Detection and measurement of benzimidazole resistance alleles in Haemonchus contortus using real-time PCR with locked nucleic acid Taqman probes

Benzimidazole resistance is a common problem in parasitic nematodes of ruminants and early detection is vital if its spread is to be monitored and controlled. Real time PCR offers a fast and reliable method for rapid detection and measurement of resistance allele frequencies. In Haemonchus contortus a single nucleotide polymorphism at codon 200 of the beta-tubulin gene (TTC to TAC), causing a phenylalanine to tyrosine amino acid substitution, has been shown to be involved in many cases of resistance. Locked nucleic acid (LNA) Taqman probes have been used in this work to detect and measure the frequency of resistance alleles in individual and multiple H. contortus. Detection of resistant genotypes using LNA Taqman probes in individual H. contortus is simpler and more reliable than with previously described assays. Measurement of the frequency of resistant alleles in populations of H. contortus was achieved by using the cycle threshold (C(t)) values and a standard curve derived from populations with known allele frequencies. Results using the LNA probes on individual and multiple worms gave similar results to the allele specific PCR. The sensitivity of the LNA assay on multiple nematodes allowed reliable detection of > or = 10% resistance allele frequency. Using the final fluorescence method, it was possible to differentiate populations with approximately 0, 5 and 10% resistance allele frequencies.     

A highly sensitive,non‐invasive qPCR‐based strategy for direct quantification of Yersinia ruckeri in fish faeces

Finfish with asymptomatic Yersinia ruckeri infections pose a major risk as they can transmit the pathogen and cause clinical outbreaks in stock populations. Current tools have insufficient quantitative ability for accurately detecting the trace levels of Y. ruckeri typically associated with asymptomatic infection, necessitate invasive or lethal sampling, or require long processing times. This study presents a highly sensitive qPCR‐based method, targeting part of the Y. ruckeri 16S rRNA sequence, that is capable of detecting extremely low levels of Y. ruckeri in noninvasively collected faecal samples. Quantitative precision and accuracy of faecal sample analysis was consistent, despite the complexity of the faecal matrix. The assay demonstrated linearity over a six log‐wide dynamic range. Its limit of detection (LOD) and limit of quantification (LOQ) were 4 and 10 copies of the target sequence, respectively. Sensitivity of the assay was comparable to other qPCR‐based methods without requiring invasive or lethal sampling. Applicability as a screening strategy was tested using passively collected faecal samples. Asymptomatic Y. ruckeri infection was detected in all samples, although none of the fish exhibited overt infection. This method will be beneficial for finfish disease management if developed further as a noninvasive, screening tool against asymptomatic Y. ruckeri infection.     

Biomechanical comparison of cortical radial graft versus ulnar transposition graft limb-sparing techniques for the distal radial site in dogs

OBJECTIVE: To compare the biomechanical properties of radial graft (RG) versus ulnar transposition graft (UTG) limb-sparing techniques in the dog. STUDY DESIGN: Cadaveric study. ANIMALS: Six pairs of normal canine thoracic limbs. METHODS: In each pair of limbs, 1 limb was subjected to the RG technique and the other to the UTG technique. Limbs were tested in axial loading until failure. Modes of failure and biomechanical properties were compared between the 2 groups. Percent coverage of the metacarpal bone by the plate was retrospectively compared between the limbs that failed by fracture of the metacarpal bones and those that did not. RESULTS: RG limbs had significantly greater stiffness, yield load, maximum load, maximum energy, and post-yield energy. All UTG limbs failed by cranial bending of the plate. Half of the RG limbs failed by caudal bending of the plate and half by fracture of the third metacarpal bone at the distal end of the plate. Limbs with <80% plate coverage of the metacarpal bone were significantly more likely to fail by metacarpal fracture. CONCLUSIONS: The RG technique was biomechanically superior to the UTG technique under the conditions studied. At least 80% plate coverage of the metacarpal bone should be achieved. CLINICAL RELEVANCE: Exercise restriction and coaptation bandages should be considered for patients undergoing the UTG technique to minimize potential biomechanical complications.     

Aflatoxin B1 induced alterations in the stability of the lysosomal membrane in Oreochromis mossambicus (Peters, 1852)

This article elucidates the effect of crude dietary aflatoxin B₁ (AFB₁) on the stability of the lysosomal membrane in tilapia (Oreochromis mossambicus), and the analysis was made at the biochemical and cellular level. The liver lysosomal acid phosphatase activity was used to evaluate the functional integrity of the lysosome. Three experimental groups were fed on diets containing 0.375, 2.5 and 6 mg kg^?1 of AFB₁ for a time period of 6 weeks, whereas a fourth group fed on the semi‐purified diet formed the control. Significant changes were observed in the stability of the lysosomal membrane in aflatoxin exposed groups. This is the first report, so far, of changes induced by AFB₁ on the stability of lysosomal membrane in tilapia (O. mossambicus).     

李栗谷天人观研究——以人的命运问题为中心

李栗谷在对天、命观的研究中,从天人关系的角度思考了人性命运的问题。他以圣人与天的经历为基础,力图把传统的重要经典联成一个整体,证明了天与人,天道与人事之间存在着密切的关系:天与道遥相呼应,人对于人性有积极作用;人所应循之道是通过圣人来传递的,圣人与天相通。另外,在关于命运的看法上,儒学和基督教的观点体现出更多的互补性,而不是相异性。     

DORSAL PLANE COMPUTED TOMOGRAPHIC IMAGING OF THE ETHMOID REGION TO EVALUATE CHRONIC NASAL DISEASE IN THE DOG

Computed tomographic anatomy of the normal canine ethmoid region using both transverse and dorsal imaging plane is described. In the transverse plane, the cribriform plate appears as an irregular heart-shaped to rectangular structure that is generally visualized on no more than one to two contiguous 4-mm slices. In the dorsal plane, the cribriform plate appears as a discrete "V"-shaped structure and can be visualized on four to five contiguous slices. Successful visualization of the cribriform was less dependent on patient positioning for the dorsal plane imaging studies. Computed tomographic findings in four dogs with chronic nasal disease and complete postmortem examination of the ethmoid region indicate that the dorsal imaging plane allows a more accurate assessment of cribriform plate involvement than the transverse imaging plane.     

MAGNETIC RESONANCE IMAGING OF THE NORMAL FELINE ABDOMEN: AN ANATOMIC REFERENCE

Magnetic resonance images of two adult domestic short-haired cats were obtained with a whole body scanner. Images of the abdomen were compared with cross-sectional anatomy cadaver specimens from the same two cats. Anatomic structures were first identified on the cadaver specimens with the aid of anatomy texts and references and were then identified and labeled on the magnetic resonance images. Results from this project provide an atlas of normal cross-sectional MRI anatomy of the feline abdomen.     

Modelling individual variability in growth of bull trout in the Walla Walla River Basin using a hierarchical von Bertalanffy growth model

We examined growth in length of fluvial bull trout (Salvelinus confluentus) in the Walla Walla River Basin, Washington and Oregon. Our objectives were to quantify individual variability in growth; examine growth within and among years, life history forms, life stages and sexes; and estimate von Bertalanffy growth parameters. Individual variability was evaluated by modelling asymptotic length (L_∞) and the growth coefficient (k) as random variables. All models were fit with Bayesian methods and were evaluated for fit by the deviance information criterion. By incorporating individual variability, population‐level estimates of L_∞ and k appeared appropriate and estimated growth trajectories for specific bull trout fit individual observed patterns in growth. Growth trajectories and positive correlation between individual estimates of L_∞ and k suggest that some individuals grow at a faster rate and reach a larger maximum size than other individuals and those differences are maintained throughout life. Selected models suggest that fluvial migrants have higher estimates of L_∞ and k than residents, but there were only slight differences in parameter estimates among migrants from two adjacent spawning populations in the Walla Walla River Basin, as well as between males and females. Growth rates increased for fluvial migrants after subadult emigration. Individual variability in growth is consistent with the life history diversity assumed essential for bull trout population persistence. Quantifying this variability is important for modelling population dynamics and viability to conserve this threatened species.     

Tritrichomonas foetus infection in purebred cats in Germany: prevalence of clinical signs and the role of co-infection with other enteroparasites

The aim of this study was to determine the prevalence of Tritrichomonas foetus infection and associated clinical signs in purebred cats in Germany, to investigate the role of co-infection, and identify determinants of infection. Faecal specimens accompanied by epidemiological questionnaires were scored and collected from 230 purebred cats. Faeces were examined for trichomonads and other enteroparasites. The prevalence of T foetus was 15.7% among cats and 18.5% among catteries. An abnormal faecal score and history of diarrhoea were observed in 64% and 61% of T foetus-positive cats, respectively, and correlated significantly with infection. Co-infection, observed in 36% of T foetus-infected cats, was not associated with diarrhoea. Norwegian Forest cats were infected significantly more often than other breeds. No association was found with any environmental factors. This study demonstrated a high prevalence of symptomatic T foetus infections in purebred cats in Germany. Co-infection with other enteroparasites did not worsen clinical signs of trichomonosis.     

Fine‐scale habitat partitioning of Chilean and Peale's dolphins and their overlap with aquaculture

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