Antimicrobial effect of essential oils against several pathogens and their influence on inhibition of salmonella enteritidis, listeria monocytogenes and staphylococcus aureus in paste of “alheira” during storage

Fermented meat products are part of the daily diet in rural areas of Portugal and have become very popular in urban centers. “Alheiras" are traditional, slightly smoked, naturally fermented meat sausages typical of the Northern regions (Trás-os-Montes) in Portugal. Essential oils (EOs), traditionally used as flavoring agents, have been revealing good antimicrobial properties, becoming a good natural alternative to the use of chemical preservatives. The aim of this study was to investigate the antimicrobial effect of some EOs against several pathogens and their influence on inhibition of Salmonella Enteritidis, Listeria monocytogenes and Staphylococcus aureus in “alheira” during storage. First, the in vitro antimicrobial effect of 23 EOs against 41 foodborne and spoilage microorganisms was screened by the disc diffusion assay method (21 Gram-positive bacteria, 18 Gram-negative bacteria and two yeasts). Then, the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) was studied for the EOs that displayed a better antimicrobial activity (i.e. halo > 10 mm) by the Microtiter plate assay. The antimicrobial effect of different concentrations of oregano EO (the EO with the lower MIC) (4%, 1.5%, 0.5%, 0.195% and 0.0975%) was evaluated in paste of “alheira” against Salmonella Enteritidis, L. monocytogenes and St. aureus along 21 days of storage at 4 ºC. The pH, water activity values and lactic acid bacteria were also evaluated. At last, sensory assessment was performed. Results showed that antimicrobial activity was variable, according to EOs used and microorganism. In general, oregano and thyme were the essential oils that showed highest antimicrobial activity and anise, fennel, garlic and ginger were the EOs with lower activity. The lowest minimum inhibitory concentration (0.0244%) against L. innocua was observed for Oregano. Oregano and thyme also showed higher MICs, against all microorganisms. Utilization of oregano EO in paste of “alheira” along 21 days of storage at 4 ºC resulted in a natural strategy to improve its safety against S. Enteritidis, L. monocytogenes and St. aureus. Although, oregano EO possesses antibacterial properties in vitro, their utilization as food antimicrobial agents must be assessed in the food product, in particular in a complex matrix as “alheira”. The antibacterial effect varies according to the oregano EO concentration and pathogen used. The results showed that 4% and 1.5% of oregano EO demonstrated the best antimicrobial activity against all the pathogens tested. 0.5% was able to result in ~ 2 log reduction for S. Enteritidis, whereas the lowest concentrations used (0.195% and 0.0975%) resulted in ~ 2-3 log reduction after 21 days Fermented meat products are part of the daily diet in rural areas of Portugal and have become very popular in urban centers. “Alheiras" are traditional, slightly smoked, naturally fermented meat sausages typical of the Northern regions (Trás-os-Montes) in Portugal. Essential oils (EOs), traditionally used as flavoring agents, have been revealing good antimicrobial properties, becoming a good natural alternative to the use of chemical preservatives. The aim of this study was to investigate the antimicrobial effect of some EOs against several pathogens and their influence on inhibition of Salmonella Enteritidis, Listeria monocytogenes and Staphylococcus aureus in “alheira” during storage. First, the in vitro antimicrobial effect of 23 EOs against 41 foodborne and spoilage microorganisms was screened by the disc diffusion assay method (21 Gram-positive bacteria, 18 Gram-negative bacteria and two yeasts). Then, the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) was studied for the EOs that displayed a better antimicrobial activity (i.e. halo > 10 mm) by the Microtiter plate assay. The antimicrobial effect of different concentrations of oregano EO (the EO with the lower MIC) (4%, 1.5%, 0.5%, 0.195% and 0.0975%) was evaluated in paste of “alheira” against Salmonella Enteritidis, L. monocytogenes and St. aureus along 21 days of storage at 4 ºC. The pH, water activity values and lactic acid bacteria were also evaluated. At last, sensory assessment was performed. Results showed that antimicrobial activity was variable, according to EOs used and microorganism. In general, oregano and thyme were the essential oils that showed highest antimicrobial activity and anise, fennel, garlic and ginger were the EOs with lower activity. The lowest minimum inhibitory concentration (0.0244%) against L. innocua was observed for Oregano. Oregano and thyme also showed higher MICs, against all microorganisms. Utilization of oregano EO in paste of “alheira” along 21 days of storage at 4 ºC resulted in a natural strategy to improve its safety against S. Enteritidis, L. monocytogenes and St. aureus. Although, oregano EO possesses antibacterial properties in vitro, their utilization as food antimicrobial agents must be assessed in the food product, in particular in a complex matrix as “alheira”. The antibacterial effect varies according to the oregano EO concentration and pathogen used. The results showed that 4% and 1.5% of oregano EO demonstrated the best antimicrobial activity against all the pathogens tested. 0.5% was able to result in ~ 2 log reduction for S. Enteritidis, whereas the lowest concentrations used (0.195% and 0.0975%) resulted in ~ 2-3 log reduction after 21 daysOs produtos à base de carne fermentada fazem parte da dieta quotidiana nas zonas rurais de Portugal e tornaram-se muito populares nos centros urbanos. Este estudo teve como objetivo avaliar o efeito antimicrobiano de alguns óleos essenciais (OEs) contra vários agentes patogénicos e a sua influência na inibição de Salmonella Enteritidis, Listeria monocytogenes e Staphylococcus aureus. O efeito antimicrobiano in vitro, de 23 OEs contra 41 microrganismos isolados de alimentos (21 bactérias Gram-positivas, 18 bactérias Gram-negativas e duas leveduras) foi avaliado por ensaios de difusão do disco. De seguida, a concentração mínima inibitória (CMI) e a concentração mínima bactericida (CBM) foram determinadas para os OEs que mostraram uma maior atividade antimicrobiana (ou seja, halo > 10 mm) através de um ensaio realizado em microplaca. O efeito antimicrobiano de diferentes concentrações de OEs (OE com CMI mais baixo) (4%, 1,5%, 0,5%, 0,195% e 0,0975%) foi avaliado em pasta de "alheira" contra S. Enteritidis, L. monocytogenes e St. aureus ao longo de 21 dias de armazenamento a 4 ° C. O pH, os valores da atividade da água e o nível de bactérias do ácido lático também foram monitorizados. Por fim, foi avaliada a qualidade sensorial da pasta de “alheira” contendo OEs. Os resultados mostraram que a atividade antimicrobiana foi variável, de acordo com os OEs utilizados e os microrganismos alvo. Em geral, os OEs de orégãos e de tomilho foram os que apresentaram maior atividade antimicrobiana, e os OEs de anis, de funcho, de alho e de gengibre a menor atividade. A menor concentração mínima inibitória (0.0244%) contra L. innocua foi observada para o OE de orégão. Os OEs de orégãos e de tomilho também apresentaram maiores CMI contra todos os microrganismos. A utilização do OE de orégãos em pasta de "alheira" ao longo de 21 dias de armazenamento a 4 ° C resultou numa estratégia natural para melhorar sua segurança por redução dos níveis de S. Enteritidis, L. monocytogenes e St. aureus. Embora, o EO de orégãos possua propriedades antibacterianas in vitro, a sua utilização como agente antimicrobiano alimentar deve ser avaliada no produto alimentar, em particular numa matriz complexa como a "alheira". O efeito antibacteriano varia de acordo com a concentração de OE de orégãos e patogénico utilizado. Os resultados mostraram que 4% e 1.5% de OE de orégãos apresentaram a maior atividade antimicrobiana contra todos os patogénicos testados. Para a concentração de 0.5% obteve-se uma redução de 2 log para S. Enteritidis, enquanto que as concentrações mais baixas utilizadas (0.195% e 0.0975%) resultaram numa redução de ~ 2-3 log para L. monocytogenes após 21 dias. Embora o OE de orégãos tenha mostrado propriedades anti-listeria em baixas concentrações, o mesmo não foi observado para S. Enteritidis e St. aureus. As contagens de bactérias do ácido lático foram ~109 CFU / ml para todas as amostras e não foram detetadas diferenças nos valores de pH e aw entre as amostras. O impacto sensorial do OE de orégãos a 0.195% na "alheira" foi avaliado por testes de aceitabilidade geral, numa escala just-about-right (JAR). A tendência de consumo e o potencial de consumo foi avaliado, já que o impacto sensorial dos OEs em géneros alimentícios foi descrito como uma restrição ao consumo. Os resultados mostraram que a concentração teve um impacto negativo na aceitação de "alheira", devido ao seu sabor muito intenso. Estes resultados podem ser interessantes para a indústria da carne, ajudando a garantir a segurança microbiológica dos produtos, atendendo às necessidades naturais do consumidor. No entanto, é necessário validar esses resultados na produção in situ de "alheira", adicionando o OE de orégãos como ingrediente e avaliar sua aceitabilidade pelo consumidor. Será também de explorar a utilização dos EOs, em concentrações mais baixas, em combinação com outros tratamentos

Energy Estimation of Cosmic Rays with the Engineering Radio Array of the Pierre Auger Observatory

The Auger Engineering Radio Array (AERA) is part of the Pierre Auger Observatory and is used to detect the radio emission of cosmic-ray air showers. These observations are compared to the data of the surface detector stations of the Observatory, which provide well-calibrated information on the cosmic-ray energies and arrival directions. The response of the radio stations in the 30 to 80 MHz regime has been thoroughly calibrated to enable the reconstruction of the incoming electric field. For the latter, the energy deposit per area is determined from the radio pulses at each observer position and is interpolated using a two-dimensional function that takes into account signal asymmetries due to interference between the geomagnetic and charge-excess emission components. The spatial integral over the signal distribution gives a direct measurement of the energy transferred from the primary cosmic ray into radio emission in the AERA frequency range. We measure 15.8 MeV of radiation energy for a 1 EeV air shower arriving perpendicularly to the geomagnetic field. This radiation energy -- corrected for geometrical effects -- is used as a cosmic-ray energy estimator. Performing an absolute energy calibration against the surface-detector information, we observe that this radio-energy estimator scales quadratically with the cosmic-ray energy as expected for coherent emission. We find an energy resolution of the radio reconstruction of 22% for the data set and 17% for a high-quality subset containing only events with at least five radio stations with signal.Comment: Replaced with published version. Added journal reference and DO

Measurement of the Radiation Energy in the Radio Signal of Extensive Air Showers as a Universal Estimator of Cosmic-Ray Energy

We measure the energy emitted by extensive air showers in the form of radio emission in the frequency range from 30 to 80 MHz. Exploiting the accurate energy scale of the Pierre Auger Observatory, we obtain a radiation energy of 15.8 \pm 0.7 (stat) \pm 6.7 (sys) MeV for cosmic rays with an energy of 1 EeV arriving perpendicularly to a geomagnetic field of 0.24 G, scaling quadratically with the cosmic-ray energy. A comparison with predictions from state-of-the-art first-principle calculations shows agreement with our measurement. The radiation energy provides direct access to the calorimetric energy in the electromagnetic cascade of extensive air showers. Comparison with our result thus allows the direct calibration of any cosmic-ray radio detector against the well-established energy scale of the Pierre Auger Observatory.Comment: Replaced with published version. Added journal reference and DOI. Supplemental material in the ancillary file

Studies of the mass composition of cosmic rays and proton-proton interaction cross-sections at ultra-high energies with the Pierre Auger Observatory

In this work, we present an estimate of the cosmic-ray mass composition from the distributions of the depth of the shower maximum (Xmax) measured by the fluorescence detector of the Pierre Auger Observatory. We discuss the sensitivity of the mass composition measurements to the uncertainties in the properties of the hadronic interactions, particularly in the predictions of the particle interaction cross-sections. For this purpose, we adjust the fractions of cosmic-ray mass groups to fit the data with Xmax distributions from air shower simulations. We modify the proton-proton cross-sections at ultra-high energies, and the corresponding air shower simulations with rescaled nucleus-air cross-sections are obtained via Glauber theory. We compare the energy-dependent composition of ultra-high-energy cosmic rays obtained for the different extrapolations of the proton-proton cross-sections from low-energy accelerator data

Study of downward Terrestrial Gamma-ray Flashes with the surface detector of the Pierre Auger Observatory

The surface detector (SD) of the Pierre Auger Observatory, consisting of 1660 water-Cherenkov detectors (WCDs), covers 3000 km2 in the Argentinian pampa. Thanks to the high efficiency of WCDs in detecting gamma rays, it represents a unique instrument for studying downward Terrestrial Gamma-ray Flashes (TGFs) over a large area. Peculiar events, likely related to downward TGFs, were detected at the Auger Observatory. Their experimental signature and time evolution are very different from those of a shower produced by an ultrahigh-energy cosmic ray. They happen in coincidence with low thunderclouds and lightning, and their large deposited energy at the ground is compatible with that of a standard downward TGF with the source a few kilometers above the ground. A new trigger algorithm to increase the TGF-like event statistics was installed in the whole array. The study of the performance of the new trigger system during the lightning season is ongoing and will provide a handle to develop improved algorithms to implement in the Auger upgraded electronic boards. The available data sample, even if small, can give important clues about the TGF production models, in particular, the shape of WCD signals. Moreover, the SD allows us to observe more than one point in the TGF beam, providing information on the emission angle

Investigating multiple elves and halos above strong lightning with the fluorescence detectors of the Pierre Auger Observatory

ELVES are being studied since 2013 with the twenty-four FD Telescopes of the Pierre Auger Observatory, in the province of Mendoza (Argentina), the world’s largest facility for the study of ultra-high energy cosmic rays. This study exploits a dedicated trigger and extended readout. Since December 2020, this trigger has been extended to the three High levation Auger Telescopes (HEAT), which observe the night sky at elevation angles between 30 and 60 degrees, allowing a study of ELVES from closer lightning. The high time resolution of the Auger telescopes allows us to upgrade reconstruction algorithms and to do detailed studies on multiple ELVES. The origin of multiple elves can be studied by analyzing the time difference and the amplitude ratio between flashes and comparing them with the properties of radio signals detected by the ENTLN lightning network since 2018. A fraction of multi-ELVES can also be interpreted as halos following ELVES. Halos are disc-shaped light transients emitted at 70-80 km altitudes, appearing at the center of the ELVES rings, due to the rearrangement of electric charges at the base of the ionosphere after a strong lightning event