RESUMEN
Intercellular communication in the nervous system occurs through the release of neurotransmitters into the synaptic cleft between neurons. In the presynaptic neuron, the proton pumping vesicular- or vacuolar-type ATPase (V-ATPase) powers neurotransmitter loading into synaptic vesicles (SVs), with the V1 complex dissociating from the membrane region of the enzyme before exocytosis. We isolated SVs from rat brain using SidK, a V-ATPase-binding bacterial effector protein. Single-particle electron cryomicroscopy allowed high-resolution structure determination of V-ATPase within the native SV membrane. In the structure, regularly spaced cholesterol molecules decorate the enzyme's rotor and the abundant SV protein synaptophysin binds the complex stoichiometrically. ATP hydrolysis during vesicle loading results in a loss of the V1 region of V-ATPase from the SV membrane, suggesting that loading is sufficient to induce dissociation of the enzyme.
Asunto(s)
Vesículas Sinápticas , ATPasas de Translocación de Protón Vacuolares , Animales , Ratas , Proteínas Bacterianas/química , Encéfalo/ultraestructura , Encéfalo/enzimología , Colesterol/química , Microscopía por Crioelectrón , Hidrólisis , Vesículas Sinápticas/enzimología , Vesículas Sinápticas/ultraestructura , Sinaptofisina/metabolismo , ATPasas de Translocación de Protón Vacuolares/química , ATPasas de Translocación de Protón Vacuolares/aislamiento & purificación , ATPasas de Translocación de Protón Vacuolares/ultraestructura , Conformación ProteicaRESUMEN
Oxidative phosphorylation, the combined activities of the electron transport chain (ETC) and ATP synthase, has emerged as a valuable target for antibiotics to treat infection with Mycobacterium tuberculosis and related pathogens. In oxidative phosphorylation, the ETC establishes a transmembrane electrochemical proton gradient that powers ATP synthesis. Monitoring oxidative phosphorylation with luciferase-based detection of ATP synthesis or measurement of oxygen consumption can be technically challenging and expensive. These limitations reduce the utility of these methods for characterization of mycobacterial oxidative phosphorylation inhibitors. Here, we show that fluorescence-based measurement of acidification of inverted membrane vesicles (IMVs) can detect and distinguish between inhibition of the ETC, inhibition of ATP synthase, and nonspecific membrane uncoupling. In this assay, IMVs from Mycobacterium smegmatis are acidified either through the activity of the ETC or ATP synthase, the latter modified genetically to allow it to serve as an ATP-driven proton pump. Acidification is monitored by fluorescence from 9-amino-6-chloro-2-methoxyacridine, which accumulates and quenches in acidified IMVs. Nonspecific membrane uncouplers prevent both succinate- and ATP-driven IMV acidification. In contrast, the ETC Complex III2IV2 inhibitor telacebec (Q203) prevents succinate-driven acidification but not ATP-driven acidification, and the ATP synthase inhibitor bedaquiline prevents ATP-driven acidification but not succinate-driven acidification. We use the assay to show that, as proposed previously, lansoprazole sulfide is an inhibitor of Complex III2IV2, whereas thioridazine uncouples the mycobacterial membrane nonspecifically. Overall, the assay is simple, low cost, and scalable, which will make it useful for identifying and characterizing new mycobacterial oxidative phosphorylation inhibitors.
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Antibacterianos , Descubrimiento de Drogas , Mycobacterium tuberculosis , Fosforilación Oxidativa , Adenosina Trifosfato/antagonistas & inhibidores , Adenosina Trifosfato/metabolismo , Complejo III de Transporte de Electrones/efectos de los fármacos , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/metabolismo , Fosforilación Oxidativa/efectos de los fármacos , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Descubrimiento de Drogas/métodosRESUMEN
Mycobacteria, such as Mycobacterium tuberculosis, depend on the activity of adenosine triphosphate (ATP) synthase for growth. The diarylquinoline bedaquiline (BDQ), a mycobacterial ATP synthase inhibitor, is an important medication for treatment of drug-resistant tuberculosis but suffers from off-target effects and is susceptible to resistance mutations. Consequently, both new and improved mycobacterial ATP synthase inhibitors are needed. We used electron cryomicroscopy and biochemical assays to study the interaction of Mycobacterium smegmatis ATP synthase with the second generation diarylquinoline TBAJ-876 and the squaramide inhibitor SQ31f. The aryl groups of TBAJ-876 improve binding compared with BDQ, while SQ31f, which blocks ATP synthesis ~10 times more potently than ATP hydrolysis, binds a previously unknown site in the enzyme's proton-conducting channel. Remarkably, BDQ, TBAJ-876, and SQ31f all induce similar conformational changes in ATP synthase, suggesting that the resulting conformation is particularly suited for drug binding. Further, high concentrations of the diarylquinolines uncouple the transmembrane proton motive force while for SQ31f they do not, which may explain why high concentrations of diarylquinolines, but not SQ31f, have been reported to kill mycobacteria.
Asunto(s)
Diarilquinolinas , Mycobacterium tuberculosis , Diarilquinolinas/farmacología , Antituberculosos/farmacología , Antituberculosos/química , Antituberculosos/metabolismo , Adenosina Trifosfato/metabolismo , Mycobacterium tuberculosis/genéticaRESUMEN
The purpose of this article is to remind the importance of the inverse square law in radiotherapy and especially in brachytherapy. Indeed, beyond the impact in radiation therapy with high energy beam, there is the use of radionuclides and low energy photons with short FSD where it is still more important. Comparisons between Iridium Brachytherapy and low energy X-rays brachytherapy show equivalent dose distributions in the first few centimeters. If the inverse square law is not the only element influencing the dose distributions calculations, it must not be forgotten. And it is playing a major role in brachytherapy with short FSD (<6cm).
Asunto(s)
Braquiterapia , Oncología por Radiación , Humanos , Fotones/uso terapéutico , Radioisótopos/uso terapéutico , Rayos X , Dosificación Radioterapéutica , Radioisótopos de Iridio/uso terapéuticoRESUMEN
During respiration, adenosine triphosphate (ATP) synthases harness the electrochemical proton motive force (PMF) generated by the electron transport chain (ETC) to synthesize ATP. These macromolecular machines operate by a remarkable rotary catalytic mechanism that couples transmembrane proton translocation to rotation of a rotor subcomplex, and rotation to ATP synthesis. Initially, x-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, and cross-linking were the only ways to gain insights into the three-dimensional (3D) structures of ATP synthases and, in particular, provided ground-breaking insights into the soluble parts of the complex that explained the catalytic mechanism by which rotation is coupled to ATP synthesis. In contrast, early electron microscopy was limited to studying the overall shape of the assembly. However, advances in electron cryomicroscopy (cryoEM) have allowed determination of high-resolution structures, including the membrane regions of ATP synthases. These studies revealed the high-resolution structures of the remaining ATP synthase subunits and showed how these subunits work together in the intact macromolecular machine. CryoEM continues to uncover the diversity of ATP synthase structures across species and has begun to show how ATP synthases can be targeted by therapies to treat human diseases.
RESUMEN
Phosphorus (P) plays essential roles in crops growth. Natural mineral sources of phosphate are non-renewable, overexploited and unevenly distributed worldwide, making P a strategic resource for agricultural systems. The search for sustainable ways to secure P supply for fertilizer production has therefore become a critical issue worldwide. Sewage sludge (SS) is an organic waste material considered as a key alternative source of P. Switzerland and the European Union are about to make it mandatory to recover P from SS or its treatment residues. Among the many technical options to achieve this objective, SS thermochemical treatments spiked with Cl-donors appear as a promising approach to recover P from SS and separate it from mineral pollutants such as trace metal elements (TME). The purpose of Cl-donor additives is to fix P within the mineral residues, possibly in bioavailable P species forms, while promoting TME vaporization by chlorination mechanisms. This review paper compares the various thermochemical treatments investigated worldwide over the past two decades. The influence of process conditions and Cl-donor nature is discussed. The presented results show that, except for nickel and chromium, most TME can be significantly vaporized during a high temperature treatment (over 900⯰C) with Cl addition. In addition, the fixation rate and solubility of P is increased when a Cl-donor such as MgCl2 is added.
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Fósforo , Aguas del Alcantarillado , Cromo , Fertilizantes , VolatilizaciónRESUMEN
In order to provide more convenient irradiation regimens for patient comfort, radiation facility organization and health expenses, new hypofractionated protocols have been evaluated. Moderately (dose/fraction: 2.3 to 3Gy), then ultra (dose/fraction: 5.2 to 6.1Gy) hypofractionated irradiations were first validated. The current question is: is it possible to go forward using extreme hypofractionated regimens (EHR) based on 1 to 3 fractions. Different irradiation techniques are under investigation. However, brachytherapy remains the smartest way to deliver a high dose in a small volume. We report prospective and retrospective study results which evaluated EHR for breast and prostate brachytherapy. While oncological outcome and toxicity profile appear extremely encouraging for low-risk breast cancer after a 1 to 4 fractions (6.25 to 20Gy/fraction), the use of a single fraction of 19 to 23Gy appears debatable for prostate cancer. Brachytherapy represents an emblematic example of EHR but longer follow-up and more mature results are awaited in order to specify the right indications and refine the EQD2 calculation method including new biological and technical factors.
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Braquiterapia , Neoplasias de la Próstata , Braquiterapia/efectos adversos , Braquiterapia/métodos , Humanos , Masculino , Estudios Prospectivos , Neoplasias de la Próstata/radioterapia , Hipofraccionamiento de la Dosis de Radiación , Estudios RetrospectivosRESUMEN
Two protoporphyrin IX (PpIX) adamantane derivatives were synthesized and then metallated with zinc. The Zn-PpIX derivatives, exhibiting a high singlet oxygen quantum yield, were tested for their photodynamic activity against the HT-29 cell line. In order to enhance their water-solubility and their cellular bioavailability, these photosensitizers were encapsulated into the hydrophobic cavity of cyclodextrins (CD) previously attached to cellulose nanocrystals (CNCs) via electrostatic interactions. Under illumination, the encapsulated adamantanyl-porphyrins exerted an enhanced in vitro cytotoxicity, as compared with the corresponding free photosensitizers.
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Adamantano/farmacología , Antineoplásicos/farmacología , Celulosa/farmacología , Ciclodextrinas/farmacología , Fotoquimioterapia , Fármacos Fotosensibilizantes/farmacología , Protoporfirinas/farmacología , Adamantano/química , Antineoplásicos/síntesis química , Antineoplásicos/química , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Celulosa/química , Ciclodextrinas/química , Relación Dosis-Respuesta a Droga , Diseño de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Células HT29 , Humanos , Estructura Molecular , Nanopartículas/química , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/química , Protoporfirinas/química , Relación Estructura-ActividadRESUMEN
Tuberculosis-the world's leading cause of death by infectious disease-is increasingly resistant to current first-line antibiotics1. The bacterium Mycobacterium tuberculosis (which causes tuberculosis) can survive low-energy conditions, allowing infections to remain dormant and decreasing their susceptibility to many antibiotics2. Bedaquiline was developed in 2005 from a lead compound identified in a phenotypic screen against Mycobacterium smegmatis3. This drug can sterilize even latent M. tuberculosis infections4 and has become a cornerstone of treatment for multidrug-resistant and extensively drug-resistant tuberculosis1,5,6. Bedaquiline targets the mycobacterial ATP synthase3, which is an essential enzyme in the obligate aerobic Mycobacterium genus3,7, but how it binds the intact enzyme is unknown. Here we determined cryo-electron microscopy structures of M. smegmatis ATP synthase alone and in complex with bedaquiline. The drug-free structure suggests that hook-like extensions from the α-subunits prevent the enzyme from running in reverse, inhibiting ATP hydrolysis and preserving energy in hypoxic conditions. Bedaquiline binding induces large conformational changes in the ATP synthase, creating tight binding pockets at the interface of subunits a and c that explain the potency of this drug as an antibiotic for tuberculosis.
Asunto(s)
Complejos de ATP Sintetasa/química , Antituberculosos/química , Microscopía por Crioelectrón , Diarilquinolinas/química , Mycobacterium smegmatis/enzimología , Tuberculosis/tratamiento farmacológico , Tuberculosis/microbiología , Complejos de ATP Sintetasa/antagonistas & inhibidores , Complejos de ATP Sintetasa/metabolismo , Adenosina Trifosfato/metabolismo , Antituberculosos/metabolismo , Antituberculosos/farmacología , Diarilquinolinas/metabolismo , Diarilquinolinas/farmacología , Hidrólisis/efectos de los fármacos , Modelos Moleculares , Mycobacterium smegmatis/efectos de los fármacos , RotaciónRESUMEN
The most prevalent form of Charcot-Marie-Tooth disease (CMT type 1A) is characterized by duplication of the PMP22 gene, peripheral dysmyelination and decreased nerve conduction velocities leading to muscle weakness. Recently, oxidative stress was reported as a feature in CMT1A patients. Curcumin exhibits antioxidant activities and has shown beneficial properties on peripheral nerves. However, curcumin presents unfavorable pharmacokinetics. We developed curcumin-cyclodextrin/cellulose nanocrystals (Nano-Cur) to bypass this limitation. The present study investigated the therapeutic potential of Nano-Cur in vitro in Schwann cells (SCs) and in vivo in the transgenic CMT1A rat model. In vitro, Nano-Cur treatment (0.01⯵M for 8â¯h) reduced reactive oxygen species and improved mitochondrial membrane potential in CMT1A SCs. Moreover, Nano-Cur treatment (0.01⯵M for 1 week) increased the expression of myelin basic protein in SC/neuron co-cultures. Preliminary in vivo experiments carried out in WT rats showed that intraperitoneal (i.p.) injection of Nano-Cur treatment containing 0.2â¯mg/kg of curcumin strongly enhanced the bioavailability of curcumin. Afterwards, in 1-month-old male CMT1A rats, Nano-Cur treatment (0.2â¯mg/kg/day, i.p. for 8 weeks) significantly improved sensori-motor functions (grip strength, balance performance, and mechanical and thermal sensitivities). Importantly, sensory and motor nerve conduction velocities were improved. Further histological and biochemical analyses indicated that myelin sheath thickness and myelin protein expression (myelin protein zero and PMP22) were increased. In addition, oxidative stress markers were decreased in the sciatic nerve and gastrocnemius muscle. Finally, Nrf2 expression and some major antioxidant enzymes were increased in sciatic nerve. Therefore, Nano-Cur significantly improved cellular, electrophysiological, and functional features of CMT1A rats.
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Enfermedad de Charcot-Marie-Tooth , Curcumina , Ciclodextrinas , Nanopartículas , Animales , Celulosa , Enfermedad de Charcot-Marie-Tooth/tratamiento farmacológico , Enfermedad de Charcot-Marie-Tooth/genética , Curcumina/farmacología , Humanos , Masculino , Estrés Oxidativo , Fenotipo , Ratas , Ratas TransgénicasRESUMEN
ß-Hemoglobinopathies can trigger rapid production of red blood cells in a process known as stress erythropoiesis. Cellular stress prompts differentiating erythroid precursors to express high levels of fetal γ-globin. However, the mechanisms underlying γ-globin production during cellular stress are still poorly defined. Here, we use CRISPR-Cas genome editing to model the stress caused by reduced levels of adult ß-globin. We find that decreased ß-globin is sufficient to induce robust re-expression of γ-globin, and RNA sequencing (RNA-seq) of differentiating isogenic erythroid precursors implicates ATF4 as a causal regulator of this response. ATF4 binds within the HBS1L-MYB intergenic enhancer and regulates expression of MYB, a known γ-globin regulator. Overall, the reduction of ATF4 upon ß-globin knockout decreases the levels of MYB and BCL11A. Identification of ATF4 as a key regulator of globin compensation adds mechanistic insight to the poorly understood phenomenon of stress-induced globin compensation and could inform strategies to treat hemoglobinopathies.
Asunto(s)
Factor de Transcripción Activador 4/metabolismo , Regulación de la Expresión Génica , Proteínas Proto-Oncogénicas c-myb/genética , Globinas beta/metabolismo , gamma-Globinas/genética , Factor de Transcripción Activador 4/genética , Secuencia de Bases , Diferenciación Celular/genética , Línea Celular , ADN Intergénico/genética , Regulación hacia Abajo/genética , Elementos de Facilitación Genéticos/genética , Hemoglobina Fetal/genética , Células Madre Hematopoyéticas/metabolismo , Humanos , Mutación/genética , Unión Proteica , Proteínas Proto-Oncogénicas c-myb/metabolismo , Proteínas Represoras/metabolismo , Factores de Tiempo , Transcripción Genética , Transcriptoma/genética , Regulación hacia Arriba/genética , gamma-Globinas/metabolismoRESUMEN
AIMS: To assess the bacterial diversity in the French egg processing industry and to explore the adhesion and spoilage potential of selected bacteria. METHODS AND RESULTS: Sterile stainless steel chips were suspended for 2 months inside the pipelines of seven egg processing companies, before and after the pasteurizer, at warm and cold seasons. After exposure, the bacterial diversity was assessed by 16S rDNA sequencing. The 231 collected isolates were mainly facultative anaerobic Gram positive bacteria, such as Streptococcus, Staphylococcus, Bacillus and Kocuria. Sixty-five representative isolates were further characterized in vitro regarding the potential for adhesion and egg product spoilage. A high diversity was observed from one genus to another. Kocuria and Rothia isolates showed significantly higher adhesion than the isolates of the other genera. Only the isolates belonging to the genera Bacillus and Lysinibacillus, associated with high enzymatic activities on a solid egg-based medium, were able to induce spoilage of liquid whole egg. CONCLUSIONS: Bacteria collected on stainless steel surfaces placed in egg processing industries could be associated to liquid egg product spoilage. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new insights on the bacterial contamination in egg processing companies and represents a first step for the effective control of undesirable bacteria in liquid egg products.
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Bacterias/aislamiento & purificación , Biodiversidad , Huevos/microbiología , Contaminación de Alimentos/análisis , Manipulación de Alimentos/instrumentación , Acero Inoxidable/análisis , Bacterias/clasificación , Bacterias/genética , Huevos/análisis , Microbiología de Alimentos , PasteurizaciónRESUMEN
This paper describes the preparation of two chalcone/ß-cyclodextrin/cellulose-nanocrystals complexes and the study of their antiproliferative activities against two colorectal and two prostatic cancer cell lines. The aim of this work was to enhance hydrosolubility of chalcones thanks to the hydrophilic character of cellulose nanocrystals. These latter were linked, through ionic interactions, to a cationic derivative of ß-cyclodextrins whose lipophilic cavity allowed the encapsulation of hydrophobic chalcones: 3-hydroxy-3',4,4',5'-tetramethoxychalcone (1) and 3',4,4',5'-tetramethoxychalcone (2). First, we showed that encapsulation allowed hydrosolubilization of chalcones. Then, chalcone/ß-cyclodextrin/cellulose-nanocrystals complexes demonstrated enhanced in vitro antiproliferative activities, compared to the corresponding free-chalcones.
Asunto(s)
Celulosa/química , Chalconas/química , Nanopartículas/química , beta-Ciclodextrinas/química , HumanosRESUMEN
Due to their design and mode of operation, French Vertical Flow Constructed Wetlands (VFCWs) accumulate suspended solids from the inflow wastewater in the form of a sludge layer at the surface of the first filter. In order to maintain the treatment performance over the long term, the characteristics of the sludge deposits and their evolution have to be well described. In this objective, a panel of sludge deposit samples taken from 14 French VFCW sewage treatment plants was investigated. Elemental composition and organic matter content, nature and reactivity were analyzed. Results clearly revealed two categories of sludge deposits, namely the "young-age plants" type (1â¯year of operation and less) and the "mature plants" type (3â¯years of operation and more). Sludge deposits from the "mature plants" exhibited same biological, physical and chemical properties. Their organic matter was globally less abundant, more humified and less biodegradable than in the young-age plants type. Their overall contents in trace metals were also higher, although in a limited manner. The effect of additional treatments, particularly FeCl3 injection for phosphorus precipitation, was observable in the "young-age plants" group. Finally, the sludge deposits sampled from one particular plant with specific operating conditions were found to exhibit very different characteristics from those of either groups identified. This observation underlined the influence of local conditions on the typology of the sludge deposits.
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Eliminación de Residuos Líquidos/métodos , Humedales , Filtración , Francia , Fósforo , Aguas del Alcantarillado , Aguas ResidualesRESUMEN
French vertical flow constructed wetlands (French VFCWs) are widely used for the treatment of wastewaters from small communities. In the system, unsettled wastewater is percolated through two successive stages of filter-cells planted with reeds. This causes the formation of a surface sludge layer. This layer plays positive roles in the treatment performance, but also leads to clogging. The objective of this study was to contribute to the description of the sludge deposits characteristics and their dynamics of evolution, which may control the development of clogging. Representative samples of sludge deposits were taken from 14 French VFCWs full-scale plants and analyzed for particle size, dynamic vapor sorption and other parameters of composition to compare their structure and evaluate the factors of influence. Results showed that ageing of the surface deposits layer over the years of operation in each plant induced the formation and integration of microaggregates within the initial macrostructure of fresh organic matter (OM). The humification process of the OM was found to play a key role in the aggregation process. The injection of FeCl3 operated to precipitate phosphates before filtration was found to accelerate the aggregation process in the early phase (<1 year) of operation of the sludge.
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Sustancias Húmicas/análisis , Aguas del Alcantarillado/química , Vapor/análisis , Aguas Residuales/química , Purificación del Agua/métodos , Humedales , Filtración , Francia , Modelos Teóricos , Tamaño de la PartículaRESUMEN
The pine processionary moth Thaumetopoea pityocampa (Lepidoptera: Notodontidae) is the main pine defoliator in the Mediterranean region. Its urticating larvae cause severe human and animal health concerns in the invaded areas. This species shows a high phenotypic variability for various traits, such as phenology, fecundity and tolerance to extreme temperatures. This study presents the construction and analysis of extensive genomic and transcriptomic resources, which are an obligate prerequisite to understand their underlying genetic architecture. Using a well-studied population from Portugal with peculiar phenological characteristics, the karyotype was first determined and a first draft genome of 537 Mb total length was assembled into 68,292 scaffolds (N50 = 164 kb). From this genome assembly, 29,415 coding genes were predicted. To circumvent some limitations for fine-scale physical mapping of genomic regions of interest, a 3X coverage BAC library was also developed. In particular, 11 BACs from this library were individually sequenced to assess the assembly quality. Additionally, de novo transcriptomic resources were generated from various developmental stages sequenced with HiSeq and MiSeq Illumina technologies. The reads were de novo assembled into 62,376 and 63,175 transcripts, respectively. Then, a robust subset of the genome-predicted coding genes, the de novo transcriptome assemblies and previously published 454/Sanger data were clustered to obtain a high-quality and comprehensive reference transcriptome consisting of 29,701 bona fide unigenes. These sequences covered 99% of the cegma and 88% of the busco highly conserved eukaryotic genes and 84% of the busco arthropod gene set. Moreover, 90% of these transcripts could be localized on the draft genome. The described information is available via a genome annotation portal (http://bipaa.genouest.org/sp/thaumetopoea_pityocampa/).
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Genoma de los Insectos , Lepidópteros/genética , Animales , Cromosomas Artificiales Bacterianos , Bases de Datos Genéticas , Perfilación de la Expresión Génica , Biblioteca de Genes , Cariotipificación , TranscriptomaRESUMEN
Chlamydia and Chlamydia-like bacteria are well known to infect several organisms and may cause a wide range of diseases, particularly in ruminants. To gain insight into the prevalence and diversity of these intracellular bacteria, we applied a pan-Chlamydiales real-time PCR to 1,134 veterinary samples taken from 130 Tunisian ruminant herds. The true adjusted animal population-level prevalence was 12.9% in cattle, against 8.7% in sheep. In addition, the true adjusted herd-level prevalence of Chlamydiae was 80% in cattle and 25.5% in sheep. Chlamydiales from three family-level lineages were detected indicating a high biodiversity of Chlamydiales in ruminant herds. Our results showed that Parachlamydia acanthamoebae could be responsible for bovine and ovine chlamydiosis in central-eastern Tunisia. Multivariable logistic regression analysis at the animal population level indicated that strata and digestive disorders variables were the important risk factors of bovine and ovine chlamydiosis. However, origin and age variables were found to be associated with bovine and ovine chlamydiosis, respectively. At the herd level, risk factors for Chlamydia positivity were as follows: abortion and herd size for cattle against breeding system, cleaning frequency, quarantine, use of disinfectant and floor type for sheep. Paying attention to these risk factors will help improvement of control programs against this harmful zoonotic disease.
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Animales Domésticos , Enfermedades de los Bovinos/epidemiología , Infecciones por Chlamydia/veterinaria , Chlamydia/aislamiento & purificación , Chlamydiales/aislamiento & purificación , Enfermedades de las Ovejas/epidemiología , Aborto Veterinario/microbiología , Animales , Bovinos , Enfermedades de los Bovinos/microbiología , Infecciones por Chlamydia/epidemiología , Infecciones por Chlamydia/microbiología , ADN Bacteriano/genética , Granjas , Femenino , Prevalencia , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Factores de Riesgo , Ovinos , Enfermedades de las Ovejas/microbiología , TúnezRESUMEN
The design of French VFCWs leads to the formation of a sludge layer at the surface of the first filters due to the retention of suspended solids from the percolation of unsettled wastewater. This layer plays a major role in the system but still little is known on its characteristics and evolutions. In this study, suspended solids and sludge deposits sampled from two French VFCW plants were analyzed by different methods in the objective to assess the evolution of particulate organic matter (POM) along the treatment chain and within the sludge layer, and identify relevant analytical indicators of these phenomena. The treatment chain included an aerobic trickling filter followed by FeCl3 injection and two successive stages of filters. Thermal analyses showed that OM contents of suspended solids decreased along the treatment chain. POM in inflow suspended solids was predominantly composed of reactive, biodegradable compounds which were partly hydrolyzed and mineralized notably at the trickling filter stage. 3D fluorescence spectra collected from aqueous POM extracts confirmed the evolution of organic matter from low-molecular reactive compounds to more complex and stable structures such as humic-like substances. FTIR confirmed the mineralization of POM's reactive constituents along the treatment chain by the decrease in the intensities of the characteristics bands of aliphatic compounds or proteins, and its humification in the sludge deposits through the relative increase of the bands at 1634cm-1 (vC=O) and 1238cm-1 (δC=O and/or δOH). Isotopic ratios δ2H/1H and δ15N/14N were found to be good indicators of POM evolutions. The higher values of δ2H/1H and δ15N/14N ratios measured in sludge deposits as compared to inflow suspended solids were related to POM humification and to microbial processes of POM hydrolysis and mineralization, respectively.
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Understanding the processes of adaptive divergence, which may ultimately lead to speciation, is a major question in evolutionary biology. Allochronic differentiation refers to a particular situation where gene flow is primarily impeded by temporal isolation between early and late reproducers. This process has been suggested to occur in a large array of organisms, even though it is still overlooked in the literature. We here focused on a well-documented case of incipient allochronic speciation in the winter pine processionary moth Thaumetopoea pityocampa. This species typically reproduces in summer and larval development occurs throughout autumn and winter. A unique, phenologically shifted population (SP) was discovered in 1997 in Portugal. It was proved to be strongly differentiated from the sympatric "winter population" (WP), but its evolutionary history could only now be explored. We took advantage of the recent assembly of a draft genome and of the development of pan-genomic RAD-seq markers to decipher the demographic history of the differentiating populations and develop genome scans of adaptive differentiation. We showed that the SP diverged relatively recently, that is, few hundred years ago, and went through two successive bottlenecks followed by population size expansions, while the sympatric WP is currently experiencing a population decline. We identified outlier SNPs that were mapped onto the genome, but none were associated with the phenological shift or with subsequent adaptations. The strong genetic drift that occurred along the SP lineage certainly challenged our capacity to reveal functionally important loci.
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Flujo Génico , Mariposas Nocturnas/genética , Mariposas Nocturnas/fisiología , Adaptación Fisiológica/genética , Animales , Demografía , Frecuencia de los Genes , Variación Genética , Genética de Población , Genoma de los Insectos , Filogenia , Polimorfismo de Nucleótido Simple/genética , Análisis de Componente Principal , Estaciones del Año , Factores de TiempoRESUMEN
Different breeding systems associated with specific bovine genetic resources have coexisted in Burundi. To prepare for the development of a national action plan for the improvement of bovine genetic resources in Burundi, we aimed at performing genetic characterization of Ankole and Ankole × European crossbred individuals and assessing the effect of European ancestry on milk productivity of cows kept under the mixed crops livestock system. To that end, we genotyped 37 Ankole and 138 crossbred individuals on 42 636 SNPs and combined these genotypes with those from 21 cattle breeds, representative of the bovine genetic diversity. We also measured milk yield not suckled and estimated suckled milk. Given the results, we confirmed the indicine × African taurine admixed origin of the Ankole in Burundi and showed that crossbred individuals present a high proportion of European ancestry (i.e. 57% on average). As the proportion of European ancestry increased, milk yield increased by 0.03 ± 0.01 l/day, at a lower extent than expected. We also observed that breeders were unable to correctly evaluate the European proportion in their livestock. Our results may provide useful information for objective dairy breeding in Burundi. As an example, an ex-situ conservation program of Ankole within the framework of value chains is proposed as an accompanying strategy to improve the sustainability of the crossbreeding program.