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Probe-free real-time reverse transcription polymerase chain reaction assays for the detection and typing of porcine reproductive and respiratory syndrome virus in Canada
2015
Eschbaumer, Michael | Li, Wansi (May) | Wernike, Kerstin | Marshall, Frank | Czub, Markus
Porcine reproductive and respiratory syndrome (PRRS) has tremendous impact on the pork industry in North America. The molecular diagnosis of infection with PRRS virus (PRRSV) is hampered by its considerable strain diversity. In this study, 43 previously published or newly developed primers for probe-free real-time reverse transcription polymerase chain reaction (RT-PCR) were evaluated on their sensitivity, specificity, reproducibility, and repeatability, using a diverse panel of 36 PRRSV strains as well as other arteriviruses and unrelated porcine viruses. Three primer pairs had excellent diagnostic and analytical sensitivity on par with a probe-based reference assay, absolute specificity to virus genotype and species, as well as over 95% reproducibility and repeatability across a wide dynamic range.
Mostrar más [+] Menos [-]Relationship between intracranial pressure as measured by an epidural intracranial pressure monitoring system and optic nerve sheath diameter in healthy dogs
2015
llie, Laura A. | Thomovsky, Elizabeth J. | Johnson, Paula A. | Bentley, Tim | Heng, Hock Gan | Lee, Hee C. | Moore, George E.
OBJECTIVE To evaluate the association between ultrasonographically measured optic nerve sheath diameter (ONSD) and acute increases in intracranial pressure (ICP) as measured by an epidural intracranial pressure monitoring system (EICPMS) in healthy dogs. ANIMALS 6 young healthy dogs. PROCEDURES An EICPMS connected to a pressure monitor was used to generate a continuous pressure waveform in each anesthetized dog. A 22-gauge IV catheter was inserted into the brain parenchyma through the contralateral parietal bone, and 0.5 to 2.0 mL of anticoagulated autologous blood was injected at predetermined intervals. At baseline (immediately after EICPMS placement) and following each injection, the ICP as indicated by EICPMS was recorded, and 3 ultrasonographic images of the optic nerve sheath of each eye were obtained. The ONSD was measured at maximum diameter and at 5 mm caudal to the optic disk. RESULTS In linear models, the maximum ONSD was positively associated with increasing ICP. Specifically, the rate of maximum ONSD increase was greater for pressures ≤ 20 mm Hg above baseline (0.0534 mm/1 mm Hg ICP increase) than for pressures > 40 mm Hg above baseline (0.0087 mm/1 mm Hg ICP increase). The relationship of ICP to maximum ONSD was slightly nonlinear and best explained by comparison of fractional polynomial regression models. CONCLUSIONS AND CLINICAL RELEVANCE ICP was positively and nonlinearly associated with increasing maximum ONSD, especially when ICP was ≤ 20 mm Hg above baseline, supporting the conclusion that ultrasonographic measurement of maximum ONSD may provide a noninvasive monitoring tool for evaluation of ICP in dogs. Further research is needed to assess the utility of these measurements in clinical patients.
Mostrar más [+] Menos [-]Discrimination of healthy versus sick steers by means of continuous remote monitoring of animal activity
2015
Smith, Jacqueline L. | Vanzant, Eric S. | Carter, Craig N. | Jackson, Carney B.
OBJECTIVE To test a unique electronic ear tag designed to collect movement data to determine whether physical activity of sick steers differed from that of healthy steers. ANIMALS 206 steers. PROCEDURES Physical activity in 2 groups of steers during November and December of 2010 (101 steers; the tag of 1 steer failed, and thus that steer was removed from the study, which resulted in data for 100 steers) and 2011 (105 steers) was monitored with an electronic ear tag device with an on-board triple-axis accelerometer. The accelerometer recorded motion in all 3 axes in the form of counts per minute. A radio-frequency transmitter on the ear tag delivered serial packets of motion data to a local server. An algorithm was developed to analyze the activity data to determine whether this technique could be used to assess health status with high accuracy. RESULTS Steers that became sick had significantly fewer activity counts (approx 25% fewer), compared with the activity counts of steers that remained healthy the entire time. CONCLUSIONS AND CLINICAL RELEVANCE In this study, automated detection of health status in growing cattle was feasible through remote monitoring of animal activity. Early identification of sick animals should lead to improved health outcomes, increased marketability, and improved animal well-being and help to minimize the use of antimicrobials that could contribute to resistant bacteria.
Mostrar más [+] Menos [-]Tear film osmolality and electrolyte composition in healthy horses
2015
Best, Lori J. | Hendrix, Diane V. H. | Ward, Daniel A.
OBJECTIVE To evaluate the tear film osmolality and electrolyte composition in healthy horses. ANIMALS 15 healthy adult horses. PROCEDURES Each horse was manually restrained, and an ophthalmic examination, which included slit-lamp biomicroscopy, indirect ophthalmoscopy, and a Schirmer tear test, was performed. Tear samples were collected from both eyes with microcapillary tubes 3 times at 5-minute intervals. The tear samples for each horse were pooled, and the osmolality and electrolyte concentrations were measured. The mean (SD) was calculated for each variable to establish preliminary guidelines for tear film osmolality and electrolyte composition in healthy horses. RESULTS The mean (SD) tear film osmolality was 283.51 (9.33) mmol/kg, and the mean (SD) sodium, potassium, magnesium, and calcium concentrations were 134.75 (10), 16.3 (5.77), 3.48 (1.97), and 1.06 (0.42) mmol/L, respectively. The sodium concentration in the tear film was similar to that in serum, whereas the potassium concentration in the tear film was approximately 4.75 times that of serum. CONCLUSIONS AND CLINICAL RELEVANCE Results provided preliminary guidelines with which tear samples obtained from horses with keratopathies can be compared. Measurement of tear film osmolality in these horses was easy and noninvasive. The tear film concentration of divalent cations was greater than expected and was higher than the divalent cation concentrations in the tear films of rabbits and humans. These data may be clinically useful for the diagnosis and monitoring of hyperosmolar ocular surface disease in horses.
Mostrar más [+] Menos [-]Enhanced inactivation of avian influenza virus at −20°C by disinfectants supplemented with calcium chloride or other antifreeze agents
2015
Guan, Jiewen | Chan, Maria | Brooks, Brian W. | Rohonczy, Elizabeth
Avian influenza outbreaks have occurred during winter months, and effective disinfection of poultry premises at freezing temperatures is needed. The commercial disinfectants Virkon and Accel, supplemented with an antifreeze agent [propylene glycol (PG), methanol (MeOH), or calcium chloride (CaCl2)], were evaluated for their effectiveness in killing avian influenza virus (AIV) at −20°C or 21°C. An AIV suspension was applied to stainless steel disks, air-dried, and covered with a disinfectant or antifreeze agent for 5 to 30 min. Virkon (2%) and Accel (6.25%) with 30% PG, 20% MeOH, or 20% CaCl2 inactivated 6 log10 AIV within 5 min at −20°C and 21°C. At these temperatures PG and MeOH alone did not kill AIV, but the 20% CaCl2 solution alone inactivated 5 log10 AIV within 10 min. The results suggested that CaCl2 is potentially useful to enhance the effectiveness of disinfection of poultry facilities after outbreaks of AIV infection in warm and cold seasons.
Mostrar más [+] Menos [-]Role of the capsular polysaccharide as a virulence factor for Streptococcus suis serotype 14
2015
Roy, David | Auger, Jean-Philippe | Segura, Mariela | Fittipaldi, Nahuel | Takamatsu, Daisuke | Okura, Masatoshi | Gottschalk, Marcelo
Streptococcus suis is an important swine pathogen and a zoonotic agent causing meningitis and septicemia. Although serotype 2 is the most virulent type, serotype 14 is emerging, and understanding of its pathogenesis is limited. To study the role of the capsular polysaccharide (CPS) of serotype 14 as a virulence factor, we constructed knockout mutants devoid of either cps14B, a highly conserved regulatory gene, or neu14C, a gene coding for uridine diphospho-N-acetylglucosamine 2-epimerase, which is involved in sialic acid synthesis. The mutants showed total loss of the CPS with coagglutination assays and electron microscopy. Phagocytosis assays showed high susceptibility of mutant Δcps14B. An in vivo murine model was used to demonstrate attenuated virulence of this non-encapsulated mutant. Despite the difference in the CPS composition of different serotypes, this study has demonstrated for the first time that the CPS of a serotype other than 2 is also an important antiphagocytic factor and a critical virulence factor.
Mostrar más [+] Menos [-]Comparison of a live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit with a commercial vaccine for efficacy of protection against internal egg contamination by Salmonella in hens
2015
Nandre, Rahul M. | Eo, Seong Kug | Park, Sang Youel | Lee, John Hwa
This study compared a new live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit (SE-LTB) with a commercial Salmonella Enteritidis (SE) vaccine for efficacy of protection against SE infection in laying hens. Chickens were divided into 3 groups of 20 each. Group A chickens were inoculated orally with phosphate-buffered saline and served as controls, group B chickens were inoculated orally with the vaccine candidate, and group C chickens were inoculated intramuscularly with a commercial vaccine, the primary inoculation in groups B and C being at 10 wk of age and the booster at 16 wk. Groups B and C showed significantly higher titers of plasma immunoglobulin G, intestinal secretory immunoglobulin A, and egg yolk immunoglobulin Y antibodies compared with the control group, and both vaccinated groups showed a significantly elevated cellular immune response. After virulent challenge, group B had significantly lower production of thin-shelled and/or malformed eggs and a significantly lower rate of SE contamination of eggs compared with the control group. Furthermore, the challenge strain was detected significantly less in all of the examined organs of group B compared with the control group. Group C had lower gross lesion scores only in the spleen and had lower bacterial counts only in the spleen, ceca, and ovary. These findings indicate that vaccination with the SE-LTB vaccine candidate can efficiently reduce internal egg and internal organ contamination by Salmonella and has advantages over the commercial vaccine.
Mostrar más [+] Menos [-]Single nucleotide polymorphisms in the bovine Histophilus somni genome; a comparison of new and old isolates
2015
Madampage, Claudia Avis | Rawlyk, Neil | Crockford, Gordon | Van Donkersgoed, Joyce | Dorin, Craig | Potter, Andrew
Histophilus somni, a causative agent of the bovine respiratory disease complex, can also cause a variety of systemic disorders, including bronchopneumonia, myocarditis, pericarditis, arthritis, pleuritis, and infectious thrombotic meningoencephalitis. The purpose of this study was to determine if currently circulating strains differ from those of the 1980s by identifying genomic changes. Single nucleotide polymorphisms (SNPs) and insertion and deletion (INDEL) sites were examined by whole-genome sequencing in 12 samples, 6 old and 6 new. The 31 028 SNP/INDELs recorded were compared against the reference genome sequence of the pathogenic H. somni strain 2336. The distribution of about 75% of these SNPs within a specified gene differed between old and new isolates and did not follow any particular pattern. The other 25% clustered into 2 groups containing the same SNPs in various genes: group I included 5 old isolates and 1 new isolate; group II included 5 new isolates and 1 old isolate. For putative virulence genes there were more SNPs in group I compared with strain 2336, itself an older isolate, than in group II. Although only 25% of all the SNPs formed 2 clusters, the results suggest some genetic difference in various genes between old and new strains.
Mostrar más [+] Menos [-]Pharmacological effects of a C-phycocyanin-based multicomponent nutraceutical in an in-vitro canine chondrocyte model of osteoarthritis
2015
Martinez, Stephanie E. | Chen, Yufei | Ho, Emmanuel A. | Martinez, Steven A. | Davies, Neal A.
Multicomponent nutraceuticals are becoming increasingly popular treatments or adjunctive therapies for osteoarthritis in veterinary medicine despite lack of evidence of efficacy for many products. The objective of this study was to evaluate the anti-inflammatory and antioxidant activities of a commercially available C-phycocyanin-based nutraceutical and select constituent ingredients in an in-vitro model of canine osteoarthritis. Normal canine articular chondrocytes were used in an in-vitro model of osteoarthritis. Inflammatory conditions were induced using interleukin-1β. The nutraceutical preparation as a whole, its individual constituents, as well as carprofen were evaluated at concentrations of 0 to 250 μg/mL for reduction of the following inflammatory mediators and indicators of catabolism of the extracellular matrix: prostaglandin E2 (PGE2), tumor necrosis factor-α (TFN-α), interleukin-6 (IL-6), metalloproteinase-3 (MMP-3), nitric oxide, and sulfated glycosaminoglycans (sGAGs). Validated, commercially available assay kits were used for quantitation of inflammatory mediators. The antioxidant capacities, as well as cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and lipoxygenase (LOX) inhibitory activities of the whole nutraceutical preparation and select constituents, were also assessed using validated commercially available assay kits. The antioxidant capacity of the nutraceutical and constituents was concentration-dependent. The nutraceutical and constituents appear to display anti-inflammatory activity primarily through the inhibition of COX-2. The nutraceutical displayed similar strength to carprofen in reducing TNF-α, IL-6, MMP-3, nitric oxide, and sGAGs at select concentration ranges. The C-phycocyanin (CPC)-based nutraceutical and constituents may be able to mediate 3 primary pathogenic mechanisms of osteoarthritis: inflammation, chondral degeneration, and oxidative stress in vitro. The nutraceutical may be clinically useful in veterinary medicine and its efficacy should be further investigated in vivo.
Mostrar más [+] Menos [-]Prevalence of internal parasites in beef cows in the United States: Results of the National Animal Health Monitoring System’s (NAHMS) beef study, 2007‐2008
2015
Stromberg, Bert E. | Gasbarre, Louis C. | Ballweber, Lora R. | Dargatz, David A. | Rodriguez, Judith M. | Kopral, Christine A. | Zarlenga, Dante S.
During the United States Department of Agriculture (USDA) National Animal Health Monitoring System’s (NAHMS) 2007‐2008 beef study, 567 producers from 24 US States were offered the opportunity to collect fecal samples from weaned beef calves and have them evaluated for the presence of parasite eggs (Phase 1). Participating producers were provided with instructions and materials for sample collection. Up to 20 fresh fecal samples were collected from each of the 99 participating operations. Fresh fecal samples were submitted to one of 3 randomly assigned laboratories for evaluation. Upon arrival at the laboratories, all samples were processed for the enumeration of strongyle, Nematodirus, and Trichuris eggs using the modified Wisconsin technique. The presence or absence of coccidian oocysts and tapeworm eggs was also noted. In submissions where the strongyle eggs per gram exceeded 30, aliquots from 2 to 6 animals were pooled for DNA extraction. Extracted DNA was subjected to genus level polymerase chain reaction (PCR) identification for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. In this study, 85.6% of the samples had strongyle type, Nematodirus, and Trichuris eggs. Among the samples evaluated, 91% had Cooperia, 79% Ostertagia, 53% Haemonchus, 38% Oesophagostomum, 18% Nematodirus, 7% Trichuris, and 3% Trichostrongylus. The prevalence of coccidia and tapeworm eggs was 59.9% and 13.7%, respectively.
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