RESUMO
Two sequential batch reactors (R1 and R2) of aerobic granular sludge (AGS) were inoculated with activated sludge of different origins. The objective was to investigate the granulation and the consistency between the structure of the microbial communities (16S rRNA amplicon sequencing) in each reactor and their metabolic performance (removal of C, N, and P). Both reactors were fed with acetate-based synthetic wastewater, targeting an anaerobic-aerobic cycle reputed to favor the phosphorus- and glycogen-accumulating organisms (PAO and GAO). Stable granulation was achieved in both reactors, where, instead of PAO, the dominant genera were ordinary heterotrophic organisms (OHO) such as Thauera, Paracoccus, and Flavobacterium known for their high capacity of aerobic storage of polyhydroxyalkanoates (PHA). Generally, there was good consistency between the metabolic behavior of each reactor and the bacterial genera detected. Both reactors showed high removals of C and complete nitrification (Nitrosomonas and Nitrospira detected) but a low level of simultaneous nitrification-denitrification (SND) during the aerated phase. The latter causes that nitrates were recycled to the initial phase, in detriment of PAO selection. Meanwhile, the study showed that selecting slow-growing OHOs (with aerobic storage capacity) favors stable granulation, revealing an alternative AGS technology for C and N removal.
Assuntos
Esgotos , Eliminação de Resíduos Líquidos , Esgotos/química , Reatores Biológicos/microbiologia , RNA Ribossômico 16S , Fósforo/metabolismo , Bactérias/genética , Bactérias/metabolismo , Nitrogênio/análise , DesnitrificaçãoRESUMO
Two one-step real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of Zika virus (ZIKV) were developed, based on two different primer design approaches: (1) open source, based on a combination of sequence diversity clustering (phylogeny and principal component analysis) and LAVA algorithm, using 45 whole genome ZIKV sequences retrieved from the National Center for Biotechnology Information (NCBI) database; (2) standard software for LAMP primer design (Primer Explorer V4), using 59 sequences of the ZIKV 3' UTR. The assays were firstly evaluated with External Quality Assessment panels from INSTAND e.V. (Germany) and EVD-LabNet (The Netherlands) including 4 and 12 unknown samples, respectively, and secondly, with 9 human, mosquito, and monkey ZIKV isolates from Africa (Senegal, Ivory Coast, and Uganda) and America (Brazil). The limit of detection as determined by probit analysis was 181 molecules for both RT-LAMP assays, and 100% reproducibility in the assays was obtained for 103 molecules (4/8 repetitions were positive for 102 molecules). Both assays were specific, amplifying only ZIKV RNA and not cross-detecting other arboviruses included in this study.
Assuntos
Algoritmos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Software , Infecção por Zika virus/diagnóstico , Zika virus/genética , África , Animais , Brasil , Células Cultivadas , Culicidae/virologia , Alemanha , Haplorrinos , Humanos , Ensaio de Proficiência Laboratorial , Limite de Detecção , Técnicas de Diagnóstico Molecular/normas , Países Baixos , Técnicas de Amplificação de Ácido Nucleico/normas , RNA Viral/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Zika virus/isolamento & purificação , Infecção por Zika virus/veterináriaRESUMO
Molecular detection of Zika virus (ZIKV) is a key element of outbreak management. Multiple PCR and isothermal ZIKV assays targeting different ZIKV sequences have been published. In this study, we compared a qRT-PCR, 2 RT-LAMP assays (based on different primer design approaches), and an RT-RPA for the detection of African and Asian/American lineages of ZIKV isolates from human, mosquito, and monkey. Results showed that RT-LAMP detected 100% of samples with a time threshold (Tt) of 18.01 ± 11.71 min while qRT-PCR detected 88.88% of samples with a Tt of 58.30 ± 16.58 min and RT-RPA 50% of samples with a Tt of 3.70 ± 0.44 min.
Assuntos
Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecção por Zika virus/diagnóstico , Zika virus/genética , Animais , Brasil , Células Cultivadas , Côte d'Ivoire , Culicidae , Haplorrinos , Humanos , Recombinases/metabolismo , Reprodutibilidade dos Testes , Senegal , Uganda , Zika virus/isolamento & purificação , Infecção por Zika virus/virologiaRESUMO
This paper focuses on environmental concerns regarding used antifreeze. It shows the results of laboratory respirometric tests conducted to evaluate the potential impact of antifreeze on activated sludge. In this first step of the study, only the components of virgin antifreeze were addressed, i.e. ethylene glycol, and antifoaming and corrosion inhibitory substances. The plateau resulting from oxygen uptake rate measurements of mixed liquor taken at the end of an aeration tank is used as a reference to evaluate the response of biomass to pulse injection of increasing dosages of new antifreeze, compared to that of formaldehyde, a known toxic compound. While 200 mg/L of formaldehyde steadily decreased the biomass activity (7 mg O2/Lh), there were no toxicity effects for doses up to 5000 mg/L of ethylene glycol from new antifreeze coolant (23 mg O2/Lh, compared to the initial 15 mg O2/Lh for the mixed liquor). It is estimated that ethylene glycol concentration in municipal wastewaters should be several order of magnitude lower then the 1000 mg/L shown to be tolerated by the biomass in this work, but at the present stage, the research does not provide enough information to recommend a definitive management scheme for antifreeze discharge. Many other management issues for used antifreeze must be addressed in future studies before any decision on tolerating sewer discharge in Mexico, as a transitory management option that will conciliate the actual practices with legality, can be made.