Transfer of verocytotoxigenic Escherichia coli O157, O26, O111, O103 and O145 from fleece to carcass during sheep slaughter in an Irish export abattoir.

The purpose of this study was to investigate carriage and transfer of verocytotoxigenic Escherichia coli (VTEC) O157, O26, O111, O103 and O145 from fleece to dressed carcasses of 500 sheep, and to establish the virulence potential of recovered VTEC. Individual sheep were tracked and sampled (10 g fleece, full carcass swab) through the slaughter process. Samples were examined for the presence of verotoxin (vt1 and vt2) genes using a duplex real-time PCR assay and positive samples were further screened for the presence of the above five serogroups by real-time PCR. VTEC cells were recovered from PCR positive samples by serogroup specific immunomagnetic separation and confirmed by serogroup specific latex agglutination and PCR. Isolates were subject to a virulence screen (vt1, vt2, eaeA and hlyA) by PCR and isolates carrying vt genes were examined by Pulsed-Field Gel Electrophoresis (PFGE). VTEC O26 was recovered from 5/500 (1.0%) fleece and 2/500 (0.4%) carcass samples. VTEC O157 was isolated from 4/500 (0.8%) fleece samples and 3/500 (0.6%) carcass samples. E. coli O103 was recovered from 84/500 (16.8%) fleece and 68/500 (13.6%) carcasses, but only one E. coli O103 isolate (0.2%) carried vt genes. E. coli O145 was recovered from one fleece sample, but did not carry vt genes. E. coli O111 was not detected in any samples. For the four serogroups recovered, the direct transfer from fleece to carcass was not observed with PFGE showing that VTEC O26 isolates from a matched fleece/carcass "pair" were not identical. This study shows that while VTEC O157 are being carried by sheep presented for slaughter in Ireland, other potentially clinically significant verotoxin producing strains (particularly VTEC O26) are emerging.

Tracking verocytotoxigenic Escherichia coli O157, O26, O111, O103 and O145 in Irish cattle.

The purpose of this study was to investigate carriage and transfer of verocytotoxigenic Escherichia coli (VTEC) O157, O26, O111, O103 and O145 from faeces and hide to dressed carcasses of Irish cattle as well as establishing the virulence potential of VTEC carried by these cattle. Individual cattle was tracked and faecal samples, hide and carcass (pre-evisceration and post-wash) swabs were analysed for verotoxin (vt1 and vt2) genes using a duplex real-time PCR assay. Positive samples were screened for the five serogroups of interest by real-time PCR. Isolates were recovered from PCR positive samples using immunomagnetic separation and confirmed by latex agglutination and PCR. Isolates were subject to a virulence screen (vt1, vt2, eaeA and hlyA) by PCR. Isolates carrying vt genes were examined by Pulsed-Field Gel Electrophoresis (PFGE). Of the VTEC isolated, E. coli O157 was the most frequently recovered from hide (17.6%), faeces (2.3%) and pre-evisceration/post-wash carcass (0.7%) samples. VTEC O26 was isolated from 0.2% of hide swabs and 1.5% of faeces samples. VTEC O145 was isolated from 0.7% of faeces samples. VTEC O26 and VTEC O145 were not recovered from carcass swabs. Non-VTEC O103 was recovered from all sample types (27.1% hide, 8.5% faeces, 5.5% pre-evisceration carcass, 2.2% post-wash carcass), with 0.2% of hide swabs and 1.0% of faeces samples found to be positive for VTEC O103 isolates. E. coli O111 was not detected in any samples. For the four serogroups recovered, the direct transfer from hide to carcass was not observed. This study shows that while VTEC O157 are being carried by cattle presented for slaughter in Ireland, a number of other verotoxin producing strains are beginning to emerge.

Cholinergic differentiation factor/leukemia inhibitory factor enhances functional effects of adrenal medulla grafts after hippocampal lesions in rats.

We recently found that adrenal medulla grafts implanted into the hippocampus of rats survived for several months and significantly decreased the deficits produced by hippocampal lesions in the radial maze test [Jousselin-Hossaja et al. (1994) Neuroscience 59, 275-284]. These grafts contained choline acetyltransferase immunopositive chromaffin cells and received cholinergic innervation. In the experiments reported here, adrenal medulla grafts implanted in lesioned hippocampus were treated with cholinergic differentiation factor/leukemia inhibitory factor. In the presence of this factor, the number of chromaffin cells with cholinergic phenotypes increased as well as the beneficial effects of the grafts on the performances of rats in the radial maze. These results suggest that the functional effects of adrenal medulla grafted into the hippocampus set into play cholinergic mechanisms. The cholinergic differentiation factor/leukemia inhibitory factor may also have facilitated the survival and recovery of cholinergic neurons in the host tissue. However, due to the large range of action of this cytokine and the richness of the adrenal medulla contents, non cholinergic factors are also probably involved. Our results may help to elucidate the functions of the cholinergic differentiation factor/leukemia inhibitory factor since they provide the first indication that its intracerebral injection may have behavioral effects. Moreover, our data confirm the possibility of improving the efficiency of adrenal medulla implants in the central nervous system by appropriate treatments, not only by facilitating survival but also by selectively amplifying some potential factors of the graft. This might greatly enlarge the field of this grafting technique for analysing the normal functioning of the brain and for repairing it.

Effects of adrenal medulla grafts on memory capacities of rats after hippocampal lesions.

Behavioral and immunocytochemical techniques were used to study the effects of adrenal medulla grafts implanted in hippocampus--after lesion of this structure--on the memory capacities of rats. Performances of the grafted rats in the radial maze test were significantly improved and, in some aspects, fully restored. On the other hand, grafts had no significant effects on a one-trial spatial recognition test and impaired object recognition. Immunocytochemical identification showed that the grafts contained chromaffin cells with a choline acetyltransferase stainings while, in parallel, phenylethanolamine-N-methyltransferase stainings seemed to be decreased. Cholinergic innervation was established between the graft and the host hippocampus. A likely interpretation of this complex pattern of results is that the functional effects of the grafts depended on the arousal level induced by the behavioral task. At the neurobiological level, these effects probably set into play an interaction between opioid, catecholaminergic and cholinergic factors. Our results may contribute to the clarification of the problem of specificity of functional effects of intracerebral grafts as well as the problem of hippocampal role in learning and memory.