Mitochondrial cytochrome c oxidase subunit 1 (cox1) gene sequence of Spirocerca lupi (Nematoda, Spirurida): avenues for potential implications

Canine spirocercosis is a life-threatening parasitosis caused by Spirocerca lupi (Nematoda, Spirurida) that is presently emerging in several countries. This study characterised an informative region within the mitochondrial (mtDNA) gene encoding for the cytochrome c oxidase subunit 1 (cox1) of S. lupi by Polymerase Chain Reaction (PCR)-coupled sequencing. Specimens from five different countries in Europe, Asia and Africa were examined and two different sequence variants of cox1 (i.e. haplotypes) were determined, displaying nucleotidic variation at 6 of 689 positions. All of these positions were invariable among all the parasite individuals from Europe (haplotype 1) and among the African and Asian individuals (haplotype 2), but differed between Europe and Asia/Africa. The S. lupi cox1 sequences were consistent with those of other common Spirurida previously reported at both nucleotidic and phylogenetic levels. This study provides molecular information essential for identification of the nematode, irrespective of its life cycle stage. Crucial implications for the specific molecular diagnosis of clinical spirocercosis and investigation of the evolution, population genetics, ecology and epidemiology of S. lupi are discussed.     

RADIOGRAPHIC AND COMPUTED TOMOGRAPHIC CHANGES AND CLINICAL PRESENTATION OF SPIROCERCOSIS IN THE DOG

A retrospective study of 39 dogs with spirocercosis is described, emphasizing radiographic and computed tomographic aspects and clinical presentation. Dogs were classified as complicated or uncomplicated, both clinically and radiographically. Besides the expected upper gastrointestinal signs, a high incidence of respiratory (77%) and locomotor (23%) complications were present. All dogs had thoracic radiographs. Esophageal masses were radiographically classified as typical or atypical according to their location. Twenty-seven dogs had a typical caudal esophageal mass. Six dogs had a mass atypically located in the hilar region. These masses were smaller and more difficult to visualize radiographically. The remaining 6 dogs did not have a radiographically detectable esophageal mass. Radiology as an initial diagnostic tool was effective in detecting and localizing the mass and to detect early respiratory abnormalities such as pleuritis, mediastinitis, pneumonia, and bronchial displacement. Endoscopy was the modality of choice to confirm antemortem esophageal masses. In dogs where the mass filled the whole esophageal lumen, endoscopy failed to give essential information necessary for surgical excision of neoplastic masses, such as the extent of esophageal wall attachment. Caudal esophageal sphincter involvement was difficult to determine endoscopically with large caudal esophageal masses. Computed tomography was performed on 3 dogs and did not address the latter problems completely, but was found to be a sensitive tool to detect focal aortic mineralization and early spondylitis, both typical for the disease, and essential in the diagnosis of non- or extramural esophageal abnormalities.     

Seroprevalence of infectious bovine rhinotracheitis and bovine viral diarrhea virus type 1 and type 2 in non-vaccinated cattle herds in the Pacific Region of Central Costa Rica

The objectives of this cross-sectional study were to estimate the seroprevalence of infectious bovine rhinotracheitis (IBR, BHV-1) and bovine viral diarrhea virus (BVDV) in a population of non-vaccinated, double purpose, dairy and beef herds in the Pacific Region of Central Costa Rica. Blood samples were collected from a total of 496 animals from 35 herds. Sera were tested for antibodies against BHV-1(IBR) and BVDV types 1 and 2 using serum neutralization test. The average number of animals tested in each herd for each of the viruses was 14. Overall individual seroprevalence was 48%, 27%, and 19% for IBR, BVDV type 1, and BVDV type 2, respectively. Median within-herd seroprevalence for IBR, BVDV type 1 and type 2 were 43%, 27%, and 24%, respectively.     

A comparison between spatial clustering models for determining N-fertilization management zones in orchards

Precision Agriculture - Site-specific agricultural management (SSM) relies on identifying within-field spatial variability and is used for variable rate input of resources. Precision agricultural...     

Evaluation of selected growth factor expression in canine spirocercosis (Spirocerca lupi)-associated non-neoplastic nodules and sarcomas

The study aims to assess the expression of vascular endothelial growth factor (VEGF)-A, fibroblast growth factor (FGF) and platelet-derived growth factor (PDGF) in the progression of the spirocercosis-induced oesophageal nodule in the dog from early, non-neoplastic, inflammatory nodule to sarcomatous neoplasia. Triplicate 4 μm-thick sections from 62 paraffin blocks containing Spirocerca-induced nodules were treated according to the steptavidin-biotin (LSAB) immunohistochemical method using polyclonal goat-anti-canine VEGF and anti-human FGF and PDGF antibodies. The nodules were classified as early inflammatory (n=15), pre-neoplastic (n=27) and neoplastic (n=20). Additionally, 10 sections of normal distal third of the oesophagus and 21 non-spirocercosis-related sarcomas were evaluated and compared with the Spirocerca-induced nodules. Five non-overlapping high power fields per case were evaluated under the light microscope and the fibroblasts were evaluated for percentage of labelled cells. The intensity of labelling was further classified as weak (score 1) or strong (score 2). The intensity score was multiplied by the percentage of labelled fibroblasts to yield a field score and the final score was obtained by calculating an average of the five fields. Antigen labelling was compared between the different histological grades and the controls using the Kruskal-Wallis test followed by the Mann-Whitney test for comparison between specific groups. The level of significance was set at 0.05. There were significant differences between the groups' score in all the growth factors that were examined. The normal oesophagus showed no labelling for any of the growth factors. FGF scored highest in the non-spirocercosis-related sarcoma group (median 118, 3-194) followed by the spirocercosis-induced sarcoma (34.5, 0-138), pre-neoplastic nodule (8, 0-99) and early nodule (0, 0-30) groups. All the differences among the groups were significant. VEGF scored highest in the non-spirocercosis-related sarcoma group (median 47, 1-110) followed by the spirocercosis-induced sarcoma (26, 0-136), pre-neoplastic nodule (0, 0-62) and early nodule (0, 0-35) groups. PDGF scored highest in the non-spirocercosis-related sarcoma group (median 29.2, 0-70) followed by the pre-neoplastic nodule (23.4, 0-95), early nodule (13.6, 0-132) and spirocercosis-induced sarcoma (0, 0-47) groups. The expression of VEGF and FGF increased as the nodule progressed from early inflammation to sarcoma, but it was not limited to spirocercosis-induced sarcomas. The expression of PDGF in spirocercosis was restricted to the early stages of nodule progression. Further investigation is warranted to establish whether FGF, VEGF or PDGF play a role in the pathogenesis of the malignant transformation in canine spirocercosis or are they simply integral to angiogenesis induction?     

Botryllus schlosseri as a Unique Colonial Chordate Model for the Study and Modulation of Innate Immune Activity

Understanding the mechanisms that sustain immunological nonreactivity is essential for maintaining tissue in syngeneic and allogeneic settings, such as transplantation and pregnancy tolerance. While most transplantation rejections occur due to the adaptive immune response, the proinflammatory response of innate immunity is necessary for the activation of adaptive immunity. Botryllus schlosseri, a colonial tunicate, which is the nearest invertebrate group to the vertebrates, is devoid of T- and B-cell-based adaptive immunity. It has unique characteristics that make it a valuable model system for studying innate immunity mechanisms: (i) a natural allogeneic transplantation phenomenon that results in either fusion or rejection; (ii) whole animal regeneration and noninflammatory resorption on a weekly basis; (iii) allogeneic resorption which is comparable to human chronic rejection. Recent studies in B. schlosseri have led to the recognition of a molecular and cellular framework underlying the innate immunity loss of tolerance to allogeneic tissues. Additionally, B. schlosseri was developed as a model for studying hematopoietic stem cell (HSC) transplantation, and it provides further insights into the similarities between the HSC niches of human and B. schlosseri. In this review, we discuss why studying the molecular and cellular pathways that direct successful innate immune tolerance in B. schlosseri can provide novel insights into and potential modulations of these immune processes in humans.     

Season-induced changes in bovine sperm motility following a freeze-thaw procedure

Decreased conception rate of dairy cows in the summer is mainly associated with the deleterious effects of environmental thermal stress on the female reproductive tract. Here, we suggest that decreased reproductive performance might be partially due to inferior-quality semen. Semen from five representative bulls was collected in summer (August to September) and winter (December to January) and evaluated with a computerized sperm-quality analyzer for bulls (SQA-Vb). No seasonal effect was found in fresh ejaculate, but sperm examined post-thawing showed lower velocity, motility and progressive motility (P<0.04) in summer vs. winter samples. Element concentrations in the seminal plasma, determined by inductively coupled plasma-atomic emission spectrometry, differed between seasons, with higher (P<0.01) concentration values of K, Mg, Na and S elements in winter vs. summer samples. Therefore, season-induced alterations in seminal plasma element concentration should be taken into account when using an extender for cryopreservation. Acrosome integrity was assessed by a triple-fluorescence test using Hoechst 33342, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and propidium iodide. Acrosome reaction was examined by a one-step staining method using FITC-PSA. The proportion of sperm cells with a damaged acrosome post-thawing tended to be higher (P<0.07) in semen collected during the summer vs. winter. Such alterations suggest that seasonal reductions in sperm function might also be involved in the decreased conception rate of dairy cows in summer.     

A gene-coexpression network for global discovery of conserved genetic modules

To elucidate gene function on a global scale, we identified pairs of genes that are coexpressed over 3182 DNA microarrays from humans, flies, worms, and yeast. We found 22,163 such coexpression relationships, each of which has been conserved across evolution. This conservation implies that the coexpression of these gene pairs confers a selective advantage and therefore that these genes are functionally related. Many of these relationships provide strong evidence for the involvement of new genes in core biological functions such as the cell cycle, secretion, and protein expression. We experimentally confirmed the predictions implied by some of these links and identified cell proliferation functions for several genes. By assembling these links into a gene-coexpression network, we found several components that were animal-specific as well as interrelationships between newly evolved and ancient modules.     

Mycobacterium avium subsp. paratuberculosis from free-ranging deer and rabbits surrounding Minnesota dairy herds

The objectives of this study were to estimate the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) among deer and rabbits surrounding infected and noninfected Minnesota dairy farms using fecal culture, and to describe the frequency that farm management practices were used that could potentially lead to transmission of infection between these species. Fecal samples from cows and the cow environment were collected from 108 Minnesota dairy herds, and fecal pellets from free-ranging white-tailed deer and eastern cottontail rabbits were collected from locations surrounding 114 farms; all samples were tested using bacterial culture. In addition, a questionnaire was administered to 114 herd owners. Sixty-two percent of the dairy herds had at least 1 positive fecal pool or environmental sample. A total of 218 rabbit samples were collected from 90% of the herds, and 309 deer samples were collected from 47% of the herds. On 2 (4%) of the farms sampled, 1 deer fecal sample was MAP positive. Both farms had samples from the cow fecal pool and cow environment that were positive by culture. On 2 (2%) other farms, 1 rabbit fecal sample was positive by culture to MAP, with one of these farms having positive cow fecal pools and cow environmental samples. Pasture was used on 79% of the study farms as a grazing area for cattle, mainly for dry cows (75%) and bred or prebred heifers (87%). Of the 114 farms, 88 (77%) provided access to drylot for their cattle, mainly for milking cows (77/88; 88%) and bred heifers (87%). Of all study farms, 90 (79%) used some solid manure broadcasting on their crop fields. Of all 114 farms, the estimated probability of daily physical contact between cattle manure and deer or rabbits was 20% and 25%, respectively. Possible contact between cattle manure and deer or rabbits was estimated to occur primarily from March through December. The frequency of pasture or drylot use and manure spreading on crop fields may be important risk factors for transmission of MAP among dairy cattle, deer, and rabbits. Although the MAP prevalence among rabbits and deer is low, their role as MAP reservoirs should be considered.     

Changes in blood pressure following escalating doses of phenylpropanolamine and a suggested protocol for monitoring

This prospective, cross-over, blinded study evaluated the effect of various doses of phenylpropanolamine (PPA) on blood pressure in dogs. Dogs were randomized to receive a placebo or 1 of 3 dosages of immediate release PPA, q12h for 7 days [1 mg/kg body weight (BW), 2 mg/kg BW, or 4 mg/kg BW] in a cross-over design. Blood pressure was recorded every 2 h, for 12 h, on days 1 and 7. There were significant increases in systolic, diastolic, and mean blood pressure following administration of PPA at 2 mg/kg BW and 4 mg/kg BW. A significant decrease in heart rate was also noted at all PPA dosages, but not in the placebo. Administration of PPA was associated with a dose response increase in blood pressure. Dosages of up to 2 mg/kg BW should be considered safe in healthy dogs.