Characteristics of the Shiga-like toxin produced by Escherichia coli associated with porcine edema disease

Shiga-like toxin (SLT-IIv) from Escherichia coli strains associated with edema disease of pigs was characterized and compared with SLT-I, SLT-II, and the SLT of E. coli strain HI8 (SLT-HI8). SLT-IIv from an E. coli K12 in which the genes for SLT-IIv had been cloned was indistinguishable from SLT-IIv of wild strains of E. coli from edema disease. There was cross-neutralization among all SLTs except SLT-I. The different SLTs could be distinguished by heat lability, with the descending order of heat lability being SLT-IIv, SLT-II, SLT-I, and SLT-HI8. SLT-IIv and SLT-HI8 had lower cytotoxic titers on HeLa cells compared with Vero cells and were more active on MDBK cells than were the other SLTs. All SLTs were enterotoxic in rabbit but not in pig intestine and SLT-IIv was less enterotoxic than SLT-I. SLT-IIv had a lower LD50 in mice than did the other SLTs.     

The effects of antigenic competition on the efficacy of multivalent footrot vaccines

SUMMARY A multivalent footrot vaccine has been developed, containing pilus antigens produced in recombinant Pseudomonas aeruginosa and representing all nine serogroups of Dichelobacter (Bacteroides) nodosus commonly recognised in the field. The responses of sheep to the multivalent vaccine have been compared with those to monovalent vaccines representing only a single serogroup. Antigenie competition between serogroups occurred in sheep immunised with the multivalent formulation, but high levels of protection were still achieved. The study showed that in multivalent footrot vaccines, antigenic competition is predominantly due to the presence of a family of immunologically-related pilus antigens rather than to interference by extraneous proteins.     

Isolation and characterization of nine bacteriophages that lyse O149 enterotoxigenic Escherichia coli

The goal of this study was to isolate and characterize phages that might be used in prevention and treatment of porcine post-weaning diarrhea due to O149 enterotoxigenic E. coli (ETEC). Serotype O149:H10:F4 was especially targeted because this is the dominant ETEC serotype. Mixtures of 10 strains of O149:H10:F4 ETEC and of 10 O149:H43:F4 ETEC were used as hosts for isolation of phages in sewage from 38 Ontario pig farms. Six phages (GJ1-GJ6) that lysed O149:H10:F4 ETEC and three (GJ7-GJ9) that lysed O149:H43:F4 ETEC were isolated. All phages produced large, clear plaques. All nine phages had necks and contractile tails and therefore belonged to the Myoviridae. Their estimated genome sizes were 48.3-50.7kb and their restriction enzyme fragments suggested that they were closely related. Phages GJ1-GJ6 lysed 99-100% of 85 O149:H10:F4 ETEC, 0-12% of 42 O149:H43:F4 ETEC, 3-35% of 37 non-O149 porcine ETEC, and 6-68% of the 72 strains of the ECOR collection. Phages GJ7-GJ9 lysed 86-98% of the O149:H43:F4 ETEC, 2-53% of the O149:H10:F4 ETEC, and 24-41% of the non-O149 porcine ETEC. Titres of the nine phages were unaffected by exposure for 16h to pH 5-9. Among phages GJ1-GJ6, resistance of O149:H10:F4 ETEC to one phage was generally not accompanied by resistance to other phages. It is concluded that the nine phages are suitable candidates for prophylaxis and therapy of porcine post-weaning diarrhea due to O149 ETEC.     

Contribution of AIDA-I to the pathogenicity of a porcine diarrheagenic Escherichia coli and to intestinal colonization through biofilm formation in pigs

In order to evaluate the role of the AIDA-I of porcine diarrheagenic Escherichia coli strain PD20 serogroup O143 (AIDA-I⁺, STb⁺), a mutant strain PD20M (AIDA-I⁻, STb⁺) was generated from strain PD20 by an allelic exchange procedure. In addition, the full-length aidA gene was reintroduced into strain PD20M to generate the complemented strain PD20C (pTaidA, AIDA-I⁺, STb⁺). A non-pathogenic E. coli strain PD71 was used as negative control. Each strain was inoculated to newborn pigs via stomach tube. Severity of diarrhea was evaluated clinically and intestinal colonization was assessed by histology, immunohistochemistry (IHC), and transmission electron microscopy (TEM) including immunogold electron microscopy (IGEM). The adhesion pattern to HeLa cells, bacterial auto-aggregation and biofilm formation were evaluated in vitro. Pigs infected with strains PD20 or PD20C developed diarrhea 16 and 28 h after inoculation, respectively, in contrast to pigs infected with strains PD20M or PD71. Histology, IHC, TEM and IGEM examinations showed heavy bacterial colonization with biofilm formation in the large intestine, and marked in vivo expression of AIDA-I protein in pigs infected with strains PD20 or PD20C in contrast to pigs infected with strains PD20M or PD71. The in vitro assays showed marked diffuse adherence to HeLa cells, enhanced bacterial auto-aggregation and significant biofilm formation (p < 0.05) by the AIDA-I⁺ strains, when compared to AIDA-I⁻ strains. These results demonstrate that expression of AIDA-I is essential for intestinal colonization and in vitro bacterial autoaggregation and biofilm formation. Thus, AIDA-I may be considered a significant virulence determinant in development of diarrhea caused by porcine diarrheagenic AIDA-I⁺ E. coli PD20 in piglets.     

Impact of biodiversity-climate futures on primary production and metabolism in a model benthic estuarine system

Background

A territory as a prerequisite for breeding limits the maximum number of breeders in a given area, and thus lowers the proportion of breeders if population size increases. However, some territorially breeding animals can have dramatic density fluctuations and little is known about the change from density-dependent processes to density-independence of breeding during a population increase or an outbreak. We suggest that territoriality, breeding suppression and its break-down can be understood with an incomplete-control model, developed for social breeders and social suppression.

Results

We studied density dependence in an arvicoline species, the bank vole, known as a territorial breeder with cyclic and non-cyclic density fluctuations and periodically high densities in different parts of its range. Our long-term data base from 38 experimental populations in large enclosures in boreal grassland confirms that breeding rates are density-regulated at moderate densities, probably by social suppression of subordinate potential breeders. We conducted an experiment, were we doubled and tripled this moderate density under otherwise the same conditions and measured space use, mortality, reproduction and faecal stress hormone levels (FGM) of adult females. We found that mortality did not differ among the densities, but the regulation of the breeding rate broke down: at double and triple densities all females were breeding, while at the low density the breeding rate was regulated as observed before. Spatial overlap among females increased with density, while a minimum territory size was maintained. Mean stress hormone levels were higher in double and triple densities than at moderate density.

Conclusions

At low and moderate densities, breeding suppression by the dominant breeders, But above a density-threshold (similar to a competition point), the dominance of breeders could not be sustained (incomplete control). In our experiment, this point was reached after territories could not shrink any further, while the number of intruders continued to increase with increasing density. Probably suppression becomes too costly for the dominants, and increasing number of other breeders reduces the effectiveness of threats. In wild populations, crossing this threshold would allow for a rapid density increase or population outbreaks, enabling territorial species to escape density-dependency.     

Shiga toxin genes in avian Escherichia coli

The purpose of this study was to determine the presence of stx genes in avian pathogenic Escherichia coli (APEC). We examined 97 APEC isolates: 34 from lesions of avian cellulitis, 31 from avian septicemia, 13 from swollen head syndrome (SHS) in chickens, and 19 from diseased turkeys. We also examined five isolates from the feces of healthy chickens. All 102 E. coli isolates were tested for the presence of stx genes by PCR amplification and by colony blots using probes specific for stx1 and stx2. Fifty-three percent (52) of the 97 APEC carried stx gene sequences: one isolate carried stx2 sequences, two carried both stx1 and stx2 sequences, and the remaining 49 isolates carried only stx1 sequences. Twenty-six isolates were positive by both hybridization and PCR amplification, 10 were positive by PCR only, and 16 were positive by hybridization only. All the stx-positive isolates were negative by PCR for the eae and E-hlyA genes. The five isolates from healthy chickens were all negative for stx. All 13 SHS isolates were positive for the stx1 gene and had low titres for cytotoxicity in the Vero cell assay (VCA). Other stx-positive isolates were negative in the VCA. The stx1 gene from one SHS E. coli isolate was cloned and sequenced and shown to be identical to that of the stx gene of Shigella dysenteriae. The observations indicate that stx1 gene sequences are widespread among APEC but that cytotoxicity on Vero cells is uncommon.     

A comparison of anesthetic risk factors and outcomes in light and draft horses

As a result of anatomic and physiologic differences, draft breeds may be at greater risk of developing anesthetic complications. The aim of the study was to evaluate and compare anesthetic management of draft (DR) and light (LT) horses. A case‐matched retrospective study of 371 clinical case records of DR (124 cases) and LT (247 cases) horses presented for general anesthesia between 1991 and 1998 was performed. Data were tabulated and comparisons were made using Student's t‐test (significance p < 0.05). Prior to induction, there were significant differences in mean body weight, rectal temperature, PCV, RBC, and serum TP concentration between DR and LT breeds. There were differences in mean doses of pre‐operative butorphanol (LT 21 µg kg^?1; DR 17 µg kg^?1), induction guaifenesin (LT 99 mg kg^?1; DR 88 mg kg^?1), and intraoperative ketamine (LT 0.35 mg kg^?1; DR 0.56 mg kg^?1) required. There were no significant differences in the mean doses of pre‐operative xylazine, detomidine, or induction barbiturate administered. The mean, average, and maximum concentrations of inspired halothane were significantly higher for DR than for LT horses. Draft horses received 33% less intraoperative IV fluids (8.2 mL kg^?1 hour^?1) than LT horses. Mean anesthetic duration, time to extubation, and standing recovery were not significantly different. Induction complications were not reported for either group. Rates of occurrence of intraoperative bradycardia, hypercarbia, hypoxemia, and metabolic acidosis (SBE, TCO₂, and bicarbonate concentration) did not differ significantly. Average MAP was greater in DR horses, but neither the degree nor the mean duration of hypotension differed between DR and LT horses. Mean PaO₂ was significantly lower in DR (246 mm Hg, 32.8 kPa) than in LT (305 mm Hg, 40.7 kPa) breeds. Draft horses were at greater relative risk of hypoventilation than LT horses. The greater MAP and requirement for halothane and intraoperative ketamine may indicate problems in achieving and maintaining a surgical plane of anesthesia. Draft horses may be at a greater risk of ventilation–perfusion mismatching.     

Combining chlorophyll meter readings and high spatial resolution remote sensing images for in-season site-specific nitrogen management of corn

The chlorophyll meter (CM) has been commonly used for in-season nitrogen (N) management of corn (Zea mays L.). Nevertheless, it has limited potential for site-specific N management in large fields due to difficulties in using it to generate N status maps. The objective of this study was to determine how well CM readings can be estimated using aerial hyper-spectral and simulated multi-spectral remote sensing images at different corn growth stages. Two field experiments were conducted in Minnesota, USA during 2005 involving different N application rates and timings on a corn-soybean [Glycine max (L.) Merr.] rotation field and a corn-corn rotation field. Four flights were made during the growing season using the AISA Eagle Hyper-spectral Imager and CM readings were collected at four or five different growth stages. The results indicated that single multi-spectral and hyper-spectral band or vegetation index could explain 64–86% and 73–88% of the variability in CM readings, respectively, except at growth stage V9 in the corn-soybean rotation field where no band or vegetation index could explain more than 37% of the variability in CM readings. Multiple regression analysis demonstrated that the combination of 2–4 broad-bands or 3–8 narrow-bands could explain 41–92% or 61–94% of the variability in CM readings across the two fields and different corn growth stages investigated. It was concluded that the combination of CM readings with high spatial resolution hyper-spectral or multi-spectral remote sensing images can overcome the limitations of using them individually, thus offering a practical solution to N deficiency detection and possibly in-season site-specific N management in large continuous corn fields or at later stages in corn-soybean rotation fields.