Validation of the specificity and sensitivity of species-specific primers that provide a reliable molecular diagnostic for <Emphasis Type="Italic">Xiphinema diversicaudatum</Emphasis>, <Emphasis Type="Italic">X. index</Emphasis> and <Emphasis Type="Italic">X. vuittenezi</Emphasis>

Xiphinema diversicaudatum and X. index are vector nematode species of economic importance in viticulture regions as they can transmit Arabis Mosaic, Grapevine Fanleaf and Strawberry Latent Ringspot viruses to grapevine. Wang et al. (2003) designed species-specific diagnostic primers from ribosomal genes for both these vector species as well as a vector and a non-vector species X. italiae and X. vuittenezi, respectively. Our study aimed to confirm the specificity and determine the sensitivity and reliability of the primers for the two vector species, X. diversicaudatumand X. indexwhen challenged with closely related longidorid species and general nematode communities typical of vineyard soil. With one exception, no PCR product was observed when the primers were tested against six Longidorus, one Paralongidorus and one Xiphinema non-target species. Occasionally (three out of eight replicate PCR reactions) a weak PCR product was noted when primers for X. index were tested with L. elongatus. Furthermore, when challenged with a range of non-target nematode species comprising the nematode community typical of viticulture soil, no PCR product was amplified. An experimental dilution series of extracted DNA rigorously demonstrated that DNA from an equivalent single specimen of the target virus-vector species, X. diversicaudatum and/or X. index, could be detected amongst 1000 equivalent non-targetX. vuittenezi. Also, extracted DNA from an equivalent single target specimen was detected when added to DNA extracted from the overall soil nematode community. The primers were assessed further by using serial mixtures of actual nematodes rather than extracted DNA to simulate field soil. Using this method, a single target nematode could be detected amongst 200 non-target specimens. Given their specificity, sensitivity and reliability, it appears that these diagnostic primers will be of great benefit to phytosanitary/quarantine services related to the viticulture industry.     

Treatment of contaminated radial fracture in Sprague-Dawley rats by application of a degradable polymer releasing gentamicin

Fracture-related infections remain a leading cause of morbidity and mortality. We aimed to establish a simple contaminated radial osteotomy model to assess the efficacy of a biodegradable polymer poly(sebacic-co-ricinoleic acid) [p(SA-RA)] containing 20% w/w gentamicin. A unilateral transverse osteotomy was induced in Sprague-Dawley (SD) rats, followed by application of Staphylococcus aureus suspension over the fracture. After successfully establishing the contaminated open fracture model, we treated the rats either systemically (intraperitoneal cefuroxime), locally with p(SA-RA) containing gentamicin, or both. Control groups included non-contaminated group and contaminated groups that were either untreated or treated with the polymer alone. After 4 weeks, the bones were subjected to micro-CT scanning and microbiological and histopathology evaluations. Micro-CT analysis revealed similar changes in the group subjected to both local and systemic treatment as in the non-contaminated control group. Lack of detectable bacterial growth was noted in most animals of the group subjected to both local and systemic treatment, and all samples were negative for S. aureus. Histopathological evaluation revealed that all treatment modalities containing antibiotics were highly effective in reducing infection and promoting callus repair, resulting in early bone healing. While p(SA-RA) containing gentamicin treatment showed better results than cefuroxime, the combination of local and systemic treatment displayed the highest therapeutic potential in this model.     

An outbreak of tick paralysis in free-ranging mouflon (Ovis ammon musimon)

During November 2006, two live and one dead mouflon (Ovis ammon musimon) were presented with a history of weakness, tremors, and paralysis. After a detailed gross and histologic examination and a bacteriologic, parasitologic, and rabies evaluation, a preliminary diagnosis of tick paralysis was established. A thorough field search revealed 13 affected mouflons found in the open hunting ground "Sveti Juraj" near the town of Senj (Croatia), along with an additional 35 mouflon carcasses. All 13 mouflons were placed in a quiet, semidark stable. All detectable ticks were removed manually, and the animals were topically treated with 250 ppm of Amitraz water emulsion (Taktic 12.5% EC, Intervet International, 5830 Boxmeer, Netherlands). The collected ticks were identified as Ixodes ricinus, Dermacentor marginatus, and Haemaphysalis punctata. In the following 24 hr, all treated animals recovered fully. This report describes a naturally occurring outbreak of tick paralysis in free-ranging mouflons from a karst habitat.     

Adjuvant immunotherapy of feline fibrosarcoma with recombinant feline interferon-omega

BACKGROUND: Recombinant feline interferon-omega (rFeIFN-omega) was tested as a treatment option for cats with fibrosarcoma to assess safety and feasibility. HYPOTHESIS: Treatment with rFeIFN-omega in cats with fibrosarcoma is safe and feasible. ANIMALS: Twenty domestic cats. METHODS: In an open-labeled uncontrolled clinical trial 12 injections of 1 x 10(6) U/kg rFeIFN-omega were administered over a 5-week period: the 1st through 4th injections were given intratumorally, and the 5th through 12th injections were administered subcutaneously at the tumor excision site. Wide surgical excision of the tumors was carried out after the 4th injection and before the 5th injection of rFeIFN-omega. A Common Terminology Criteria for Adverse Events (CTCAE) analysis was conducted. Flow cytometry of fibrosarcoma cells after incubation with rFeIFN-omega and recombinant feline interferon-gamma was performed to assess the biological effect of rFeIFN-omega. RESULTS: Changes in blood cell count, increases in serum aspartate-amino-transferase activity, serum bilirubin concentration, serum creatinine and serum electrolyte concentrations, weight loss, anorexia, increased body temperature, and reduced general condition were observed but were mostly minor (grade 1 and 2) and self limiting. Eosinophilia (P = .025), neutropenia (P = .021), and weight loss (P < .001) were statistically correlated with rFeIFN-omega-treatment (analysis of parameters before treatment and after 3 injections of rFeIFN-omega). Flow cytometry of 5 unrelated feline fibrosarcoma cell lines showed increased expression of major histocompatibility complex (MHC) class I molecules (P = .026) in response to in vitro incubation with rFeIFN-omega, whereas expression of MHC class II molecules was not affected significantly. CONCLUSIONS AND CLINICAL IMPORTANCE: RFeIFN-omega for the treatment of feline fibrosarcoma is safe, well tolerated, and can be easily performed in practice. To assess the efficacy of the treatment, it should be tested in a placebo-controlled trial.     

How to consider history in landscape ecology: patterns,processes, and pathways

Landscape Ecology - Landscape ecology early on developed the awareness that central objects of investigation are not stable over time and therefore the historical dimension must be included, or at...     

The combined effects of temperature and GnRHa treatment on the final stages of sexual maturation in Atlantic salmon (Salmo salar L.) females

Atlantic salmon (Salmo salar L.) females (2 SW), maturing for the first time, were reared under one of three temperature regimes (high: 14.3 ± 0.5°C; natural: 10.6 ± 1.0°C; and cold: 6.9 ± 1.0°C) in combination with one of two experimental treatments; an injection of GnRH analogue (GnRHa) contained in biodegradable microspheres, or a sham injection (microspheres only). The six experimental groups were then reared under simulated natural photoperiod for 4 weeks. Blood samples were drawn for analysis of plasma steroid levels and the fish were inspected for ovulation weekly. Batches of stripped eggs were incubated in triplicate incubators in raceways until the eyed stage. Treatment with GnRHa resulted in a substantial advancement and synchronization of ovulation at all temperatures, while exposure to cold water also appeared to advance ovulation slightly. While 75% (warm and cold) to 90% (natural) of GnRHa fish ovulated during the 4-week trial, only 30% of sham-treated females exposed to cold water, and none of the sham-treated fish held at higher temperatures, ovulated during this period. Survival rates of embryos to the eyed-stage were significantly higher for broodstock exposed to cold water. Plasma levels of testosterone (T), 17β-oestradiol (E₂), and 17α,20β-dihydroxy-4-pregnen-3-one (17,20βP) were all significantly affected by treatment with GnRHa and, to a lesser extent, temperature. The efficiency of GnRHa in counteracting the negative effects of high temperature on ovulation and the associated changes in circulating sex steroids suggest that temperature inhibition operates at least in part at the brain or pituitary.     

Effect of mineral nitrogen fertilizer forms on N2O emissions from arable soils in winter wheat production

Nitrogen fertilizers are supposed to be a major source of nitrous oxide (N₂O) emissions from arable soils. The objective of this study was to compare the effect of N forms on N₂O emissions from arable fields cropped with winter wheat (Triticum aestivum L.). In three field trials in North‐West Germany (two trials in 2011/2012, one trial in 2012/2013), direct N₂O emissions during a one‐year measurement period, starting after application of either urea, ammonium sulfate (AS) or calcium ammonium nitrate (CAN), were compared at an application rate of 220 kg N ha^?1. During the growth season (March to August) of winter wheat, N₂O emission rates were significantly higher in all three field experiments and in all treatments receiving N fertilizer than from the non‐fertilized treatments (control). At two of the three sites, cumulative N₂O emissions from N fertilizer decreased in the order of urea > AS > CAN, with emissions ranging from 522–617 g N ha^?1 (0.24–0.28% of applied fertilizer) for urea, 368–554 g N ha^?1 (0.17–0.25%) for AS, and 242–264 g N ha^?1 (0.11–0.12%) for CAN during March to August. These results suggest that mineral nitrogen forms can differ in N₂O emissions during the growth period of winter wheat. Strong variations in the seasonal dynamics of N₂O emissions between sites were observed which could partly be related to weather events (e.g., precipitation). Between harvest and the following spring (post‐harvest period) no significant differences in N₂O emissions between fertilized and non‐fertilized treatments were detected on two of three fields. Only on one site post‐harvest emissions from the AS treatment were significantly higher than all other fertilizer forms as well as compared to the control treatment. The cumulative one‐year emissions varied depending on fertilizer form across the three field sites from 0.05% to 0.51% with one exception at one field site (AS: 0.94%). The calculated overall fertilizer induced emission averaged for the three fields was 0.38% which was only about 1/3 of the IPCC default value of 1.0%.