Alan Warren

Investigation of Escherichia coli removal in various designs of subsurface flow wetlands used for wastewater treatment.

Comparative studies were carried out on both pilot-scale and laboratory systems (microcosms) in order to determine the Escherichia coli removal rates and to investigate the E. coli removal kinetics in various designs of constructed wetlands employing horizontal subsurface flow through the root zone of emergent macrophytes (RZM). Parameters investigated include the presence or absence of hydrophytes, in this case the common reed Phragmites australis, and the type of planting medium (pea gravel or soil). The average E. coli removal rates were 41–72% for microcosms, and 96.6–98.9% for pilot-scale systems. The poorer removal rates achieved by the microcosms might be explained by a combination of lower retention time and possibly less efficient microbial communities. Removal kinetics studies show that most E. coli removal occurs in the first third of the system, however there was a clear difference in the removal kinetics between the planted gravel bed and the unplanted soil bed with the population of E. coli decreasing faster in the gravel bed. A comparison of two hybrid microcosms revealed that the system comprising a planted soil followed by a planted gravel gave better E. coli removal than the system comprising a planted gravel followed by a planted soil.

Effect of bone meal (calcium phosphate) amendments on metal release from contaminated soils - a leaching column study

Metal-contaminated soil may be remediated in situ by the formation of highly insoluble metal phosphates if an appropriate phosphorus (P) source can be found. Leaching column experiments have been carried out to assess the suitability of bone meal as such a source. Bone meal additions reduced metal release from a contaminated soil, increased soil and leachate pH and decreased soil leachate toxicity. Minimal P leaching occurred from the soil. The data are consistent with a proton consuming bone meal (calcium phosphate) dissolution reaction followed by the formation of metal phosphates. Although, no metal phosphates were observed to form using X-ray diffraction of scanning electron microscopy this could be due to their low concentration. Relatively low (1:50 bone meal:soil) concentrations of fine (90-500 microns) bone meal would appear to be an effective treatment for metal-contaminated soils.

The All-Data-Based Evolutionary Hypothesis of Ciliated Protists with a Revised Classification of the Phylum Ciliophora (Eukaryota, Alveolata).

The phylum Ciliophora plays important roles in a wide range of biological studies. However, the evolutionary relationships of many groups remain unclear due to a lack of sufficient molecular data. In this study, molecular dataset was expanded with representatives from 55 orders and all major lineages. The main findings are: (1) 14 classes were recovered including one new class, Protocruziea n. cl.; (2) in addition to the two main branches, Postciliodesmatophora and Intramacronucleata, a third branch, the Mesodiniea, is identified as being basal to the other two subphyla; (3) the newly defined order Discocephalida is revealed to be a sister clade to the euplotids, strongly suggesting the separation of discocephalids from the hypotrichs; (4) the separation of mobilids from the peritrichs is not supported; (5) Loxocephalida is basal to the main scuticociliate assemblage, whereas the thigmotrichs are placed within the order Pleuronematida; (6) the monophyly of classes Phyllopharyngea, Karyorelictea, Armophorea, Prostomatea, Plagiopylea, Colpodea and Heterotrichea are confirmed; (7) ambiguous genera Askenasia, CyclotrichiumParaspathidium and Plagiocampa show close affiliation to the well known plagiopyleans; (8) validity of the subclass Rhynchostomatia is supported, and (9) the systematic positions of Halteriida and Linconophoria remain unresolved and are thus regarded as incertae sedis within Spirotrichea.

Bacterivory in ciliates isolated from constructed wetlands (reed beds) used for wastewater treatment

The bacterivorous activity of ciliates, isolated from constructed wetlands employing the root zone method of wastewater treatment, was investigated by measuring their grazing rates upon fluorescently-labelled bacteria (FLB), specifically Escherichia coli. The highest mean grazing rates were recorded for Paramecium spp (1.85 FLB/cell/min), which was the largest ciliate used in the study, followed by oxytrichids (1.104 FLB/cell/min), Halteria (0.648 FLB/cell/min) and scuticociliates (0.433 FLB/cell/min), the smallest ciliates used in the study. Lowest feeding rates were found in the anaerobes Plagiopyla and Caenomorpha, although the experimental conditions are likely to have adversely affected these forms. Large variations in grazing rates between individual cells from the same populations were observed. The proportion of empty cells, i.e. those without ingested FLB, and the decrease in the proportion of empty cells with incubation time, showed large differences between taxa. The present study indicates that ciliates are capable of all observed E. coli removal from wastewaters treated in constructed wetlands using the root zone method. However, it is unlikely that ciliates in situ continuously maintain maximum feeding rates. A variety of other processes, biotic and abiotic, are thought to be involved in the removal of pathogenic and indicator bacteria such as E. coli.

An approach to analyzing spatial patterns of planktonic ciliate communities for monitoring water quality in Jiaozhou Bay, northern China.

Spatial patterns of planktonic ciliate communities for assessment of marine environmental status were studied from June 2007 to May 2008 in Jiaozhou Bay, Qingdao, northern China. Ciliate communities were sampled biweekly at five sampling sites with a spatial gradient of environmental stress. Multivariate/univariate analyses demonstrated that: (1) the planktonic ciliate community structures represented significant differences among the five sites; (2) spatial patterns of the ciliate communities were significantly correlated with environmental variables, especially the nutrients nitrate nitrogen (NO₃-N) and soluble reactive phosphates (SRP); (3) five dominant species (e.g., Rimostrombidium veniliae, Strombidium capitatum, Mesodinium pupula and Strombidinopsis acutum) were significantly correlated with nitrogen and/or SRP; and (4) both species richness and species diversity indices were correlated with NO₃-N and salinity. These results suggest that planktonic ciliated protozoa might be used as a robust bioindicator of marine water quality.

Taxonomic Redescriptions of Two Ciliates, Protogastrostyla pulchra n. g., n. comb. and Hemigastrostyla enigmatica (Ciliophora: Spirotrichea, Stichotrichia), with Phylogenetic Analyses Based on 18S and 28S rRNA Gene Sequences

ABSTRACT. The morphology and infraciliature of two stichotrichid ciliates, Gastrostyla pulchra (Perejaslawzewa 1886) Kahl, 1932 and Hemigastrostyla enigmatica (Dragesco and Dragesco‐Kernéis 1986) Song & Wilbert, 1997 , collected from marine and brackish sediments, were investigated by using living observations and protargol impregnations. Both 18S and 28S rRNA genes of these two species were sequenced. The 18S rDNA show high similarities (98.4%–99.7%) among populations of each species. There is about 94% similarity in 18S rDNA genes between G. pulchra and Gastrostyla steinii, the type species of the genus, which has been confirmed to be an oxytrichid by previous studies. In the phylogenetic trees of 18S, 28S, and combined 18S and 28S rDNA, both G. pulchra and H. enigmatica are consistently placed outside the well‐established oxytrichid clade. Based on our analyses and previous ontogenetic data, we conclude that these two species may represent some lower groups in the subclass Stichotrichia, and that G. pulchra should represent a new genus, Protogastrostyla n. g. This new genus, which is morphologically similar to Gastrostyla, differs in its morphogenesis: the apical part of the old AZM is retained combining with the newly built membranelles that develop from the proters oral primordium; the primary primordia of the dorsal kinety; and marginal primordia commence de novo without a definite contribution from the old structure.

Removal of pathogens from wastewaters by the root zone method (RZM)

Full-scale and laboratory-scale artificially constructed reed beds utilising the root zone method (RZM) of wastewater treatment were investigated for their removal efficiencies of a range of pathogenic microorganisms. Performances were compared for RZMs employing different hydrophytes, planting substrates and climatic conditions. All pathogens were removed by the RZM to some extent. Planted beds generally performed better than unplanted beds although the type of hydrophyte used was not significant. Gravel beds were more efficient than soil beds in the removal of protozoan pathogens and of helminth eggs; numbers of these pathogens were below levels of detection in gravel bed effluents. For faecal coliforms the type of planting substrate was not significant, providing hydrophytes were present. Climatic conditions made little difference to treatment efficiency although seasonal variation was observed in temperature climates, where the removal of faecal coliforms declined in the winter months, and in subtropical climates where higher numbers of free-living amoebae were recorded during the summer. It was concluded that the RZM is a suitable method for the treatment of domestic wastewaters in tropical and subtropical regions of Mexico.

An approach to determining the sampling effort for analyzing biofilm-dwelling ciliate colonization using an artificial substratum in coastal waters

A new approach to determining sampling effort for analyzing biofilm-dwelling ciliate colonization was studied in the coastal waters of the Yellow Sea, northern China, from May to June 2010. The optimal sample size for evaluating biofilm-dwelling ciliate colonization increased with shortening exposure time, and can be determined according to the probability of recovering those species with a specified cumulative contribution to communities. More slide-replicates were required at a depth of 3 m than at 1 m to recover equivalent proportions of the ciliate communities. For routine colonization dynamics analyses, 10 slide-replicates (175 cm2) were sufficient to achieve a 95% probability of recovering those species with a cumulative contribution of >90% to the ciliate communities at a depth of 1 m. These results suggest that 10 slide-replicates immersed at a depth of 1 m may be an optimal sampling strategy for analyzing the colonization dynamics of biofilm-dwelling ciliate communities in marine habitats.

Planktonic protist communities in a semi-enclosed mariculture pond: structural variation and correlation with environmental conditions

In order to evaluate the environmental status within a mariculture pond, temporal variations of physico-chemical factors, protist community structure and interactions between biota and environmental conditions were investigated during a complete cycle in semi-enclosed shrimp-farming waters near Qingdao, north China. Results revealed that: (1) a total of 54 protist taxa with ten dominant species was present, comprising 4 chlorophyceans, 2 chrysophyceans, 5 cryptophyceans, 10 dinoflagellates, 3 euglenophyceans, 10 diatoms, 18 ciliates and 2 sarcodines; (2) a single peak of protist abundance occurred in October, mainly due to the chlorophyceans, diatoms and chrysophyceans, while the bimodal peaks of biomass in July and October were mainly due to the ciliates, dinoflagellates and diatoms; (3) the succession of protist communities significantly correlated with the changes of nutrients, salinity and temperature, especially phosphate, either alone or in combination with NO 3 ; (4) species diversity and evenness indices were found to be relatively independent of physico-chemical factors, whereas species richness and the ratio of biomass to abundance were strongly correlated with water temperature and abundances of bacteria. It was concluded that planktonic protists are potentially useful bioindicators of water quality in a semi-enclosed mariculture system.

Small Subunit rRNA Phylogenies Show that the Class Nassophorea is Not Monophyletic (Phylum Ciliophora)

ABSTRACT. The hypostome ciliates have been generally classified into two classes, Phyllopharyngea and Nassophorea. The status of Nassophorea and its relationship with Phyllopharyngea is one of the most controversial issues in ciliate systematics. Here we focus on the phylogenetic interrelationships of Nassophorea and Phyllopharyngea based on small subunit ribosomal RNA gene sequences. The three nassophorean subgroups, synhymeniids, microthoracids, and nassulids, each emerged as monophyletic, with synhymeniids as a sister group of Phyllopharyngea, and microthoracids as a sister of the synhymeniids+Phyllopharyngea clade in all phylogenies. The exact placement of the nassulids, however, remains uncertain. Following a detailed analysis of phenotypic characters, we hypothesize that: (1) the Phyllopharyngea could have evolved from synhymeniids, with the further development of their subkinetal microtubules as one of the major events; and (2) the development of monokinetid structures, as well as the reduction and specialization of the cyrtos and cortex, might have occurred during the diversifications of the microthoracids, synhymeniids, and Phyllopharyngea from a common ancestor. Expanding the class Phyllopharyngea to include the synhymeniids as a subclass, and designating a new subclass Subkinetalia n. subcl. for the group comprising cyrtophorians, chonotrichians, rhynchodians, and suctorians, are proposed.