Influence of processing steps in cold-smoked salmon production on survival and growth of persistent and presumed non-persistent Listeria monocytogenes

Cold-smoked salmon is a ready-to-eat product in which Listeria monocytogenes sometimes can grow to high numbers. The bacterium can colonize the processing environment and it is believed to survive or even grow during the processing steps. The purpose of the present study was to determine if the steps in the processing of cold-smoked salmon affect survival and subsequent growth of a persistent strain of L. monocytogenes to a lesser degree than presumed non-persistent strains. We used a sequence of experiments increasing in complexity: (i) small salmon blocks salted, smoked or dried under model conditions, (ii) fillets of salmon cold-smoked in a pilot plant and finally, (iii) assessment of the bacterial levels before and after processing during commercial scale production. L. monocytogenes proliferated on salmon blocks that were brined or dipped in liquid smoke and left at 25 degrees C in a humidity chamber for 24 h. However, combining brining and liquid smoke with a drying (25 degrees C) step reduced the bacterium 10-100 fold over a 24 h period. Non-salted, brine injected or dry salted salmon fillets were surface inoculated with L. monocytogenes and cold-smoked in a pilot plant. L. monocytogenes was reduced from 10(3) to 10-10(2) CFU/cm(2) immediately after cold-smoking. The greatest reductions were observed in dry salted and brine injected fillets as compared to cold-smoking of non-salted fresh fillets. Levels of L. monocytogenes decreased further when the cold-smoked fish was vacuum-packed and stored at 5 degrees C. A similar decline was seen when inoculating brine injected fillets after cold-smoking. High phenol concentrations are a likely cause of this marked growth inhibition. In a commercial production facility, the total viable count of salmon fillets was reduced 10-1000 fold by salting, cold-smoking and process-freezing (a freezing step after smoking and before slicing). The prevalence of L. monocytogenes in the commercial production facility was too low to determine any quantitative effects, however, one of nine samples was positive before processing and none after. Taken together, the processing steps involved in cold-smoking of salmon are bactericidal and reduce, but do not eliminate L. monocytogenes. A persistent strain was no less sensitive to the processing steps than a clinical strain or strain EGD.     

Ad libitum fluid intake leads to no leg swelling in male Ironman triathlet an observational field study

ABSTRACT: BACKGROUND: An association between fluid intake and limb swelling has been described for 100-km ultra-marathoners. We investigated a potential development of peripheral oedemata in Ironman triathletes competing over 3.8 km swimming, 180 km cycling and 42.2 km running. METHODS: In 15 male Ironman triathletes, fluid intake, changes in body mass, fat mass, skeletal muscle mass, limb volumes and skinfold thickness were measured. Changes in renal function, parameters of skeletal muscle damage, hematologic parameters and osmolality in both serum and urine were determined. Skinfold thicknesses at hands and feet were measured using LIPOMETER(R) and changes of limb volumes were measured using plethysmography. RESULTS: The athletes consumed a total of 8.6 +/- 4.4 L of fluids, equal to 0.79 +/- 0.43 L/h. Body mass, skeletal muscle mass and the volume of the lower leg decreased (p <0.05), fat mass, skinfold thicknesses and the volume of the arm remained unchanged (p >0.05). The decrease in skeletal muscle mass was associated with the decrease in body mass (p >0.05). The decrease in the lower leg volume was unrelated to fluid intake (p >0.05). Haemoglobin, haematocrit and serum sodium remained unchanged (p >0.05). Osmolality in serum and urine increased (p <0.05). The change in body mass was related to post-race serum sodium concentration ([Na+]) (r = -0.52, p <0.05) and post-race serum osmolality (r = -0.60, p <0.05). CONCLUSIONS: In these Ironman triathletes, ad libitum fluid intake maintained plasma [Na+] and plasma osmolality and led to no peripheral oedemata. The volume of the lower leg decreased and the decrease was unrelated to fluid intake. Future studies may investigate ultra-triathletes competing in a Triple Iron triathlon over 11.4 km swimming, 540 km cycling and 126.6 km running to find an association between fluid intake and the development of peripheral oedemata.     

A robust protein host for anchoring chelating ligands and organocatalysts

In order to put the previously proposed concept of directed evolution of hybrid catalysts (proteins that harbor synthetic transition-metal catalysts or organocatalysts) into practice, several prerequisites must be met. The availability of a robust host protein that can be expressed in sufficiently large amounts, and that can be purified in a simple manner is crucial. The thermostable enzyme tHisF from Thermotoga maritima, which constitutes the synthase subunit of a bi-enzyme complex that is instrumental in the biosynthesis of histidine, fulfills these requirements. In the present study, fermentation has been miniaturized and parallelized, as has purification of the protein by simple heat treatment. Several mutants with strategically placed cysteines for subsequent bioconjugation have been produced. One of the tHisF mutants, Cys9Ala/Asp11Cys, was subjected to bioconjugation by the introduction of a variety of ligands for potential metal ligation, of a ligand/metal moiety, and of several organocatalytic entities that comprise a flavin or thiazolium salts. Characterization by mass spectrometry and tryptic digestion was achieved. As a result of this study, a platform for performing future directed evolution of these hybrid catalysts is now available.     

Estimating the nitrogen nutrition index in grass seed crops using a UAV-mounted multispectral camera

The application of adequate nitrogen (N) fertilizers to grass seed crops is important to achieve high seed yield. Application of N will inevitably result in over-fertilization on some fields and, concomitantly, an increased risk of adverse environmental impacts, such as ground- and/or surface-water contamination. This study was designed to estimate the N status of two grass seed crops: red fescue (Festuca rubra L.) and perennial ryegrass (Lolium perenne L.) using images captured with an unmanned aerial vehicle (UAV) mounted multispectral camera. Two types of UAV, a fixed-wing UAV and a multi-rotor UAV, operating at two different heights and mounted with the same multispectral camera, were used in different field experiments at the same location in Denmark in the period from 432 to 861 growing degree-days. Seven vegetation indices, calculated from multispectral images with four bands: red, green, red edge and near infrared (NIR), were evaluated for their relationship to dry matter (DM), N concentration, N uptake and N nutrition index (NNI). The results showed a better prediction of N concentration, N uptake and NNI, than DM using vegetation indices. Furthermore, among all vegetation indices, two red-edge-based indices, normalized difference red edge (NDRE) and red edge chlorophyll index (CI_RE), performed best in estimating N concentration (R² = 0.69–0.88), N uptake (R² = 0.41–0.84) and NNI (R² = 0.47–0.86). In addition, there was no effect from the choice of UAV, and thereby flight height, on the estimation of NNI. The choice of UAV type therefore seems not to influence the possibility of diagnosing N status in grass seed crops. We conclude that it is possible to estimate NNI based on multispectral images from drone-mounted cameras, and the method could guide farmers as to whether they should apply additional N to the field. We also conclude that further research should focus on estimating the quantity of N to apply and on further developing the method to include more grass species.     

Regioselective lipase-catalyzed transesterification of tributyrin. Influence of salt hydrates on acyl migration

The influence of the nature of solid salt hydrates on the rate of hexanoic acid catalyzed acyl migration in 1,2-dibutyrin has been examined in hexane. The results show that the rate of acyl migration is faster when hydrogen phosphate salts are included compared to sulfate salts. The rate is essentially the same in the presence of sulfate salts and without salts. In regioselective lipase-catalyzed acidolysis of tributyrin with hexanoic acid, the use of hydrogen phosphate salts to control water activity leads to a higher rate of migration than the use of sulfate salts. Minor differences are observed in interesterification with ethyl hexanoate.     

Functional roles and subsite locations of Leu177, Trp178 and Asn182 of Aspergillus awamori glucoamylase determined by site-directed mutagenesis

Fungal glucoamylases contain four conserved regions. One regionfrom the Aspergillus niger enzyme contains three key carboxylicacid residues, the general acid catalytic group, Glu179, alongwith Asp176 and Glu180. Three site-directed mutations, Leu177– His, Trp178 – Arg and Asn182 – Ala, wereconstructed near these acidic groups to reveal the functionof other conserved residues in this region. Leu177 and Trp178are strictly conserved among fungal glucoamylases, while anamide, predominantly Asn, always occurs at position 182. Substitutionsof Leu177 or Trp178 cause significant decreases in k_cat withthe substrates tested. Similar increases in activation energiesobtained with Leu177 – His with both -(1,4)- and -(1,6)-linkedsubstrates indicate Leu177 is located in subsite 1. K_M valuesobtained with the Trp178 – Arg mutation increase for an-(1,6)-linked substrate, but not for -(1,4)-linked substrates.Calculated differences in activation energy between substratesindicate Trp178 interacts specifically with subsite 2. The Asn182 Ala mutation did not change k_cat or K_M values, indicating thatAsn182 is not crucial for activity. These results support amechanism for glucoamylase catalytic activity consisting ofa fast substrate binding step followed by a conformational changeat subsite 1 to stabilize the transition state complex.     

The Hamburg Metaphor Database project: issues in resource creation

This paper concerns metaphor resource creation. It provides an account of methods used, problems discovered, and insights gained at the Hamburg Metaphor Database project, intended to inform similar resource creation initiatives, as well as future metaphor processing algorithms. After introducing the project, the theoretical underpinnings that motivate the subdivision of represented information into a conceptual and a lexical level are laid out. The acquisition of metaphor attestations from electronic corpora is explained, and annotation practices as well as database contents are evaluated. The paper concludes with an overview of related projects and an outline of possible future work.