Identification by means of molecular tools of the microbiota responsible for the formation of histamine accumulated in commercial cheeses in Spain

Histamine intoxication is an important food safety and public health concern. Ripened cheeses are the most common dairy products in which histamine can accumulate. Histamine is formed by the microbiota present in cheese by decarboxylation of histidine, due to the action of the histidine decarboxylase. This study's objective was to identify the responsible for the formation of histamine accumulated in commercial cheeses. The content of histamine of 39 different types of cheeses marketed in Spain, of varying milk origin, was assessed. About one third of the cheeses analysed contained more than 200 mg/kg histamine; two cheeses exceeded 500 mg/kg histamine, the consumption of such cheeses can be harmful or even toxic for consumers. The five cheeses with the highest histamine concentrations were selected for in-depth molecular analysis. Firstly, bacterial and yeast isolates were obtained, and then the total genetic material from the cheeses was analysed, in order to verify the putative presence of the hdc histidine decarboxylase gene. In order to identify the histamine producing microorganisms, the nucleotide sequences of the histidine decarboxylase genes from the cheeses were amplified, and subjected them to Sanger sequencing. In four of the five selected cheeses, the main histamine producer was identified as Lentilactobacillus parabuchneri, whereas in the remaining cheese it was Tetragenococcus halophilus. The hdc gene was located in an unstable plasmid, only present in that cheese sample. Since all histamine producing microorganisms identified in this study are not part of the species used in cheese starter cultures, an improvement of hygienic manufacturing practices and/or thermal treatments for microbial inactivation in milk may be considered to prevent histamine accumulation in cheeses during ripening

Updated NNLO QCD predictions for the weak radiative B-meson decays

Weak radiative decays of the B mesons belong to the most important flavor changing processes that provide constraints on physics at the TeV scale. In the derivation of such constraints, accurate standard model predictions for the inclusive branching ratios play a crucial role. In the current Letter we present an update of these predictions, incorporating all our results for the O(alpha_s^2) and lower-order perturbative corrections that have been calculated after 2006. New estimates of nonperturbative effects are taken into account, too. For the CP- and isospin-averaged branching ratios, we find B_{s gamma} = (3.36 +_ 0.23) * 10^-4 and B_{d gamma} = 1.73^{+0.12}_{-0.22} * 10^-5, for E_gamma > 1.6GeV. Both results remain in agreement with the current experimental averages. Normalizing their sum to the inclusive semileptonic branching ratio, we obtain R_gamma = ( B_{s gamma} + B_{d gamma})/B_{c l nu} = (3.31 +_ 0.22) * 10^-3. A new bound from B_{s gamma} on the charged Higgs boson mass in the two-Higgs-doublet-model II reads M_{H^+} > 480 GeV at 95%C.L.Comment: journal version, 5 pages, no figure

Potential of histamine-degrading microorganisms and diamine oxidase (DAO) for the reduction of histamine accumulation along the cheese ripening process

Lentilactobacillus parabuchneri is the main bacteria responsible for the accumulation of histamine in cheese. The goal of this study was to assess the efficiency of potential histamine-degrading microbial strains or, alternatively, the action of the diamine oxidase (DAO) enzyme in the reduction of histamine accumulation along the ripening process in cheese. A total of 8 cheese variants of cow milk cheese were manufactured, all of them containing L. parabuchneri Deutsche Sammlung von Mikroorganismen 5987 (except for the negative control cheese variant) along with histamine–degrading strains (Lacticaseibacillus casei 4a and 18b; Lactobacillus delbrueckii subsp. bulgaricus Colección Española de Cultivos Tipo (CECT) 4005 and Streptococcus salivarius subsp. thermophilus CECT 7207; two commercial yogurt starter cultures; or Debaryomyces hansenii), or DAO enzyme, tested in each cheese variant. Histamine was quantified along 100 days of cheese ripening. All the degrading measures tested significantly reduced the concentration of histamine. The highest degree of degradation was observed in the cheese variant containing D. hansenii, where the histamine content decreased up to 45.32 %. Cheese variants with L. casei, or L. bulgaricus and S. thermophilus strains, also decreased in terms of histamine content by 43.05 % and 42.31 %, respectively. No significant physicochemical changes (weight, pH, water activity, color, or texture) were observed as a consequence of the addition of potential histamine-degrading adjunct cultures or DAO in cheeses. However, the addition of histamine-degrading microorganisms was associated with a particular, not unpleasant aroma. Altogether, these results suggest that the use of certain histamine-degrading microorganisms could be proposed as a suitable measure in order to decrease the amount of histamine accumulated in cheeses. © 2022 The Author

The significance of cheese sampling in the determination of histamine concentration: Distribution pattern of histamine in ripened cheeses

Cheeses are becoming a major safety and public health concern: cheeses available in supermarkets occasionally contain high histamine concentrations that can have negative effects on consumer health. In this study, we have attempted to assess the histamine distribution pattern in ripened cheeses, with the purpose of establishing a correct cheese sampling strategy for the quantification of histamine. To this aim, histamine was determined in four distinct areas of twelve long-ripened hard cheeses: the external and internal rind, along with the outer and inner core of the wedge. The concentrations measured were remarkably different: histamine accumulated in the central core, whereas the lowest amount was found in the peripheral rind. To explain this heterogenous distribution, histamine producers were determined in the four areas by identifying the hdc sequences obtained from cheese samples. Non-starter bacteria were identified as main histamine producers; however, these microbiota were homogeneously distributed throughout the wedge. Nevertheless, the analysis of psychochemical properties of the different areas revealed an observable trend: histamine tended to accumulate in the saltier, more humid, and less oxidized areas in a wedge. Overall, this study highlights the significance of a correct sampling strategy when histamine is quantified in cheese

Flavour constraints on scenarios with two or three heavy squark generations

We re-assess constraints from flavour-changing neutral currents in the kaon system on supersymmetric scenarios with a light gluino, two heavy generations of squarks and a lighter third generation. We compute for the first time limits in scenarios with three heavy squark families, taking into account QCD corrections at the next-to-leading order. We compare our limits with those in the case of two heavy families. We use the mass insertion approximation and consider contributions from gluino exchange to constrain the mixing between the first and second squark generation. While it is not possible to perform a general analysis, we assess the relevance of each kind of flavour- and CP-violating parameters. We also provide ready to use magic numbers for the computation of the Wilson coefficients at 2 GeV for these scenarios.Comment: 23 pages, 14 figures; v3: matches published version (contains improvements in the presentation and clarifications

Supersymmetric Contributions to Bs->K+K-

Inspired by the existing calculation of B->piK decays in supersymmetry (SUSY), we evaluate the dominant SUSY contributions to Bs->K+K-. We show that the observables of this process can be significantly modified in the presence of SUSY. In particular, the branching ratio can be increased considerably compared to the prediction of the standard model (SM). The effect is even more dramatic for the CP-violating asymmetries A_dir and A_mix. These asymmetries, expected to be small in the SM (A_dir is predicted to take only positive values), change drastically with SUSY contributions. The measurement of these observables can therefore be used to detect the presence of physics beyond the SM, and put constraints on its parameters.Comment: 17 pages, 2 figures. This new version contains one added reference and some minor style change

Multi-modelling predictions show high uncertainty of required carbon input changes to reach a 4‰ target

Soils store vast amounts of carbon (C) on land, and increasing soil organic carbon (SOC) stocks in already managed soils such as croplands may be one way to remove C from the atmosphere, thereby limiting subsequent warming. The main objective of this study was to estimate the amount of additional C input needed to annually increase SOC stocks by 4%(0) at 16 long-term agricultural experiments in Europe, including exogenous organic matter (EOM) additions. We used an ensemble of six SOC models and ran them under two configurations: (1) with default parametrization and (2) with parameters calibrated site-by-site to fit the evolution of SOC stocks in the control treatments (without EOM). We compared model simulations and analysed the factors generating variability across models. The calibrated ensemble was able to reproduce the SOC stock evolution in the unfertilised control treatments. We found that, on average, the experimental sites needed an additional 1.5 +/- 1.2 Mg C ha(-)(1) year(-1) to increase SOC stocks by 4%(0) per year over 30 years, compared to the C input in the control treatments (multi-model median +/- median standard deviation across sites). That is, a 119% increase compared to the control. While mean annual temperature, initial SOC stocks and initial C input had a significant effect on the variability of the predicted C input in the default configuration (i.e., the relative standard deviation of the predicted C input from the mean), only water-related variables (i.e., mean annual precipitation and potential evapotranspiration) explained the divergence between models when calibrated. Our work highlights the challenge of increasing SOC stocks in agriculture and accentuates the need to increasingly lean on multi-model ensembles when predicting SOC stock trends and related processes. To increase the reliability of SOC models under future climate change, we suggest model developers to better constrain the effect of water-related variables on SOC decomposition

Implications from clean observables for the binned analysis of B -> K*ll at large recoil

We perform a frequentist analysis of q^2-dependent B-> K*(->Kpi)ll angular observables at large recoil, aiming at bridging the gap between current theoretical analyses and the actual experimental measurements. We focus on the most appropriate set of observables to measure and on the role of the q^2-binning. We highlight the importance of the observables P_i exhibiting a limited sensitivity to soft form factors for the search for New Physics contributions. We compute predictions for these binned observables in the Standard Model, and we compare them with their experimental determination extracted from recent LHCb data. Analyzing b->s and b->sll transitions within four different New Physics scenarios, we identify several New Physics benchmark points which can be discriminated through the measurement of P_i observables with a fine q^2-binning. We emphasise the importance (and risks) of using observables with (un)suppressed dependence on soft form factors for the search of New Physics, which we illustrate by the different size of hadronic uncertainties attached to two related observables (P_1 and S_3). We illustrate how the q^2-dependent angular observables measured in several bins can help to unravel New Physics contributions to B-> K*(->Kpi)ll, and show the extraordinary constraining power that the clean observables will have in the near future. We provide semi-numerical expressions for these observables as functions of the relevant Wilson coefficients at the low scale.Comment: 50 pages, 21 figures. Improved form factor analysis, conclusions unchanged. Plots with full resolution. Version published in JHE

Encapsulation of Lactobacillus plantarum in casein-chitosan microparticles facilitates the arrival to the colon and develops an immunomodulatory effect

The current work describes the capability of casein-chitosan microparticles to encapsulate Lactobacillus plantarum (CECT 220 and WCFS1 strains) and evaluates their ability to target the distal areas of the gut and to stimulate the immune system. Microparticles were prepared by complex coacervation, between sodium caseinate and chitosan in an aqueous suspension of the bacteria, and dried by spray-drying. In order to increase the survival rate of the loaded bacteria, microparticles were cross-linked with one of the following cross-linkers: tripolyphosphate, calcium salts or vanillin. Overall, microparticles displayed a mean size of about 7.5 μm with a bacteria loading of about 11 Log CFU/g, when cross-linked with vanillin (MP-LP-V). For conventional microparticles, the payload was 10.12 Log CFU/g. The storage stability study at 25 ◦C/60% RH, MP-LP-V offered the highest degree of protection without signif- icant modification of the payload in 260 days. Compared with control (aqueous suspension of bacteria), MP-LP-V also displayed a significantly higher degree of protection against probiotic inactivation in simulated gastric and intestinal fluids. In vivo results evidenced that microparticles, orally administered to rats, were able to reach the distal ileum and colon in about 4 h post-administration. Additionally, the effect of the daily administration of 107 CFU/mouse of MP-LP-V, for 3 weeks, induced an immunomodulatory effect characterized by an important enhancement of Th1 and Th17 responses. In conclusion, these microparticles seem to be a promising strategy for increasing survival and efficacy of probiotics, allowing the formulation of cost-effective and more stable and effective probiotic-based nutraceuticals