Spirometra erinacei / S. erinaceieuropaei in a feral cat in Manawatu with chronic intermittent diarrhoea

CASE HISTORY: A feral cat captured in the Manawatu region of New Zealand was treated for worms and fleas, and kept confined in a metabolic cage. It showed good appetite and weight gain but had intermittent watery, yellow diarrhoea.

CLINICAL FINDINGS: Clinical examination under sedation was unremarkable and routine blood tests showed no significant abnormalities. The cat was negative for feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). Different canned cat foods did not alter the course of the diarrhoea, and the cat was euthanised 6 months after capture. At necropsy, two sections of adult Spirometra tapeworms were found in the jejunum and typical Spirometra eggs were found in colonic contents. Molecular identification of the parasite was undertaken, using the cytochrome- c oxidase subunit-1 gene (cox1) sequence.

DIAGNOSIS: Chronic intermittent diarrhoea associated with Spirometra erinacei / S. erinaceieuropaei infection.

CLINICAL RELEVANCE: Spirometra has not been reported in New Zealand before but has been associated with gastrointestinal disease in cats in other parts of the world. It requires speciestargeted treatment to be eliminated effectively, and is zoonotic. Diagnosis could be diffi cult for clinicians who are not familiar with the parasite and its life cycle.     

Pharmacokinetics of hemoglobin-based oxygen carrier hemoglobin-glutamer-200 bovine (oxyglobin) in the horse

Oxyglobin (OXY) is a hemoglobin‐based oxygen carrier (HBOC) made of glutaraldehyde‐polymerized bovine hemoglobin (bHb). Products similar to OXY are under development for use as temporary blood substitutes in trauma, shock and anemia. Since they all may increase blood O₂‐carrying capacity and thus, possibly tissue oxygenation, they may also be used to enhance performance of both equine and human athletes. That is why HBOCs are banned from use in athletic competition. Our goal was to determine the pharmacokinetics of OXY after intravenous (IV) infusion to horses. Blood and urine samples were collected from adult horses that received an IV dose of 32.5 g of OXY. Concentrations of OXY in plasma and urine were quantified using a newly developed LC/Q‐TOF‐MS/MS detection technique. Level of quantification (LOQ) was 50 μg mL^–1. The decline of the plasma concentration‐time curve of the HBOC was described by a 2‐compartment model (C1 and C2). The median distribution alpha (t1/2k1,0) and elimination beta (t1/2k2,0) half‐lives were 1.3 and 12.0 hours, respectively. The bHb molecules in OXY are not of uniform size and vary substantially in molecular weight (MW). Of the OXY molecules 53% were eliminated in C1, which represented the smaller MW molecules and 47% in C2, which represented the larger MW bHb. The maximal 0‐time plasma concentration was 662.0 μg/mL and declined to 97.1 μg mL^–1 at 24 h. The area below the plasma concentration‐time curve was 5143 μg h^–1 mL^–1. The volumes of C₁ and C₂ were 86.9 and 63.9 mL kg^–1, respectively. Oxyglobin was not detected in urine. This study shows the detection and quantification in equine plasma of a HBOC following IV infusion and demonstrates the short half‐life of about 50% of infused bHb molecules.     

Molecular epidemiology of clinical isolates of Pseudomonas aeruginosa isolated from horses in Ireland

Clinical isolates (n = 63) of Pseudomonas aeruginosa obtained from various sites in 63 horses were compared using ERIC2 RAPD PCR to determine their genetic relatedness. Resulting banding patterns (n = 24 genotypes) showed a high degree of genetic heterogeneity amongst all isolates examined, indicating a relative non-clonal relationship between isolates from these patients, employing this genotyping technique. This study characterised 63 clinical isolates into 24 distinct genotypes, with the largest cluster (genotype E) accounting for 10/63 (15.9%) of the isolates. ERIC2 RAPD PCR proved to be a highly discriminatory molecular typing tool of P. aeruginosa in isolates recovered from horses. With the adoption of several controls to aid reproducibility, this technique may be useful as an alternative to PFGE, particularly in epidemiological investigations of outbreaks where speed may be a significant parameter. This is the first report of clonal heterogeneity amongst P. aeruginosa from horses and demonstrated that ERIC RAPD PCR is a rapid method for the examination of this species in horses, which may be useful in outbreak analysis.     

Early Development and Putative Primordial Germ Cells Characterization in Dogs

Previously, three distinct populations of putative primordial germ cells (PGCs), namely gonocytes, intermediate cells and pre‐spermatogonia, have been described in the human foetal testis. According to our knowledge, these PGCs have not been studied in any other species. The aim of our study was to identify similar PGC populations in canine embryos. First, we develop a protocol for canine embryo isolation. Following our protocol, 15 canine embryos at 21–25 days of pregnancy were isolated by ovaryhysterectomy surgery. Our data indicate that dramatic changes occur in canine embryo development and PGCs specification between 21 to 25 days of gestation. At that moment, only two PGC populations with distinct morphology can be identified by histological analyses. Cell population 1 presented round nuclei with prominent nucleolus and a high nuclear to cytoplasm ratio, showing gonocyte morphology. Cell population 2 was often localized at the periphery of the testicular cords and presented typical features of PGC. Both germ cell populations were positively immunostained with anti‐human OCT‐4 antibody. However, at day 25, all cells of population 1 reacted positively with OCT‐4, whereas in population 2, fewer cells were positive for this marker. These two PGCs populations present morphological features similar to gonocytes and intermediate cells from human foetal testis. It is expected that a population of pre‐spermatogonia would be observed at later stages of canine foetus development. We also showed that anti‐human OCT‐4 antibody can be useful to identify canine PGC in vivo.     

Primary structure and biochemical properties of an M2 muscarinic receptor

A partial amino acid sequence obtained for porcine atrial muscarinic acetylcholine receptor was used to isolate complementary DNA clones containing the complete receptor coding region. The deduced 466-amino acid polypeptide exhibits extensive structural and sequence homology with other receptors coupled to guanine nucleotide binding (G) proteins (for example, the beta-adrenergic receptor and rhodopsins); this similarity predicts a structure of seven membrane-spanning regions distinguished by the disposition of a large cytoplasmic domain. Stable transfection of the Chinese hamster ovary cell line with the atrial receptor complementary DNA leads to the binding of muscarinic antagonists in these cells with affinities characteristic of the M2 receptor subtype. The atrial muscarinic receptor is encoded by a unique gene consisting of a single coding exon and multiple, alternatively spliced 5' noncoding regions. The atrial receptor is distinct from the cerebral muscarinic receptor gene product, sharing only 38% overall amino acid homology and possessing a completely nonhomologous large cytoplasmic domain, suggesting a role for the latter region in differential effector coupling.     

Spectral Detection of Emergence in Corn and Cotton

Multispectral reflectance of emerging cotton (Gossypium hirsutum) and corn (Zeamays) seedlings was measured during the 2000 and 2001 growing seasons. Reflectance in blue, green, red, and near infrared (NIR) wave lengths was used to detect seedling emergence, to monitor leaf area growth, and to measure the effect of bare soil reflectance on scene (bare soil and seedlings) reflectance. Cotton and corn seedlings were detected 1 day after initial emergence (1 DAE) in 2000 by the red band. The red band detected seedlings in 2001 at 9 and 8 DAE in early and late planted corn, respectively, and on 0 DAE for cotton. The normalized difference vegetative index (NDVI) detected seedlings at 1 DAE or 2 DAE in both years. Seedling ground cover in 2000 on the initial detection date in the target areas averaged 1.3% and 0.9%, respectively, for cotton and corn; comparable values in 2001 for cotton, early planted corn, and late planted corn, were 1.4%, 0.4%, and 0.8%, respectively. The red wave band was the most sensitive single band for detecting the presence of seedlings, but NDVI was the most sensitive spectral indicator, which was apparently due to the red band since the NIR band did not always detect seedlings. Seedling leaf area was linearly correlated with NDVI values beginning at 1 or 2 DAE. Bare soil was the major component of the scene during stand establishment and dominated single band reflectance and NDVI values. A dry soil surface that was smoothed and sealed by rain usually caused single band reflectance to increase. The high variability in spectral characteristics of bare soil restricted the interpretation of the spectral data to concluding whether or not seedlings were emerging, but without estimating numbers and seedling size.     

关于内蒙古大兴安岭林区实施森林抚育补贴项目的探讨

文章通过对内蒙古大兴安岭林区实施森林抚育补贴项目的背景、必要性的分析,论述了森林抚育是促进林区生态、经济和社会协调发展的有力措施。     

Feline B7.1 and B7.2 proteins produced from swinepox virus vectors are natively processed and biologically active: potential for use as nonchemical adjuvants

Costimulatory ligands, B7.1 and B7.2, have been incorporated into viral and DNA vectors as potential nonchemical adjuvants to enhance CTL and humoral immune responses against viral pathogens. In addition, soluble B7 proteins, minus their transmembrane and cytoplasmic domains, have been shown to block the down regulation of T-cell activation through blockade of B7/CTLA-4 interactions in mouse tumor models. Recently, we developed swinepox virus (SPV) vectors for delivery of feline leukemia antigens for vaccine use in cats [Winslow, B.J., Cochran, M.D., Holzenburg, A., Sun, J., Junker, D.E., Collisson, E.W., 2003. Replication and expression of a swinepox virus vector delivering feline leukemia virus Gag and Env to cell lines of swine and feline origin. Virus Res. 98, 1-15]. To explore the use of feline B7.1 and B7.2 ligands as nonchemical adjuvants, SPV vectors containing full-length feline B7.1 and B7.2 ligands were constructed and analyzed. Full-length feline B7.1 and B7.2 produced from SPV vectors were natively processed and costimulated Jurkat cells to produce IL-2, in vitro. In addition, we explored the feasibility of utilizing SPV as a novel expression vector to produce soluble forms of feline B7.1 (sB7.1) and B7.2 (sB7.2) in tissue culture. The transmembrane and cytoplasmic regions of the B7.1 and B7.2 genes were replaced with a poly-histidine tag and purified via a two-step chromatography procedure. Receptor binding and costimulation activity was measured. Although feline sB7.1-his and sB7.2-his proteins bound to the human homolog receptors, CTLA-4 and CD28, both soluble ligands possessed greater affinity for CTLA-4, compared to CD28. However, both retained the ability to partially block CD28-mediated costimulation in vitro. Results from these studies establish the use of SPV as a mammalian expression vector and suggest that full-length-vectored and purified soluble feline B7 ligands may be valuable, nonchemical immune-modulators.