat, Chanhee Chae (page 93)

The aim of this study was to evaluate the protective efficacy of the CIRCOQ porcine circovirus type 2 (PCV2) subunit vaccine in piglets with high maternally derived antibodies (MDAs) against disease caused by natural infection with PCV2d. A total of 130 weaned, 21-day-old healthy pigs was allocated into 3 trial groups. The signs of respiratory disorder were higher in unvaccinated pigs than in vaccinated pigs at 13 to 17 weeks old (P < 0.05), 18 to 22 weeks old (P < 0.001), and 23 to 27 weeks old (P < 0.01). The unvaccinated pigs had an early rate of dermatitis at 8 to 12 weeks old (10.0%), 13 to 17 weeks old (30.0%), 18 to 22 weeks old (46.7%), and 23 to 27 weeks old (33.3%), while there were no cases of dermatitis in vaccinated pigs. There was a significant difference (P < 0.05) in the mortality of pigs in the unvaccinated group and the 2-dosed vaccinated group. PCV2 viremia was detected in the blood and peaked at 105 days old in both unvaccinated pigs (Ct-adj = 8.40) and pigs vaccinated with 1 dose (Ct-adj = 6.37), while no detectable PCV2 virus was found in the blood of pigs vaccinated with 2 doses. At 77 and 105 days old, the PCV2 viremia load (Ct-adj) of unvaccinated pigs and those vaccinated with 1 dose was significantly higher (P < 0.05) than that of the 2-dosed vaccinated pigs. The body weight (BW), average weight gain (AWG), and average daily gain (ADG) in both groups of vaccinated pigs were significantly higher (P < 0.05) than those of unvaccinated pigs. The study vaccine was significantly efficacious in protecting vaccinated pigs against clinical symptoms, blood viral load, and mortality, as well as improving productivity, compared with unvaccinated pigs.

Impact of meloxicam on respiratory virus titers and health outcomes when administered concurrently with a modified live respiratory vaccine in abruptly weaned beef steers

Elizabeth R. Homerosky, Michael J. Jelinski, Craig Dorin (page 101)

Abruptly weaned crossbred steer calves (N = 271) were used in a randomized, blinded 2-arm clinical trial to assess the impact of a long-acting non-steroidal anti-inflammatory drug on bovine herpesvirus type 1, bovine respiratory syncytial virus, parainfluenza virus type 3, and coronavirus titers and health outcomes when administered concurrently with a modified live respiratory vaccine upon arrival at a feedlot. Treatment groups included a control (saline; n = 135) and an experimental group (injectable meloxicam; n = 136). Viral antibody titers and body weight were measured on arrival, day 7, and day 21, along with a final weight on day 45. Body weight and antibody titers for all viruses increased over time (P < 0.001); however, there were no differences by treatment group or a significant group × time interaction when evaluated using repeated measures analysis of variance. Interestingly, the use of meloxicam was associated with increased treatment risk (P < 0.05). In conclusion, the administration of meloxicam may adversely affect health; however, a decreased vaccine response is likely not a contributing factor.

Pre-warming following premedication limits hypothermia before and during anesthesia in Sprague-Dawley rats (Rattus norvegicus)

Maxime Rufiange, Vivian S.Y. Leung, Keith Simpson, Daniel S.J. Pang (page 106)

In humans and other mammals, general anesthesia impairs thermoregulation, leading to warm core blood redistributing to the periphery. This redistribution is an important contributor to hypothermia that can be reduced with pre-warming before anesthesia. Additionally, sedation following premedication has been associated with hypothermia in dogs. In a prospective, randomized, cross-over study, 8 adult male and female rats (weighing 388 to 755 g) were sedated with intramuscular ketamine-midazolam-hydromorphone, then placed in an unwarmed cage or warmed box for 14 minutes, followed by 30 minutes of isoflurane anesthesia with active warming. Core body temperature was monitored throughout. After sedation, warmed rats gained 0.28°C ± 0.13°C and unwarmed rats lost 0.19°C ± 0.43°C, a significant difference between groups (P = 0.004). After anesthesia, warmed rats maintained higher core temperatures (P < 0.0001) with 2/8 and 6/8 of warmed and unwarmed rats becoming hypothermic, respectively. Pre-warming during sedation and active warming during general anesthesia is effective in minimizing hypothermia.

Impact of selected individual dog traits on echocardiographic parameters obtained in 1-dimensional (M-mode) and 2-dimensional (2D) imaging

Oktawia Szpinda, Marta Parzeniecka-Jaworska, Michal Jank, Magdalena Garncarz, Michal Czopowicz (page 112)

The popularity and availability of echocardiography in veterinary practice for companion animals have substantially increased in recent years. The results obtained during the procedure are compared to reference values established for the general dog population or to standards developed for a specific dog breed. The aim of this study was to determine whether individual dog traits, such as body weight, chest structure, and level of physical activity and performance, affect the reference values for echocardiographic parameters. Published reference values for echocardiographic examination parameters for 32 dog breeds were analyzed and the relationship between individual echocardiographic parameters and body weight, chest structure, and level of physical activity and performance was then statistically analyzed. It was found that echocardiographic parameters are affected by the dog’s weight and physical activity. There was no significant relationship between heart size and chest structure. The great variety of dog breeds means that echocardiographic findings should be individually interpreted rather than establishing reference ranges for each breed in population studies. This will allow for a more accurate interpretation of the results obtained in the echocardiographic examination and consequently lead to earlier diagnosis of changes in myocardial morphology.

A genome-wide analysis of cardiac lesions of pigs that die during transport: Is heart failure of in-transit-loss pigs associated with a heritable cardiomyopathy?

Katherine Zurbrigg, Francesca Bertolini, Muhammed Walugembe, Toni van Dreumel, David Alves, Robert Friendship, Terri L. O’Sullivan, Max F. Rothschild (page 119)

While heart failure is a primary cause of death for many in-transit-loss (ITL) pigs, the underlying cause of these deaths is not known. Cardiomyopathies are considered a common cause of heart failure in humans and often have a genetic component. The objective of this study was to determine if genes associated with cardiomyopathies could be identified in ITL pigs. Samples from the hearts of pigs that died during transport to an abattoir in Ontario, Canada were collected and genotyped along with samples from pigs that did not die during transport (ILT hearts: n = 149; non-ITL/control hearts: n = 387). Genome-wide analyses were carried out on each of the determined phenotypes (gross cardiac lesions) using a medium density single nucleotide polymorphism (SNP) chip and 500 kb windows/regions for analysis, with 250 kb regions of overlap. The distribution derived by a multidimensional scaling (MDS) analysis of all phenotypes demonstrated a lack of complete separation between phenotypes of affected and unaffected animals, which made diagnosis difficult. Although genetic differences were small, a few genes associated with dilated cardiomyopathy (DCM) and arrhythmogenic right ventricular cardiomyopathy (ARVM) were identified. In addition, multiple genes associated with cardiac arrhythmias and ventricular hypertrophy were identified that can possibly result in heart failure. The results of this preliminary study did not provide convincing evidence that a single, heritable cardiomyopathy is the cause of heart failure in ITL pigs.

Effects of head position on internal and external carotid pressures in standing sedated horses

Stacy R. Caffey, Caleb M. Lund, Kelly D. Farnsworth, Boel A. Fransson, Claude A. Ragle (page 127)

The effects of head position on internal carotid artery (ICA) and external carotid artery (ECA) pressures in standing sedated horses were evaluated in this study. The common carotid artery (CCA) was catheterized in 6 horses using an ultrasound-guided technique to facilitate placement of a pressure transducer within the ICA and ECA at the level of the guttural pouch. Transducer position was confirmed by endoscopic visualization. Mean arterial pressure (MAP) was measured with horses in both a head-up and head-down position. The dorsal metatarsal artery was catheterized as a control. Maintaining a head-up position decreased MAP in both the ICA (median: 75.21 mmHg) and ECA (median: 79.43 mmHg), relative to the head-down position (ICA median: 104.65 mmHg; ECA median: 102.26 mmHg). Mean arterial pressure in the dorsal metatarsal artery was not affected by head position. The head-up position resulted in lower arterial pressures in both the ICA and ECA (P = 0.03) compared with the head-down position in standing sedated horses.

Retrospective analysis of use of fluorine-18 fluorodeoxyglucose-positron emission tomography/computed tomography (18F-FDG PET/CT) for detection of metastatic lymph nodes in dogs diagnosed with appendicular osteosarcoma

Powell T. Slinkard, Elissa K. Randall, Lynn R. Griffin (page 131)

The purpose of this retrospective analysis was to determine if fluorine-18 fluorodeoxyglucose-positron emission tomography/computed tomography (18F-FDG PET/CT) could potentially be an accurate staging tool for detecting metastatic lymph nodes in dogs with appendicular osteosarcoma based on the quantitative measurement of the maximum standard uptake value (SUVmax) of lymph nodes. A total of 53 dogs were identified that presented for staging via 18F-FDG PET/CT for primary appendicular osteosarcoma. Patients were categorized according to lymph node status of having either metastatic or non-metastatic nodes based on cytological or histological analysis. Maximum standard uptake (SUVmax) values of the sampled lymph node(s) were recorded and 3/77 (3.9%) of sampled lymph nodes were confirmed metastatic. A Mann-Whitney test revealed a statistical difference in the SUVmax of the metastatic versus non-metastatic lymph nodes [median: 6.6 to 95% confidence interval (CI): 2.56 to 14.37 versus 2.18 95% CI: 2.32 to 3.17, respectively, P-value = 0.05]. This retrospective analysis revealed a significant difference in the SUVmax as measured on 18F-FDG PET/CT between metastatic lymph nodes and non-metastatic lymph nodes in canine patients afflicted with appendicular osteosarcoma, in spite of the small numbers analyzed. While these results are promising, they should be interpreted with caution and further studies are justified.

Assessment of neutrophil function in canine cancer patients undergoing chemotherapy and correlation with neutrophil numbers

Arefeh Ravanbakhsh, Khawaja Ashfaque Ahmed, Valerie MacDonald-Dickinson, Nicole J. Fernandez, Melissa D. Meachem, Ryan M. Dickinson (page 137)

Decreased neutrophil function following administration of chemotherapy has been reported in dogs with lymphoma. The first objective of our study was to determine if neutrophil oxidative burst and phagocytic activity are affected by chemotherapy 7 to 10 days following initiation of treatment in dogs with lymphoma and non-lymphoma malignancies. The second objective was to determine if there is a correlation between neutrophil numbers and neutrophil function before or after initiation of chemotherapy. Flow cytometric assessment of neutrophil oxidative burst and phagocytosis following stimulation with Escherichia coli was performed in 9 dogs diagnosed with lymphoma and 17 non-lymphoma tumor-bearing dogs pre- and post-chemotherapy, as well as 14 tumor-free control dogs. Spearman rank correlation was performed to determine if blood neutrophil numbers and neutrophil function were significantly correlated. Lymphoma patients showed significantly reduced percentage neutrophil oxidative burst post-chemotherapy compared to healthy controls as well as compared to pre-chemotherapy values (P = 0.0022 and P = 0.0020, respectively). Lymphoma patients also exhibited significantly reduced neutrophil phagocytosis activity post-chemotherapy compared to controls and pre-chemotherapy values (P = 0.0016 and P = 0.014, respectively). Dogs with non-lymphoma malignancies also showed a significant decrease in both percentage oxidative burst and phagocytosis post-chemotherapy compared to pre-chemotherapy values (P = 0.00040 and P = 0.029, respectively). Neutrophil numbers and function were not significantly correlated. The results of the study suggest that chemotherapeutic treatment decreases neutrophil oxidative burst and phagocytic activity 7 to 10 days post-treatment in dogs with various malignancies. Furthermore, neutrophil numbers cannot be used to predict neutrophil function.

Brief Communication

Development of a SYBR Green-based real-time quantitative polymerase chain reaction assay to detect enzootic nasal tumor virus in goats

Rongze He, Yulan Du, Linli Gan, Muhammad Ali Mohsin, Bao-Xiang He (page 145)

Enzootic nasal adenocarcinoma is a contagious respiratory disease in goats that is caused by the enzootic nasal tumor virus 2 (ENTV-2). In order to increase the number of available detection methods for ENTV-2, we developed a SYBR Green real-time polymerase chain reaction (SGrPCR) assay that targets the gag gene of ENTV-2. The low limit of detection of the assay was 3.68 × 101 copies/µL, a hundredfold more sensitive than conventional PCR. The melt curve showed a single sharp melt peak at 83°C, which indicated that there was no non-specific amplification or primer dimer formation. The intra-assay and inter-assay coefficients of variation were 1.58% and 1.82%, respectively. There was no cross-reactivity with closely related goat viruses (i.e., orf virus, peste des petits ruminants virus, goatpox virus, foot-and-mouth disease virus) and endogenous retroviruses. In conclusion, the SGrPCR assay is specific for the gag gene of ENTV-2 and provides a rapid and sensitive approach for detecting ENTV-2 in clinical samples.

Comparison of the bacteriostatic effects of quaternary ammonium compounds and their combinations on a dairy farm environment and the microbial contamination of dairy products

Nannan Chen, Shanshan Liu, Yu Liu, Tongtong Bai, Lihua Jia, Li Wang, Shan Yue, Chenhua Wu, Wenjing Huang, Li Gao, Jinlei Fu, Siyu Liu, Tong Zhao, Juan Li, Yuntao Zhang, Zhanbo Zhu (page 151)

Disinfection is key for controlling microbial contamination and ensuring the safe production of milk and dairy products. In this study, we developed a new disinfection method using quaternary ammonium surfactant N-dodecyl-2-(pyridin-1-yl) acetamide chloride as the main component to form a bactericidal complex with either chlorhexidine acetate or glutaraldehyde, and we evaluated the bactericidal effects, safety, and clinical application value of the compound disinfectants. An in vivo acute oral toxicity assay in mice showed an LD50 > 5000 mg/kg body weight without abnormality in pathological tissue sections. Comparison with commercially available products also showed that they have outstanding bactericidal effects. Clinical trials proved that the compound disinfectants have excellent bactericidal effects on the air and ground of the dairy farm and on the skin of cattle, especially in a dairy farm environment. Our findings confirm that the new compound disinfectants have excellent bactericidal performance and are safe to use as disinfectants to prevent mastitis and contamination of the cattle farm environment.

Preliminary study of urinary excretion of liver-type fatty acid-binding protein in a cat model of chronic kidney disease

Akiko Watanabe, Keiichi Ohata, Tsuyoshi Oikawa, Takeshi Sugaya, Masao Miyazaki, Hiroshi Satoh, Masaaki Katayama (page 156)

Urinary liver-type fatty acid-binding protein (uL-FABP) is a clinically useful biomarker for monitoring chronic kidney disease (CKD) in humans. However, long-term monitoring of uL-FABP in CKD cats has not been reported. The objective of this preliminary study was to investigate whether the urinary excretion of L-FABP could predict the deterioration of renal function in 2 CKD model cats. Urinary liver-type fatty acid-binding protein (uL-FABP) increased before standard renal biomarkers, including serum creatinine, blood urea nitrogen, and symmetric dimethylarginine, in 1 cat with deteriorating renal function, but remained low and relatively stable in another cat with stable renal function. Our results suggest that uL-FABP is a potential clinical biomarker for predicting the progression of CKD in cats, as it is in humans.