Using a gel-free PCR-ELISA for the molecular identification of wheat genotypes carrying wheat–rye translocations

Current techniques to identify wheat lines possessing the 1RS chromosome are generally unsuitable in relation to the speed and cost needs in modern wheat breeding programmes. A gel-free, direct amplicon capture PCR-ELISA assay was developed and evaluated, aiming at speeding the identification of wheat genotypes possessing the 1RS chromosome arm in breeding programmes. The chosen target sequence was the repetitive, interspersed rye-specific element RIS-1. Primers were end-labelled with digoxigenin and biotin, and amplicons captured on to straptavidine-coated microplates. Subsequent immunodetection of the digoxigenin moiety readily distinguished 1RS from non-1RS control genotypes tested. When a nursery consisting of 120 winter and spring wheat lines was screened by PCR-gel electrophoresis and the PCR-ELISA, a perfect agreement between both techniques was observed. Test robustness, as measured by the tolerance to variations in DNA input, was better for PCR-ELISA than PCR-gel electrophoresis. In conclusion, a simple, robust, fast and scalable technique for the detection of 1RS chromosome carriers in wheat breeding programmes is now available.     

Substituting fishmeal with mixtures of wheat,corn and soya bean meals in diets for the white leg shrimp,Litopenaeus vannamei (Boone): effect on production parameters and preliminary economic assessment

We analysed the effect on production and economic performance of juvenile Litopenaeus vannamei of isoproteic diets substituting fishmeal by 25%, 50%, 75% and 100% with mixtures of wheat, soya bean and cornmeals. In a laboratory trial, 10 juveniles m^?2 (1.1 ± 0.1 g) were reared in 60‐L plastic containers using a recirculation system for 90 days. Three replicates were used to test each diet. A commercial diet serving as a reference and the diet with the highest content of essential amino acids (50% substitution, 6.46 ± 1.1 g) produced significantly higher shrimp final weight (7.12 ± 0.9 g, P < 0.05). There were not significant differences in specific growth rate and mean survival (85.9 ± 0.2%, P > 0.05). In a pond trial, 10 shrimp m^?2 (1.08 ± 0.3 g) were cultivated in 1.5 m^?3 cages for 35 days, testing the diets in triplicate. Final weight was significantly higher (P < 0.05) when 100% substitution was used (10.89 ± 0.24 g), while survival did not differ significantly among diets (P > 0.05). The optimal level of substitution was estimated at 86.0%. Apparently, nutrients contained in the diets combined well with natural feed available in the pond, up to a point where 7.3% of fishmeal inclusion is recommended. An economic analysis showed that 100% substitution produced the best results. We conclude that mixtures of wheat, corn and soya meals are potential alternatives to replace fishmeal effectively in diets for L. vannamei.     

Design and validation of a wireless sensor network architecture for precision horticulture applications

This paper proposes a general wireless sensor network architecture for monitoring horticultural crops that are distributed among small plots scattered at distances of up to 10 km from one another. The technology used for the real implementation of the architecture is based on the B-MAC (Berkeley Medium Access Control) medium access protocol to assure a high degree of sensor node power autonomy. To resolve this issue, a series of specialized sensor nodes (Soil-Mote, Environmental-Mote and Water-Mote) have been developed along with a gateway to interconnect them with the farm central offices. Before starting device development, simulations were conducted to ensure that acceptable performance would be achieved with the selected technology in terms of node autonomy, achieved throughput and delays. To that end, it was necessary to implement the selected B-MAC protocol in the ns-2 (Network Simulator-2) simulation framework. The final system was deployed on a real crop to check and validate the simulation results against experimental results.     

Postpartum follicular development in Brahman cows under two stocking rates

The aim was to study postpartum follicular population dynamics in Brahman cows in the Mexican humid tropics and under different stocking rates. Lactating adult cows (n = 104) grazed native grammas (Paspalum spp., Axonopus spp.) under two stocking rates: 2.8 (high) or 1.7 (low) cows/ha. Follicle numbers and their size (millimetres) were measured by ovarian ultrasound scanning. Animals detected in oestrus and inseminated were taken out of the study. Both stocking rates produced similar (P < 0.05) body condition scores (BCS) and follicular dynamics. The percent of follicles, averaged over stocking rates, of <4 mm diminished and those of 5–8 mm increased with time, whilst those of >9 mm remained constant; respective values for the previously mentioned sizes were 60 days postpartum (n = 101), 49%, 56% and 5%; 120 days (n = 70), 28%, 66% and 6%; and 180 days (n = 15), 9%, 79% and 12%. There was a positive correlation between BCS and follicle number for size <4 mm (r = 0.73, P < 0.01), whilst for size 5–8 mm, this was negative (r = −0.51, P < 0.05), for follicles >9 mm, no difference was found (r = −0.08, P > 0.05). It was concluded that stocking rate did not affect follicular population dynamics of Brahman cows, but BCS dictated the number and categories of follicles present regardless of the stocking rate utilised.     

Behavioral switching in Magellanic woodpeckers reveals perception of habitat quality at different spatial scales

Context

The switching pattern between behavioral modes provides a mechanistic basis for understanding how animals perceive and memorize the habitat quality in their home ranges.

Objectives

We assessed if Magellanic woodpeckers (Campephilus magellanicus) move based on habitat quality at local (neighboring trees) and home range scales.

Methods

We used state-space models to examine the relationship between remotely-sensed estimates of habitat quality (tree decay) and movement of adult woodpeckers tracked with GPS telemetry in southern Chile.

Results

Woodpeckers spent most time (>?80%) in the area-restricted search (ARS) mode in contrast to the exploratory transient mode, characterized by frequent directional displacements (>?50 m). The extent to which individuals switched between behavioral modes was related to habitat quality at different scales. Woodpeckers switched to and remained in the ARS mode when encountering moderate levels of heterogeneity in habitat quality. At very low or high heterogeneity levels, however, individuals switched to and remained in the transient mode, respectively. Likewise, as habitat quality declined locally and across home range, woodpeckers were more likely to adopt a transient mode.

Conclusions

Although woodpeckers seemed to easily perceive and memorize habitat quality at different spatial scales, our results suggest that spatial memory will less effective under extreme levels of habitat heterogeneity.

     

A genome‐wide association study for partial resistance to southern corn rust in tropical maize

Southern corn rust (SCR) is a fungal disease found on corn in several countries worldwide. In Brazil, the disease can result in productivity losses of 65%, especially in areas with a history of the disease. In this study, the genetic architecture and identification of genomic regions associated with SCR resistance was investigated by performing a genome‐wide association study. Genotyping‐by‐sequencing was performed to carry out the association between single nucleotide polymorphism (SNP) markers and phenotypic data from two environments on a panel of 164 maize inbred lines. Eight SNPs were identified as significant for SCR resistance. These SNPs were colocalized with QTL regions, some of which underlie candidate resistance genes with functions that play an important role in the stress response during pathogen recognition. These candidate genes, involved in plant defense pathways, could be associated with partial resistance to SCR and provide a partial comprehensive insight into the genetic architecture of this trait. After validation of the SNPs, they will be useful for marker–assisted selection and for a better understanding of maize resistance to SCR.     

Edwardsiella ictaluri as the causative agent of mortality in cultured Nile tilapia

Edwardsiella ictaluri was consistently isolated from the spleens, livers, and head kidneys of diseased Nile tilapia Oreochromis niloticus from a farm experiencing mortality events in several culture ponds. We describe the first published outbreak of E. ictaluri-induced edwardsiellosis in Nile tilapia. Pure cultures of the isolated bacteria were characterized both biochemically and molecularly. Biochemical analysis was performed using the API-20E and RapID One systems, and antimicrobial susceptibility was determined by the broth microdilution method. Molecular analysis involved sequencing of the 16S rRNA gene, species-specific real-time polymerase chain reaction (PCR), and PCR-mediated genomic fingerprinting (rep-PCR). Pairwise sequence analysis of the 16S rRNA gene identified the case isolates to be a 100% match to E. ictaluri cultured from channel catfish in the southeastern United States. However, rep-PCR analysis identified the case isolates to be genetically different from representative strains isolated from disease outbreaks in cultured channel catfish in Mississippi. Infectivity challenges (intraperitoneal injection and immersion) demonstrated that a representative E. ictaluri strain isolated from tilapia was pathogenic to naive tilapia, reproducing clinical signs and mortality, thereby establishing Koch's postulates.     

Genetic analysis and antimicrobial susceptibility of Francisella noatunensis subsp. orientalis (syn. F. asiatica) isolates from fish

Francisella noatunensis subsp. orientalis (syn. F. asiatica) (Fno) is an emergent fish pathogen that causes acute to chronic disease in a wide variety of freshwater, brackish and marine fish. Due to the emergent nature of this bacterium, established protocols to measure antimicrobial susceptibility are lacking. In this project we compare three different methods to examine the antimicrobial susceptibility (Etest, broth microdilution and disk diffusion) of 10 different isolates of Fno from two different fish species and four different geographic outbreaks from 2006 to 2010. PCR mediated genomic fingerprinting (rep-PCR) performed on the different isolates confirmed genetic homogeneity amongst the different isolates. The in vitro susceptibility data presented here provides important baseline data for future research monitoring the development of antibiotic resistance among Fno isolates as well as provides invaluable data for the development of potential therapeutics.