High-Resolution Mass Spectrometry Non-Targeted Detection of Per- and Polyfluoroalkyl Substances in Roe Deer (Capreolus capreolus).

Among wildlife species, roe deer stands out as a valuable indicator of environmental pollution due to its ecological significance and role as a game animal. The assessment of poly- and perfluoro substances (PFASs) bioaccumulation is of the utmost importance, relying on the liver and muscles as the main organs of interest. The study concerned the identification of 60 PFAS through a non-target workflow analysis based on HPLC Q-Exactive Orbitrap High-Resolution Mass Spectrometry in a homogeneous group of 18 female roe deer species. The developed strategy allowed us to individuate the 60 PFAS compounds with different levels of confirmation. Apart from seven PFASs identified via analytical standards, the remaining fifty-three features were identified with CL 2 or 3. Moreover, by applying a differential statistic approach, it was possible to distinguish the bioaccumulation patterns in the liver and muscle, identifying 12 PFAS upregulated in the muscle and 20 in the liver. The analysis reveals that specific PFAS compounds present exclusively in either the muscle or in the liver. The study emphasises the specificity of the liver and muscle as significant bioaccumulation sites for PFAS, raising questions about the underlying mechanisms of this process. In conclusion, the presented non-targeted PFAS analysis workflow evidenced promising and reliable results, successfully demonstrating its feasibility in the field of environmental research.

Influence of Area, Age and Sex on Per- and Polyfluorinated Alkyl Substances Detected in Roe Deer Muscle and Liver from Selected Areas of Northern Italy.

Due to their physicochemical properties, per- and polyfluorinated alkyl substances (PFASs) persist and bioaccumulate in living organisms, causing adverse health effects. Since exposure to xenobiotics is influenced by factors related to both the living organism and the considered compounds, biomonitoring PFASs' presence in the environment is of crucial importance. This study aimed to detect and quantify 15 PFASs in the muscle and liver of 40 roe deer from a specific area in Northern Italy by UPLC-HRMS. In the roe deer, liver PFAS concentrations were higher than those seen in muscle (p < 0.05). Although PFAS content in animals from urbanized areas was higher than those found in deer from rural areas, this difference was not statistically significant. In female roe deer, the concentration was higher than in males (p < 0.05); moreover, older animals showed higher concentrations of PFASs in the liver than younger animals (p < 0.05). In conclusion, the amount of PFASs was higher in tissues from roe deer belonging to urbanized areas, showing that this species might serve as a good bioindicator due to its territorial behavior. PFAS content was significantly higher in female roe deer, although the reason is not fully known. Finally, PFAS concentration was higher in the liver of older animals, probably due to compromised hepatic function.

First Investigation of the Physiological Distribution of Legacy and Emerging Perfluoroalkyl Substances in Raw Bovine Milk According to the Component Fraction.

Bovine milk is a pillar of the human diet and plays a key role in the nutrition of infants. Perfluoroalkyl substances (PFASs) are well-recognized highly stable organic compounds that are able to pollute ecosystems persistently and threaten both human and animal health. The study aimed to analyze the distribution of 14 PFASs within the milk matrix by comparing their content in whole milk, and its skimmed and creamed fractions. Raw milk samples were individually collected from 23 healthy cows (10 primiparous and 13 multiparous) reared on a farm in Northern Italy not surrounded by known point sources of PFASs. Each sample was fractioned in whole, skim, and cream components to undergo PFAS analysis using liquid chromatography-high-resolution mass spectrometry. All samples contained at least one PFAS, with perfluorobutanoic acid (PFBA) being the primary contaminant in all three fractions, followed by perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA). PFOS was shown to be significantly (p < 0.001) more concentrated in cream than in raw and skimmed milk. Multiparous cows showed a higher frequency of positive samples in all analyzed fractions. Further research is necessary to assess the risk of dairy diets and high-fat dairy products and to investigate the toxicological effects of PFASs on cattle, even in environments without known PFAS sources.

Clinical pharmacokinetics of a dexmedetomidine-methadone combination in dogs undergoing routine anaesthesia after buccal or intramuscular administration.

This study aimed to define the pharmacokinetic profiles of dexmedetomidine and methadone administered simultaneously in dogs by either an oral transmucosal route or intramuscular route and to determine the bioavailability of the oral transmucosal administration relative to the intramuscular one of both drugs, so as the applicability of this administration route in dogs. Twelve client-owned dogs, scheduled for diagnostic procedures, were treated with a combination of dexmedetomidine hydrochloride (10 µg/kg) and methadone hydrochloride (0.4 mg/kg) through an oral transmucosal route or intramuscularly. Oral transmucosal administration caused ptyalism in most subjects, and intramuscular administration caused transient peripheral vasoconstriction. The results showed reduced and delayed absorption of both dexmedetomidine and methadone when administered through an oral transmucosal route, with median (range) Cmax values of 0.82 (0.42-1.49) ng/ml and 13.22 (2.80-52.30) ng/ml, respectively. The relative bioavailability was low: 16.34% (dexmedetomidine) and 15.5% (methadone). Intramuscular administration resulted in a more efficient absorption profile, with AUC and Cmax values for both drugs approximately 10 times higher. Dexmedetomidine and methadone administered simultaneously by an oral transmucosal route using injectable formulations were not well absorbed through the oral mucosa. Nevertheless, additional studies on these drugs combination using alternative administration routes are recommended.

Socio-spatial inequalities related to tuberculosis in the city of Itaboraí, Rio de Janeiro. / Desigualdades socioespaciais relacionadas à tuberculose no município de Itaboraí, Rio de Janeiro.

INTRODUCTION: Tuberculosis remains as a global public health problem and its occurrence and distribution is associated with social inequalities. Itaboraí is among the priority Brazilian municipalities for tuberculosis control. The study aimed to combine social indicators into a composite indicator to measure differences in living conditions of the population in Itaboraí, as well as evidence of health inequalities related to tuberculosis. METHODS: An ecological study, with spatial analysis of tuberculosis and its relation to socioeconomic and demographic status of households. Data were obtained from individuals declared in the Mortality Information System (SIM), notified in the Notification of Injury Information System - SINAN or Tuberculosis Special Treatment Information System - SITETB from 2007 to 2013. RESULTS: Evidence indicates association between sociodemographic inequalities and occurrence of tuberculosis in the population. Findings point to a lower percentage of healing and a higher percentage of abandonment and death in areas with high social vulnerability, with a relative risk over twice as high than that found in areas of lower vulnerability. CONCLUSION: The study showed strong evidence that the influence of unequal sociodemographic conditions have a negative impact on health conditions of the population in Itaboraí.

INTRODUÇÃO: A tuberculose ainda é um problema de saúde pública mundial e sua ocorrência e distribuição estão associadas às desigualdades sociais. Itaboraí está entre os municípios brasileiros prioritários para o controle dessa doença. Os objetivos deste estudo foram combinar indicadores sociais em um indicador sintético para mensurar as diferenças nas condições de vida da população de Itaboraí, assim como evidenciar prováveis desigualdades em saúde relacionadas à tuberculose. MÉTODOS: Estudo ecológico, com análise espacial da tuberculose e sua relação com a situação socioeconômica e demográfica das famílias residentes. Os dados foram obtidos de indivíduos residentes, declarados no Sistema de Informação sobre Mortalidade (SIM), notificados no Sistema de Informação de Agravos de Notificação (SINAN) ou registrados no Sistema de Informação de Tratamentos Especiais de Tuberculose (SITETB), entre 2007 a 2013. RESULTADOS: Evidências indicam associação entre desigualdades sociodemográficas e ocorrência de tuberculose na população. Achados apontam menor percentual de cura e maior percentagem de abandono e óbito em áreas com maior vulnerabilidade social, com risco relativo mais de duas vezes superior ao encontrado nas áreas de menor vulnerabilidade. CONCLUSÃO: O estudo demonstrou fortes evidências de que a influência de condições sociodemográficas desiguais tem efeitos perniciosos nas condições de saúde da população em Itaboraí.

Transcriptomic responses of a simplified soil microcosm to a plant pathogen and its biocontrol agent reveal a complex reaction to harsh habitat.

BACKGROUND: Soil microorganisms are key determinants of soil fertility and plant health. Soil phytopathogenic fungi are one of the most important causes of crop losses worldwide. Microbial biocontrol agents have been extensively studied as alternatives for controlling phytopathogenic soil microorganisms, but molecular interactions between them have mainly been characterised in dual cultures, without taking into account the soil microbial community. We used an RNA sequencing approach to elucidate the molecular interplay of a soil microbial community in response to a plant pathogen and its biocontrol agent, in order to examine the molecular patterns activated by the microorganisms. RESULTS: A simplified soil microcosm containing 11 soil microorganisms was incubated with a plant root pathogen (Armillaria mellea) and its biocontrol agent (Trichoderma atroviride) for 24 h under controlled conditions. More than 46 million paired-end reads were obtained for each replicate and 28,309 differentially expressed genes were identified in total. Pathway analysis revealed complex adaptations of soil microorganisms to the harsh conditions of the soil matrix and to reciprocal microbial competition/cooperation relationships. Both the phytopathogen and its biocontrol agent were specifically recognised by the simplified soil microcosm: defence reaction mechanisms and neutral adaptation processes were activated in response to competitive (T. atroviride) or non-competitive (A. mellea) microorganisms, respectively. Moreover, activation of resistance mechanisms dominated in the simplified soil microcosm in the presence of both A. mellea and T. atroviride. Biocontrol processes of T. atroviride were already activated during incubation in the simplified soil microcosm, possibly to occupy niches in a competitive ecosystem, and they were not further enhanced by the introduction of A. mellea. CONCLUSIONS: This work represents an additional step towards understanding molecular interactions between plant pathogens and biocontrol agents within a soil ecosystem. Global transcriptional analysis of the simplified soil microcosm revealed complex metabolic adaptation in the soil environment and specific responses to antagonistic or neutral intruders.

Moderate warming in microcosm experiment does not affect microbial communities in temperate vineyard soils.

Changes in the soil microbial community structure can lead to dramatic changes in the soil ecosystem. Temperature, which is projected to increase with climate change, is commonly assumed to affect microbial communities, but its effects on agricultural soils are not fully understood. We collected soil samples from six vineyards characterised by a difference of about 2 °C in daily soil temperature over the year and simulated in a microcosm experiment different temperature regimes over a period of 1 year: seasonal fluctuations in soil temperature based on the average daily soil temperature measured in the field; soil temperature warming (2 °C above the normal seasonal temperatures); and constant temperatures normally registered in these temperate soils in winter (3 °C) and in summer (20 °C). Changes in the soil bacterial and fungal community structures were analysed by automated ribosomal intergenic spacer analysis (ARISA). We did not find any effect of warming on soil bacterial and fungal communities, while stable temperatures affected the fungal more than the bacterial communities, although this effect was soil dependent. The soil bacterial community exhibited soil-dependent seasonal fluctuations, while the fungal community was mainly stable. Each soil harbours different microbial communities that respond differently to seasonal temperature fluctuations; therefore, any generalization regarding the effect of climate change on soil communities should be made carefully.

Microbial community structure in vineyard soils across altitudinal gradients and in different seasons.

Microbial communities living in nine vineyards distributed over three altitudinal transects were studied over 2 years. Fungal and bacterial community dynamics were explored using automated ribosomal intergenic spacer analysis (ARISA) and by determining bacterial cells and fungal colony-forming units (CFUs). Moreover, extensive chemical and physical analyses of the soils were carried out. Multivariate analyses demonstrated that bacterial and fungal communities are affected by altitude, which acts as a complex physicochemical gradient. In fact, soil moisture, Al, Mg, Mn and clay content are changing with altitude and influencing the bacterial genetic structure, while in the case of fungi, soil moisture, B and clay content are found to be the main drivers of the community. Moreover, other exchangeable cations and heavy metals, not correlating with altitude, are involved in the ordination of the sites, especially Cu. Qualitative ARISA revealed the presence of a stable core microbiome of operational taxonomic units (OTUs) within each transect, which ranged between 57% and 68% of total OTUs in the case of fungi and between 63% and 72% for bacteria. No seasonal effect on the composition of microbial communities was found, demonstrating that bacterial and fungal communities in vineyards are mostly stable over the considered seasons.

Studying trophic interactions between a plant pathogen and two different antagonistic microorganisms using a 13C-labeled compound and isotope ratio mass spectrometry.

RATIONALE: The study of the interactions among microorganisms, especially between pathogens and other microorganisms, is a very useful way to identify possible biocontrol agents (BCAs). In this study we verified the capability of δ(13)C analysis using isotope ratio mass spectrometry (IRMS) to detect active parasitism or metabolic assimilation of (13)C-labeled Armillaria mellea (plant pathogen) by Trichoderma atroviride and Pseudomonas fluorescens (two BCAs). METHODS: The three microorganisms were labeled in pure-culture using a specific medium to which D-glucose (13)C was added. The δ(13)C analysis of mycelia/cells and DNA was undertaken using IRMS at different times, to study the uptake kinetics of (13)C. The mechanisms of interaction were studied by implementing dual-culture tests and measuring the δ(13)C values of the two BCAs after 29 days of contact with the labeled pathogen. RESULTS: A. mellea absorbed (13)C more slowly (plateau at 21 days) than T. atroviride and P. fluorescens (3 and 1 day, respectively) in pure-culture. The maximum δ(13)C values were higher in A. mellea and T. atroviride mycelia (8,019.9‰ and 10,383.7‰, respectively) than in P. fluorescens (953.4‰ in cells). In dual-culture the mycelia of T. atroviride which remained in direct contact with labeled A. mellea showed an increased δ(13)C value with respect to the unlabeled treatment (66.4‰ and -26.6‰, respectively), due to active interaction. Lower assimilation of (13)C was detected in P. fluorescens. CONCLUSIONS: This work demonstrates that IRMS can be used for the in-depth study of direct parasitism and interaction process between biocontrol agents and labeled pathogens, allowing the screening of potential new BCAs.

Producción de artículos científicos sobre salud en seis países de América Latina, 1973 a 1992 / Production of scientific articles about health in six Latin American countries, 1973-1992

Con objeto de observar las tendencias de la investigación sobre salud en América Latina, se analizó la producción de artículos resultantes de investigaciones biomédicas, clínicas y de salud pública originadas en Argentina, Brasil, Chile, Cuba, México y Venezuela de 1973 a 1992, inclusive. De la base de datos del Institute for Scientific Information (ISI), se recuperaron 41 238 artículos con primeros autores residentes en esos países. Esta producción se analizó por área de estudio, tipo de investigación, país, número de autores e instituciones que participaron en la investigación, y citaciones recibidas por cada artículo. Se hizo también un análisis de 95 artículos en epidemiología seleccionados de 570 publicados por autores de los seis países en 11 revistas de salud pública de gran prestigio internacional. Los resultados muestran que el número de trabajos publicados aumentó 117% entre el primero y último quinquenios. La investigación clínica tuvo la distribución más homogénea entre los países y la de salud pública, la más concentrada (60,7% se originó en el Brasil). Los artículos de investigación biomédica y de salud pública tuvieron un crecimiento relativamente mayor que los de investigación clínica a lo largo del período. Se observó una disminución relativa de artículos de un solo autor ­lo que sugiere trabajo en equipo más frecuente­ y unaumento de artículos con autores vinculados a dos o más instituciones nacionales o extranjeras, lo cual indica mayor cooperación entre instituciones y países. El promedio de citacionesrecibidas por artículo fue 3, lo que corresponde a menos de la mitad de las que reciben los artículos de la base del ISI (7,78). En cuanto al subconjunto de los 95 artículos de epidemiología,la gran mayoría (96%) trataban de enfermedades infecciosas o del área maternoinfantil, mientras que en la literatura internacional 78% de los artículos estaban dedicados a las enfermedades crónicas. En este conjunto se observó más cooperación con instituciones extranjeras y un índice de 4,36 citaciones por artículo.Se concluye que, pese a las limitaciones inherentes a este tipo de estudio, se pudieron notar algunas tendencias generales del desarrollo de la investigación en los seis países con mayor producción científica de América Latina y formular hipótesis sobre los factores que determinan esas tendencias. Tomados con cautela, los resultados de estudios de este tipo pueden ser de gran valor al definir políticas de ciencia y tecnología en salud

The production of articles resulting from biomedical, clinical, and public health studies that originated in Argentina, Brazil, Chile, Cuba, Mexico, and Venezuela from 1973 through 1992 was analyzed to discover trends in health research in Latin America. From the database of the Institute for Scientific Information (ISI), 41 238 articles with first authors who resided in those countries were extracted. These articles were analyzed by subject area, type of study, country, number of authors and institutions that participated in the investigation, and citations received by each article. Also analyzed were 95 articles in epidemiology selected from a pool of 570 published by authors from the six countries in 11 public health journals that enjoy international prestige. The results showed that the number of published works increased by 117% between the first and last five-year periods within the study period. Clinical research was distributed the most evenly among the countries, and public health research was the most concentrated (60.7% originated in Brazil). The numbers of biomedical and public health research articles showed relatively more growth than those reporting on clinical research throughout the period. A relative decrease was found in articles by only one author, which suggests a greater frequency of team efforts, and an increase was seen in articles with authors tied to two or more national or foreign institutions, which indicates greater cooperation between institutions and countries. The average number of citations received by each article was 3, which was less than half the number received by the articles in the ISI database (7.78). Regarding the subset of 95 articles in epidemiology, the great majority (96%) dealt with infectious diseases or maternal and child health, while in the international literature 78% of such articles were about chronic diseases. This group of articles gave evidence of more cooperation with international institutions and had a citation index of 4.36 per article. It is concluded that, despite the inherent limitations, this type of study reveals some general trends in the development of research in the six Latin American countries with the greatest scientific production and makes it possible to formulate hypotheses on the factors that influence these trends. Taken with the proper caution, the results of studies like this one can be of great value in defining health science and technology policies