Child well-being in early childhood education and care during COVID-19: Child sensitivity in small, fixed groups.

The article explores child well-being in Danish early childhood education and care (ECEC) during the time of COVID-19. A phased reopening of Denmark occurred in spring 2020 under strict health guidelines. Two ECEC institutions were followed first-hand to observe the impact of the pandemic on pedagogy and child well-being. Observations and interviews were conducted with follow-up interviews and an online survey a year later. The findings suggest that the pandemic caused pedagogues to work in a more child-sensitive way with elevated staff/child ratios and children in small, fixed groups; however, child well-being was not negatively affected, despite the acute situation.

The use of smoke as a strategy for masking boar taint in sausages and bacon.

Smoke has often been recommended as a masking agent for boar taint. However, guidelines on how much smoke is necessary have not been established. We compared different smoking parameters in bacon (smoking times) and sausages (smoking times and use of liquid smoke). In streaky bacon from entire male pigs with skatole concentrations of up to 0.6 µg/g and androstenone concentrations of up to 5.8 µg/g in the neck fat, three smoking times were compared: 10, 30 and 60 min. Boar taint was partially, but not fully, masked. The longer the smoking time, the better the masking effect. In sausages from entire male pigs with an average skatole concentration of up to 0.6 µg/g and androstenone concentration of up to 3.6 µg/g (the meat part) or 2.4 µg/g (the fat part) in the neck fat, smoking for 40 and 80 min fully masked the boar taint, whereas only a minor effect was seen after 10 and 20 min smoking. Liquid smoke (0.1%) did not mask boar taint when added to sausages from entire male pigs with an average skatole concentration of 0.36 µg/g (meat) or 0.38 µg/g (fat) and androstenone concentration of 2.3 µg/g (meat) and 2.9 µg/g (fat). To effectively mask boar taint, an intense smoked flavour is therefore necessary, and the longer the smoking time, the better. In contrast, the use of liquid smoke mixed into the product was not effective in the concentrations used in the current study.

The effect of salt reduction on sensory quality and microbial growth in hotdog sausages, bacon, ham and salami.

Sodium chloride (NaCl) is a multi-functional ingredient used to inhibit microbial growth and to ensure good texture and taste in processed meat. This study showed how moderately (22-25%) and greatly (43-50%) reduction of NaCl affected yield, sensory quality and microbial growth in hotdog sausages, bacon, cooked cured ham and salami. In greatly reduced products, the yield was reduced by 8% in sausages and 6% in ham, whereas the yield in bacon and salami remained unaffected. The microbial growth was generally not affected by reducing the content of NaCl to 2.0% in sausages, 2.3% in bacon, 1.7% in ham and 6.3% in salami (aqueous phase). Salt taste, juiciness and texture were the sensory parameters most affected by the NaCl reduction. In sausages and ham, reduction from 2.2% to 1.7% and from 2.3% to 1.3% (w/w), respectively, did not alter the sensory properties. In contrast, the sensory properties of bacon and salami were significantly affected already after a moderately reduction.

Listeria monocytogenes efficiently invades Caco-2 cells after low-temperature storage in broth and on deli meat.

The objective of this study was to investigate how various growth conditions influence the virulence of Listeria monocytogenes monitored by its ability to invade the epithelial cell lines Caco-2 and INT-407. The growth conditions examined were modified atmosphere-packaged deli meat and brain heart infusion broth (BHI) with and without salt. Five strains of L. monocytogenes were selected to investigate their invasiveness and all strains invaded Caco-2 cells at higher levels than INT-407 cells. Further, the clinical strains (3443 and 3734) were more invasive (p < 0.05) than the strains isolated from meat and food-processing environments (3008, 3126, and 4140) after grown in BHI at 30 degrees C. This attenuation could not be ascribed to a defective Internalin A as all strains encoded an intact inlA gene. To determine the influence of food products on virulence, the ability of L. monocytogenes to invade Caco-2 cells was compared after growth on a fermented sausage and on cured cooked ham to that of bacteria grown in BHI broth supplemented with salt. Samples were stored under chilling conditions for up to 4 weeks. The results showed no difference (p > 0.05) in invasiveness after 7 days at 10 degrees C in BHI broth or on sausage, whereas a slight increase (p < 0.05) was observed after incubation on ham for 2 and 4 weeks compared to that in BHI broth. Most importantly, our results show that L. monocytogenes efficiently invade Caco-2 cells even after 4 weeks of storage at chilled temperature. This is highly relevant for safety assessment of this organism in food as these conditions reflect storage of ready-to-eat food products in domestic refrigerators.

Effect of modified atmosphere and temperature abuse on the growth from spores and cereulide production of Bacillus weihenstephanensis in a cooked chilled meat sausage.

The effect of modified atmosphere packaging (MAP) on the germination and growth of toxin producing psychrotolerant Bacillus spp is not well described. A model agar system mimicking a cooked meat product was used in initial experiments. Incubation at refrigeration temperature of 8 degrees C for 5 weeks of 26 Bacillus weihenstephanensis including two emetic toxin (cereulide) producing strains showed that B. weihenstephanensis is sensitive to MAP containing CO2. The sensitivity to 20% CO2 was dependent on strain and oxygen level, being increased when oxygen was excluded from the MAP. Growth from spores was observed at the earliest within 2 weeks when 20% CO2 was combined with 2% O2 and in 3 weeks when combined with "0"% O2 (the remaining atmosphere was made up from N2). Results were validated in a cooked meat sausage model for two non-emetic and one emetic B. weihenstephanensis strain. The packaging film oxygen transfer rates (OTR) were 1.3 and 40 ml/m(2)/24 h and the atmospheres were 2% O2/20% CO2 and "0"% O2/20% CO2. Oxygen availability had a large impact on the growth from spores in the MAP meat sausage, only the most oxygen restricted condition (OTR of 1.3 ml/m(2)/24 h and "0"% O2/20 % CO2 inhibited growth of the three strains during 4 weeks storage at 8 degrees C. Cereulide production was undetectable during storage at 8 degrees C irrespective of choice of the MAP (quantified by liquid chromatography mass spectrometry/mass spectrometry). MAP storage at 8 degrees C for 1 and 3 weeks followed by opening of packages and temperature abuse for 1.5 h daily at 20 degrees C during 1 week resulted in increased cell counts and variable cereulide production in the meat sausage. A pre-history at 8 degrees C for 1 week in MAP with OTR of 1.3 or 40 ml/m(2)/24 h and 2% O2 resulted in cereulide concentrations of 0.816-1.353 microg/g meat sausage, while a pre-history under the most oxygen restricted condition (OTR of 1.3 ml/m(2)/24 h, "0"% O2/20 % CO2 resulted in minimal cereulide production (0.004 microg/g meat sausage) at abuse condition. Extension of MAP storage at 8 degrees C for 3 weeks followed by abuse resulted in a substantially reduced cereulide production. Data demonstrates that MAP can be used to inhibit growth of a psychrotolerant toxin producing Bacillus spp. during chill storage at 8 degrees C, and substantially reduce the risk of emetic food poisoning at abuse condition. Results are of relevance for improving safety of ready to eat processed chilled foods of extended durability.

Determination of mycophenolic acid in meat products using mixed mode reversed phase-anion exchange clean-up and liquid chromatography-high-resolution mass spectrometry.

A method for determination of mycophenolic acid (MPA) in dry-cured ham, fermented sausage and liver pâté is described. MPA was extracted from meat with bicarbonate-acetonitrile, further cleaned-up by mixed mode reversed phase-anion exchange and detected using a LC-MS system with electrospray ionisation-time-of-flight detection. The limit of detection was 4 microg/kg in sausage and 6 microg/kg in ham and pâté. The method was successfully used for quantification of MPA in dry-cured ham and liver pâté artificially inoculated with Penicillium brevicompactum. Levels ranged from 190 microg/kg in centre to 11 mg/kg in surface of ham and from 150 microg/kg in bottom to 14 mg/kg in surface of pâté.

Intracellular pH homeostasis plays a role in the tolerance of Debaryomyces hansenii and Candida zeylanoides to acidified nitrite.

The effects of acidified-nitrite stress on the growth initiation and intracellular pH (pH(i)) of individual cells of Debaryomyces hansenii and Candida zeylanoides were investigated. Our results show that 200 microg/ml of nitrite caused pronounced growth inhibition and intracellular acidification of D. hansenii at an external pH (pH(ex)) value of 4.5 but did not at pH(ex) 5.5. These results indicate that nitrous acid as such plays an important role in the antifungal effect of acidified nitrite. Furthermore, both yeast species experienced severe growth inhibition and a pH(i) decrease at pH(ex) 4.5, suggesting that at least some of the antifungal effects of acidified nitrite may be due to intracellular acidification. For C. zeylanoides, this phenomenon could be explained in part by the uncoupling effect of energy generation from growth. Debaryomyces hansenii was more tolerant to acidified nitrite at pH(ex) 5.5 than C. zeylanoides, as determined by the rate of growth initiation. In combination with the fact that D. hansenii was able to maintain pH(i) homeostasis at pH(ex) 5.5 but C. zeylanoides was not, our results suggest that the ability to maintain pH(i) homeostasis plays a role in the acidified-nitrite tolerance of D. hansenii and C. zeylanoides. Possible mechanisms underlying the different abilities of the two yeast species to maintain their pH(i) homeostasis during acidified-nitrite stress, comprising the intracellular buffer capacity and the plasma membrane ATPase activity, were investigated, but none of these mechanisms could explain the difference.

Mycobiota in the processing areas of two different meat products.

Mould growth is not accepted on most types of North European meat products and is considered as both an economic and aesthetic problem for the producers. In order to determine the mycobiota in processing areas of fermented sausage and liver pâté, filamentous fungi were isolated from air, equipment and raw materials in the processing areas of two fermented sausage processing plants and two liver pâté processing plants. A total of 336 samples were examined. The diversity of filamentous fungi in the processing areas was high; at least 17 different genera were identified. The main isolated genera were identified as Aspergillus, Botrytis, Cladosporium, Epicoccum, Eurotium, Penicillium, Phaeoacremonium and Phoma. Of these, Penicillium and Eurotium were the most important for contamination of fermented sausage, whereas Penicillium and Cladosporium were most important for liver pâté. Cladosporium was isolated more frequently in the processing plants examined in the autumn than in the spring. The seasonal variation indicates that outdoor air is an important source for this contamination. Eurotium was isolated frequently at one of the fermented sausage plants. Penicillium was isolated frequently at all four processing plants and was in addition found on moulded meat products. Sixteen Penicillium species were identified. The most frequently isolated were P. brevicompactum and the closely related P. bialowiezense, P. solitum, P. palitans, P. fagi and a new, not described species named P. "milanense" (ined.; Frisvad, 2007 personal com.). Isolation of a new species illustrates that the mycobiota in the processing areas of North European meat products has not yet been intensively investigated. Several mycotoxin producing species were isolated; the most prevalent were P. brevicompactum/P. bialowiezense and P. palitans. A screening for secondary metabolites showed that isolates of these species consistently produced mycophenolic acid and cyclopiazonic acid, respectively. Presence of these toxinogenic species in the processing areas implies a risk of mycotoxin contamination of the products if they are or has been subjected to mould growth. The ochratoxin A producing species P. nordicum and P. verrucosum were not isolated during the study. It was concluded that Penicillium species are the most important contaminants of the meat products because of their high prevalence in the production environment, their presence on meat products and their toxinogenic properties.

Occurrence and growth of yeasts in processed meat products - Implications for potential spoilage.

Spoilage of meat products is in general attributed to bacteria but new processing and storage techniques inhibiting growth of bacteria may provide opportunities for yeasts to dominate the microflora and cause spoilage of the product. With the aim of obtaining a deeper understanding of the potential role of yeast in spoilage of five different processed meat products (bacon, ham, salami and two different liver patés), yeasts were isolated, enumerated and identified during processing, in the final product and in the final product at the end of shelf life. Yeasts were isolated along the bacon production line in numbers up to 4.2 log (CFU/g). Smoking of the bacon reduced the yeast counts to lower than 1.0 log (CFU/g) or non-detectable levels. In general, yeasts were only isolated in low numbers during the production of salami, cooked ham and liver paté. In the final products yeasts were detected in low numbers in a few samples (3 out of 30) samples, 1.0-1.3 log (CFU/g). By the end of storage, yeasts were only detected in 1 out of 25 investigated samples 1.8 log (CFU/g). A combination of phenotypic and genotypic methods was used to identify the yeast microflora present during production of the processed meat products. The yeast microflora was complex with 4-12 different species isolated from the different production sites. In general, Candida zeylanoides, Debaryomyces hansenii and the newly described Candida alimentaria were found to be the dominant yeast species. In addition, three putatively previously undescribed yeast species were isolated. Fourteen isolates, representing seven different species isolated during the production of the processed meat products and one species isolated from spoiled, modified atmosphere packed, sliced ham, were screened for their ability to grow in a meat model substrate under a low oxygen/high carbon-dioxide atmosphere (0.5% O(2), 20% CO(2), 79.5% N(2)) at two different temperatures (5 and 8°C). Eleven out of the tested 14 strains were able to grow in the meat model substrate with C. zeylanoides, D. hansenii, Pichia guilliermondii and Candida sake reaching levels of 10(5)-5×10(6) log (CFU/g), where sensoryical changes appear.

Influence of different histories of the inoculum on lag phase and growth of Listeria monocytogenes in meat models.

The aim of this study was to determine the effect of history of inoculum and preservatives on the lag phase and growth rate of Listeria monocytogenes strains in meat products packaged under modified atmosphere conditions. Inocula with different histories were added to meat models, and growth rate and lag phase of two strains of L. monocytogenes were measured at 5 and 10 degrees C. The meat model stored at 10 degrees C contained sodium lactate, but the model stored at 5 degrees C did not. The five different histories of the inocula included cold propagation, biofilm formation, and starvation. The lag phase ranged from 1 to 10 days and was affected by the history of the inoculum, whereas the growth rate was constant except for one combination of history of inoculum and strain, where growth did not start during the incubation period. In a second series of experiments, the growth rate and lag phase of the two Listeria strains and the effects of two different histories of inoculum were tested in meat models with pH 5.7 or 6.5 and increasing amounts of NaCl. The growth rate depended on salt concentration, bacterial strain, and pH, whereas lag phase duration depended on history of inoculum, salt concentration, and pH. The lag phase duration was highly dependent on the history of the inoculum, and higher amounts of preservative (NaCl) made these effects even more noticeable. The results of this study underline the importance of the effects of the history of the inoculum on lag phase duration and could be used to predict lag phase in industrial meat products.

Changes in growth, rRNA content, and cell morphology of Listeria monocytogenes induced by CO2 up- and downshift.

Cell morphology, rRNA content, and growth were examined for Listeria monocytogenes LO28 and EGD, respectively, grown in brain-heart infusion (BHI) and on slices of sausage at 10 degrees C in 100% CO2, 100% N2, and air. In CO2, filamentous cells were formed by both strains on sausage slices and by L. monocytogenes EGD in BHI. Filamentation was not induced by anaerobiosis only. Fluorescent in situ rRNA hybridization (FISH) of cells grown in BHI showed that the L. monocytogenes EGD filaments consisted of chains of individual slightly elongated cells. The rods formed by L. monocytogenes LO28 had the same size in air and CO2. Septation and cell division were induced in the filaments after a CO2 downshift (i.e., exposure to air). In BHI, the number of colony forming units increased rapidly when L. monocytogenes EGD grown in CO2 was exposed to air whereas the number of L. monocytogenes LO28 remained almost unchanged. On sausage slices, the number of colony forming units also increased rapidly for both strains in response to CO2 downshift. Large variations in rRNA content of individual cells were observed in the tested scenarios. The results demonstrate the risk of underestimating the number of infectious units under circumstances where filamentation may occur. Furthermore, the study illustrates the lack of residual inhibitory effect of CO2 in this type of products after opening.

Leuconostoc carnosum 4010 has the potential for use as a protective culture for vacuum-packed meats: culture isolation, bacteriocin identification, and meat application experiments.

A new culture, Leuconostoc carnosum 4010, for biopreservation of vacuum-packed meats is described. The culture originated from bacteriocin-producing lactic acid bacteria (LAB) naturally present in vacuum-packed meat products. Approximately, 72,000 colonies were isolated from 48 different vacuum-packed meat products and examined for antibacterial activity. Bacteriocin-producing colonies were isolated from 46% of the packages examined. Leuc. carnosum was the predominant bacteriocin-producing strain and Leuc. carnosum 4010 was selected for further experiments because it showed strong antilisterial activity without producing any undesirable flavour components in meat products. For identification of the bacteriocins produced, partial purification was carried out by ammonium sulphate precipitation, dialysis, and cation exchange chromatography. SDS-PAGE analysis revealed two bands with inhibitory activity corresponding to molecular sizes of 4.6 and 5.3 kDa. N-terminal amino acid sequencing showed that Leuc. carnosum 4010 produced two bacteriocins highly similar or identical to leucocin A and leucocin C. Application experiments showed that the addition of 10(7) cfu/g Leuc. carnosum 4010 to a vacuum-packaged meat sausage immediately reduced the number of viable Listeria monocytogenes cells to a level below the detection limit and no increase of L. monocytogenes was observed during storage at 5 degrees C for 21 days. The results presented demonstrate that Leuc. carnosum 4010 is suitable as a new protective culture for cold-stored, cooked, sliced, and vacuum-packed meat products.

6-Chloro-3-alkylamino-4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide derivatives potently and selectively activate ATP sensitive potassium channels of pancreatic beta-cells.

6-Chloro-3-alkylamino-4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide derivatives were synthesized and characterized as activators of adenosine 5'-triphosphate (ATP) sensitive potassium (K(ATP)) channels in the beta-cells by measuring effects on membrane potential and insulin release in vitro. The effects on vascular tissue in vitro were measured on rat aorta and small mesenteric vessels. Selected compounds were characterized as competitive inhibitors of [(3)H]glibenclamide binding to membranes of HEK293 cells expressing human SUR1/Kir6.2 and as potent inhibitors of insulin release in isolated rat islets. 6-Chloro-3-(1-methylcyclobutyl)amino-4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide (54) was found to bind and activate the SUR1/Kir6.2 K(ATP) channels in the low nanomolar range and to be at least 1000 times more potent than the reference compound diazoxide with respect to inhibition of insulin release from rat islets. Several compounds, e.g., 3-propylamino- (30), 3-isopropylamino- (34), 3-(S)-sec-butylamino- (37), and 3-(1-methylcyclopropyl)amino-4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide (53), which were found to be potent and beta-cell selective activators of K(ATP) channels in vitro, were found to inhibit insulin secretion in rats with minimal effects on blood pressure and to exhibit good oral pharmacokinetic properties.