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.     

Application and field validation of a PCR assay for the detection of Mycoplasma hyopneumoniae from swine lung tissue samples.

A PCR assay was validated for the detection of Mycoplasma hyopneumoniae in porcine lung tissue. The detection limit of the assay was 0.18 colony-forming units/g of lung sample spiked with M. hyopneumoniae. In field validation, 426 pigs from 220 cases were examined for M. hyopneumoniae infection by M. hyopneumoniae PCR and a fluorescent antibody (FA) test. In total, 103 pig lungs (24.2%) were positive in the PCR test, and 69 pig lungs (16.2%) were positive in the FA test, among which, 62 pigs were positive for both PCR and FA test. Most of the PCR-positive but FA test-negative cases had lesions compatible with M. hyopneumoniae infection. With Bayesian modeling, the diagnostic sensitivity and specificity of the PCR were determined to be 97.3% and 93.0%, respectively.     

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.

     

A Markov chain approach to crop yield forecasting

This study proposes a methodology for forecasting crop yields at intermediate times in the growing season using Markov chain theory. A Markov chain is constructed, based on historical data, to provide forecast distributions of crop yield for various crop and soil moisture condition classes at selected times prior to harvest. Expected yield and the associated standard error are obtained for each condition class. The methodology is compared to a regression approach in which the independent variables are the various crop and soil moisture conditions. The Markov chain approach requires less stringent assumptions and provides more information than the regression approach. However, the potential loss of precision in the forecast using this approach requires separate evaluation for each application. A data base created by the CERES-Maize model, which simulates the growth and development of a corn crop, is used to demonstrate the development of the forecast yield distributions using the Markov chain approach.     

Severe Neutropenia Associated With Griseofulvin Therapy in Cats With Feline Immunodeficiency Virus Infection

Griseofulvin administration was associated with the development of absolute neutropenia in six of seven (86%) cats with feline immunodeficiency virus (FIV) infection. The neutropenia was severe (less than 400 neutrophils/microliter) in four of the six affected cats, and one cat died from sepsis. Neutrophil counts returned to baseline values within 15 days after drug withdrawal in all surviving cats. No symptoms or hematologic abnormalities were observed in four normal (FIV-seronegative) cats treated with the same lot of griseofulvin at equivalent doses. Neutropenia recurred in two of two FIV-seropositive cats upon griseofulvin rechallenge. Cats with FIV infections appear to be at increased risk for griseofulvin-associated neutropenia. This phenomenon may be analogous to the increased frequency of antibiotic-induced neutropenias observed in humans infected with the human immunodeficiency virus.     

Modeling topographic effects on net ecosystem productivity of boreal black spruce forests

Current regional estimates of net primary productivity (NPP) of boreal black spruce overlook the large variation in NPP caused by small-scale topographic effects on soil water, temperature and nutrient availability. Topographic effects on black spruce NPP could likely be modeled by simulating the lateral and vertical movement of water, and its effects on soil nutrient transformation and uptake, through three-dimensional watersheds defined by aspects and slopes of their topographic positions. To examine this likelihood, the ecosystem model 'ecosys' was run for 120 years on a transect that included upper- and lower-slope positions and a basin in which a basal water table was set 0.5 m below the soil surface. For the run, we used soil properties and weather conditions recorded at the 115-year-old BOREAS Southern Old Black Spruce site. Short-term model performance was tested by comparing diurnal and annual carbon (C) transfers simulated under 1994 weather conditions during the 115th year of the model run with those measured at this site during 1994 by eddy covariance, surface chambers and allometry. After 115 years, annual spruce NPP simulated at the upper-slope positions was twice that at the basin (350 versus 170 g C m-2), whereas accumulated wood C was almost three times as large (6.8 versus 2.4 kg C m-2). In the model, increases in NPP and wood growth in upper-slope positions were caused by lower soil water contents, higher soil temperatures, and more rapid O2 uptake that accelerated heterotrophic respiration and hence nutrient mineralization and uptake. Modeled differences in wood growth with topographic position were quantitatively consistent with measurements of boreal black spruce at several research sites differing in water table depth. Modeled differences also agreed with differences in wood growth rates derived from allometric measurements at boreal black spruce sites differing in productivity indices as a result of differences in subsurface hydrology. The magnitude of these differences clearly indicates the importance of accounting for subsurface hydrology in regional estimates of boreal forest productivity.     

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.