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.     

A multi-method approach to delineate and validate migratory corridors

Context

Managers are faced with numerous methods for delineating wildlife movement corridors, and often must make decisions with limited data. Delineated corridors should be robust to different data and models.

Objectives

We present a multi-method approach for delineating and validating wildlife corridors using multiple data sources, which can be used conserve landscape connectivity. We used this approach to delineate and validate migration corridors for wildebeest (Connochaetes taurinus) in the Tarangire Ecosystem of northern Tanzania.

Methods

We used two types of locational data (distance sampling detections and GPS collar locations), and three modeling methods (negative binomial regression, logistic regression, and Maxent), to generate resource selection functions (RSFs) and define resistance surfaces. We compared two corridor detection algorithms (cost-distance and circuit theory), to delineate corridors. We validated corridors by comparing random and wildebeest locations that fell within corridors, and cross-validated by data type.

Results

Both data types produced similar RSFs. Wildebeest consistently selected migration habitat in flatter terrain farther from human settlements. Validation indicated three of the combinations of data type, modeling, and corridor detection algorithms (detection data with Maxent modeling, GPS collar data with logistic regression modeling, and GPS collar data with Maxent modeling, all using cost-distance) far outperformed the other seven. We merged the predictive corridors from these three data-method combinations to reveal habitat with highest probability of use.

Conclusions

The use of multiple methods ensures that planning is able to prioritize conservation of migration corridors based on all available information.

     

Assessment of pulse co-oximetry technology after in vivo adjustment in anaesthetized dogs

ObjectiveTo compare values of haemoglobin concentration (SpHb), arterial haemoglobin saturation (SpO₂) and calculated arterial oxygen content (SpOC), measured noninvasively with a pulse co-oximeter before and after in vivo adjustment (via calibration of the device using a measured haemoglobin concentration) with those measured invasively using a spectrophotometric-based blood gas analyser in anaesthetized dogs.Study designProspective observational clinical study.AnimalsA group of 39 adult dogs.MethodsIn all dogs after standard instrumentation, the dorsal metatarsal artery was catheterised for blood sampling, and a pulse co-oximeter probe was applied to the tongue for noninvasive measurements. Paired data for SpHb, SpO₂ and SpOC from the pulse co-oximeter and haemoglobin arterial oxygen saturation (SaO₂) and arterial oxygen content (CaO₂) from the blood gas analyser were obtained before and after in vivo adjustment. Bland–Altman analysis for repeated measurements was used to evaluate the bias, precision and agreement between the pulse co-oximeter and the blood gas analyser. Data are presented as mean differences and 95% limits of agreement (LoA).ResultsA total of 39 data pairs were obtained before in vivo adjustment. The mean invasively measured haemoglobin–SpHb difference was –2.7 g dL^?1 with LoA of –4.9 to –0.5 g dL^?1. After in vivo adjustment, 104 data pairs were obtained. The mean invasively measured haemoglobin–SpHb difference was –0.2 g dL^?1 with LoA of –1.1 to 0.6 g dL^?1. The mean SaO₂–SpO₂ difference was 0.86% with LoA of –0.8% to 2.5% and that between CaO₂–SpOC was 0.66 mL dL^–1 with LoA of –2.59 to 3.91 mL dL^–1.ConclusionsBefore in vivo adjustment, pulse co-oximeter derived values overestimated the spectrophotometric-based blood gas analyser haemoglobin and CaO₂ values. After in vivo adjustment, the accuracy, precision and LoA markedly improved. Therefore, in vivo adjustment is recommended when using this device to monitor SpHb in anaesthetised dogs.     

Primary intraocular primitive neuroectodermal tumor (retinoblastoma) causing unilateral blindness in a gelding

A 14-year-old gray gelding was presented for investigation of a visible, pale-colored ocular mass in the right eye. An intraocular mass was identified clinically and ultrasonographically as originating from the superior nasal quadrant of the ciliary body and retina. The mass occupied the majority of the vitreous chamber and some of the superior anterior chamber of the eye. The affected eye was blind. Following exenteration, a primary intraocular primitive neuroectodermal tumor (i.e. a retinoblastoma/medulloepithelioma), a rarely described intraocular mass in adult horses, was identified by pathologic examination. The gelding returned to normal use following a short recovery period.     

Microscopic analysis of the effect of azoxystrobin treatments on Mycosphaerella graminicola infection using green fluorescent protein (GFP)‐expressing transformants

Green fluorescent protein (GFP)‐expressing transformants were used to investigate the effects of strobilurin fungicide azoxystrobin on Mycosphaerella graminicola infection. Azoxystrobin treatments (125 or 250 g AI ha^?1) were applied at various stages of the infection process under controlled conditions. GFP transformants showed conserved in vitro sensitivity to azoxystrobin and pathogenicity. Azoxystrobin controlled over 90% of M graminicola infections when applied before or during penetration of the pathogen (15% of the incubation phase). Azoxystrobin also impaired the growth of intercellular hyphae in M graminicola post‐penetration infection stages when applied at up to 50% of the incubation phase. Incubating infections observed in treated leaves were viable, but their growth was impaired and they did not induce necrosis under controlled conditions. Reduction by half of azoxystrobin dosage had little or no effect on azoxystrobin efficiency in controlling M graminicola. The contribution of post‐penetration fungistatic effect to azoxystrobin curative properties toward M graminicola in a field situation is discussed. © 2001 Society of Chemical Industry     

The transmission by nematodes of tobraviruses is not determined exclusively by the virus coat protein

The coat protein gene of the nematode non-transmissible, SP5 isolate of pea early-browning tobravius was replaced with that of the highly nematode transmissible, PPK20 isolate of tobacco rattle tobravirus. Plants were infected with the recombinant virus when mechanically inoculated and the virus invaded the plants systemically. However, although the PPK20 isolate of TRV was transmitted by nematodes from these plants, the recombinant virus was not transmitted. Therefore, the virus coat protein is not the exclusive determinant of nematode transmission.     

Quantitative PCR monitoring of the effect of azoxystrobin treatments on Mycosphaerella graminicola epidemics in the field

Quantitative PCR and visual monitoring of Mycosphaerella graminicola epidemics were performed to investigate the effect of curative and preventative applications of azoxystrobin in wheat field crops. A non-systemic protectant and a systemic curative fungicide, chlorothalonil and epoxiconazole, respectively, were used as references. PCR diagnosis detected leaf infection by M graminicola 3 weeks before symptom appearance, thereby allowing a clear distinction between curative and preventative treatments. When applied 1 week after the beginning of infection, azoxystrobin curative activity was intermediate between chlorothalonil (low effect) and epoxiconazole. When applied preventatively, none of the fungicides completely prevented leaf infection. There was some indication that azoxystrobin preventative treatments may delay fungal DNA increase more than epoxiconazole at the beginning of leaf infection. Both curative and preventative treatments increased the time lapse between the earliest PCR detection and the measurement of a 10% necrotic leaf area. Azoxystrobin only slightly decreased the speed of necrotic area increase compared with epoxiconazole. Hence, azoxystrobin activity toward M graminicola mainly resides in lengthening the time lapse between the earliest PCR detection and the measurement of a 10% necrotic leaf area. Information generated in this way is useful for optimal positioning of azoxystrobin treatments on M graminicola.