Colonisation of conventional weaned pigs by enteropathogenic Escherichia coli (EPEC) and its hazard potential for human health

Enteropathogenic Escherichia coli (EPEC) bacteria frequently cause severe enteric diseases primarily in children and in young rabbits. Their pathogenicity for pigs has been tested by oral infection of colostrum-deprived newborn, and of severely immunosuppressed weaned pigs, but colonisation of conventional weaned pigs by porcine EPEC has not been experimentally studied. EPEC show similarities to enterohaemorrhagic E. coli (EHEC) additionally carrying shiga toxin genes integrated into the chromosome by lambdoid phages. We have demonstrated earlier that the porcine EPEC prototype strain P86-1390 (O45) could be transduced in vivo (in ligated loops of weaned pigs), by Stx2 phage derived from a human EHEC. Thus, the ability of this porcine EPEC strain to colonise conventional weaned pigs under farming conditions became a question of relevance to human health. To clarify this question, four intragastric infection experiments were performed on a total of 95 conventional weaned pigs. The EPEC P86-1390 and other well-characterised porcine EPEC strains were applied to 54 pigs, leaving 41 weaned pigs as negative controls. In three experiments moderate predispositions were applied: coinfections with enterotoxigenic E. coli (ETEC) or with low-virulence TGE coronavirus, application of fumonisin B1 with a normal therapeutic dose of dexamethasone, and the increase of soybean protein concentration in the feed. A total of 41 weaned pigs served as negative controls inoculated with a commensal porcine E. coli. Housing conditions simulated the farm environment. As an overall result, ileal segments of 18.5% of infected pigs were shown to be colonised by EPEC, while no EPEC were detected in the ilea of controls. Among predisposing factors occurring on farms, feed protein content increased by 20% (26.3% crude protein, provided by 48% soybean meal) seemed to enhance EPEC colonisation and resulted in the mobilisation of spontaneous latent EPEC/ETEC infection. The results indicate that under normal farm conditions porcine EPEC may colonise conventional weaned pigs by inducing ileal attaching effacing (AE) lesions with reasonable frequency, without clinical signs. The results also suggest that conventional weaned pigs may represent undetected reservoirs of porcine EPEC, potentially giving rise to the emergence of new types of EHEC due to natural transduction by Stx phages.     

Microarray based comparative genotyping of gentamicin resistant Escherichia coli strains from food animals and humans

Recent data from the European and Hungarian Antimicrobial Resistance Monitoring Systems have indicated that the routine use of gentamicin in human and veterinary medicine frequently leads to the selection of gentamicin resistance in Escherichia coli. The aim of this study was to provide molecular characterization of gentamicin resistance in clinical and commensal E. coli strains representing humans and food producing animals by genotyping for antimicrobial resistance and virulence using a miniaturized microarray. All 50 strains tested proved to be multidrug resistant defined as resistance to three or more antimicrobial classes. Antimicrobial resistances genes such as aadA1-like, strB, bla(TEM), sul1 and tet(A) or tet(B), and corresponding phenotypes (streptomycin-, ampicillin-, sulfamethoxazole- and tetracycline resistance) were detected in >50% of isolates regardless of the host or clinical background. However, certain genes encoding gentamicin resistance such as aac(6')-Ib and ant(2″)-Ia as well as catB3-like genes for phenicol resistance were only detected in human isolates. Among virulence genes, the increased serum survival gene iss was predominant in all host groups. Although the majority of gentamicin resistant E. coli strains were characterized by diverse antimicrobial resistance, and virulence gene patterns, accentuated links between catB3-like, aac(6')-Ib, bla(CTX-M-1) and sat genes could be detected in human strains. Further resistance/virulence gene associations (tet(A) with iroN and iss) were detected in poultry strains. In conclusion, the simultaneous characterization of antimicrobial resistance and virulence genotypes of representative clinical and commensal strains of E. coli should be useful for the identification of emerging genotypes with human and or animal health implications.     

Breed identity and leadership in a mixed flock of sheep

The aim of this study was to analyze the breed identity and leadership in a mixed flock of sheep. The flock consisted of 89 Suffolk adult ewes and 250 Columbia ewes and 45 Columbia rams. The animals were kept in the pasture during the day. Each hour the flock was allowed to feed for 20 minutes, and then was moved for 40 minutes to a small paddock near the grazing area. In the afternoon, all sheep were taken to the night pen. The animals were observed for 120 days, 4 times a day (480 leadership breed positions), with respect to the following conditions: (1) leave pen at front of flock, (2) return to pen at front of flock, (3) first to move to the grassland, (4) return to the paddock, (5) drink first (on arrival time at the night pen), and (6) preference for an area in the night barn. The Suffolk breed were first in leaving (94% of the times) and in returning to the night pen (65%) (P < 0.05), and 35% returned with the Columbia sheep. The Suffolk breed were first to move to the grazing area (90%) and 10% moved with the other breed (P < 0.05). When returning to the rest area, the Columbia sheep were first to reach 54% of the time, the 2 breeds were together 29% of the time, and the Suffolk sheep were alone 17% of the time (P < 0.05). When comparing who drank first, 50% of the time both breeds went together, 28% of the time the Columbia sheep were first, and 22% of the time the Suffolk sheep were first (P < 0.05). At night, most of the Suffolk sheep had a preference for 1 barn area (P < 0.05). The results of our study suggest that most animals of each breed performed activities together. Suffolk sheep were leaders when moving to the grazing area and when selecting the place to rest.     

Dating branch growth units in a tropical tree using morphological and anatomical markers: the case of Parkia velutina Benoist (Mimoso?deae)

Context

In tropical areas, studies based on the retrospective analysis of tree development have focused principally on growth ring research. The interpretation of primary growth markers is overlooked although it opens perspectives to provide long time-series on tree-crown development.

Aims

This study focused on Parkia velutina, an emergent tree of neotropical rain forests. Our objectives were (1) to characterize the phenological cycle of this species, and (2) to identify temporally interpretable morphological and anatomical markers.

Methods

We collected dominant branches in 14 adult trees and identified growth markers that limit longitudinal and radial increments. We coupled this approach with a 2-year phenological survey of 20 trees.

Results

Leaf shedding, growth unit elongation and growth ring formation define the phenological cycle. At tree scale, this cycle is synchronous and affects all axes. At population scale, trees can be desynchronized. This cycle is annual despite some slight variability. Successive growth units and growth rings are easily identifiable.

Conclusion

Dating a branch by counting the number of growth units or growth rings is possible in many years with a reasonable error. Nevertheless, estimating their precise month of formation in order to study climatic influences remains difficult.     

Phenotypic plasticity and climatic adaptation in an Atlantic maritime pine breeding population

Context

Pinus pinaster Ait. is found in the Iberian Peninsula under Mediterranean and Atlantic conditions. Both climates encounter each other in Galicia (NW Spain), where two bioclimatic regions can be differentiated: coastal and inland. A breeding program was launched in the coastal area, with two breeding and deployment areas delimited.

Aims

We analyse plasticity patterns across regions in a coastal breeding population to assess the suitability of current breeding areas and how genetic material will likely respond to future climate.

Methods

Total height at ages 3 and 8?years was assessed in 16 trials established along the coast and in inner Galicia. Clustering of environments with similar genotypic performance, family sensitivities to climatic factors and stability analyses were performed.

Results

Sizeable genetic variation in plasticity was found among families, and crossover genotype-by-environment interactions were detected within and between regions. It was unfeasible to regionalize Galicia into alternative areas of stable genotypic performance. Only the cold regime was found to noticeably underlie the array of phenotypic responses to changing environmental conditions.

Conclusions

Results suggest that previous delimitation in two breeding areas is pointless and indicate reduced effects of a changing climate towards Mediterranean conditions on decreasing population fitness.     

The genetic diversity associated with seed proteins in a collection of Spanish underground vetches (<Emphasis Type="Italic">Vicia sativa</Emphasis> L. subsp. <Emphasis Type="Italic">amphicarpa</Emphasis> (Dorthes) Asch. et Graebn.)

The characterization of plant genetic resources is the first step towards improving their use. The Spanish Plant Genetic Resources Centre, which belongs to the National Institute for Agricultural and Food Technology Research (CRF-INIA), conserves accessions of wild underground vetches collected in Spain. In the present work, 26 underground vetch accessions were characterized in terms of their seed storage proteins (separated by SDS-PAGE) as a means of assessing the genetic variation of these plants and their agronomic value. Vicia sativa cv. Vereda was used as control. A total of 54 bands were detected, of which 49 were polymorphic. Fifty eight different electrophoretic patterns were observed in total. Protein bands were scored in terms of their presence (1) or absence (0) for all the seeds studied, and two matrices constructed, one with all the bands present in each accession, the other with the different patterns for each accession. Dendrograms based on the Jaccard similarity index and the UPGMA clustering method were produced from these matrices, and the degree of genetic variation between and within accessions was calculated. The groups obtained were compared with the chromosome number for each accession. The results reflect the great diversity of underground vetch seed storage proteins. The aerial and underground seeds of 16 accessions were then analysed separately. In some cases, the aerial and underground flowers of the same plant produced different proteins.     

Physicochemical and Enzyme Characterization of Small and Large Starch Granules Isolated from Two Maize Cultivars

In this study, some morphological, physicochemical, and the initial characterization of the starch granule enzymes of blue and white maizes of small and large granules are described. Starch was isolated from blue and white maizes, and the small and large granules were separated. The efficiency of separation was higher in large granules than small of the blue maize starch. The amylose content was slightly greater (1.3%) in large granules than smaller in both starches studied. No differences in the average gelatinization temperature were found between large and small granules, but the enthalpy of gelatinization value was higher in the small granules. The electrophoretic pattern showed that the granule‐bound starch synthase (GBSSI) had higher expression in large than small granules and that explain the higher amylose content in the former granules. The differences showed in the starch biosynthesis enzymes in small and large granules might explain partially the physicochemical and functional properties of maize.     

Characterization of the functional domain of major surface protein 1a involved in adhesion of the rickettsia Anaplasma marginale to host cells

The major surface protein (MSP) 1a of the genus type species Anaplasma marginale (Rickettsiales: Anaplasmataceae) has been shown to mediate adhesion, infection and transmission of the organism, as well as to contribute to protective immunity in cattle. MSP1a contains a variable number of tandemly repeated peptides in the amino-terminal region, while the remainder of the protein is highly conserved among isolates. The number of repeats varies among geographic isolates of A. marginale but is constant within an isolate and has been used as a stable genetic marker of isolate identity. Because the sequence of the tandem repeats is the most variable part of the protein among isolates, this region of the protein is most likely to be involved in adhesion to host cells, a prerequisite to infection. The purpose of this study was to characterize the organization and function of the MSP1a tandem repeats of A. marginale in adhesion to host cells. We demonstrated by use of recombinant mutant proteins that the tandemly repeated region of MSP1a was necessary and sufficient to mediate adhesion of MSP1a to tick cells and bovine erythrocytes. Synthetic peptides representing the predominant sequences of individual repeats were tested for their adhesive capacity for tick cell extract (TCE). Peptides containing acidic amino acids D or E at position 20 bound to TCE, while peptides with a G as the 20th amino acid were not adhesive to TCE. Antibodies produced in rabbits against a synthetic repeat peptide neutralized A. marginale infection of cultured tick cells, and the neutralization observed was similar to that effected by antibodies produced against the whole MSP1a recombinant protein. Analysis of tandemly repeated MSP1a peptides of several geographic isolates of A. marginale revealed a complex relationship between the msp1alpha genotype and the tick-transmissible phenotype of the isolate and suggested that both the sequence and conformation of the repeated peptides influenced the adhesive properties of MSP1a. These studies demonstrated that the tandemly repeated region of the protein mediates the adhesive function of MSP1a.     

Mapping of QTLs in tomato line FLA456 associated with resistance to a virus causing tomato yellow leaf curl disease

Tomato (yellow) leaf curl disease (TYLCD) is a serious threat to tomato production in the tropics and subtropics. The genetics of resistance to Tomato yellow leaf curl Thailand virus Taiwan strain (TYLCTHV-[TW]) in a highly resistant tomato line FLA456 was studied through quantitative trait loci (QTL) analysis. Four QTLs named qTy4.1, qTy6.1, qTy10.1 and qTy11.1 were detected on chromosomes 4, 6, 10, and 11, respectively, through evaluation of an F₆ recombinant inbred line (RIL) population derived from a cross between FLA456 (resistant) and CLN1621L (susceptible). Gene action of all QTLs was recessive based on disease reaction of the F₁. The markers SINAC1 and SLM4-34 flanked qTy4.1 on chromosome 4, and SLM11-12 and SLM11-17 defined qTy11.1, which co-located with the previously identified Ty-5 and Ty-2 loci, respectively. qTy6.1 was flanked by the markers SLM6-55 and TES-0014, and qTy10.1 by the markers SLM10-80-SLM10-46 on chromosomes 6 and 10. The LOD values of the putative QTLs ranged from 2.79 to 13.76. The phenotypic variance explained by each QTL ranged from 7.1 to 31.9 %. The four QTLs collectively contributed about 60.5 % of the phenotypic variation in resistance against TYLCTHV-[TW]. Group mean severity scores of those RILs possessing three or four qTy were generally lower than RIL groups with only one or no qTy. Given the diversity of begomoviruses that cause TYLCD across the regions, the new QTLs from FLA456 would be valuable in tomato breeding for developing varieties with durable resistance. Two QTL intervals (qTy4.1 and qTy10.1) contained virus resistance candidate genes such as CTV.22 and eukaryotic translation initiation factor 4E.