The impact of cattle dung pats on earthworm distribution in grazed pastures.

BACKGROUND: Grazed grassland management regimes can have various effects on soil fauna. For example, effects on earthworms can be negative through compaction induced by grazing animals, or positive mediated by increases in sward productivity and cattle dung pats providing a food source. Knowledge gaps exist in relation to the behaviour of different earthworm species i.e. their movement towards and aggregation under dung pats, the legacy effects of pats and the spatial area of recruitment. The present study addressed these knowledge gaps in field experiments, over 2 years, using natural and simulated dung pats on two permanent, intensively grazed pastures in Ireland. RESULTS: Dung pats strongly affected spatial earthworm distribution, with up to four times more earthworms aggregating beneath pats, than in the control locations away from pats. In these earthworm communities comprising 11 species, temporally different aggregation and dispersal patterns were observed, including absence of individual species from control locations, but no clear successional responses. Epigeic species in general, but also certain species of the anecic and endogeic groups were aggregating under dung. Sampling after complete dung pat disappearance (27 weeks after application) suggested an absence of a dung pat legacy effect on earthworm communities. Based on species distributions, the maximum size of the recruitment area from which earthworms moved to pats was estimated to be 3.8 m2 per dung pat. Since actual grazing over 6 weeks would result in the deposition of about 300 dung pats per ha, it is estimated that a surface area of 1140 m2 or about 11% of the total grazing area can be influenced by dung pats in a given grazing period. CONCLUSIONS: This study showed that the presence of dung pats in pastures creates temporary hot spots in spatial earthworm species distribution, which changes over time. The findings highlight the importance of considering dung pats, temporally and spatially, when sampling earthworms in grazed pastures. Published comparisons of grazed and cut grasslands probably reached incorrect conclusions by ignoring or deliberately avoiding dung pats. Furthermore, the observed intense aggregation of earthworms beneath dung pats suggests that earthworm functions need to be assessed separately at these hot spots.

Improving the identification of hydrologically sensitive areas using LiDAR DEMs for the delineation and mitigation of critical source areas of diffuse pollution.

Identifying critical source areas (CSAs) of diffuse pollution in agricultural catchments requires the accurate identification of hydrologically sensitive areas (HSAs) at highest propensity for generating surface runoff and transporting pollutants. A new GIS-based HSA Index is presented that improves the identification of HSAs at the sub-field scale by accounting for microtopographic controls. The Index is based on high resolution LiDAR data and a soil topographic index (STI) and also considers the hydrological disconnection of overland flow via topographic impediment from flow sinks. The HSA Index was applied to four intensive agricultural catchments (~7.5-12km(2)) with contrasting topography and soil types, and validated using rainfall-quickflow measurements during saturated winter storm events in 2009-2014. Total flow sink volume capacities ranged from 8298 to 59,584m(3) and caused 8.5-24.2% of overland-flow-generating-areas and 16.8-33.4% of catchment areas to become hydrologically disconnected from the open drainage channel network. HSA maps identified 'breakthrough points' and 'delivery points' along surface runoff pathways as vulnerable points where diffuse pollutants could be transported between fields or delivered to the open drainage network, respectively. Using these as proposed locations for targeting mitigation measures such as riparian buffer strips reduced potential costs compared to blanket implementation within an example agri-environment scheme by 66% and 91% over 1 and 5years respectively, which included LiDAR DEM acquisition costs. The HSA Index can be used as a hydrologically realistic transport component within a fully evolved sub-field scale CSA model, and can also be used to guide the implementation of 'treatment-train' mitigation strategies concurrent with sustainable agricultural intensification.

Consequences of varied soil hydraulic and meteorological complexity on unsaturated zone time lag estimates.

The true efficacy of a programme of agricultural mitigation measures within a catchment to improve water quality can be determined only after a certain hydrologic time lag period (subsequent to implementation) has elapsed. As the biophysical response to policy is not synchronous, accurate estimates of total time lag (unsaturated and saturated) become critical to manage the expectations of policy makers. The estimation of the vertical unsaturated zone component of time lag is vital as it indicates early trends (initial breakthrough), bulk (centre of mass) and total (Exit) travel times. Typically, estimation of time lag through the unsaturated zone is poor, due to the lack of site specific soil physical data, or by assuming saturated conditions. Numerical models (e.g. Hydrus 1D) enable estimates of time lag with varied levels of input data. The current study examines the consequences of varied soil hydraulic and meteorological complexity on unsaturated zone time lag estimates using simulated and actual soil profiles. Results indicated that: greater temporal resolution (from daily to hourly) of meteorological data was more critical as the saturated hydraulic conductivity of the soil decreased; high clay content soils failed to converge reflecting prevalence of lateral component as a contaminant pathway; elucidation of soil hydraulic properties was influenced by the complexity of soil physical data employed (textural menu, ROSETTA, full and partial soil water characteristic curves), which consequently affected time lag ranges; as the importance of the unsaturated zone increases with respect to total travel times the requirements for high complexity/resolution input data become greater. The methodology presented herein demonstrates that decisions made regarding input data and landscape position will have consequences for the estimated range of vertical travel times. Insufficiencies or inaccuracies regarding such input data can therefore mislead policy makers regarding the achievability of water quality targets.

First Report of Beet mild curly top virus Infection of Chile Pepper in North-Central Mexico.

During the 2005 growing season, widespread virus-like symptoms were observed in pepper (Capsicum annuum) fields in north-central Mexico. Early in the season, plants were chlorotic and stunted with thickened, elongated leaves. From mid to late season, the affected plants showed severe yellowing, upwardly rolled, small leaves, and a few deformed fruits. Symptoms were similar to those described for curtoviruses in pepper (1). The leafhopper vector of curtoviruses, Circulifer tenellus, was first reported in the area in 1953 (3) (its presence was confirmed again in January 2008). Pepper fields were sampled in the states of Aguascalientes and Zacatecas, and five symptomatic plants from Zacatecas tested positive for the presence of curtoviruses by PCR using primers to the coat protein (CP) coding region (2). PCR amplicons from three samples of Ancho and Mirasol pepper types from Zacatecas, which also tested positive by PCR using the rep coding region (2), were sequenced and compared with reported curtoviruses. The samples showed 91% identity with the CP coding region and 93% identity with the rep coding region of Beet mild curly top virus (formerly the Worland strain). A survey of pepper fields from Aguascalientes and Zacatecas based on symptomatic plants was conducted from July to August of 2005. Forty-three fields of different types of pepper, including those growing under mulch and drip irrigation, were surveyed. Twenty-five plants in each of five contiguous rows were inspected for the symptoms described above. Disease symptoms were noted in Mirasol, Ancho, Pasilla, and Guajillo pepper types, and the average disease incidence was 9.87% (range: 1.6 to 48%), 15.2% (range: 6.4 to 25.6%), 7.85% (range: 2.4 to 15.2%), and 20.8% (range: 8 to 33.6%), respectively. To our knowledge, this is the first report of curtovirus infection of chile pepper in this region of Mexico. The moderate level of curtovirus infection found here suggests the need to initiate management strategies for this disease. References: (1) L. L. Black et al. Page 98 in: Pepper Diseases. A Field Guide. AVRDC, Taiwan, 1991. (2) R. Creamer et al. Plant Dis. 89:480, 2005. (3) D. A. Young and N. W. Frazier. Hilgardia 23:25, 1954.

Relationship between the endophyte Embellisia spp. and the toxic alkaloid swainsonine in major locoweed species (Astragalus and Oxytropis).

Locoweeds (Astragalus and Oxytropis spp. that contain the toxic alkaloid swainsonine) cause widespread poisoning of livestock on western rangelands. There are 354 species of Astragalus and 22 species of Oxytropis in the US and Canada. Recently, a fungal endophyte, Embellisia spp., was isolated from Astragalus and Oxytropis spp. and shown to produce swainsonine. We conducted a survey of the major locoweeds from areas where locoweed poisoning has occurred to verify the presence of the endophyte and to relate endophyte infection with swainsonine concentrations. Species found to contain the fungal endophyte and produce substantial amounts of swainsonine were A. wootoni, A. pubentissimus, A. mollissimus, A. lentiginosus, and O. sericea. Astragalus species generally had higher concentrations of swainsonine than Oxytropis. Swainsonine was not detected in A. alpinus, A. cibarius, A. coltonii, A. filipes, or O. campestris. The endophyte could not be cultured from A. mollissimus var. thompsonii or A. amphioxys, but was detected by polymerase chain reaction, and only 30% of these samples contained trace levels of swainsonine. Further research is necessary to determine if the endophyte is able to colonize these and other species of Astragalus and Oxytropis and determine environmental influences on its growth and synthesis of swainsonine.

Curtovirus Infection of Chile Pepper in New Mexico.

Chile pepper-producing areas of southern New Mexico (NM) were surveyed during 2001 and 2002 to identify which curtoviruses were infecting chile peppers and to determine the distribution of the viruses among fields. Plants with symptoms resembling Beet curly top virus (BCTV) were collected from 10 fields and tested for the presence of curtoviruses by polymerase chain reaction using primers designed to detect a portion of the coat protein (cp) gene, and tested for specific curtoviruses using primers designed to detect to a portion of the replication-associated protein (rep) gene. All amplicons were sequenced and compared with curtoviruses for which complete sequences were available. Amplification was successful from 79.5% of the chile pepper samples. Analysis of the CP sequences showed that more than 92% of the NM field isolates shared high (98 to 100%) amino acid identity with well-characterized curtoviruses. However, eight NM isolates displayed a distinct CP sequence that shared only 86 to 88% amino acid identity with those curtoviruses. Comparison of the rep gene sequence showed that 18.5% of the NM field isolates tested shared 98 to 100% amino acid identity with Beet mild curly top virus (BMCTV), 48% shared 96 to 97% amino acid identity with Beet severe curly top virus (BSCTV), and 32% shared 93 to 97% amino acid identity with BMCTV and BSCTV. Although the distribution of curtoviruses was not identical among all fields sampled, little or no spatial patterns were found among the field isolates. This study revealed the complexity of curtoviruses in a single crop and limited geographical area.

The toxicosis of Embellisia fungi from locoweed (Oxytropis lambertii) is similar to locoweed toxicosis in rats.

Locoweeds cause significant livestock poisoning and economic loss in the western United States. The toxicity of Embellisia sp. fungi isolated from locoweed was compared with locoweed toxicity using the rat as a model. Rats were fed diets containing locoweed, fungus and alfalfa, or alfalfa. Locoweed- and fungus-fed rats consumed swainsonine-containing food at approximately 1.3 mg x kg(-1) x d(-1), gained less weight (P = 0.001) and ate less than controls. Swainsonine is the principal agent responsible for inducing locoism in animals. The concentrations of alkaline phosphatase and aspartate aminotransferase enzymes were greater (P < 0.05) in serum of locoweed- and fungus-fed rats compared with control rats. Similar intracellular vacuolation was observed in renal, pancreatic, and hepatic tissues of rats that consumed either locoweed or fungus. Rats that ate locoweed or Embellisia fungi displayed indistinguishable toxicity symptoms. The Embellisia fungi from locoweed can induce toxicity without the plants. Locoism management strategies need to involve management of the Embellisia fungi.

Iris yellow spot virus on Onion in New Mexico.

Onions are an important crop for New Mexico with 7,700 acres (3,116 ha) harvested in the state in 2003 (3). In 2002, onions of several cultivars were first noticed with diamond-shaped chlorotic or bleached lesions on seed stalks or leaves, typical of those reported for Iris yellow spot virus (IYSV). A more widespread survey of breeding stocks and commercial onion fields revealed similar symptoms on thrips-infested onions in Dona Ana and Rio Arriba counties. Incidence of disease symptoms ranged from <0.5 to nearly 30%. Symptomatic leaves were assayed for the presence of IYSV using enzyme-linked immunosorbent assay (ELISA; Agdia, Elkhart, IN) and antisera acquired from Agdia. Symptomatic leaves from breeding and commercial fields tested positive for IYSV. The virus was transmitted by Thrips tabaci from symptomatic onions to three onion cvs. New Mex Mesa, New Mex Vado, and New Mex Cryspy in growth chamber tests. All three cultivars showed symptoms of IYSV and tested positive for the disease using ELISA. However, New Mex Vado and New Mex Cryspy cultivars each showed 24% infection (4 infected plants of 17 tested) compared with 59% infection (10 infected plants of 17 tested) for New Mex Mesa, suggesting that not all cultivars are equally susceptible to the virus. To our knowledge, this is the first report of IYSV in onions in New Mexico, which has also been reported in the western United States in Idaho, Oregon, Colorado, and Washington (1,2,4). References: (1) L. J. du Toit et al. Plant Dis. 88:222, 2004. (2) J. M. Hall et al. Plant Dis. 77:952, 1993. (3) National Agricultural Statistics Service, On-line publication. USDA, 2004. (4) H. F. Schwartz et al. Plant Dis. 86:560, 2002.

Assessing soil biodiversity across Great Britain: national trends in the occurrence of heterotrophic bacteria and invertebrates in soil.

An assessment of the biodiversity of soils was a component of the Countryside Survey 2000 (CS2000). This was the first integrated survey of soil biota and chemical properties at a national scale. A total of 1052 soil samples were collected across Great Britain during CS2000 and analysed for a range of soil microbial and invertebrate characteristics resulting in the production of a series of robust datasets. A principal objective was to use these datasets to investigate relationships between soil biota and environmental factors such as geographical location, vegetation, land use, land cover, soil type and pollutant levels as first stages in characterising the inherent biodiversity of British soils and investigating the potential of soil biodiversity as indicators of soil health at a regional or national scale. Preliminary results for culturable heterotrophic, invertebrate taxa, Acari, Collembola and Oribatid mites are presented here to illustrate the nature of the data collected and the patterns of soil biodiversity in relation to large-scale regional, vegetation and soil characteristics across the British countryside.

Maize fine streak virus, a New Leafhopper-Transmitted Rhabdovirus.

ABSTRACT A previously uncharacterized virus was isolated from fall-planted sweet corn (Zea mays L., Syngenta GSS 0966) leaves showing fine chlorotic streaks. Symptomatic plants were negative in enzyme-linked immunosorbent assay against many maize viruses, but reacted weakly with antisera to Sorghum stunt mosaic virus suggesting a distant relationship between the viruses. The virus was readily transmitted by vascular puncture inoculation (VPI), but not by leaf-rub inoculation. Symptoms on maize included dwarfing and fine chlorotic streaks along intermediate and small veins that developed 12 to 17 days post-VPI. The isolated virus was bacilliform (231 +/- 5 nm long and 71 +/- 2 nm wide), with a knobby surface, and obvious helical structure typical of rhabdovirus morphology. Nucleorhabdovirus virions were observed by transmission electron microscopy of infected maize leaf tissue sections. Proteins unique to infected plants were observed in extracts of infected leaves, and the isolated virion contained three proteins with molecular masses 82 +/- 2, 50 +/- 3, and 32 +/- 2 kDa. Preliminary sequence analysis indicated the virus had similarity to members of the family Rhabdoviridae. The virus was transmitted by Graminella nigrifrons under persistent conditions. The data indicate the virus, provisionally designated Maize fine streak virus, is a new species in the genus Nucleorhabdovirus.

Pepper mottle virus Causing Disease in Chile Peppers in Southern New Mexico.

The primary pepper producing areas of southern New Mexico were surveyed to identify the viruses causing severe disease in chile peppers over a 2-year period. The survey included weeds commonly found in and around pepper fields. Using indirect enzyme-linked immunosorbent assay (ELISA), Pepper mottle virus (PepMoV) was associated with plants showing mosaic and distortion of foliage and fruit deformation. PepMoV and Cucumber mosaic virus (CMV) were determined based on ELISA to be infecting chile peppers and weeds singly or in combination. Four perennial plant species were infected with PepMoV and CMV, including Solanum elaeagnifolium (silverleaf nightshade), Convolvulus arvensis (field bindweed), and Chamysuraces sp. (small groundcherry), which had not previously been identified as hosts for PepMoV. Some peppers and weeds surveyed were also infected at a lower level by several other plant viruses.

Cell-to-cell movement and assembly of a plant closterovirus: roles for the capsid proteins and Hsp70 homolog.

Diverse animal and plant viruses are able to translocate their virions between neighboring cells via intercellular connections. In this work, we analyze the virion assembly and cell-to-cell movement of a plant closterovirus and reveal a strong correlation between these two processes. The filamentous virions of a closterovirus possess a long body formed by the major capsid protein (CP) and a short tail formed by the minor capsid protein (CPm). Genetic and biochemical analyses show that the functions of these virion components are distinct. A virion body is required primarily for genome protection, whereas a tail represents a specialized device for cell-to-cell movement. Furthermore, tail assembly is mediated by the viral Hsp70 homolog (Hsp70h) that becomes an integral part of the virion. Inactivation of the ATPase domain of Hsp70h results in assembly of tailless virions that are incapable of translocation. A dual role for the viral molecular chaperone Hsp70h in virion assembly and transport, combined with the previous finding of this protein in intercellular channels, allowed us to propose a model of closteroviral movement from cell to cell.

Serological characterization of the 3'-proximal encoded proteins of beet yellows closterovirus.

The 3'-proximal open reading frames (ORFs) of beet yellows closterovirus, California isolate (BYV-CA), were sequenced and the expression of the corresponding proteins analyzed. The nucleotide sequence of ORF 5 (coding for p24) was most conserved compared with ORF 7 (coding for p20) and ORF 8 (coding for p21) among the isolates analyzed. Polyclonal antisera were produced to GST fusion proteins of p24, p20, and p21. Accumulation of p24, CP, p20 and p21 was studied in infected Tetragonia expansa plants and Chenopodium quinoa protoplasts. All four proteins were expressed in all tissues (old leaves, young leaves and stems), and most abundantly in young leaves. The subcellular localization of each protein in different tissues showed that compared with p24, CP and p21, p20 accumulated less in transfected protoplasts. Immunogold labeling in sugarbeet with p24 and CP antisera demonstrated co-localization of p24 and CP in vascular petiole tissues. In infectivity neutralization tests, antisera against p24 and CP greatly reduced transmission of BYV by viruliferous aphids compared with viruliferous aphids fed on preimmune serum or antiserum to p21.

Characterization of Beet Yellows Closterovirus-Specific RNAs in Infected Plants and Protoplasts.

ABSTRACT Tetragonia expansa plants infected with a California isolate of beet yellows virus (BYV-60) contained multiple BYV-specific RNAs identified by Northern blot hybridization. These RNAs were identified by cDNA probes specific to six open reading frames (ORFs). One genomic RNA and five subgenomic (sg) RNAs representing the p65/p6.4, p64, p24, p22, and p21 ORFs were identified. A probe derived from the 3'-terminal ORF (p21) hybridized to each of the sgRNAs, indicating the RNAs are 3' coterminal. Hybridization with 5'- and 3'-end probes indicated that preparations of BYV particles contained the genomic RNA as well as two additional RNA molecules corresponding in size to the coat protein (CP) sgRNA and an unidentified RNA. A Chenopodium quinoa protoplast system also was used to study BYV replication. The temporal accumulation of BYV-specific RNAs and CP was investigated in protoplasts transfected with purified virion RNA. Accumulation of genomic plus-strand RNA was evident as early as 15 h postinoculation. The development of this protoplast system is significant for studies of closterovirus replication.

Transmission of Sorghum Stunt Mosaic Rhabdovirus by the Leafhopper Vector, Graminella sonora (Homoptera: Cicadellidae).

The transmission parameters of sorghum stunt mosaic rhabdovirus (SSMV) by the leafhopper Graminella sonora were determined. The minimum acquisition and inoculation times were 6 and 1 h, respectively, while 100% transmission was obtained from a 48-h acquisition access or a 24-h inoculation access period. Transmission efficiency was highest at temperatures of 24 to 36°C. The minimum latent period prior to transmission was 9 days at 30°C, with an 11-day incubation period necessary for 100% transmission. After a 14-day incubation period at 16°C, no transmission was obtained; however, doubling the incubation period to 28 days gave high levels of transmission. Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) of individual leafhoppers held at 16°C for a 14-day incubation period detected no virus in leafhoppers, whereas over 50% of the leafhoppers had detectable virus after a 28-day incubation period. Graminella nigrifrons and Peregrinus maidis were not able to transmit SSMV.