Profiling pro-inflammatory cytokine and chemokine mRNA expression levels as a novel method for selection of increased innate immune responsiveness

Previous studies using F1 reciprocal crosses and two parental lines of broilers show the sire is instrumental in determining the in vitro leukocyte function and cytokine/chemokine profile. Since the innate immune response is the primary means young chickens have to protect themselves, we hypothesize utilizing a novel genomics approach to select sires based on an elevated pro-inflammatory cytokine and chemokine profile. By identifying sires with increased pro-inflammatory cytokine (interleukin [IL]-1beta and IL-6) and chemokine (CXCLi2 and CCLi2) mRNA expression levels, we expect the progeny will also have elevated profiles. We characterized the pro-inflammatory cytokine and chemokine profile of 119 sires using quantitative real-time RT-PCR (qRT-PCR) and identified two populations with inherently high and low mRNA expression levels of IL-1beta, IL-6, CXCLi2, and CCLi2. Select high and low sires were then used to produce progeny for the second phase of the trial. Blood samples were collected from 214 progeny and the cytokine and chemokine mRNA expression levels determined. Progeny from high sires had significantly (P相似文献   

Interleukin-2 and interferon-gamma mRNA expression in canine anal furunculosis lesions and the effect of ciclosporin therapy

German Shepherd Dogs have an increased incidence of anal furunculosis (AF), which is a disease characterised by inflammation and ulceration of the perianal tissues. Ciclosporin, an immunosuppressive drug, has been successfully used to treat AF, suggesting that the pathogenesis of disease is likely to have an immune-mediated component. Previous research has shown that the cytokine mRNA profile in AF lesions is consistent with T cell-mediated inflammation. The aims of the current study were to quantify IL-2 and IFNgamma mRNA expression in AF biopsies taken before and after treatment with ciclosporin and to compare cytokine expression with lesion severity. Twenty-two dogs with AF were recruited into the study and lesional biopsies were taken prior to ciclosporin therapy. Lesion severity was graded using a visual analogue scale. All dogs were evaluated after 4 weeks of ciclosporin therapy and, in 10 dogs with persistent disease, residual lesions were resected. RNA was extracted from AF-lesional tissue and control perianal tissue samples (n=10), which was used as the template for RT-PCR. Analysis of IL-2 and IFNgamma mRNA expression was performed using real-time PCR. IL-2 and IFNgamma mRNA was consistently detected in pre-treatment AF biopsies and, when quantified, this was significantly increased compared to control tissue (P<0.05). However, no correlation was seen between lesion severity and pre-treatment cytokine mRNA expression. In the ten paired pre- and post-treatment samples, IL-2 mRNA expression was significantly reduced in residual disease tissue following ciclosporin therapy (P=0.013). Treatment with ciclosporin seemed to result in decreased expression of IFNgamma mRNA in AF lesions but this was not statistically significant. In six of the 10 dogs with persistent disease, T cell cytokine mRNA could still be detected in the tissues, suggesting that there was inadequate immunosuppression. The absence of a correlation between T cell cytokine expression and the severity of disease suggests that tissue destruction observed in AF might be a consequence of other inflammatory mediators or downstream effects of T cell activation.     

Comparison of three methods of surveillance with application to the detection of Johne's disease seropositivity in Texas cattle

Surveillance and monitoring are important for measuring the level of disease in a population, documenting changes in prevalence over time, determining high-risk areas for implementation of control measures, eradicating disease, and documenting freedom from disease. The documentation of freedom from disease has importance for international trade and the resumption of production after an outbreak. Johne's disease (JD) is an example of an endemic disease of cattle that has variable prevalence related to environmental and animal-level factors. Three methods of sample collection were used to describe the prevalence and distribution of JD seropositivity in Texas. Sampled cattle were: (1) extensively managed herds, (2) market cattle, and (3) clinically ill cattle examined by practicing veterinarians throughout Texas. Samples were evaluated for JD using a commercially available serum ELISA. Proportion of seropositive samples was compared and spatial distributions were evaluated for clustering. Difference of JD seropositivity was observed among the three sample populations suggesting that estimation of disease prevalence is dependant upon the source of samples.     

Transplacental toxoplasmosis in a wild southern sea otter (Enhydra lutris nereis)

In September 2004, a neonatal sea otter pup was found alive on the beach in northern Monterey Bay, CA. Efforts to locate the mother were unsuccessful. Due to a poor prognosis for successful rehabilitation, the pup was euthanized. Postmortem examination revealed emaciation, systemic lymphadenopathy and a malformation of the left cerebral temporal lobe. On histopathology, free tachyzoites and tissue cysts compatible with Toxoplasma gondii were observed in the brain, heart, thymus, liver, lymph nodes and peri-umbilical adipose. The presence of T. gondii within host tissues was associated with lymphoplasmacytic inflammation and tissue necrosis. Immunofluorescent antibody tests using postmortem serum were positive for anti-T. gondii IgM and IgG (at 1:320 and 1:1280 serum dilution, respectively), but were negative for IgG directed against Sarcocystis neurona and Neospora caninum (<1:40 each). Brain immunohistochemistry revealed positive staining for tachyzoites and tissue cysts using antiserum raised to T. gondii, but not S. neurona or N. caninum. T. gondii parasite DNA was obtained from extracts of brain and muscle by PCR amplification using the diagnostic B1 locus. Restriction enzyme digestion followed by gel electrophoresis and DNA sequencing confirmed the presence of Type X T. gondii, the strain identified in the majority of southern sea otter infections.     

Bone marrow lesions of the distal condyles of the third metacarpal bone are common and not always related to lameness in sports and pleasure horses

The metacarpophalangeal joint is a common site of pain in horses. Little information is available about bone marrow lesions in the distal condyles of the third metacarpal bone in sports and pleasure horses. Aims of this prospective, retrospective, observational study were to evaluate the prevalence of bone marrow lesions in the distal condyles of the third metacarpal bone, to describe their anatomic distribution and to correlate lesions with the presence of lameness and the level and type of activity. All sports and pleasure horses undergoing standing low‐field MRI of the front fetlock region between 2009 and 2016 were included and divided into three lameness groups according to the results of diagnostic analgesia. Bone marrow lesions were analyzed and graded by one reader. Grades were compared between anatomical locations and between lameness groups. A total of 166 horses were sampled. The prevalence of bone marrow lesions was 76.5% (127/166). The dorsal aspects of the medial condyle (31%, 39/127) and of the sagittal ridge (28%, 36/127) were the most commonly affected locations. There was no significant difference between both forelimbs (lame limb and non‐lame limb) of the same horse. Lesion severity was neither significantly associated with the lameness group nor with the type or level of activity. The prevalence of bone marrow lesions in the distal condyles of the third metacarpal bone is high in this population and the clinical significance is not always clear. Further studies are required to elucidate the clinical significance of this finding in sports and pleasure horses.     

Cytology and the cell block method in diagnostic characterization of canine lymphadenopathy and in the immunophenotyping of nodal lymphoma

Minimally invasive techniques used to evaluate canine peripheral lymphadenopathy (PLN), including fine needle aspiration biopsy with cytological evaluation (FNAB‐C) and flow cytometry (FC), have benefits and limitations. The cell block (CB) method is an alternate processing technique in which fine needle aspirate biopsy samples are concentrated, fixed, and embedded in paraffin for routine histological processing/staining. Utilizing three observers, we determined the diagnostic value of the CB in evaluating canine PLN across six categories (non‐diagnostic, reactive, inflammatory/infectious, probable lymphoma and lymphoma, metastatic neoplasia) and correlated findings to immunophenotypic and clonal antigen receptor rearrangement results in canine nodal lymphoma. Eighty‐five paired FNAB‐C and CB samples were evaluated from canine patients presenting to the University of Minnesota Veterinary Oncology or Internal Medicine services. Diagnostic quality samples were obtained in 55/85 (65%) CB and 81/85 (95%) FNAB‐C samples, respectively, and nodal pathology impacted CB diagnostic yield. Overall percent agreement between diagnostic‐quality FNAB‐C and CB samples was 86%, but increased to 95% if the categories of lymphoma and probable lymphoma were combined. There was 100% agreement for both the diagnoses of metastatic neoplasia and reactive lymph nodes and 92% agreement for the diagnosis of lymphoma/probable lymphoma. Using immunohistochemistry (IHC), CB samples correctly immunophenotyped 22/23 (96%) cases of B‐cell lymphoma, but only 1/6 (17%) cases of T‐cell lymphoma. IHC was not completed on nine cases of lymphoproliferative disease because of insufficient cellularity. When the CB method (CBM) yielded diagnostic quality samples there was good to excellent agreement with FNAB‐C samples and CB samples were suitable for some IHC tests.     

Radiotherapy isocenters verified by matching to bony landmarks of the canine and feline head differ when localized using volumetric versus planar imaging

The “gold standard” for verification of patient positioning before linear accelerator‐based stereotactic radiation therapy is kilovoltage cone‐beam computed tomography (kV‐CBCT), which is not uniformly available or utilized; planar imaging is sometimes used instead. The primary aim of this study was to determine if the position of the bony skull, when used as a surrogate for isocenter verification, is different when orthogonal megavoltage (MV) portal or kilovoltage (kV/kV) radiographs are used for image guidance, rather than kV‐CBCT. A secondary aim was to determine the influence of intra‐observer variability, body size and skull conformation on positioning, as determined using these three imaging modalities. Dogs and cats receiving radiotherapy of the head were recruited for this prospective analytical study. Planar (MV portal and kV/kV images) and volumetric (kV‐CBCT) images were acquired before treatment, and manually coregistered with reference images. Differences in skull position when matched based on MV portal, kV/kV images and kV‐CBCT were compared. A total of 65 subjects and 148 unique datasets were evaluated. The Wilcoxon rank‐sum test was used to evaluate effects of transitioning between imaging modalities. When comparing magnitude of shifts in MV to kV‐CBCT, MV to kV/kV and kV/kV to kV‐CBCT, there were statistically significant differences. Results were not measurably impacted by body size, skull conformation or interobserver differences. Based on shift magnitude and direction, an isotropic setup margin of at least 1 mm should be incorporated within the planning target volume when MV or kV planar imaging is used for position verification.