Sex-Related Embryotoxicity of Pulp Mill Effluent Extracts in Medaka (Oryzias latipes) Female Leucophore-free FLFII Strain.

The aim of the present study was to evaluate the effects of exposure to Chilean pulp mill effluent extracts on developing postfertilized medaka embryos before and after sex definition relative to sex steroids (testosterone and 17beta-estradiol) and a wood phytoestrogen (beta-sitosterol). Our study included 2 waterborne semichronic exposure experiments, using a 24-h post fertilization (hpf) unknown-sex FLFII (female leucophore free) group and a second 72-hpf FLFII phenotypic sex-identified group (male autofluorescence leucophore) strain of medaka embryos. Chronic exposure of both FLFII strain embryo groups showed similar delay in time to hatch and decreased hatchability. Teratogenic responses such as vertebral malformation (fusion, incomplete formation, and lack of vertebral formation process) and pericardial edema were observed in both experiments, with a high percentage related to FLFII fluorescent leucophore-identified males. In addition, high mortality associated with severe malformations was observed in male and female embryos exposed to testosterone. Our research has demonstrated that exposure to Chilean mill effluent extracts caused severe male medaka embryotoxicity (in postfertilized embryos) before and after sex definition and, irrespective of the experimental group and effluent treatment, suggests partial removal following secondary treatment. Furthermore, differences in the severity and type of teratogenic effects with previous experiments (d-rR medaka strain), are associated with the unique phenotypes of this medaka mutant strain. Environ Toxicol Chem 2021;40:2297-2305. © 2021 SETAC.

Endocrine Disruptor Impacts on Fish From Chile: The Influence of Wastewaters.

Industrial wastewaters and urban discharges contain complex mixtures of chemicals capable of impacting reproductive performance in freshwater fish, called endocrine-disrupting compounds (EDCs). In Chile, the issue was highlighted by our group beginning over 15 years ago, by analyzing the impacts of pulp and paper mill effluents (PPME) in the Biobio, Itata, and Cruces River basins. All of the rivers studied are important freshwater ecosystems located in the Mediterranean region of Central Chile, each with a unique fish biodiversity. Sequentially, we developed a strategy based on laboratory assays, semicontrolled-field experiments (e.g., caging) and wild fish population assessments to explore the issue of reproductive impacts on both introduced and native fish in Chile. The integration of watershed, field, and laboratory studies was effective at understanding the endocrine responses in Chilean freshwater systems. The studies demonstrated that regardless of the type of treatment, pulp mill effluents can contain compounds capable of impacting endocrine systems. Urban wastewater treatment plant effluents (WWTP) were also investigated using the same integrated strategy. Although not directly compared, PPME and WWTP effluent seem to cause similar estrogenic effects in fish after waterborne exposure, with differing intensities. This body of work underscores the urgent need for further studies on the basic biology of Chilean native fish species, and an improved understanding on reproductive development and variability across Chilean ecosystems. The lack of knowledge of the ontogeny of Chilean fish, especially maturation and sexual development, with an emphasis on associated habitats and landscapes, are impediment factors for their conservation and protection against the threat of EDCs. The assessment of effects on native species in the receiving environment is critical for supporting and designing protective regulations and remediation strategies, and for conserving the unique Chilean fish biodiversity.

Red-legged cormorant uses plastic as nest material in an artificial breeding colony of Atacama Desert coast.

Plastic debris affects the survival of numerous marine species such as seabirds. Several studies have report ingestion and entanglement of seabird species in plastic debris but just few describe plastic presence in nests. Here we report the first evidence of a novel artificial nesting colony of red-legged cormorant (Phalacrocorax gaimardi) using plastic as nesting material in two major seaport structures in Northern Chile. We sampled 18 abandoned nests and determined that overall nests contained plastic: industrial polypropylene bulk bags (35%), plastic bags (33%) and fishing gear (10%). Plastic types differ among seaports, ENAEX (National Company of Explosives) seaport located near the main city contained plastic bags as predominant material while GNL (Liquid Natural Gas Company) seaport located near the industrial area contained bulkbag fibers. Further research should analyze debris in beaches nearby the nesting colony in order to assess whether red-legged cormorants have any plastic preference or if they use the most available in the nature.

A propósito de la contingencia COVID-19. ECMO en el adulto: Oxigenación por membrana extracorpórea. A quién, cómo y cuándo / With regard to COVID-19 contingency. ECMO in adults. Extracorporeal Membrane Oxygenation). To whom, how and when

ECMO (Extracorporeal Membrane Oxygenation) is an extracorporeal life support system in catastrophic lung failure, shock and cardiopulmonary resuscitation, in different age groups, with multiple physiologic features. When the candidate to be submitted is too unstable to be transported to a hospital with ECMO, cannulation before transfer allows stabilization and subsequent transport. The aim of this article is to review the current concepts of extracorporeal support, its indications, national and international experience, and its possible role in the SARS-Cov2 pandemic.

Assessing wild fish exposure to ligands for sex steroid receptors from pulp and paper mill effluents in the Biobio River Basin, Central Chile.

Bioactive substances in the Biobio River Basin in Chile were examined by deploying Semi Permeable Membrane Devices (SPMDs) upstream and downstream of 4 pulp mill effluent discharges. Androgenic and estrogenic activity of SPMD extracts were then evaluated using in vitro fish sex steroid receptor binding assays. The results indicated the occurrence of estrogenic type compounds associated with one of the mill discharges. A significant correlation among the presence of these compounds, an increase in gonadosomatic index GSI and induction of hepatic EROD activity of two native fish species was observed. However, no significant presence of mature oocytes in female gonads was detected. Although EROD induction was observed in sites impacted by mill effluents, an increase of its activity occurred towards the downstream areas, suggesting other non-mill sources. More research is needed to understand the environmental changes in context of the new technological improvements in treatment systems to MBBR (Moving Bed Biofilm Reactor) recently implemented by the pulp mill industries.

Reanimación cardiopulmonar extracorpórea: caso clínico de paro cardiaco extra hospitalario / Extracorporeal cardiopulmonay resuscitation: case report on an out-of-hospital cardiopulmonary arrest

Out-of-hospital cardiopulmonary arrest (OHCA) is highly lethal. Although overall survival is increasing, hospital discharge with good neurological prognosis remains low and highly variable. In some countries, protocols are being implemented, which include techniques in cardiopulmonary resuscitation, allowing a better neurological prognosis for those patients who undergo an OHCA. Following these new techniques and the incorporation of these new protocols already accepted in the guidelines of advanced cardiopulmonary resuscitation, we report a 54 years old male who presented an OHCA and received advanced cardiopulmonary by a professional team in situ. He was transferred to the emergency department, where optimal advanced resuscitation was continued, until the connection to extracorporeal cardiopulmonary support, with the aim of reestablishing blood flow, a technique known as cardiopulmonary resuscitation (ECPR: extracorporeal cardiopulmonary resuscitation). The patient was discharged from the hospital 25 days later.

Evaluating the potential of effluent extracts from pulp and paper mills in Canada, Brazil, and New Zealand to affect fish reproduction: Estrogenic effects in fish.

The authors examined the potential of pulp mill effluent from pulp-producing countries (Canada, Brazil, New Zealand) to affect fish reproduction. Specifically, the estrogenic effects in juvenile rainbow trout (Oncorhynchus mykiss) pulse-exposed to 11 different mill effluent extracts (intraperitoneal injections of solid-phase extraction-dichloromethane nonpolar fraction). The results indicated that effluent extracts were estrogenic in juvenile trout irrespective of the gender, as reflected by increasing level of plasma vitellogenin (VTG; Brazil > New Zealand > Canada). Despite the high variability observed among mills, differences in VTG levels were related to the type of mill process (kraft > elementary chlorine-free kraft > thermomechanical pulping). Moreover, effluent treatments did not appear to significantly decrease VTG induction. A consistent estrogenic effect was observed in those mills that process a combination of feedstocks (softwood and hardwood), with the highest increase in VTG related to eucalyptus feedstock. The results demonstrate significant estrogenic effects of pulp mill effluents on chronically exposed juvenile trout, suggesting that in vivo metabolic activation of precursors is necessary to cause the observed increases in VTG levels. This molecular estrogenic response provides a useful starting point for predicting population-level impacts through the adverse outcome pathway methodology. Environ Toxicol Chem 2017;36:1547-1555. © 2016 SETAC.

Evaluating the potential of effluents and wood feedstocks from pulp and paper mills in Brazil, Canada, and New Zealand to affect fish reproduction: chemical profiling and in vitro assessments.

This study investigates factors affecting reproduction in fish exposed to pulp and paper mill effluents by comparing effluents from countries with varying levels of documented effects. To explore the hypothesis of wood as a common source of endocrine disrupting compounds, feedstocks from each country were analyzed. Analyses included in vitro assays for androgenic activity (binding to goldfish testis androgen receptors), estrogenic activity (yeast estrogen screen), and neurotransmitter enzyme inhibition (monoamine oxidase and glutamic acid decarboxylase). Chemical analyses included conventional extractives, known androgens, and gas chromatograph index (GCI) profiles. All effluents and wood contained androgenic activity, particularly in nonpolar fractions, although known androgens were undetected. Effluents with low suspended solids, having undergone conventional biotreatment had lower androgenic activities. Estrogenic activity was only associated with Brazilian effluents and undetected in wood. All effluents and wood inhibited neurotransmitter enzymes, predominantly in polar fractions. Kraft elemental chlorine free mills were associated with the greatest neurotransmitter inhibition. Effluent and wood GCI profiles were correlated with androgenic activity and neurotransmitter enzyme inhibition. Differences in feedstock bioactivities were not reflected in effluents, implying mill factors mitigate bioactive wood components. No differences in bioactivities could be discerned on the basis of country of origin, thus we predict effluents in regions lacking monitoring would affect fish reproduction and therefore recommend implementing such programs.

Estrogenic and anti-estrogenic effects of wood extractives present in pulp and paper mill effluents on rainbow trout.

Wood extractives present in pulp and paper mill effluents may cause reproductive disturbances in fish. A chronic-exposure toxicity experiment using immature rainbow trout (Oncorhynchus mykiss) was conducted in order to assess the endocrine disrupting effects of two Chilean pulp and paper mill specific extracts (solid phase extraction, SPE) obtained from primary and secondary treated effluents. The (anti)estrogenic potencies and toxicity of the wood extractives regularly present in pulp mill effluent such as dehydroabietic acid (DHAA), beta-sitosterol (BS), and model estrogen 17beta-estradiol (E2) were evaluated by analysis of plasma vitellogenin (VTG) levels, gonadal somatic index (GSI) and liver ethoxyresorufin-O-deethylase (EROD) activity, respectively. The protocol involved the use of multiple intra-peritoneal injections (1 injection every 7 days for a total exposure period of 28 days). Analysis of variance/covariance, demonstrated no differences associated with fish gender other than GSI. The phytosterol BS, E2 and both pulp mill effluent extracts showed significant inductions of EROD and increased VTG levels after 4, 7, 14, 21 and 28 of exposure. While fish injected with secondary treated effluent extract showed a delayed induction in VTG levels compared to primary effluent injected fish, no effects on VTG and EROD levels were observed in DHAA injected fish. Moreover simultaneous injection of DHAA+E2 reduced the VTG levels found in E2 injected fish, indicating a potential indirect anti-estrogenic effect of this resin acid. The results of this study indicate that Chilean pulp and paper mill effluent extracts are estrogenic in rainbow trout males and females.

Effects of pulp and paper mill effluent extractives on aromatase CYP19a gene expression and sex steroid levels in juvenile triploid rainbow trout.

We evaluated plasma testosterone (T) and 17beta-estradiol (E2) levels and ovarian aromatase CYP19a gene expression following a single intraperitoneal injection of Chilean pulp and paper mill effluent extracts into juvenile triploid rainbow trout. Fish injected with untreated effluent extracts had increased plasma T after 4 days, while plasma E2 concentration was increased in fish injected with both primary and secondary treated effluent extracts at the same sampling period. Ovarian CYP19a gene expression as measured by qRT-PCR was significantly induced in fish injected with the untreated, primary and secondary treated pulp and paper mill effluent extracts. Similar induction of CYP19a expression was found in fish injected with the androgens androstenedione (ADD) and T. A Principal Component Analysis (PCA) was conducted in order to identify structure in relationships between all measured variables and identifying which factors were most responsible for the variance observed within the plasma steroid levels, upregulation of ovarian CYP19a gene expression and the final estrogenic effect of increased plasma VTG levels. This analysis indicated a cluster correlation between plasma T levels and CYP19a gene expression (Factor 1, explaining 27.2% of total variance), a cluster including condition factor and liver somatic index (Factor 2, explaining 17.3%) and an additional cluster including plasma E2 and vitellogenin levels (Factor 3, explaining an additional 15.8%). The present results indicate that Chilean pulp and paper mill effluent extracts cause estrogenic effects in triploid rainbow trout. These effects could be related to the compounds present in the effluent that act as estrogen receptor agonists, or that induce changes leading to increased amounts of endogenous estrogens, reflected by increased E2 levels and induced aromatase expression/activity.

El bloqueo interescalénico provee mejor calidad e índice de satisfacción que la analgesia intra-articular en el manejo del dolor postoperatorio en las cirugías de hombro / Interscalenic block provides for higher quality and improved satisfaction indexes than intraarticular analgesia in managing post surgery shoulder pain

Introducción: La analgesia intra-articular (AI) y el Bloqueo Interescalénico Continuo (BIEC) se utilizan para el manejo del dolor postoperatorio de las cirugías de hombro. La utilidad e índice de satisfacción obtenidos por su empleo son controversiales. Objetivo: Evaluar la utilidad de la AI versus el BIEC, comparando la calidad de la analgesia postoperatoria e índice de satisfacción e incidencia de complicaciones asociadas en pacientes beneficiarios de una cirugía del hombro asistido por artroscopia. Material y Método: Estudio prospectivo controlado de 47 pacientes ASA I-II, divididos en dos grupos. Grupop 1 AI: (n=23)Ropivacaína 0,20 por ciento 25 ml + Morfina 2mg.; Grupo 2: BIEC (n=24) Bupivacaína 0,0625 por ciento + Sufentanyl 1 up/ml con un débito de 7 ml/hr en IC. El dolor posquirúrgico se registro entre las 2 y 48 horas mediante Escala Visual Análoga (EVA). Se utilizó morfina endovenosa postoperatoria. Se registro el consumo de morfina, la incidencia de efectos adversos, el grado de satisfacción experimentados por los pacientes a las 48 hrs. Resultados: No hubo diferencias estadísticamente significativas en los parámetros demográficos de ambos grupos . El EVA postoperatorio y el consumo de Morfina (CM) entre las 4 y 12 horas fueron similares entre los dos grupos. El EVA entre las 24 y 48 hrs. fue significativamente menor en el grupo BIEC (25 +/- 4 mm) versus el grupo AI (47 +/-6 mm), (p=0,001). el CM entre las 24 y 48 hrs.fue de 13,5 +/- 3 mg en el grupo AI versus 6 +/- 3 mg en el grupo 2 (p=0,001). El grado de satisfacción experimentado por los pacientes fue superior en el grupo BIEC (p=0,03). HUbo mayor incidencia de náuseas y/o vómitos en el grupo AI. Conclusiones: El manejo del dolor postoperatorio mediante las técnicas de AI y BIEC es adecuado en cirugías de hombro. El BIEC se demuestra más efectivo que la AI después de las 12 hrs. postoperatorias, presentando los pacientes menor dolor y NVPO y un mayor índice de satisfacción.

Introduction: Intraarticular analgesia (IA) and Continuous Interscalenic Block (CIB) methods are used as analgesia in post shoulder surgery. The results obtained and the satisfaction index are rather controversial. Objective: To evaluate the usefulness of IA vs. CIB by comparing the post surgery analgesia and degree of satisfaction to the complications associated to arthroscopic shoulder surgery (ASS). Materials and Method: Controlled clinical trial with 47 ASS I and II patients separated in two groups. Group one, IA method: (n=23): 25ml of 0.2 percent ropivacaine plus 2 mgs of morphine; Group two, CIB method: (n=24) 0.0625 percent bupivacaine and 1 up/ml continuous sufentatil supply at 7ml/hr. Post surgery pain was recorded after 2 to 48 hours by means of a Visual Analogue Scale (VAS) and intravenous morphine was used for pain management. The use of morphine was recorded as well as adverse effects and the degree of satisfactions of patients after 48 hours. Results: There were no significant differences in demographic data between these two groups. Post surgery VAS and morphine use after 4 and 12 hours were similar between the two groups. VAS recorded after 24 and 48 hours was significantly lower in Group CIB (25+- 4mm) compared to Group IA (47+- 6mm), (p=0.001). The use of morphine from 24 to 48 hours was 13.5 +- 3mg in Group IA while use in Group CIB was 6 +- 3mg (p=0.01).The degree of satisfaction of patients was higher in Group CIB (p=0.03). Patients in Group IA experienced nausea and/or vomiting. Conclusions: The IA and CIB methods for pain management are adequate in post shoulder surgery. CIB proves to be more effective than IA after 12 hours with patients feeling less pain, a lower nvpo and a higher degree of satisfaction.

Basal and 3,3',4,4',5-pentachlorobiphenyl-induced expression of cytochrome P450 1A, 1B and 1C genes in zebrafish.

The cytochrome P4501C (CYP1C) gene subfamily was recently discovered in fish, and zebrafish (Danio rerio) CYP1C1 transcript has been cloned. Here we cloned the paralogous CYP1C2, showing that the amino acid sequence is 78% identical to CYP1C1, and examined gene structure and expression of CYP1A, CYP1B1, CYP1C1, and CYP1C2. Xenobiotic response elements were observed upstream of the coding regions in all four genes. Zebrafish adults and embryos were exposed (24 h) to 100 nM 3,3',4,4',5-polychlorinated biphenyl (PCB126) or 20 ppm acetone and subsequently held in clean water for 24 h (adults) or 48 h (embryos). All adult organs examined (eye, gill, heart, liver, kidney, brain, gut, and gonads) and embryos showed basal expression of the four genes. CYP1A was most strongly expressed in liver, whereas CYP1B1, CYP1C1, and CYP1C2 were most strongly expressed in heart and eye. CYP1B1 and the CYP1C genes showed an expression pattern similar to one another and to mammalian CYP1B1. In embryos CYP1C1 and CYP1C2 tended to have a higher basal expression than CYP1A and CYP1B1. PCB126 induced CYP1A in all organs, and CYP1B1 and CYP1C1 in all organs except gonads, or gonads and brain, respectively. CYP1C2 induction was significant only in the liver. However, in embryos all four genes were induced strongly by PCB126. The results are consistent with CYP1C1 and CYP1C2, as well as CYP1A and CYP1B1, being regulated by the aryl hydrocarbon receptor. While CYP1A may have a protective role against AHR agonists in liver and gut, CYP1B1, CYP1C1, and CYP1C2 may also play endogenous roles in eye and heart and possibly other organs, as well as during development.