Environmentally-friendly technology for rapid identification and quantification of emerging pollutants from wastewater using infrared spectroscopy.

The monitoring of emerging pollutants in wastewaters is nowadays an issue of special concern, with the classical quantification methods being time and reagent consuming. In this sense, a FTIR transmission spectroscopy based chemometric methodology was developed for the determination of eight of these pollutants. A total of 456 samples were, therefore, obtained, from an activated sludge wastewater treatment process spiked with the studied pollutants, and analysed in the range of 200 cm-1 to 14,000 cm-1. Then, a k-nearest neighbour (kNN) analysis aiming at identifying each sample pollutant was employed. Next, partial least squares (PLS) and ordinary least squares (OLS) modelling approaches were employed in order to obtain suitable prediction models. This procedure resulted in good prediction abilities regarding the estimation of atrazine, desloratadine, paracetamol, ß-estradiol, ibuprofen, carbamazepine, sulfamethoxazole and ethynylestradiol concentrations in wastewaters. These promising results suggest this technology as a fast, eco-friendly and reagent free alternative methodology for the quantification of emerging pollutants in wastewaters.

Quantification of pharmaceutical compounds in wastewater samples by near infrared spectroscopy (NIR).

The quantification of pollutants, as pharmaceuticals, in wastewater is an issue of special concern. Usually, typical methods to quantify these products are time and reagent consuming. This paper describes the development and validation of a Fourier transform near-infrared (FT-NIR) spectroscopy methodology for the quantification of pharmaceuticals in wastewaters. For this purpose, 276 samples obtained from an activated sludge wastewater treatment process were analysed in the range of 200 cm-1 to 14,000 cm-1, and further treated by chemometric techniques to develop and validate the quantification models. The obtained results were found adequate for the prediction of ibuprofen, sulfamethoxazole, 17ß-estradiol and carbamazepine with coefficients of determination (R2) around 0.95 and residual prediction deviation (RPD) values above four, for the overall (training and validation) data points. These results are very promising and confirm that this technology can be seen as an alternative for the quantification of pharmaceuticals in wastewater.

New PLS analysis approach to wine volatile compounds characterization by near infrared spectroscopy (NIR).

This work aims to explore the potential of near infrared (NIR) spectroscopy to quantify volatile compounds in Vinho Verde wines, commonly determined by gas chromatography. For this purpose, 105 Vinho Verde wine samples were analyzed using Fourier transform near infrared (FT-NIR) transmission spectroscopy in the range of 5435 cm-1 to 6357 cm-1. Boxplot and principal components analysis (PCA) were performed for clusters identification and outliers removal. A partial least square (PLS) regression was then applied to develop the calibration models, by a new iterative approach. The predictive ability of the models was confirmed by an external validation procedure with an independent sample set. The obtained results could be considered as quite good with coefficients of determination (R2) varying from 0.94 to 0.97. The current methodology, using NIR spectroscopy and chemometrics, can be seen as a promising rapid tool to determine volatile compounds in Vinho Verde wines.

Genetic diversity of Saccharum complex using ISSR markers.

Sugarcane (Saccharum sp, Poaceae) is native to Southeast Asia, and due to growing demand as raw material, its cultivation recently expanded to new frontiers. The genetic diversity analysis is essential for targeting strategies in the formation and maintenance of a germplasm. This study aimed to assess the genetic diversity of 26 accessions of sugarcane from the Active Germplasm Bank of Embrapa Coastal Tablelands, using inter-simple sequence repeat (ISSR) molecular markers. Sixteen primers were used, resulting in 87 fragments with 91.13% of polymorphism. The similarity of the individuals ranged between 0.22 and 0.87. Individuals RB867515 and RB92579 were closer genetically, and the most distant ones were PI240785 and NSL 291970. Four distinct clusters were formed, using UPGMA. This information can be used to prioritize the selection of accessions for the conduction of hybridization in breeding and germplasm exchange actions.

Occurrence and Probability Maps of Lutzomyia longipalpis and Lutzomyia cruzi (Diptera: Psychodidae: Phlebotominae) in Brazil.

Leishmaniases are serious diseases caused by trypanosomatid protozoans of the genus Leishmania transmitted by the bite of phlebotomine sand flies. We analyzed records pertaining to Lutzomyia longipalpis (Lutz and Neiva, 1912) and Lutzomyia cruzi (Mangabeira, 1938) in Brazil from the following sources: the collection of phlebotomine sand flies of the Centro de Pesquisas René Rachou/Fiocruz (FIOCRUZ-COLFLEB), the "SpeciesLink" (CRIA) database, from systematic surveys of scientific articles and gray literature (dissertations, theses, and communications), and disease data obtained from the Information System for Notifiable Diseases/Ministry of Health (SINAN/MS). Environmental data and ecological niche modeling (ESMS) using the approach of MaxEnt algorithm produced maps of occurrence probability for both Lu. longipalpis and Lu. cruzi. Lutzomyia longipalpis was found in 229 Brazilian municipalities and Lu. cruzi in 27. The species were sympatric in 16 municipalities of the Central-West region of Brazil. Our results show that Lu. longipalpis is widely distributed and associated with the high number of cases of visceral leishmaniasis reported in Brazil.

Quantitative image analysis as a tool for Yarrowia lipolytica dimorphic growth evaluation in different culture media.

Yarrowia lipolytica, a yeast strain with a huge biotechnological potential, capable to produce metabolites such as γ-decalactone, citric acid, intracellular lipids and enzymes, possesses the ability to change its morphology in response to environmental conditions. In the present study, a quantitative image analysis (QIA) procedure was developed for the identification and quantification of Y. lipolytica W29 and MTLY40-2P strains dimorphic growth, cultivated in batch cultures on hydrophilic (glucose and N-acetylglucosamine (GlcNAc) and hydrophobic (olive oil and castor oil) media. The morphological characterization of yeast cells by QIA techniques revealed that hydrophobic carbon sources, namely castor oil, should be preferred for both strains growth in the yeast single cell morphotype. On the other hand, hydrophilic sugars, namely glucose and GlcNAc caused a dimorphic transition growth towards the hyphae morphotype. Experiments for γ-decalactone production with MTLY40-2P strain in two distinct morphotypes (yeast single cells and hyphae cells) were also performed. The obtained results showed the adequacy of the proposed morphology monitoring tool in relation to each morphotype on the aroma production ability. The present work allowed establishing that QIA techniques can be a valuable tool for the identification of the best culture conditions for industrial processes implementation.

Avaliação histopatológica de órgãos reprodutivos e bexiga de fêmeas suínas descartadas / Histopathology of reproductive organs and bladder of culled sows

Em um sistema intensivo de produção de suínos, as falhas reprodutivas são uma das principais razões de descarte de matrizes e queda nos índices produtivos. A infecção urinária (cistite) e as endometrites são consideradas importantes causas de descarte em fêmeas suínas, por terem consequências reprodutivas relevantes e elevarem a taxa de reposição do plantel. O presente estudo teve o objetivo de avaliar o aparelho reprodutivo e a bexiga de fêmeas suínas de descarte normal de granjas, bem como investigar a existência de relação entre as patologias encontradas. Foram examinadas 79 matrizes suínas oriundas de 20 rebanhos localizados no Estado de Santa Catarina. De cada fêmea foram coletados os ovários, fragmentos de útero e bexiga. Dentre as fêmeas avaliadas, 32 (40,5%) tinham diferentes graduações de cistite, 24 (30,4%) tinham algum tipo de inflamação uterina, e 9 (11,4%) estavam em anestro, com ovários inativos. Contudo, não foi observada dependência significativa entre cistite e endometrite nas amostras analisadas.

Reproductive failures are the major reasons for removal of sows and decrease of production rates in an intensive swine production system. Urinary infection and endometritis are considered important causes for culling of sows, due to relevant reproductive consequences and increase of the replacement rates. The present study aimed to evaluate the reproductive and urinary system of culled sows, as well as investigate the occurrence of cystitis and endometritis in analyzed sows. Samples, such as ovaries, uterus fragments and bladder were collected from 79 sows originated from 20 farms of Santa Catarina State. Results showed that, 32 (40,5%) analyzed sows presented cystitis in different levels, 24 (30,4%) had some class of uterine inflammation, and 9 (11,4%) were in anestrous, with inactive ovaries. However, unsignificative dependence between cystitis and endometritis in analyzed samples was observed.

Quantitative image analysis for the characterization of microbial aggregates in biological wastewater treatment: a review.

Quantitative image analysis techniques have gained an undeniable role in several fields of research during the last decade. In the field of biological wastewater treatment (WWT) processes, several computer applications have been developed for monitoring microbial entities, either as individual cells or in different types of aggregates. New descriptors have been defined that are more reliable, objective, and useful than the subjective and time-consuming parameters classically used to monitor biological WWT processes. Examples of this application include the objective prediction of filamentous bulking, known to be one of the most problematic phenomena occurring in activated sludge technology. It also demonstrated its usefulness in classifying protozoa and metazoa populations. In high-rate anaerobic processes, based on granular sludge, aggregation times and fragmentation phenomena could be detected during critical events, e.g., toxic and organic overloads. Currently, the major efforts and needs are in the development of quantitative image analysis techniques focusing on its application coupled with stained samples, either by classical or fluorescent-based techniques. The use of quantitative morphological parameters in process control and online applications is also being investigated. This work reviews the major advances of quantitative image analysis applied to biological WWT processes.

Characterization of activated sludge abnormalities by image analysis and chemometric techniques.

This work focuses on the use of chemometric techniques for identifying activated sludge process abnormalities. Chemometric methods combined with image analysis can improve activated sludge systems monitoring and minimize the need of analytical measurements. For that purpose data was collected from aggregated and filamentous biomass, biomass composition on Gram-positive/Gram-negative bacteria and viable/damaged bacteria, and operational parameters. Principal component analysis (PCA) was subsequently applied to identify activated sludge abnormalities, allowing the identification of several disturbances, namely filamentous bulking, pinpoint flocs formation, and zoogleal bulking as well as normal conditions by grouping the collected samples in corresponding clusters.

Identifying different types of bulking in an activated sludge system through quantitative image analysis.

The present study proposes an image analysis methodology for the identification of different types of disturbances in wastewater treatment activated sludge systems. Up to date, most reported image analysis methodologies have been used in activated sludge processes with the aim of filamentous bulking detection, however, other disturbances could be foreseen in wastewater treatment plants. Such disturbances can lead to fluctuations in the biomass contents, affecting the mixed liquor suspended solids (MLSS), and in the sludge settling ability, affecting the sludge volume index (SVI). Therefore, this work focuses on predicting the MLSS and SVI parameters for different types of disturbances affecting an activated sludge system. Four experiments were conducted simulating filamentous bulking, zoogleal or viscous bulking, pinpoint floc formation, and normal operating conditions. Alongside the MLSS and SVI determination, the aggregated and filamentous biomass contents and morphology were studied as well as the biomass Gram and viability status, by means of image analysis.

Dilution and magnification effects on image analysis applications in activated sludge characterization.

The properties of activated sludge systems can be characterized using image analysis procedures. When these systems operate with high biomass content, accurate sludge characterization requires samples to be diluted. Selection of the best image acquisition magnification is directly related to the amount of biomass screened. The aim of the present study was to survey the effects of dilution and magnification on the assessment of aggregated and filamentous bacterial content and structure using image analysis procedures. Assessments of biomass content and structure were affected by dilutions. Therefore, the correct operating dilution requires careful consideration. Moreover, the acquisition methodology comprising a 100× magnification allowed data on aggregated and filamentous biomass to be determined and smaller aggregates to be identified and characterized, without affecting the accuracy of lower magnifications regarding biomass representativeness.

A Comparison between bright field and phase-contrast image analysis techniques in activated sludge morphological characterization.

Different approaches using microscopy image analysis procedures were employed for characterization of activated sludge systems. The approaches varied mainly on the type of visualization and acquisition method used for collection of data. In this context, this study focused on the comparison of the two most common acquisition methods: bright field and phase-contrast microscopy. Images were acquired from seven different wastewater treatment plants for a combined period of two years. Advantages and disadvantages of each acquisition technique and the results are discussed. Bright field microscopy proved to be more simple and inexpensive and provided the best overall results.

Correlation between sludge settling ability and image analysis information using partial least squares.

In the last years there has been an increase on the research of the activated sludge processes, and mainly on the solid-liquid separation stage, considered of critical importance, due to the different problems that may arise affecting the compaction and the settling of the sludge. Furthermore, image analysis procedures are, nowadays considered to be an adequate method to characterize both aggregated and filamentous bacteria, and increasingly used to monitor bulking events in pilot plants. As a result of that, in this work, image analysis routines were developed in Matlab environment, allowing the identification and characterization of microbial aggregates and protruding filaments. Moreover, the large amount of activated sludge data collected with the image analysis implementation can be subsequently treated by multivariate statistical procedures such as PLS. In the current work the implementation of image analysis and PLS techniques has shown to provide important information for better understanding the behavior of activated sludge processes, and to predict, at some extent, the sludge volume index. As a matter of fact, the obtained results allowed explaining the strong relationships between the sludge settling properties and the free filamentous bacteria contents, aggregates size and aggregates morphology, establishing relevant relationships between macroscopic and microscopic properties of the biological system.

Monitoring of activated sludge settling ability through image analysis: validation on full-scale wastewater treatment plants.

In recent years, a great deal of attention has been focused on the research of activated sludge processes, where the solid-liquid separation phase is frequently considered of critical importance, due to the different problems that severely affect the compaction and the settling of the sludge. Bearing that in mind, in this work, image analysis routines were developed in Matlab environment, allowing the identification and characterization of microbial aggregates and protruding filaments in eight different wastewater treatment plants, for a combined period of 2 years. The monitoring of the activated sludge contents allowed for the detection of bulking events proving that the developed image analysis methodology is adequate for a continuous examination of the morphological changes in microbial aggregates and subsequent estimation of the sludge volume index. In fact, the obtained results proved that the developed image analysis methodology is a feasible method for the continuous monitoring of activated sludge systems and identification of disturbances.

Stalked protozoa identification by image analysis and multivariable statistical techniques.

Protozoa are considered good indicators of the treatment quality in activated sludge systems as they are sensitive to physical, chemical and operational processes. Therefore, it is possible to correlate the predominance of certain species or groups and several operational parameters of the plant. This work presents a semiautomatic image analysis procedure for the recognition of the stalked protozoa species most frequently found in wastewater treatment plants by determining the geometrical, morphological and signature data and subsequent processing by discriminant analysis and neural network techniques. Geometrical descriptors were found to be responsible for the best identification ability and the identification of the crucial Opercularia and Vorticella microstoma microorganisms provided some degree of confidence to establish their presence in wastewater treatment plants.

Disseminação do vírus da doença de Aujeszky, envolvendo o comércio de reprodutores suínos de reposição / Spread of virus of Aujeszky's disease by the trade of replacement swine breeders

Relata-se a disseminação da infeção pelo vírus da doença de Aujeszky (VDA) a partir da comercialização de reprodutores suínos oriundos de duas granjas de reprodutores suídeos certificada (GRSC) que tiveram surto da DA. Após a confirmação do diagnóstico de DA, os suínos comercializados 37 a 45 dias anteriores aos surtos foram recolhidos, submetidos ao exame sorológico para o VDA e enviados ao abate. Nos rebanhos-destino foram realizados exames sorológicos para o VDA 17 a 37 dias após o recolhimento, naqueles que estavam lojados nas baias vizinhas àquelas onde haviam sido alojados os suínos comprados; seis meses mais tarde, foi realizada outra sorologia por amostragem dos reprodutores. No total, 52 rebanhos compraram suínos das duas granjas, e, destes, 37 (69,8 por cento) receberam, pelo menos, um animal com sorologia positiva para o VDA. Somente sete (18,9 por cento) deles apresentaram sorologia positiva para o VDA, e em 30 (81,1 por cento), não houve indícios de disseminação da infecção. A contaminação pelo VDA de granjas GRSC representa enorme potencial para a disseminação da infecção, por meio do comércio de suínos de reposição. A rastreabilidade dos animais comercializados em um período anterior ao diagnóstico, com imediata remoção dos lotes de suínos dos rebanhos-destino, evitou a disseminação da infecção

It was reported the spreading of the infection caused by the virus of Aujesky's disease (VAD) by the commercialization of breeders originated from two pigs farms GRSC (Farms of Certified Swine Breeders - FCSB) which had an outbreak of Aujeszky's disease. After the positive diagnosis of Aujesky's disease, the pigs traded from 37 to 45 day before the outbreaks were removed from the herd, bled for serological exams for the VAD and sent to slaughterhouses. The herds which received these pigs were serologically tested for the VAD, 17 to 37 days after the removal of the animals. Serological tests were also performed in pigs lodged at neighboring cages to those which had been lodged with the pigs bought and a sampling test was done six months later. Thus, 52 flocks bought pigs from the farms 1 and 2. From those, 37 (69.8 percent) received at least one serum-positive pig for the VAD. Only seven (18.9 percent) of them were infected and 30 (81.1 percent) pigs showed no indications of infection by the VAD. The occurrence of VAD in FCSB farms represents huge potential for spreading of the VAD, by the trade of replacement pigs. The traceability investigation and removal of the animals traded before the diagnosis, with immediate removal of the positive lots of pigs at the destiny herd, avoided the spread of the infection

Uso do plasma suíno ultrafiltrado na recuperação de leitões com sinais clínicos de circovirose / The use of ultra-filtered swine plasma in the recovery of pigs with clinical signs of porcine circovirus infection

Testou-se o efeito do plasma suíno ultrafiltrado spray-dried, associado a um acidificante comercial na água de bebida para a recuperação de leitões com sinais clínicos da síndrome multissistêmica do definhamento dos suínos (SMDS). Utilizaram-se 40 leitões com sinais clínicos da SMDS, selecionados 20 dias após o alojamento em uma unidade de terminação, distribuídos em quatro tratamentos (T) de 10 leitões cada. No T1, os animais receberam água clorada à vontade (controle); no T2, solução com 2,5 por cento do plasma sangüíneo diluído em água; no T3, acidificante (Selko®) diluído em água na dosagem de 12ml/10l e, no T4, solução com 2,5 por cento do plasma sangüíneo e o acidificante na dose de 12ml/10l, diluídos em água. Os leitões não foram medicados e foram sacrificados aos 28 ou 40 dias de experimento para avaliação sorológica e patológica. Não houve diferença no ganho de peso e na situação clínica-patológica entre os tratamentos. Entretanto, os leitões do T4 estavam em melhor situação clínica-patológica. Os leitões dos quatro tratamentos tiveram boa recuperação, sem terem sido medicados. Observou-se alta freqüência de lesões compatíveis com a SMDS nos pulmões, rins e linfonodos. Concluiu-se que o plasma spray dried associado ao ácido não melhoraram o desempenho e a situação clínica-patológica de leitões com sintomas da SMDS

The effect of the ultra-filtered spray-dried porcine plasma, associated to a commercial acid in the drinking water was tested for recovering pigs with clinical signs of the porcine postweaning multisystemic wasting syndrome (PMWS). Forty piglets with clinical signs of the PMWS were used following a selection at 20 days after their housing in one finishing facility. They were divided in four treatment groups (T) of 10 pigs each: T1 - chlorine treated water ad libitum (control); T2 - solution prepared with 2.5 percent of plasma diluted in water; T3 - acid (Selko® ) diluted in water at the concentration of 12 ml/10l; T4 - solution prepared with 2.5 percent of plasma diluted and the acid (Selko® ) diluted in water at the concentration of 12 ml/10l. The pigs received no medication and were euthanized at 28 or 40 days after the beginning of the experiment for serological and pathological tests. Differences at the weight gain and in the clinical-pathological situation were not observed among the treatments. However, pigs from T4 were in better clinical-pathological situation. The pigs of all four treatments showed a good recovery, although they were not medicated. Even though, it was observed a high frequency of lesions compatible to PMWS in the lungs, kidneys and lymph nodes. It was concluded that the plasma spray-dried associated to the acid did not improve the performance and the clinical-pathological situation of pigs with clinical signs of PMWS

Recognition of Protozoa and Metazoa using image analysis tools, discriminant analysis, neural networks and decision trees.

Protozoa and metazoa are considered good indicators of the treatment quality in activated sludge systems due to the fact that these organisms are fairly sensitive to physical, chemical and operational processes. Therefore, it is possible to establish close relationships between the predominance of certain species or groups of species and several operational parameters of the plant, such as the biotic indices, namely the Sludge Biotic Index (SBI). This procedure requires the identification, classification and enumeration of the different species, which is usually achieved manually implying both time and expertise availability. Digital image analysis combined with multivariate statistical techniques has proved to be a useful tool to classify and quantify organisms in an automatic and not subjective way. This work presents a semi-automatic image analysis procedure for protozoa and metazoa recognition developed in Matlab language. The obtained morphological descriptors were analyzed using discriminant analysis, neural network and decision trees multivariable statistical techniques to identify and classify each protozoan or metazoan. The obtained procedure was quite adequate for distinguishing between the non-sessile protozoa classes and also for the metazoa classes, with high values for the overall species recognition with the exception of sessile protozoa. In terms of the wastewater conditions assessment the obtained results were found to be suitable for the prediction of these conditions. Finally, the discriminant analysis and neural networks results were found to be quite similar whereas the decision trees technique was less appropriate.

Análisis de imagen para la cuantificación de cambios morfológicos en biomasa granular, inducidos por la presencia de nitrato en reactores EGSB

El objetivo del trabajo fué usar el análisis de imagen para cuantificar los cambios morfológicos de granulos anaeróbios expuestos a nitratos en reactores EGSB, para complementar los resultados, se efectuaron mediciones de actividad metanogénica en presencia de acetato, propionato, butirato, etanol y una mezcla gaseosa de H2/C2 y velocidad de sedimentación

Análisis de imagen para la cuantificación de cambios morfológicos en biomasa granular, inducidos por la presencia de nitrato en reactores EGSB

El objetivo del trabajo fué usar el análisis de imagen para cuantificar los cambios morfológicos de granulos anaeróbios expuestos a nitratos en reactores EGSB, para complementar los resultados, se efectuaron mediciones de actividad metanogénica en presencia de acetato, propionato, butirato, etanol y una mezcla gaseosa de H2/C2 y velocidad de sedimentación