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Aims: To assess the prevalence of Mycobacterium bovis bacilli in faecal samples of tuberculous cattle, and to better understand the risk of environmental dissemination of bovine tuberculosis (TB) through the spreading of manure or slurry. Methods and Results: Faecal samples were collected from 72 naturally infected cattle with visible lesions of TB that had reacted to the tuberculin skin test and 12 cattle experimentally infected with M. bovis. These were examined by microbial culture and PCR to assess the presence of M. bovis bacilli. There were no positive cultures from any naturally infected test reactor animal. A single M. bovis colony was cultured from a faecal sample from one of the experimentally infected animals. A single PCR positive result was obtained from the faecal sample of one naturally infected test reactor. Conclusions: The prevalence of M. bovis in the faecal samples of TB‐infected cattle was extremely low. Significance and Impact of the Study: The results suggest that the risk of spreading TB through the use of slurry or manure as an agricultural fertilizer is lower than that suggested in some historical literature. The results could inform a reconsideration of current risk assessments and guidelines on the disposal of manure and slurry from TB‐infected herds. [ABSTRACT FROM AUTHOR]

Aims The study aimed to evaluate the effects of probiotic Lactobacillus reuteri DSM 17938 strain supplementation in patients suffering from systemic nickel allergy syndrome, in terms of modulation of faecal LAB population linked to a reduction of GI and cutaneous symptoms and to an increase of patient's quality of life. Methods and Results A preliminary double-blind randomized placebo-controlled study was planned and a culture-independent method based on denaturing gradient gel electrophoresis (DGGE) analysis coupled to the 16S rRNA gene sequencing was applied to investigate on the dynamics of faecal LAB communities before and during a low-Ni diet, supplemented with the probiotic strain. Moreover, the severity and the frequency of GI and cutaneous reactions as well as patient's clinical condition perception (VAS scores) were estimated by statistical analysis. PCR- DGGE fingerprinting obtained using LAB-specific primers revealed significant shift in faecal community with an increase in biodiversity in patients supplemented with probiotic Lact. reuteri strain. In addition, GI reactions such as symptoms related to meals and type of stools significantly improved only in patients treated with Lact. reuteri, while severity and frequency of cutaneous symptoms (urticaria, itch and eczema) and recurrent abdominal pain ( RAP) as well as VAS scores statistically decreased in both groups. Conclusions Our preliminary findings suggest that probiotic Lact. reuteri could be a useful supplementation during a low-Ni diet of patients with SNAS, to increase LAB population diversity, which could contribute to restore the intestinal homoeostasis conditions. Significance and Impact of the Study To date, no information is available on probiotics application and on their effects, in terms of intestinal microbiota modulation, on patients suffering from SNAS. Therefore, the identification of dominant LAB community and the study of its shift during the probiotic supplementation could enhance the knowledge of the SNAS syndrome. [ABSTRACT FROM AUTHOR]

Clostridium difficile is the major cause of nosocomial diarrhoea. Several detection methods are available for the laboratory diagnosis of C. difficile, but these vary in terms of sensitivity and specificity. In this study, we compared the performance of three following laboratory tests to detect C. difficile: in-house real-time PCR aiming for toxin B gene ( tcdB), EIA for detection of toxins A and B ( Premier Toxins A & B) and C. difficile culture in selective medium (bio Merieux). Our results were grouped into three categories as follows: (1) C. difficile-associated diarrhoea ( CDAD); (2) asymptomatic carriers; and (3) negative results. Among the 113 patients included in the study, 9 (8.0%) were classified as CDAD, 19 (16.8%) were asymptomatic carriers, 76 (67.2%) had negative results and 9 (8.0%) could not be categorized (positive test for C. difficile toxins only). PCR was found to be the most sensitive diagnostic test in our study, with the potential to be used as a screening method for C. difficile colonization/ CDAD. Diagnosis of CDAD would be better performed by a combination of PCR and EIA tests. [ABSTRACT FROM AUTHOR]

Aims: To develop a novel Vero cell assay that implements a real-time cell electronic sensing (RT-CES) system for the determination of the presence of verotoxin-producing Escherichia coli (VTEC). The assay overcomes the major drawbacks in conventional Vero cell assay, for example, labour-intensive and time-consuming. Methods and Results: Cells were grown onto the surfaces of microelectronic sensors that are integrated into the bottom surfaces of the microtiter plate. Cellular viability was monitored in real-time and quantified based on changes in the sensor's electrical impedance. For cell viability measurement, the data generated on the RT-CES system correlated well with those obtained by the Vero cell assay for Verotoxins. To assess cytotoxicity, test cells growing on microelectronic sensors were treated with either supernatant from pure cultures, or stool samples. The specific neutralizing antibodies of VT1 and VT2 were used to identify specific toxins in the samples. Conclusions: The RT-CES assay provides a sensitive measurement comparable to conventional crystal violet assay. The assay has been successfully and specifically used to identify VTEC in human faecal samples. Significance and Impact of the Study: The RT-CES assay significantly shortens the testing time from 48 to 72 h required by the crystal violet assay to only 15 h with automated operation. [ABSTRACT FROM AUTHOR]

Andersen, S. M., Teilmann, J., Harders, P. B., Hansen, E. H., and Hjøllund, D. 2007. Diet of harbour seals and great cormorants in Limfjord, Denmark: interspecific competition and interaction with fishery. – ICES Journal of Marine Science, 64: 1235–1245. Comparative studies on seasonal and regional variation in the diet of harbour seals and great cormorants were conducted in Limfjord, a semi-closed water system in northwest Denmark. To compare harbour seal diet from an open water system containing similar prey species, a small diet analysis from the western Baltic is included. Seal diet during spring reflected the abundance of Atlantic herring entering Limfjord to spawn (90% of the weight consumed), whereas during summer and autumn, seal diet was rather more mixed. The diet of seals in the Rødsand area and cormorants in Limfjord showed no marked seasonal trends. During spring, there was little overlap between seal and cormorant diets in Limfjord because seals fed almost exclusively on Atlantic herring, and they consumed significantly larger herring than did the cormorants. During summer and autumn, seal and cormorant diets overlapped markedly, although the fish items consumed by seals were generally larger. Few commercially targeted species were found in the stomachs and scats of seals and casts of great cormorants, but Atlantic herring were taken by the seals at a size greater than that allowed by the fishery. [ABSTRACT FROM AUTHOR]

Andersen, S. M., Teilmann, J., Harders, P. B., Hansen, E. H., and Hjøllund, D. 2007. Diet of harbour seals and great cormorants in Limfjord, Denmark: interspecific competition and interaction with fishery. – ICES Journal of Marine Science, 64: 1235–1245. Comparative studies on seasonal and regional variation in the diet of harbour seals and great cormorants were conducted in Limfjord, a semi-closed water system in northwest Denmark. To compare harbour seal diet from an open water system containing similar prey species, a small diet analysis from the western Baltic is included. Seal diet during spring reflected the abundance of Atlantic herring entering Limfjord to spawn (90% of the weight consumed), whereas during summer and autumn, seal diet was rather more mixed. The diet of seals in the Rødsand area and cormorants in Limfjord showed no marked seasonal trends. During spring, there was little overlap between seal and cormorant diets in Limfjord because seals fed almost exclusively on Atlantic herring, and they consumed significantly larger herring than did the cormorants. During summer and autumn, seal and cormorant diets overlapped markedly, although the fish items consumed by seals were generally larger. Few commercially targeted species were found in the stomachs and scats of seals and casts of great cormorants, but Atlantic herring were taken by the seals at a size greater than that allowed by the fishery. [ABSTRACT FROM PUBLISHER]

Aims: Escherichia coli O157 is considered to be one of most important human pathogens of animal origin which causes serious clinical complications. One of the most common methods to isolate E. coli O157 is the immunomagnetic separation (IMS) technique which employs specific antibodies coupled to magnetic beads to bind and extract cells from enrichment broths followed by plating onto sorbitol MacConkey agar supplemented with cefixime and potassium tellurite (CT-SMAC) plates. The aim of this study was to determine strain variation by pulsed-field gel electrophoresis (PFGE) among E. coli O157 on IMS/CT-SMAC plates. Methods and Results: Every suspect colony of E. coli O157 was tested following isolation by the IMS/CT-SMAC technique. From 124 colonies detected; six XbaI-PFGE profiles were identified. Conclusions: Our results demonstrate that mixed populations of E. coli O157 with distinguishable PFGE profiles that are simultaneously present in bovine faeces can be isolated with IMS/CT-SMAC technique. Significance and Impact of the Study: If the aim of the study was to analyse diversity of PFGE profiles of E. coli O157 in a faecal sample following isolation by the IMS/CT-SMAC technique, at least five colonies per sample should be analysed to detect different PFGE subtypes if present. [ABSTRACT FROM AUTHOR]

Aims: To determine carriage rates and densities of methanogens and sulfate-reducing bacteria in adults and children using molecular methods, and to also determine if a reciprocal relationship exists between these organisms. Methods and Results: Real-time PCR was used to detect and quantify methanogens and sulfate-reducing bacteria. Real-time PCR was more sensitive than breath methane measurements. Real-time PCR assays were applied to faecal DNA samples collected from 40 children and 12 adults. Methanogens were present in 25% of the children and 42% of the adults studied, and sulfate-reducing bacteria were detected in 15% of the children and 58% of the adults. High levels of sulfate-reducing bacteria were found in two methanogenic adults. Conclusions: Carriage rates and densities of methanogens and sulfate-reducing bacteria are greater in adults than in children. Competition does not necessarily lead to the predominance of one group in the faecal microflora. Significance and Impact of the Study: This study describes sensitive, molecular assays that could be used to monitor these organisms in gastrointestinal disease. Therapeutic exclusion of one group from the bowel would not necessarily lead to the expansion of the other, as there does not appear to be a reciprocal relationship between these groups. [ABSTRACT FROM AUTHOR]