CVMA | Table of Contents and Abstracts

Table of Contents and AbstractsJanuary 2019, Vol. 83, No. 1


Evaluation of effects of radiation therapy combined with either pamidronate or zoledronate on canine osteosarcoma cells

Katie Hoddinott, Michelle L. Oblak, Geoffrey A. Wood, Sarah Boston, Anthony J. Mutsaers (page 3)

Canine osteosarcoma is a devastating disease with an overall poor prognosis. Radiation therapy and bisphosphonates are currently used in combination for palliative treatment, despite a paucity of literature that investigates their combined use. The objectives of this study were to assess the in vitro effects of radiation therapy and bisphosphonates on canine osteosarcoma cells when used in combination. Canine osteosarcoma cell lines D17 and Dharma were treated with radiation and pamidronate or zoledronate, both alone and in combination. The effects of these treatments were assessed using clonogenic survival and cell viability assays. Dose-dependent decreases in clonogenic survival and cell viability were observed for both radiation and bisphosphonate treatment. Combination index analysis revealed antagonistic interactions when radiation and bisphosphonates were used in combination at specific doses for both D17 and Dharma osteosarcoma cells. Further investigation of the combined effects of radiation and bisphosphonates for the palliative treatment of canine osteosarcoma is warranted.

Cochleosaccular (Scheibe) dysplasia in dogs: A temporal bone study

Nevra Keskin, Hasan Albasan, Irem Gul Sancak, Michael M. Paparella, Sebahattin Cureoglu (page 11)

The objective of this study was to evaluate any otopathologic changes in temporal bone specimens from dogs with deafness related to cochleosaccular (Scheibe) dysplasia (CSD). We used the canine temporal bone collections of the Otopathology Laboratory at the University of Minnesota and of the Massachusetts Eye and Ear Infirmary at Harvard University in Boston. Our morphometric analysis included measuring the areas of the stria vascularis and the spiral ligament and counting the number of spiral ganglion cells. In addition, we noted the presence of the organ of Corti and cochlear hair cells, assessed the location of Reissner's membrane and the saccular membrane, and counted the number of both Type I and Type II vestibular hair cells in the macule of the saccule and vestibular ganglion cells. In the group of specimens from dogs with cochleosaccular dysplasia, we observed generalized degeneration in the cochlea and a significantly decreased number of Type I and Type II vestibular hair cells and vestibular ganglion cells. As hereditary deafness is presently untreatable with known therapeutic methods, dogs with cochleosaccular dysplasia should not be considered for breeding. Future therapeutic approaches, such as stem cell therapies, should be designed to target all the elements of the cochlea in addition to the saccule as it was found that both are affected in dogs with CSD.

The effect of transfixation pins on the biomechanical properties of angled acrylic connecting bars

Xavier Montasell, Gregory Herndon, David Szwec, Guy Beauchamp (page 17)

With acrylic external-fixation frames for fracture repair the acrylic columns can be contoured to allow greater versatility in the placement of transfixation pins, thus minimizing damage to the surrounding soft tissue and making mandibular and transarticular fixation easier. However, contouring affects the stiffness and ultimate strength of the construct under axial compression. In this study, polymethylmethacrylate columns 21 mm in diameter angled at 0°, 30°, 45°, 60°, or 90° with clamps were constructed. For each angulation group, pins 3.2-mm long were placed in 6 columns, 2 pins at each end, 1.5 cm from each other, and 6 columns had no pins. Each column was allowed to polymerize for a minimum of 10 min, then was placed in a biomechanical-testing machine, the load cell at the bottom end of the column and the actuator on top, with a preload of 10 to 12 N to prevent slippage. The columns underwent axial loading at a rate of 8 mm/s until catastrophic failure occurred. Data on force and deformation were collected every 0.025 s. Both stiffness and ultimate strength of the column decreased significantly (P < 0.01), up to 77% and 70%, respectively, with each increase of angulation. The columns with pins were significantly less stiff (P < 0.05) than those without pins at angulations of 45° and 60°. However, the columns with pins did not show significant differences in ultimate strength from the columns without pins at any of the angulations. The point of failure was always at the angle of the column, demonstrating that in axial compression the weakest point is not the pin–acrylic interface when pins are eccentrically located within the column.

Use of standard diagnostic techniques to determine eradication of infection in experimental equine septic arthritis

Roman V. Koziy, Seiji Yoshimura, Ryan Dickinson, Joanna M. Rybicka, Igor Moshynskyy, Musangu Ngeleka, Jose L. Bracamonte, Elemir Simko (page 24)

Septic arthritis is an important disease in horses, necessitating aggressive and prolonged therapy. In order to guide therapy, reliable methods of detecting the eradication of infection are needed. Therefore, the objective of this study was to investigate detection of eradication of infection in an experimental model of equine septic arthritis using standard diagnostic techniques. For this purpose, 17 adult horses were assigned to 3 experimental groups. The middle carpal joint of each horse was injected with Escherichia coli (Septic group, n = 8), lipopolysaccharide (LPS) (LPS group, n = 6), or sterile saline (Control group, n = 3) at day 0. Contralateral joints were not injected. Standard therapy was applied to all joints except non-injected joints in the Control group at day 1. Sequential samples of synovial fluid (SF) were collected for bacterial culture using 3 culture media [Columbia blood agar (CBA), brain heart infusion broth (BHI), and Signal blood culture medium] and for cytological evaluation [percentage neutrophils (PN), total nucleated cell count (TNCC), and total protein (TP)]. Escherichia coli-specific polymerase chain reaction (PCR) was carried out to detect E. coli DNA in synovial fluid. Culture and PCR were positive for E. coli in all joints injected with E. coli at day 1 and 1 joint was positive on BHI at day 4. Based on the results of bacterial culture, PCR, and TNCC, the elimination of infection in our experimental model occurred by day 4 post-infection in 6 out of 7 cases. Total protein (TP) and PN remained elevated at clinical threshold used for diagnosis of septic arthritis until day 14. In our experimental model of E. coli-induced arthritis, we conclude that TP and PN may not be good indicators for detecting the eradication of bacterial infection caused by E. coli from infected and subsequently treated joints.

Changes in antimicrobial susceptibility profiles of Mycoplasma bovis over time

Hugh Y. Cai, Rebeccah McDowall, Lois Parker, Emily I. Kaufman, Jeff L. Caswell (page 34)

Mycoplasma bovis is a major cause of pneumonia, arthritis, and mastitis in cattle and can lead to significant economic losses. Antimicrobial resistance is a concern and further limits the already short list of drugs effective against mycoplasmas. The objective of this study was to examine changes in in vitro minimum inhibitory concentrations (MICs) of antimicrobials of aminoglycoside, fluoroquinolone, lincosamide, macrolide, pleuromutilin, phenicol, and tetracycline classes for 210 M. bovis isolates collected from 1978 to 2009. The MIC50 values of the various antimicrobials were also compared. The MIC50 levels for enrofloxacin and danofloxacin remained low (0.25 µg/mL) across all 3 decades. MIC50 levels for tetracyclines, tilmicosin, and tylosin tartrate were low in the 1980s, then increased in the 1990s and remained high. In the 1980s, MIC50 levels were low for clindamycin, spectinomycin, and tulathromycin, increased in the 1990s to 8 µg/mL (clindamycin) and 32 µg/mL (spectinomycin and tulathromycin), then decreased again in the 2000s. Members of the fluoroquinolone class of antimicrobials had the lowest MIC50 levels across all 3 decades, which suggests in vitro susceptibility of M. bovis to this class of antimicrobials. Statistically significant associations were observed between MIC values for chlortetracycline, oxytetracycline, tylosin tartrate, and tilmicosin; between clindamycin, tulathromycin, spectinomycin, and tiamulin; and between tylosin tartrate and clindamycin. Changes in MIC levels of various antimicrobials over time show the importance of monitoring the susceptibility of mycoplasmas to antimicrobials. The number of antimicrobials that showed elevated MIC50 levels, and therefore possibly reduced in vitro effectiveness against M. bovis, supports initiatives that promote prudent use of antimicrobials in agriculture.

Comparative molecular analysis of fecal microbiota of bobcats (Lynx rufus) and domestic cats (Felis catus)

David Eshar, Charlie Lee, J. Scott Weese (page 42)

The goal of this study was to explore and describe fecal microbiota of captive and wild bobcats (Lynx rufus) and compare the results to those of domestic cats (Felis catus). Fecal samples from 27 bobcats (8 wild, 19 zoo-kept) were used for novel bacterial deoxyribonucleic acid (DNA) identification using next-generation sequencing of the V4 region of the bacterial 16S ribosomal ribonucleic acid (rRNA) gene, analyzed by Illumina sequencing, and then compared to data obtained from a colony of 10 domestic cats. In this study, the microbiota of both species was dominated by Firmicutes, followed by Proteobacteria, Actinobacteria, Bacteroidetes, and Verrucomicrobia. When compared, fecal samples from bobcats harbored more Proteobacteria and Actinobacteria than fecal samples from domestic cats. There was a remarkable inter-bobcat variation in the relative abundances of the main bacterial genera. There were no significant differences, however, between the main phyla of the microbiota of the wild and domestic bobcats. Proteobacteria in wild bobcats (P = 0.079) and Firmicutes in zoo-kept bobcats (P = 0.079) approached significance. There were no differences in predominant genera between wild and captive bobcats. The results of this study showed that there are notable differences in fecal bacterial communities between domestic cats and both captive and wild bobcats. The lack of significant differences in bacterial communities between wild and zoo-kept bobcats suggests that the varied diet provided for these felids can result in a fecal microbiota resembling that generated by a wild diet.

Peripartum metabolic profiles in a Holstein dairy herd with alarm level prevalence of subclinical ketosis detected in early lactation

Shin Oikawa, Hanan K. Elsayed, Chihoko Shibata, Kyoko Chisato, Ken Nakada (page 50)

The aim of this study was to characterize peripartum metabolic profiles, including the insulin sensitivity index, in cows diagnosed with subclinical ketosis (SCK) in the early stage of lactation. Cows that calved from January 2011 through December 2014 on a dairy farm with alarm level prevalence of SCK in Hokkaido, Japan (n = 175) were used. Blood and body condition scores (BCS) were obtained at regular health examinations in 2 consecutive periods, the first between 14 and 2 d before parturition, and the second between 3 and 14 d after parturition. Animals were divided into 3 groups at postpartum sampling: an SCK group with 35 multiparous and 15 primiparous cows having β-hydroxybutyrate (BHBA) concentrations ≥ 1.2 mM without clinical signs, a disease group of 36 multiparous and 9 primiparous cows that received treatment between parturition and postpartum sampling, and a control group consisting of 49 multiparous and 31 primiparous cows with BHBA concentrations < 1.2 mM. The prepartum revised quantitative insulin sensitivity check index was significantly lower in the multiparous SCK and disease groups than in the control group, demonstrating decreased insulin sensitivity in these cows, but not in primiparous cows. The prepartum BCS was significantly higher only in the multiparous SCK and disease groups. The prepartum apolipoprotein B-100 (ApoB-100) concentration was significantly decreased in the multiparous disease group, suggesting hepatic lipidosis. Conversely, primiparous cows had a higher prepartum ApoB-100 concentration. Prepartum decreased insulin sensitivity in the multiparous SCK and disease groups was considered to facilitate progression to SCK after calving.

Comparison of 4 commercial modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccines against heterologous Korean PRRSV-1 and PRRSV-2 challenge

Taehwan Oh, Hanjin Kim, Kee Hwan Park, Siyeon Yang, Jiwoon Jeong, Seeun Kim, Ikjae Kang, Su-Jin Park, Chanhee Chae (page 57)

The efficacy of 4 commercial porcine reproductive and respiratory syndrome virus (PRRSV) modified-live vaccines (MLV), against PRRSV-1 and PRRSV-2 was evaluated and compared in growing pigs. Two of the vaccines were based on PRRSV-1 and two on PRRSV-2. There were no significant differences between each of the two PRRSV-1 MLV vaccines and the two PRRSV-2 MLV vaccines respectively based on virology, immunological, and pathological evaluations. Vaccination with either of the PRRSV-1 MLV vaccines resulted in reduced PRRSV-1 but not PRRSV-2 viremia. Additionally, vaccination with either of the PRRSV-1 MLV vaccines resulted in reduction of lung lesions and PRRSV-1 positive cells in PRRSV-1 challenged pigs but had no significant effect in PRRSV-2 challenged pigs. In contrast, vaccination with either of the two PRRSV-2 MLV vaccines resulted in the reduction of both PRRSV-1 and PRRSV-2 viremia. The PRRSV-2 MLV vaccines were also able to effectively reduce lung lesions and PRRSV positive cells after challenge with either PRRSV-1 or PRRSV-2. Our data suggest that while vaccination with PRRSV-1 MLV vaccines can be effective against PRRSV-1, only PRRSV-2 MLV vaccines can protect against both Korean PRRSV-1 and PRRSV-2 challenges in this study.

Protective efficacy of an inactivated Brucella abortus vaccine candidate lysed by GI24 against brucellosis in Korean black goats

Wong-Kyong Kim, Ja-Young Moon, Jeong-Sang Cho, Enkhsaikhan Ochirkhuyag, Md Rashedunnabi Akanda, Byung-Yong Park, Jin Hur (page 68)

The efficacy of GI24-lysed Brucella abortus cells as a vaccine candidate against brucellosis in goats was evaluated on 2 groups of Korean black goats. Group A goats were immunized subcutaneously (SC) with sterile phosphate-buffered saline, whereas group B goats were immunized SC with approximately 3 × 109 lysed B. abortus cells. Subcutaneous immunization with the lysed cells did not cause any negative impact on the overall clinical status, such as behavior and appetite, throughout the study period. The enzyme-linked immunosorbent assay (ELISA) optical densities values for B. abortus lipopolysaccharide in serum were considerably higher in group B than those in group A. Also, the levels of the cytokines interleukin 4 (IL-4), tumor necrosis factor-alpha (TNF-α), and interferon gamma (IFN-γ) were significantly elevated in group B compared with those in group A. Following intraconjunctival challenge with B. abortus strain 544, the severity of brucellosis in terms of infection index and colonization of B. abortus in tissues was significantly lower in group B than in group A. The present study concluded that 3 of 5 goats immunized with GI24-lysed bacteria were completely protected against challenge. Future investigations are required to improve the protective efficacy offered by lysed B. abortus cells for practical applications in small ruminants.

Brief Communications

Genetic characterization of a hantavirus isolated from Heilongjiang province, China

Yulong Wang, Suya Cao, Cheng Cheng, Wendong Ju, Yuping Hua (page 75)

Hantavirus is the causative agent of hemorrhagic fever with renal syndrome (HFRS). Heilongjiang Province is experiencing an epidemic of HFRS, the main causative agent is a variant of hantavirus called Seoul virus (SEOV). In this study, the entire genome of one SEOV, the DN2 strain, was sequenced and analyzed. The alignment analysis of the sequences indicated that the DN2 strain shares the highest homology with the SEOV-LYO852 strain. The nucleotide identity is 97.6% for the S segment, 97.7% for the M segment, and 98.0% for the L segment. The corresponding amino acid sequence homologies are 99.1%, 98.9% and 99.8%. The phylogenetic analysis of the segments suggests that the DN2 strain has a high genetic relationship with SEOV strains and no genetic recombination occurs.