Effects of temperature and time in transit on polymerase chain reaction detection of feline herpesvirus DNA.

The purpose of this study was to report methods currently recommended by commercial laboratories for collecting, shipping, and processing of samples for feline herpesvirus type 1 (FHV-1) testing using polymerase chain reaction (PCR) and to determine the effect of temperature and time on the ability of 1 PCR method to detect FHV-1 DNA in experimental and clinical samples. Eleven laboratories offering FHV-1 PCR were surveyed. There was notable variation in sample types and shipping conditions recommended and PCR protocols used by these laboratories. Subsequently, using a single PCR method, FHV-1 DNA was detected in samples exposed to various temperatures within the laboratory. Finally, FHV-1 PCR was performed on paired clinical samples collected from 25 cats and shipped at ambient temperatures via US Postal Service (USPS) or with an ice pack via a courier. Samples sent by USPS were exposed to significantly longer transit time and arrived at significantly higher temperature than did samples sent by courier. Despite this, all sample pairs yielded concordant results when tested for FHV-1 DNA using this PCR method. Although it may not be necessary for samples collected for detection of FHV-1 DNA using this PCR method to be shipped under the most expedient or temperature-controlled conditions, this should be verified for a variety of PCR assays and sample types.     

Parasites of free-ranging small canids and felids in the Bolivian Chaco.

Parasite surveys of free-ranging wildlife provide important information for monitoring population health. Between March 2001 and March 2003, we sampled 10 ocelots (Leopardus pardalis), eight Geoffroy's cats (Oncifelis geoffroyi), a jaguarundi (Herpailurus yaguarondi), five pampas foxes (Pseudalopex gymnocercus), and three crab-eating foxes (Cerdocyon thous) at three sites in the Bolivian Chaco. The objective of the study was to survey the parasite fauna of these carnivores and compare prevalence of parasites among the sites. The parasite community of these carnivores was diverse, with representatives from eight genera of nematodes, two families of cestodes, two protozoan species, and six arthropod species. Fecal parasites identified from 12 of the 13 felids and five of the six canids examined included Aelurostrongylus abstrusus, Ancylostoma tubaeforme, Uncinaria sp., Crenosoma sp., Toxocara cati, Spirurida, Capillaria aerophila, Spirometra sp., Taeniidae, and Cystoisospora sp. Four tick species, Amblyomma parvum, A. tigrinum, A. ovale, and A. cajennense, and two flea species, Pulex irritans and Delostichus phyllotis, were identified. Two crab-eating foxes had serologic evidence of heartworm disease (HWD). Antibodies against Toxoplasma gondii were found in 15 of 26 animals. Although HWD was found only in canids inside the national park, parasite prevalence did not appear to differ among sites, and no evidence was found of parasite spillover from domestic to wild carnivores.     

Diagnostic cytology of fish.

Cytology is an essential part of a diagnostic workup in cases of aquatic animal diseases. It is simple to perform, inexpensive, and can yield quick and valuable results. External parasites, bacterial and fungal diseases, and gastrointestinal infestations are easily determined with wet mount cytology. Because of the relatively small number of nonlethal diagnostic techniques available for aquatic species, cytologic testing should be considered in every case. Early diagnosis can lead to more effective treatment plans, ensuring a better prognostic outcome in our patients.     

Major gene resistance in Brassica napus (oilseed rape) is overcome by changes in virulence of populations of Leptosphaeria maculans in France and Australia

Resistance of Brassica napus (oilseed rape, canola) conferred by three different major resistance genes has been overcome by changes in virulence of Leptosphaeria maculans populations in France and Australia. In South Australia where B. napus cultivars with major gene resistance derived from Brassica rapa ssp. sylvestris were grown extensively, resistance was rendered ineffective within 3 years of commercial release of the cultivar. Disease severity was higher on cultivars with sylvestris-derived resistance than cultivars with polygenic resistance. This Australian situation is compared to that in France, where resistance conferred by the Rlm1 gene was overcome nation-wide in 5 years under commercial cropping practices, and also where a source of resistance introgressed into B. napus from B. juncea was rendered inefficient in 3 years in experimental field plots near Rennes.     

Landscape attributes and life history variability shape genetic structure of trout populations in a stream network

Spatial and temporal landscape patterns have long been recognized to influence biological processes, but these processes often operate at scales that are difficult to study by conventional means. Inferences from genetic markers can overcome some of these limitations. We used a landscape genetics approach to test hypotheses concerning landscape processes influencing the demography of Lahontan cutthroat trout in a complex stream network in the Great Basin desert of the western US. Predictions were tested with population- and individual-based analyses of microsatellite DNA variation, reflecting patterns of dispersal, population stability, and local effective population sizes. Complementary genetic inferences suggested samples from migratory corridors housed a mixture of fish from tributaries, as predicted based on assumed migratory life histories in those habitats. Also as predicted, populations presumed to have greater proportions of migratory fish or from physically connected, large, or high quality habitats had higher genetic variability and reduced genetic differentiation from other populations. Populations thought to contain largely non-migratory individuals generally showed the opposite pattern, suggesting behavioral isolation. Estimated effective sizes were small, and we identified significant and severe genetic bottlenecks in several populations that were isolated, recently founded, or that inhabit streams that desiccate frequently. Overall, this work suggested that Lahontan cutthroat trout populations in stream networks are affected by a combination of landscape and metapopulation processes. Results also demonstrated that genetic patterns can reveal unexpected processes, even within a system that is well studied from a conventional ecological perspective.     

Using an individual-based model to examine the roles of habitat fragmentation and behavior on predator–prey relationships in seagrass landscapes

Seagrasses, which form critical subtidal habitats for marine organisms worldwide, are fragmented via natural processes but are increasingly being fragmented and degraded by boating, fishing, and coastal development. We constructed an individual-based model to test how habitat fragmentation and loss influenced predator–prey interactions and cohort size for a group of settling juvenile blue crabs (Callinectes sapidus Rathbun) in seagrass landscapes. Using results from field studies suggesting that strong top-down processes influence the relationship between cannibalistic blue crab populations and seagrass landscape structure, we constructed a model in which prey (juvenile blue crabs) are eaten by mesopredators (larger blue crabs) which in turn are eaten by top-level predators (e.g., large fishes). In our model, we varied the following parameters within four increasingly fragmented seagrass landscapes to test for their relative effects on cohort size: juvenile blue crab (prey) predator avoidance response, hunting ability of mesopredators and predators, the presence of a top-level predator, and prey settlement routines. Generally, prey cohort size was maximized in the presence of top-level predators and when mesopredators and predators exhibited random searching behavior vs. directed hunting. Cohort size for stationary (tethered) prey was maximized in fragmented landscapes, which corresponds to results from field experiments, whereas mobile prey able to detect and avoid predators had higher survival in continuous landscapes. Prey settlement patterns had relatively small influences on cohort size. We conclude that the effects of seagrass fragmentation and loss on organisms such as blue crabs will depend heavily on behaviors of prey and predatory organisms and how these behaviors change with landscape structure.

     

Recent pine woodland dynamics in east Glen Affric, northern Scotland, from highly resolved palaeoecological analyses

Glen Affric, a National Nature Reserve of international conservationimportance for plant and animal communities associated withScots pine (Pinus sylvestris) woodland is managed for natureconservation and woodland restoration at a landscape scale.Management plans have drawn on information on current standstructure and variation but have not used in any detailed waythe analyses of past tree population changes from sources, suchas pollen analyses. This paper reports the results of pollenanalyses from three small peat hollows at the head of Loch Beinna' Mheadhoin. The analyses demonstrate, first, that woods inthe east of the reserve several centuries ago were differentin species composition and were more varied than they are todayand, second, that the currently patchy Pinus wood in the westof the reserve, at the head of Loch Beinn a' Mheadhoin, is theresult of recent spread westward onto former heathland. Thistemporal and spatial variability in the recent past has implicationsfor the future management of the woods because future woodsmay not develop with the characteristics of the current stands,and may not be stable over time.     

Nitrosylhemoglobin as a Potential Bioactive Storage form of Nitric Oxide (NO)

Veterinary Research Communications -     

Challenges of thermal nociceptive threshold testing in the donkey

ObjectiveTo evaluate a thermal nociceptive threshold (TNT) testing device in the donkey, and the influence of potential confounding factors on TNTs.AnimalsTwo groups (Group 1 and Group 2) of eight castrated male donkeys aged 4–9 years, weighing 105–170 kg.MethodsTNTs were measured by heating a thermal probe on skin until an end-point behaviour (threshold temperature) or a cut-out temperature (51 °C) was reached. The withers and the dorsal aspect of the distal limb were used as sites for TNT testing. The effects on TNT of different confounding factors: the limb tested; rate of heating; and ambient temperature were evaluated. Data were analyzed using general linear models, and Mann-Whitney tests, p < 0.05 was considered significant.ResultsEnd-point behaviours (skin twitch or donkey looking at test device) when the thermal probe heated the withers were observed in approximately half of tests. TNT was (mean ± SD) 46.8 ± 2.85 °C. Subsequently the limb was evaluated as the test site in Group 1 followed by Group 2 donkeys; end-point behaviour being a foot-lift. In Group 1, 72% of tests ended in an end-point behaviour but the response rate was lower in Group 2 (20%), although TNTs were similar [(47.6 ± 3.3) and (47.3 ± 3.0) °C respectively] for responding animals. Rate of heating, ambient temperature and laterality (right or left) did not affect thresholds, but mean TNT was significantly higher in the forelimb (48.5 ± 2.8 °C) than the hind limb (47.4 ± 2.8 °C) (p = 0.012).ConclusionsWhen a thermal probe cut-out temperature of 51 °C was used in TNT testing in the donkey a high proportion of tests did not produce an identifiable end point behaviour. Higher cut-out temperatures damaged the skin. Under these conditions, thermal nociceptive threshold testing appears not be an appropriate analgesiometry technique in the donkey.Clinical relevanceTNT testing under these conditions is not suitable form of analgesiometry for donkeys.