Diagnostic performance measures of ELISA and quantitative PCR tests for porcine circovirus type 2 exposure using Bayesian latent class analysis

Porcine circovirus 2 (PCV2) is believed to be a necessary but not sufficient underlying cause of porcine circovirus associated disease (PCVAD) in swine (Opriessnig et al., 2007). Since the potential threat of PCVAD is dependent on the prevalence of PCV2 in swine populations, accurate diagnostic tests are important for epidemiologic surveillance. Therefore, we evaluated the diagnostic sensitivity (Se) and specificity (Sp) of a new indirect ELISA and two quantitative PCR tests for PCV2 in a series of latent class models that used Bayesian estimation procedures. A total of 4140 samples from finisher pigs were tested for evidence of PCV2 by the ELISA and a TaqMan (TM) quantitative PCR, 995 by the ELISA and a SYBR Green (SG) dye-binding PCR, 998 by both PCRs and 993 by all three tests. Overall, the median (95% probability interval) ELISA Se and Sp was 0.85 (0.83-0.87) and 0.74 (0.68-0.79), respectively, when all three tests were analyzed together at an ELISA absorbance (optical density or OD) cutoff of ≥0.3. The TM PCR Se and Sp was 0.86 (0.84-0.88) and 0.94 (0.87-0.97), respectively, and the SG PCR Se and Sp was 0.83 (0.81-0.85) and 0.98 (0.94-1.00), respectively when all three tests were analyzed together at an ELISA OD cutoff of ≥0.3. Sensitivity analysis revealed that Sp estimates in general had less stability than Se estimates, but the SG PCR(Sp) was the most stable. Limited conditional dependence between the two PCR tests was detected. We conclude that the ELISA had the highest diagnostic Se at an absorbance cutoff of ≥0.3, while the SG PCR had the highest diagnostic Sp. The prevalence levels for exposure to PCV2 in finishing swine populations across all analyses ranged from 58 to 100%.     

Canine brain tumours: a model for the human disease?

Canine brain tumours are becoming established as naturally occurring models of disease to advance diagnostic and therapeutic understanding successfully. The size and structure of the dog's brain, histopathology and molecular characteristics of canine brain tumours, as well as the presence of an intact immune system, all support the potential success of this model. The limited success of current therapeutic regimens such as surgery and radiation for dogs with intracranial tumours means that there can be tremendous mutual benefit from collaboration with our human counterparts resulting in the development of new treatments. The similarities and differences between the canine and human diseases are described in this article, emphasizing both the importance and limitations of canines in brain tumour research. Recent clinical veterinary therapeutic trials are also described to demonstrate the areas of research in which canines have already been utilized and to highlight the important potential benefits of translational research to companion dogs.     

Use of a multiplex polymerase chain reaction assay in the antemortem diagnosis of toxoplasmosis and neosporosis in the central nervous system of cats and dogs

OBJECTIVE: To develop a multiplex polymerase chain reaction (PCR) assay for the detection of Toxoplasma gondii and Neospora caninum DNA in canine and feline biological samples. SAMPLE POPULATION; Biological samples from 7 cats with systemic (n = 4) or CNS (3) toxoplasmosis, 6 dogs with neospora- or toxoplasma-associated encephalitis, and 11 animals with nonprotozoal disease. PROCEDURE: Primers for T gondii, N caninum, and the canine ferritin gene (dogs) or feline histone 3.3 gene (cats) were combined in a single PCR assay. The DNA was extracted from paraffin-embedded brain tissue, CSF, or skeletal muscle. The PCR products with positive results were cloned, and sequence identity was confirmed. RESULTS: Of 7 cats and 4 dogs with immunohistochemical or serologic evidence of toxoplasmosis, PCR results were positive for all cats and 3 dogs for T gondii, and positive for T gondii and N caninum for 1 dog. Another dog had negative PCR results for both parasites. Of 2 dogs with immunohistochemical or serologic evidence of neosporosis, PCR results were positive for 1 for N caninum and positive for the other for T gondii. All negative-control samples yielded negative results for T gondii and N caninum on the PCR assay. CONCLUSIONS AND CLINICAL RELEVANCE: Standard tests for toxoplasmosis or neosporosis associated with the CNS rely on serologic, histologic, or immunohistochemical analysis and can be difficult to interpret. The multiplex PCR assay with built-in control reactions could be a complementary clinical tool for the antemortem diagnosis of toxoplasmosis or neosporosis associated with the CNS.     

Using animal movement behavior to categorize land cover and predict consequences for connectivity and patch residence times

Context

Landscape-scale population dynamics are driven in part by movement within and dispersal among habitat patches. Predicting these processes requires information about how movement behavior varies among land cover types.

Objectives

We investigated how butterfly movement in a heterogeneous landscape varies within and between habitat and matrix land cover types, and the implications of these differences for within-patch residence times and among-patch connectivity.

Methods

We empirically measured movement behavior in the Baltimore checkerspot butterfly (Euphydryas phaeton) in three land cover classes that broadly constitute habitat and two classes that constitute matrix. We also measured habitat preference at boundaries. We predicted patch residence times and interpatch dispersal using movement parameters estimated separately for each habitat and matrix land cover subclass (5 categories), or for combined habitat and combined matrix land cover classes (2 categories). We evaluated the effects of including edge behavior on all metrics.

Results

Overall, movement was slower within habitat land cover types, and faster in matrix cover types. Butterflies at forest edges were biased to remain in open areas, and connectivity and patch residence times were most affected by behavior at structural edges. Differences in movement between matrix subclasses had a greater effect on predictions about connectivity than differences between habitat subclasses. Differences in movement among habitat subclasses had a greater effect on residence times.

Conclusions

Our findings highlight the importance of careful classification of movement and land cover in heterogeneous landscapes, and reveal how subtle differences in behavioral responses to land cover can affect landscape-scale outcomes.

     

Spondylosis deformans and diffuse idiopathic skeletal hyperostosis (dish) resulting in adjacent segment disease

Spondylosis deformans and diffuse idiopathic skeletal hyperostosis (DISH) are usually incidental findings and in most dogs are either asymptomatic or associated with mild clinical signs. Severe spondylosis deformans and DISH can result in complete bony fusion of consecutive vertebral segments. One of the recognised complications following vertebral fusion in human patients is the development of adjacent segment disease, which is defined as degenerative changes, most commonly degenerative intervertebral disc disease, in the mobile vertebral segment neighboring a region of complete vertebral fusion. A similar syndrome following cervical fusion in dogs has been termed the domino effect. The purpose of this retrospective study was to investigate the hypothesis that vertebral fusion occurring secondary to spondylosis deformans or DISH in dogs would protect fused intervertebral disc spaces from undergoing degeneration, but result in adjacent segment disease at neighbouring unfused intervertebral disc spaces. Eight dogs with clinical signs of thoracolumbar myelopathy, magnetic resonance imaging of the thoracolumbar vertebral column, and spondylosis deformans or DISH producing fusion of > or = 2 consecutive intervertebral disc spaces were evaluated. Vertebral fusion of > or = 2 consecutive intervertebral disc spaces was correlated (P = 0.0017) with adjacent segment disease at the neighbouring unfused intervertebral disc space. Vertebral fusion appeared to protect fused intervertebral disc spaces from undergoing degeneration (P < 0.0001). Adjacent segment disease should be considered in dogs with severe spondylosis deformans or DISH occurring in conjunction with a thoracolumbar myelopathy.     

Onset, duration and efficacy of four methods of local anesthesia of the horn bud in calves

ObjectiveTo determine the onset, duration and efficacy of four local anesthetic methods for the horn bud in calves.Study designCrossover study.AnimalsEight, 2 month-old Holstein Friesian bull calves.MethodsCalves were subjected to one of the four following treatments: 1) cornual nerve block (C), 2) ring block (R), 3) cornual nerve block using a percutaneous jet delivery technique (JET) all using 2% lidocaine with epinephrine (0.01 mg mL^?1), and 4) topical eutectic mixture of local anesthetics (EMLA) cream. A peripheral nerve stimulator was used to assess cutaneous sensation over the horn bud using a graded response. Onset, duration and efficacy of anesthesia were determined.ResultsThe efficacy of the blocks was as follows: C 87.5%, R 100%, JET 37.5%, EMLA 0%. The median onset time and duration of anesthesia for C and R were: 2 (range 0.5–5) and 304 (range 107–512), and 3.25 (range 1–9) and 147 (range 62–299) minutes, respectively. Three of eight JET injections had a median onset and duration of 8 (range 0.5–9) and 132 (range 101–155) minutes, respectively. The duration of the C block was significantly longer than the R block (p= 0.047).Conclusions and clinical relevanceThe relatively rapid onset and long duration of anesthesia with C or R blocks using 2% lidocaine with epinephrine validates their practical use in dehorning calves while jet injection and EMLA cream provided insufficient analgesia to be clinically useful. The efficacy of the C block requires further study.     

Genomic diversity and resistome profiles of Salmonella enterica subsp. enterica serovar Kentucky isolated from food and animal sources in Ireland

Salmonella enterica subsp. enterica serovar Kentucky is frequently isolated from poultry, dairy and beef cattle, the environment and people with clinical salmonellosis globally. However, the sources of this serovar and its diversity and antimicrobial resistance capacities remain poorly described in many regions. To further understand the genetic diversity and antimicrobial sensitivity patterns among S. Kentucky strains isolated from non-human sources in Ireland, we sequenced and analysed the genomes of 61 isolates collected from avian, bovine, canine, ovine, piscine, porcine, environmental and vegetation sources between 2000 and 2016. The majority of isolates (n = 57, 93%) were sequence type (ST) 314, while only three isolates were ST198 and one was ST152. Several isolates were multidrug-resistant (MDR) and 14 carried at least one acquired antimicrobial resistance gene. When compared to a database of publicly available ST314, four distinct clades were identified (clades I–IV), with the majority of isolates from Ireland clustering together in Clade I. Two of the three ST198 isolates were characteristic of those originating outside of the Americas (Clade ST198.2), while one was distantly clustered with isolates from South and North America (Clade ST198.1). The genomes of the two clade ST198.2 isolates encoded Salmonella Genomic Island 1 (SGI1), were multidrug-resistant and encoded polymorphisms in the DNA gyrase (gyrA) and DNA topoisomerase (parC) known to confer resistance to fluoroquinolones. The single ST152 isolate was from raw beef, clustered with isolates from food and bovine sources in North America and was pan-susceptible. Results of this study indicate that most S. Kentucky isolates from non-human sources in Ireland are closely related ST314 and only a few isolates are antimicrobial-resistant. This study also demonstrates the presence of multidrug-resistant ST198 in food sources in Ireland.