Inter-batch contamination and potential sources of vancomycin-resistant Enterococcus faecium on broiler farms.

1. Vancomycin-resistant Enterococcus faecium (VRE(fm)) has recently spread among Swedish broiler farms. The objectives were to investigate VRE(fm) persistence within barns between flocks, and to determine whether day-old chicks, feed or forklift trucks used for loading crates could be identified as a means of transmission. 2. Faeces were collected for selective culture from 12 farms (9 culture-positive, 3 culture-negative as determined by prior monitoring), and samples were collected from the barn environment before and after cleaning and disinfection, from forklift tyres, hatcheries and feed. 3. VRE(fm) was isolated only from previously known VRE(fm)-positive farms. The proportions of culture-positive environmental samples were 75% (9 out of 9 farms) prior to and 31% (7 out of 9 farms) after cleaning/disinfection. Five out of 6 samples from forklift tyres were culture-positive. No VRE(fm) were isolated from feed or hatcheries. The majority of 27 vanA gene positive isolates showed similar banding patterns by SmaI restriction digestion and pulse-field gel electrophoresis. No consistent differences were observed regarding management between VRE(fm)-positive and negative farms. 4. We conclude that VRE(fm) contaminates barns and remains present between flocks. Forklift trucks are one possible means of transmission between farms.

Effects of 24-h and 36-h sleep deprivation on human postural control and adaptation.

This study investigated whether human postural stability and adaptation were affected by sleep deprivation and the relationship between motor performance and subjective scores of sleepiness (visuo-anlogue sleepiness scores, VAS). Postural stability and subjective sleepiness were examined in 18 healthy subjects (mean age 23.8 years) following 24 and 36 h of continued wakefulness, ensured by portable EEG recordings, and compared to a control test where the assessments were made after a normal night of sleep. The responses were assessed using posturography with eyes open and closed, and vibratory proprioceptive stimulations were used to challenge postural control. Postural control was significantly affected after 24 h of sleep deprivation both in anteroposterior and in lateral directions, but less so after 36 h. Subjective VAS scores showed poor correlation with indicators of postural control performance. The clearest evidence that sleep deprivation decreased postural control was the reduction of adaptation. Also several near falls after 2-3 min during the posturographic tests showed that sleep deprivation might affect stability through momentary lapses of attention. Access to vision, somewhat, but not entirely reduced the effect of sleep deprivation. In conclusion, sleep deprivation can be a contributing factor to decreased postural control and falls.

Summary of the Swedish Campylobacter program in broilers, 2001 through 2005.

A Campylobacter monitoring program in broiler chickens was carried out in Sweden from 2001 through 2005. The objective was to reduce the occurrence of Campylobacter in the food chain through preventive measures, starting with primary production. The program involved collecting samples from all broiler flocks at slaughter and occasional additional times. The annual incidence of Campylobacter-positive slaughter batches progressively decreased from 20% in 2002 to 13% in 2005. Most of the positive batches had a high within-flock prevalence of Campylobacter. However, about 18% of the positive batches had a low-within-flock prevalence; Campylobacter spp. were isolated from at most 50% of the cloacal samples. The incidence of batches contaminated at slaughter ranged between 6 and 9% during the study period. During all 5 years, a seasonal peak of incidence was observed in the summertime. In an additional study, quantitative analyses were performed on neck skin samples and carcass rinse samples. Those results were compared with the positive and negative findings of the cloacal, cecum, and neck skin samples at slaughter. When Campylobacter was found in the cecum, there was a higher level of Campylobacter in the quantitative analyses. Those batches where Campylobacter already had been found on the farm had a higher concentration of Campylobacter than those batches in which Campylobacter was found only at slaughter. During the study period, about one-third of producers seldom delivered Campylobacter-positive batches (< 10% positive batches per year). Thus, it is possible to produce Campylobacter-free broilers in Sweden.

Genetic characterization of two gain-of-function alleles of the effector caspase DrICE in Drosophila.

Caspases are the executioners of apoptosis. Although much is known about their physiological roles and structures, detailed analyses of missense mutations of caspases are lacking. As mutations within caspases are identified in various human diseases, the study of caspase mutants will help to elucidate how caspases interact with other components of the apoptosis pathway and how they may contribute to disease. DrICE is the major effector caspase in Drosophila required for developmental and stress-induced cell death. Here, we report the isolation and characterization of six de novo drICE mutants, all of which carry point mutations affecting amino acids conserved among caspases in various species. These six mutants behave as recessive loss-of-function mutants in a homozygous condition. Surprisingly, however, two of the newly isolated drICE alleles are gain-of-function mutants in a heterozygous condition, although they are loss-of-function mutants homozygously. Interestingly, they only behave as gain-of-function mutants in the presence of an apoptotic signal. These two alleles carry missense mutations affecting conserved amino acids in close proximity to the catalytic cysteine residue. This is the first time that viable gain-of-function alleles of caspases are described in any intact organism and provides a significant exception to the expectation that mutations of conserved amino acids always abolish the pro-apoptotic activity of caspases. We discuss models about how these mutations cause the gain-of-function character of these alleles.

Surveillance programme for Campylobacter species in Swedish broilers, July 2001 to June 2002.

A total of 20,599 samples from 4133 slaughter groups representing about 71 million broilers slaughtered between July 1,2001 and June 30, 2002 were tested for Campylobacter species. From each slaughter group, 40 cloacal samples and 10 samples of neck skin were taken from the carcases on the slaughter line and the samples were pooled, 10 to a pool. Campylobacter species were detected in cloacal samples from 17 per cent of the slaughter groups and in neck skin samples from 22 per cent of the groups, and the monthly prevalence figures ranged from 6 to 32 per cent. In most of the infected groups all four pooled cloacal samples were positive, but 18 per cent of the infected groups had only one or two positive samples. More than 90 per cent of the Campylobacter-positive slaughter groups were delivered by half the producers, and one in four of the producers delivered no Campylobacter-positive slaughter groups.

Non-cell autonomous control of apoptosis by ligand-independent Hedgehog signaling in Drosophila.

Hedgehog (Hh) signaling is important for development and homeostasis in vertebrates and invertebrates. Ligand-independent, deregulated Hh signaling caused by loss of negative regulators such as Patched causes excessive cell proliferation, leading to overgrowth in Drosophila and tumors in humans, including basal-cell carcinoma and medulloblastoma. We show that in Drosophila deregulated Hh signaling also promotes cell survival by increasing the resistance to apoptosis. Surprisingly, cells with deregulated Hh activity do not protect themselves from apoptosis; instead, they promote cell survival of neighboring wild-type cells. This non-cell autonomous effect is mediated by Hh-induced Notch signaling, which elevates the protein levels of Drosophila inhibitor of apoptosis protein-1 (Diap-1), conferring resistance to apoptosis. In summary, we demonstrate that deregulated Hh signaling not only promotes proliferation but also cell survival of neighboring cells. This non-cell autonomous control of apoptosis highlights an underappreciated function of deregulated Hh signaling, which may help to generate a supportive micro-environment for tumor development.

A population-based dietary inventory of cooked meat and assessment of the daily intake of food mutagens.

Frequent consumption of meat has been associated with an increased risk of colorectal cancer. Such a risk may be due to naturally occurring compounds in the meat, substances added to the meat, or agents formed during cooking. Concerning the latter alternative, mutagenic heterocyclic amines are multi-site animal carcinogens, but their relevance to human cancer has yet to be determined. In the present study, we made a population-based inventory of cooked meat dishes consumed in the county of Stockholm, ranked dishes according to cooking method and frequency of consumption and, in addition, determined levels of mutagenic activity in six commonly consumed fried meat dishes. Meat was consumed, on average, 493 times per year, giving 1.4 daily servings. Frying was the most common way to cook meat. When ranking meat dishes according to intake frequency, the top eight dishes were as follows: sausage, steak casserole, meatballs, pork chops, pork belly, bacon, ground beef patties, and finally, mince-meat sauce. The frying sessions were performed under controlled conditions at four different temperatures, and we documented the degree of surface browning and measured mutagenic activity in six frequently eaten dishes (sausage, meatballs, pork chops, pork belly, ground beef patties, and minute beef). We found extracts from all six dishes to be mutagenic, and a mean daily dose of exposure was calculated, giving 862 revertants. This investigation leaves no doubt that a major portion of the total meat consumption is fried before ingestion and that fried meat dishes frequently consumed by an elderly population in Stockholm contain mutagenic substances. Furthermore, the study provides usable information for future epidemiological research in which it is necessary to disentangle the effect of meat per se from the effect of potentially carcinogenic heterocyclic amines.

Expression of insulin-like growth factors during bone induction in rat.

Bone formation was studied after intramuscular implantation of demineralized bone matrix. Ash weight determinations were used to verify the bone-forming ability of implants, and confirmed that no bone was formed when nonactive implants (stripped of their bone-forming ability) were used. A solution hybridization/RNase protection assay was used for the detection of specific mRNA transcripts in the implants and surrounding tissue. Analysis of insulin-like growth factor I (IGF-I) mRNA showed a transient increase peaking on day 3 following implantation. Radioimmunoassay (RIA) for IGF-I-like immunoreactivity indicated a corresponding increase of IGF-I peptide in extracts from the implants at that time point. IGF-II mRNA and alkaline phosphatase mRNA reached highest levels around day 11 following implantation. Bone formation in old rats, 50 weeks of age, was associated with lower IGF-I mRNA levels 3 days after implantation compared with young animals. IGF-II mRNA levels were also affected and tended to be higher 12 days after implantation compared with young animals. These results indicate that IGFs could be paracrine or autocrine factors in the bone-forming process. During this process, IGF-I mRNA is expressed at an early stage, in correlation with the recruitment and proliferation of surrounding mesenchymal cells, whereas IGF-II mRNA is activated significantly later, correlating to the beginning of the actual calcifying process during endochondral bone formation.