Effects of organic and inorganic additives on flotation recovery of washed cells of Saccharomyces cerevisiae resuspended in water

Separation of microbial cells by flotation recovery is usually carried out in industrial reactors or wastewater treatment systems, which contain a complex mixture of microbial nutrients and excretion products. In the present study, the separation of yeast cells by flotation recovery was carried out using a simple flotation recovery systems containing washed yeast cells resuspended in water in order to elucidate the effects of additives (defined amounts of organic and inorganic acids, ethanol, surfactants and sodium chloride) on the cellular interactions at interfaces (cell/aqueous phase and cell/air bubble). When sodium chloride, organic acids (notably propionic, succinic and acetic acids) and organic surfactants (sodium dodecyl sulphate (SDS), cetyltrimethylammonium bromide (CTAB) and Nonidet P40) were added to the flotation recovery system, significant increases in the cell recovery of yeast hydrophobic cells (Saccharomyces cerevisiae, strain FLT-01) were observed. The association of ethanol to acetic acid solution (a minor by-product of alcoholic fermentation) in the flotation recovery system, containing washed cells of strain FLT-01 resuspended in water, leading to an increased flotation recovery at pH 5.5. Thus, the association among products of the cellular metabolism (e.g., ethanol and acetic acid) can improve yeast cell recovery by flotation recovery. (c) 2006 Elsevier B.V. All rights reserved.

Flotation of blocolloids suspended in liquid phases of different compositions and pH values

The flotation capacity was determined for cells of yeasts strains belonging to the genera Hansenula, Candida and Saccharomyces. A heterogeneous group of yeasts, comprising strains from the three genera, was identified as showing high flotation capacities (degrees of flotation above 50%), which were practically not affected by variations in medium pH in both the synthetic medium and 2% molasses. Thus, the flotation capacity of the cells in this yeast group seemed strongly dependent on the liquid phase properties and/or growth medium composition, more than on the simple variation in pH of the cell suspensions. A second group of strains, belonging to the Saccharomyces genus, including also brewing yeast strains, was identified as having lower flotation capacities (degrees of flotation below 50% at pH 1.5), which showed no alterations or variations significantly affected by the medium pH. Foam volumes obtained with Saccharomyces strains were greater in synthetic media than in molasses owing to the higher air flow rates required for flotation in molasses. The flotation efficiency decreased in molasses in all cases as well as the foam volume, except in the case of Hansenula cells, which showed an increased foam volume. This was probably due to variations in product excretion by the different yeasts and/or differences in cell wall composition.

Yeast flotation viewed as the result of the interplay of supernatant composition and cell-wall hydrophobicity

Flotation is a process of cell separation based on the affinity of cells to air bubbles. In the present work, flotability and hydrophobicity were determined using cells from different yeasts (Hansenulla polymorpha, Saccharomyces cerevisiae, Candida albicans), which were propagated in different media and at different temperatures. Alterations to the supernatant of the cells were also carried out before the flotation assays. The results described here indicate that supernatants of the yeast cells can play a more important role on flotation than cell-wall hydrophobicity. For example, wall-hydrophobicity of strain FLT-01 of S. cerevisiae was high but flotation did not occur when their washed cells were resuspended in water. Additions of neopeptone to cultures of S. cerevisiae and H. polymorpha repressed flotation and increased the volume of foam. An additional task of the present work was to show that the relationship between cell-wall hydrophobicity and flotation performance was dependent on the method used for the measurement of hydrophobicity. Based on the assay procedure, two types of hydrophobicity were distinguished: (a) the apparent hydrophobicity for cells suspended in the medium and expressed by the degree of cell affinity to the organic solvent in the two-phase system supernatant/hexane; (b) the standard hydrophobicity, which was determined for cells suspended in a standard solution (acetate buffer, in the present work) within the acetate buffer/hexane system. Flotation of cells of S. cerevisiae and C albicans were best related to the degree of apparent hydrophobicity (varying with the supernatant composition at the cell/medium interface) rather than to the degree of standard hydrophobicity (varying with the alterations in the wall components, since the liquid phase was constant in the assay). However, depending on the yeast unpredictable results can be obtained. For example, cells of H. polymorpha exhibited good flotation associated to a high degree of standard hydrophobicity while having a lower degree of apparent hydrophobicity. Concerning growth temperature, flotation of cells of C albicans was strongly repressed when the temperature was raised from 30 to 38 degreesC while a similar effect was not observed in cultures of S. cerevisiae and H. polymorpha. It is difficult to understand and predict flotation of yeast cells but simple modifications made to the supernatant of cultures can activate or repress flotation. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Efficient flotation of yeast cells grown in batch culture

A fast flotation assay was used to select new floating yeast strains. The flotation ability did not seem to be directly correlated to total extracellular protein concentration of the culture. However, the hydrophobicity of the cell was definitely correlated to the flotation capacity. The Saccharomyces strains (FLT strains) were highly hydrophobic and showed an excellent flotation performance in batch cultures without additives (flotation agents) and with no need for a special flotation chamber or flotation column. A stable and well-organized structure was evident in the dried foam as shown by scanning electron microscopy which revealed its unique structure showing mummified cells (dehydrated) attached to each other. The attachment among the cells and the high protein concentration of the foams indicated that proteins might be involved in the foam formation. The floating strains (strains FLT) which were not flocculent and showed no tendency to aggregate, were capable of growing and producing ethanol in a synthetic medium containing high glucose concentration as a carbon source. The phenomenon responsible for flotation seems to be quite different from the flocculation phenomenon. (C) 1996 John Wiley & Sons, Inc.

Flotation for cell recovery from small volumes of yeast cultures

Flotation or cell recovery in foams (proportion of the total cells in the medium transferred to the foam) and flotation efficiency (proportion of the cells transferred from an initial volume of medium equal to the residual volume after flotation) are functions of time, aeration rate, initial volume of medium, and initial concentration of cells. Cell recovery reached constant values (around 96.4 +/- 6.3%) and flotation efficiency decreased (owing to increases in the liquid content of the foam), with increases in air how rate (above 6-7 ml air s(-1)) and volumes of medium (above 11 ml) added to the column. Increases in concentration of cells in the medium led to increases in the concentration of cells in the foam.

Electrocoagulation/flotation followed by fluidized bed anaerobic reactor applied to tannery effluent treatment

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Selective factors involved in oil flotation isolation of black yeasts from the environment

The oil flotation isolation technique has been successfully applied to recover chaetothyrialean black yeasts and relatives from the environment. The selective mechanisms playing a role in isolation are unknown. The fungi concerned are supposed to occupy specialized microniches in nature, taking advantage of (1) oligotrophism. Mineral oil as a main selective agent may be based on (2) hydrophobicity or on (3) assimilation. All three hypotheses are tested in this paper. Results show that cell wall hydrophobicity is unlikely to be a selective factor. Incubation under poor nutrient conditions provides competitive advantage for black yeasts, especially for Exophiala strains, which are subsequently enriched by mineral oil which enhances growth in this group of fungi. Incubation under mineral media and mineral oil can be used as selective factor.

The influence of floc size and hydraulic detention time on the performance of a dissolved air flotation (DAF) pilot unit in the light of a mathematical model

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of soil texture in the recovery of Toxocara canis eggs by a flotation method

In epidemiological surveys, the evaluation of soil contamination by Toxocara canis eggs requires a quick and easy method for the isolation of parasite eggs from soil samples. The efficiency of flotation methods is influenced by sample size, soil texture, degree of soil contamination, pretreatment, flotation solutions and time of flotation. This investigation was designed to evaluate the influence of soil texture in the recovery of T. canis eggs with the centrifugal flotation technique of Dada (Dada, B.J.O., 1979. A new technique for the recovery of Toxocara eggs from soil. J. Helminthol., 53: 141-144). Four types of soil (clay silt, sandy, silty clay and sand) were artificially contaminated with T. canis eggs (200 eggs per gram). Zinc sulphate (specific gravity 1.20) and sodium dichromate (specific gravity 1.35) were used as flotation solutions. Twenty replicated examinations were performed for each type of soil and flotation solution. There was a statistically significant difference in the results depending on soil type. The highest recovery percentages were observed in soils rich in sand (62.5% for sand and 38.0% for sandy soil). Differences were also observed with different flotation solutions. Sodium dichromate solution was more efficient for recovering T. canis eggs, regardless of the soil texture. © 1994.

A technique for the capture of free-ranging marsh deer (blastocerus dichotomus)

The marsh deer (Blastocerus dichotomus) is an endangered species froth the marshlands of central South America. Its population has declined in several regions due to the loss of available habitat caused by human activities, especially the construction of hydroelectric darns. The capture of individual deer is critical for research programs and population management. This report describes a novel live-capture technique, which uses a helicopter to drive the animals into a terrain that restricts their movement such as thick vegetation or deep water (60-120 cm in depth). Following confinement, animals are manually restrained. The short pursuit time (median = 2 thin), low mortality rate (0.82-3.28%), and the absence of injury to both the capture team and animals suggest that this method is appropriate for the safe capture of this species. Body temperature correlated with the pursuit time (R-2 = 0.15) but was not significantly altered with pursuit times <3 min.

Prevalence and Molecular Characterization of Mycoplasma ovis in Selected Free-Ranging Brazilian Deer Populations

Mycoplasma ovis is a hemoplasma that may cause anemia and mortality in small ruminants. Our aim was to determine whether M. ovis infects populations of free-ranging deer in Brazil. Bully coat samples from 64 Blastocerus dichotomus from Porto Primavera, 18 Ozotocerus bezoarticus from Pantanal, and 21 O. bezoarticus from Emas National Park were tested. Using a M. ovis PCR protocol to amplify extracted DNA, 46/64 (72%) of deer froth Porto Primavera, 10/18 (56%) from Pantanal, and 4/21 (19%) from Emas National Park were positive, giving an overall positive rate of 58% for hemoplasma in these wild deer. Sequencing and phylogenetic analysis of the 168 rRNA gene revealed 3 genetically distinct hemoplasmas including M. ovis, 'Candidatus Mycoplasma erythrocervae', and a hemoplasma most closely related to M. ovis. Phylogenetic analysis of the 23S rRNA gene from selected sequences confirmed these relationships.

OSLERUS OSLERI (COBBOLD, 1876) INFECTION IN MANED WOLF (CHRYSOCYON BRACHYURUS, ILLIGER, 1815)

Oslerus osleri is a small nematode that infects the respiratory tract of domestic and wild canids and is responsible for causing chronic nodular tracheobronchitis. This paper aims to report a case of parasitism by O. osleri in a free-living maned wolf (Chrysocyon brachyurus) that was struck by a motor vehicle. Fecal samples were collected, and the presence of spiral larvae, with S-shaped tails, was observed on flotation. This characteristic was compatible with the Filaroididae Family larvae of O. osleri. Although the animal did not show clinical signs of respiratory system impairment, a tracheobronchoscopy was performed. Semitransparent nodules, 5 mm in diameter, containing adult parasites were observed in the third distal portion of the trachea, cranial to the carina. Larval morphological characteristics and the nodular locations were compatible with an O. osleri respiratory tract infection.

Molecular characterization of Giardia duodenalis in dogs from Brazil

The intestinal protozoan parasite Giardia duodenalis (syn. Giardia intestinalis and Giardia lamblia) is a widespread enteric pathogen in human and domestic animals. This organism is one of the most common parasites in domestic dogs in Brazil. In this study, we determined the occurrence and genetic characterization of G. duodenalis isolated from dogs from south-central São Paulo state, Brazil. A total of 300 fecal samples were collected. Fecal specimens were screened for the presence of G. duodenalis using microscopy (zinc sulfate solution flotation technique) and polymerase chain reaction (PCR) targeting the small subunit ribosomal (SSU-rDNA) and glutamate dehydrogenase (GDH) genes. Genetic characterization was performed using restriction fragment length polymorphisms (RFLP) and sequencing analysis of the GDH gene. In addition, selected samples were further characterized by RFLP and sequencing of the beta-giardin gene. The overall occurrence of G. duodenalis was 17.3% (52/300). The occurrence was higher in stray dogs (28%) than in household dogs (6.25%). of the 36 PCR-positive samples that were selected for genotyping, only dog-specific genotype C (20 isolates), D (11 isolates) and mixed C+D (five isolates) isolates were detected in the study. This study provides current information on the infection rates of G. duodenalis genotypes in canine populations and describes for the first time the presence of mixed infections within host-specific C and D genotypes in dogs in Brazil. These genotypes were widespread and commonly found in domestic dogs living in urban and suburban environments of the studied area and confirmed the endemic status of Giardia in this region.

Ocorrência de larva migrans na areia de áreas de lazer das escolas municipais de ensino infantil, Araçatuba, SP, Brasil

A areia das áreas de lazer de escolas podem constituir vias de transmissão para várias zoonoses parasitárias, representando risco potencial para as crianças que brincam nesses locais. Foi avaliada a ocorrência de agentes de larva migrans em 28 escolas municipais de ensino infantil de Araçatuba, SP. Foram colhidas 535 amostras de areia das áreas de lazer dessas escolas nos meses de janeiro (verão) e julho (inverno) de 1997 para estabelecimento da freqüência de isolamento de larvas e/ou ovos de Ancylostoma spp. e de ovos de Toxocara spp., pelos métodos de centrífugo-flutuação e de Baermann, respectivamente. A presença de larvas de Ancylostoma spp. foi observada, em pelo menos uma das amostras, em 35,7% (10/28) das amostras da primeira colheita (verão) e em 46,4% (13/28) quando da segunda colheita (inverno). Ovos de Toxocara spp. não foram encontrados e a presença de ovos de Ancylostoma spp. foi observada em 0,56% (3/535) das amostras.

Prevalence of intestinal parasites in dogs from São Paulo State, Brazil

The prevalence of gastrointestinal parasites in stray dogs, and dogs with owners was investigated by fecal examinations from 271 dogs employing sedimentation, simple flotation and centrifugation-flotation methods. The centrifugation-flotation method, when compared to simple flotation or sedimentation methods was generally more accurate in the diagnosis of all intestinal parasites, but statistical differences were detected only in relation to Giardia spp. and Cystoisospora spp. (synonym Isospora spp.). The following parasites, with their respective prevalence, were diagnosed in the fecal samples: Ancylostoma spp. (23.6%); Toxocara canis (5.5%); Trichuris vulpis (4.8%); Spirocerca lupi (1.9%); Dipylidium caninum (0.7%); Giardia spp. (12.2%); Hammondia heydorni (2.6%); Cystoisospora spp. (8.5%); and Sarcocystis spp. (2.2%). The prevalence of most parasites was similar for dogs of mixed-breed and for dogs of a defined-breed, except for Cystoisospora spp. and T canis which showed a significantly higher prevalence in mixed-breed dogs. The prevalence of Ancylostoma spp. (17.1%) was significantly lower in stray dogs than in those with an owner (31.9%) and the prevalence of Giardia spp. and Cystoisospora spp. was higher in stray dogs (P < 0.05). No effect of season on the occurrence of the different parasite genera could be observed, except for Ancylostoma spp., for which an increase in the percentage of dogs shedding eggs was observed at the beginning of Summer with a peak occurrence during April and May (Autumn). The prevalence of Ancylostoma spp., T canis, T vulpis, Giardia spp. and Cystoisospora spp. was higher in adult males than in adult females, but significant differences between the two groups occurred only with Giardia spp. Young animals were found to more frequently shed Nematode eggs in feces than adult animals. (C) 2002 Elsevier B.V. B.V. All rights reserved.