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Cryptosporidium parvum infections of domestic animals can have a considerable economic impact and as oocysts are voided in the faeces of infected hosts, environmental contamination with agricultural waste has also become a matter of concern. Since only viable oocysts are potentially infectious, the numbers of oocysts excreted during infection can have important implications for both veterinary and public health. During the course of infection in experimentally infected lambs, oocyst viability was assessed by a fluorogenic vital dyes assay and by a maximized in vitro excystation assay. The excreted oocyst populations contained a higher proportion of viable oocysts 5–11 days post infection (d.p.i.) than later in the infection. Oocyst viability declined consistently 11–15 d.p.i. and coincided with periods when peaks in serum and intestinal anti-Cryptosporidium antibodies have been reported to occur. Infected lambs excreted a mean of 4·8 (standard error [S.E.]±0·4)×109 oocysts per g of faeces, of which half were non-viable and therefore of no significance for disease transmission. This study demonstrates that the numbers of viable oocysts excreted by infected lambs is smaller than previously suspected.

Coral reefs offer valuable ecosystem goods and services, such as coastal protection, erosion regulation, fishery, biodiversity, habitat and nursery grounds. However, they face threats from anthropogenic activities, including poor water quality, global warming, coastal development and unsustainable fisheries. Marine Protected Areas (MPAs) provide a structured and holistic approach in addressing these threats. Regular monitoring and assessment of these MPAs are crucial components in evaluating the MPAs design and effectiveness. Two coral reefs (i.e. Poblacion and Kadurong Reefs) were established as MPAs in Liloan, Cebu, Philippines to protect crucial habitat and biodiversity with the hope of improving fisheries by avoiding fish stock disintegration. These coral reefs provide shelter to many commercially-significant fish species, supporting subsistence and livelihood in the community. These MPAs are not only biologically rich, but they also support socio-economic stability. Hence, management and protection of the coral reefs in the MPAs of Liloan, Cebu is of paramount importance. To formulate conservation and applicable management measures, research and monitoring should be in place. This paper presents the data collected from the short term monitoring in the Poblaction and Kadurong Reefs. The paper describes an important set of data that can be used by the stakeholders to benchmark biophysical assessments for management of marine-protected areas in Liloan. This data paper provides baseline information on the health of the coral reefs of the MPAs in Liloan, Cebu. Datasets covering physico-chemical and biological parameters inclusive of water quality, coral reef cover, fish and plankton occurrence and abundance were determined using the standard protocols for surveying tropical marine resources. The results will serve as a benchmark in formulating guidelines and implementing relevant policies for the effective management and protection of the MPAs in Liloan, Cebu, Philippines.

Recent studies have revealed a high diversity of pleurostomatid ciliates in brackish habitats. Here, a novel species, Pseudolitonotus spirelis gen. et sp. n., isolated from a mangrove wetland of southern China, was investigated based on living observation, protargol staining, and molecular analyses. The new genus Pseudolitonotus gen. n. is characterized by the last left somatic kinety (LKn) being shortened and none of the right somatic kineties extending to the anterior end of the cell, thus distinguishing it from all known pleurostomatid genera. The type species, Pseudolitonotus spirelis sp. n., is characterized by the possession of two macronuclear nodules, 11–15 right and 7–9 left kineties, a single contractile vacuole subterminally located, extrusomes evenly spaced along the entire ventral margin and some forming an “apical group”, two types of cortical granules, and the bottom of the oral slit invariably being twisted. Litonotus gracilis (Pan et al. Eur J Protistol 51:494–506, 2015) is believed to be another member of this new genus as its LKn and right somatic kineties are all shortened. Hence, a new combination, Pseudolitonotus gracilis (Pan et al., 2015) comb. n., is suggested and its diagnosis is improved. Molecular phylogenetic analyses based on SSU rDNA sequence data reveal that Pseudolitonotus gen. n. is monophyletic and groups with Apolitonotus (Pan et al. J Eukaryot Microbiol 67:252–262, 2020) of the family Protolitonotidae (Wu et al. Zool Scr 46:245–253, 2017). However, the familial assignment of this new genus is uncertain based on current data.

In Europe, ticks are major vectors of both human and livestock pathogens (e.g. Lyme disease, granulocytic anaplasmosis, bovine babesiosis). Agricultural landscapes, where animal breeding is a major activity, constitute a mosaic of habitat types of various quality for tick survival and are used at different frequencies by wild and domestic hosts across seasons. This habitat heterogeneity, in time and space, conditions the dynamics of these host-vector-pathogen systems and thus drives acarological risk (defined as the density of infected ticks). The principal objective of the OSCAR project (2011-2016) was to examine the links between this heterogeneity and acarological risk for humans and their domestic animals. Here, we present the data associated with this project. This paper reports a database on the distribution and densities of I. ricinus ticks - the most common tick species in French agricultural landscapes - and the prevalence of three tick-borne pathogens (Anaplasma phagocytophilum, Borrelia spp. and Babesia spp.) in two sites in north-western (“Zone Atelier Armorique”: ZA site) and south-western (“Vallées et Coteaux de Gascogne”: VG site) France. The distribution and density of ticks along a gradient of wooded habitats, as well as biotic variables, such as the presence and abundance of their principal domestic (livestock) and wild hosts (small mammals), were measured from forest cores and edges to more or less isolated hedges, all bordering meadows. Ticks, small mammals and information on local environmental conditions were collected along 90 transects in each of the two sites in spring and autumn 2012 and 2013 and in spring 2014, corresponding to the main periods of tick activity. Local environmental conditions were recorded along each tick and small mammal transect: habitat type, vegetation type and characteristics, slope and traces of livestock presence. Samples consisted of questing ticks collected on the vegetation (mainly I. ricinus nymphs), biopsies of captured small mammals and ticks fixed on small mammals. In the VG site, livestock occurrence and abundance were recorded each week along each tick transect. A total of 29004 questing ticks and 1230 small mammals were captured during the study across the two sites and over the five field campaigns. All questing nymphs (N = 12287) and questing adults (N = 646) were identified to species. Ticks from small mammals (N = 1359) were also identified to life stage. Questing nymphs (N = 4518 I. ricinus) and trapped small mammals (N = 908) were analysed for three pathogenic agents: A. phagocytophilum, Borrelia spp. and Babesia spp. In the VG site, the average prevalence in I. ricinus nymphs for A. phagocytophilum, Borrelia spp. and Babesia spp. were, respectively 1.9% [95% CI: 1.2-2.5], 2.5% [95% CI: 1.8-3.2] and 2.7% [95% CI: 2.0-3.4]. In small mammals, no A. phagocytophilum was detected, but the prevalence for Borrelia spp. was 4.2% [95% CI: 0.9-7.5]. On this site, there was no screening of small mammals for Babesia spp. In ZA site, the average prevalence in nymphs for A. phagocytophilum, Borrelia spp. and Babesia were, respectively 2.2% [95% CI: 1.6-2.7], 3.0% [95% CI: 2.3-3.6] and 3.1% [95% CI: 2.5-3.8]. In small mammals, the prevalence of A. phagocytophilum and Borrelia spp. were, respectively 6.9% [95% CI: 4.9-8.9] and 4.1% [95% CI: 2.7-5.9]. A single animal was found positive for Babesia microti at this site amongst the 597 tested.

Permafrost, frozen ground cemented with ice, occupies about a quarter of the Earth’s hard surface and reaches up to 1000 metres depth. Due to constant subzero temperatures, permafrost represents a unique record of past epochs, whenever it comes to accumulated methane, oxygen isotope ratio or stored mummies of animals. Permafrost is also a unique environment where cryptobiotic stages of different microorganisms are trapped and stored alive for up to hundreds of thousands of years. Several protist strains and two giant protist viruses isolated from permafrost cores have been already described. In this paper, we describe a collection of 35 amoeboid protist strains isolated from the samples of Holocene and Pleistocene permanently frozen sediments. These samples are stored at −18°C in the Soil Cryology Lab, Pushchino, Russia and may be used for further studies and isolation attempts. The collection strains are maintained in liquid media and may be available upon request. The paper also presents a dataset which consists of a table describing the samples and their properties (termed "Sampling events") and a table describing the isolated strains (termed "Occurrences"). The dataset is publicly available through the GBIF portal.

Background Giardia duodenalis is a widespread protozoan parasite affecting humans and many species of animals, including dogs and cats. Due to its zoonotic potential, it is important to know the frequency of this parasite in companion animals. The aim of this study was to determine current epidemiological status of G. duodenalis in household dogs and cats. Methods In this study, 293 fecal samples from pet dogs and cats were collected from January 2017 to July 2019 and tested for G. duodenalis by PCR (using β-giardin gene). The animals were divided into groups depending on their age, breed and fecal consistency. Results The examination allowed for detection of G. duodenalis in 6.0% of canine and 3.9% of feline fecal samples. The highest frequency was revealed in young (under one-year old) dogs. Sequencing confirmed the presence of assemblages C and D in dogs and A and F in cats. Conclusion The study showed current frequency of G. duodenalis in dogs and cats and also revealed the occurrence of host-specific assemblages as well as zoonotic assemblage A.

PurposeThe aim of this study was to investigate protozoan cysts and oocyts in the faecal pellets of the German cockroach (Blattella germanica), with emphasis on the prevalence ofLophomonas blattarum.MethodsThirty adultBlattella germanica(12 males; 18 females) were trapped near Madrid, Spain. Expelled faecal pellets were collected in the laboratory over 5 days in two plastic containers. Protozoan cysts from one container were weighted and those in the other used for morphologically identification.ResultsProtozoan cysts/oocysts per gram of faecal pellet were:Nyctotherussp. (0.0019/g),Entamoeba(0.0007/g),Balantidium coli(0.0001/g),Lophomonas blattarum(0.00038/g). Observation of 189 protozoan cysts/oocysts indicated the following prevalence:Nyctotherussp. 65 (34.4%);Gregarinaspp. 34 (18%);Entamoebasp. 24 (12.7%);Cryptosporidiumsp. 17 (9%); Coccidia 16 (8.4%);Lophomonas blattarum13 (6.8%);Balantidium coli4 (2.1%); and unclassified 16 (8.4%).ConclusionLophomonas blattarumhas previously been demonstrated in the gut of cockroaches, but this is the first assessment of the prevalence inBlattella germanicain faecal pellets. The presence of protozoa in faecal pellets provides some evidence for one step in a hypothesised route of respiratory infection whereby protozoa from household insects enter the respiratory tract.

Cryptosporidium spp. are worldwide protozoan parasites that can affect to a broad range of vertebrate hosts, including rodents. In the island of Corsica (France), there are no previous data about these protozoa infecting wild rodents. To estimate the distribution and occurrence, a total of 117 wild murine rodents of the species Rattus rattus (84), Mus musculus domesticus (21), Apodemus sylvaticus (11), and Rattus norvegicus (1) were captured in 24 different biotopes. Fecal samples were screened for Cryptosporidium spp. by nested PCR to amplify an 830 bp fragment of the 18S rRNA gene. As general occurrence, 15.4% of the rodents analyzed were positive for Cryptosporidium spp., being detected widely distributed along the island in R. rattus (17.6%) and M. m. domesticus (14.3%). Cryptosporidium viatorum, Cryptosporidium sp. rat genotype II, and Cryptosporidium sp. rat genotype III were successfully identified in R. rattus. The results herein reported provide the first data on Cryptosporidium spp. in wild murine species from a Mediterranean island and constitute the first report of the zoonotic species C. viatorum in R. rattus. Although a low occurrence of Cryptosporidium spp. in murids was obtained and only in one animal the zoonotic species C. viatorum was identified, our results highlight that wild murine rodents from Corsica could mediate in the maintenance and transmission of this protozoan to the environment and other hosts including humans and animals. Further studies are required to better understand the epidemiology of Cryptosporidium spp. in wild rodents from Corsica and their possible public health repercussions.

This paper identified the dominant protozoan species in the four layers of rhizosphere soil during the six growth stages of Beta vulgaris L. and analyzed the correlations of the abundance and diversity of the dominant protozoan species with soil properties at different growth stages and soil depth. A total of 15 species of protozoa were identified; among them, Colpoda sp., Bodo sp., two kinds of Oxytricha sp., and Tachysoma sp. were the most dominant species of Beta vulgaris L. rhizosphere soil. The Colpoda sp. was eurytopic species in the Beta vulgaris L. rhizosphere soil and Tachysoma sp., Vorticella sp., Colpoda sp., Oxytricha sp.1, and Oxytricha sp. 2 were noted closely related to the acceleration function of circulation of N and P elements in soils. These dominant protozoan species were proposed to play a significant role of fertilization on N supply in rhizosphere soil during the initial growth of Beta vulgaris L.

The increase of swine production in the Pacific Islands has inevitably led to environmental pollution concerns from discharged wastewater derived from both washing and manure. The slurry accumulates in lagoons, where supernatant wastewater containing high levels of pathogens and nutrients becomes nonpoint source water pollution that deteriorates the quality of receiving water bodies. Soil filtration is a promising cost-effective technology for removing pollutants from swine wastewater, however, the excessive growth of bacteria in soil media often accompanies the filtration process. This study investigates soil filtration mediated by protozoa activities to remove Escherichia coli (E. coli) in synthetic swine wastewater. The experiment used plastic columns packed with Leilehua soil from Oahu Island, Hawaii. The soil physicochemical adsorption was seen to reduce 95.52–96.47% of E. coli. However, the average removal efficiencies were increased to 98.17% in a single stage, and 99.99% in two sequential columns, under predation conditions. The filtration media containing naturally established bacterivores with the prey, provided a bioactive means to remove E. coli from the influent. The proper design of Leilehua soil filters potentially removes E. coli from the influent to meet the standard level of recycled water.

We performed a cross-sectional parasitological survey to assess the prevalence of protozoa and helminth infection among 275 school-age children (SAC) living in rural and peri-urban areas located in different ecological zones of the Department of La Paz, Bolivia. Public health activities for the control of STH, based on the biannual administration of mebendazole to preschool and school children are implemented at national level since 1986. We found an overall prevalence of 82.2% for intestinal parasites, including protozoa (80%) and helminths (23.3%). Blastocystis and Entamoeba coli were the most prevalent protozoa (found in 44% and 20.7% of the SAC enrolled); as for helminths, Ascaris lumbricoides and Hymenolepis nana were diagnosed in 14.5% and 3.3% of the children, respectively, followed by Trichuris trichiura 1.4%, Enterobius vermicularis 1.4%, Strongyloides stercoralis 0.7% and hookworms 0.7%. Molecular characterization of Blastocystis positive samples evidenced three different subtypes (ST1, ST2, ST3) highlighting the risk of transmission also from animal reservoir. We found a significant difference in the distribution of intestinal parasitic infection (IPIs) by ecological zone (44/74. 59% in Andean highlands, 94/170, 88% in tropical lowlands and 88/94, 94% in the Yungas, p, Graphic abstractUnlabelled Image, Highlights • In the Department of La Paz, the current overall STH among school-age children prevalence is high (17.1%), with a patchy distribution throughout the region. • The prevalence of STH infections of moderate and heavy intensity shows that the morbidity due to these parasites has not been eliminated yet. • The preventive chemotherapy coverage should be significantly improved in all the at-risk populations, and strategy of PC delivery should be tailored to local conditions. • Updated epidemiological information is crucial to inform policy for STH control. • Significant efforts are still needed to reduce transmission and to improve health and sanitation in this area. Interventions should be inspired by the WASH principles, endorsed by the WHO.

The aerobic granular sludge (AGS) process is an effective wastewater treatment technology for organic matter and nutrient removal that has been introduced in the market rapidly. Until now, limited information is available on AGS regarding the removal of bacterial and viral pathogenic organisms present in sewage. This study focussed on determining the relation between reactor operational conditions (plug flow feeding, turbulent aeration and settling) and physical and biological mechanisms on removing two faecal surrogates, Escherichia coli and MS2 bacteriophages. Two AGS laboratory-scale systems were separately fed with influent spiked with 1.0 × 106 CFU/100 mL of E. coli and 1.3 × 108 PFU/100 mL of MS2 bacteriophages and followed during the different operational phases. The reactors contained only granular sludge and no flocculent sludge. Both systems showed reductions in the liquid phase of 0.3 Log10 during anaerobic feeding caused by a dilution factor and attachment of the organisms on the granules. Higher removal efficiencies were achieved during aeration, approximately 1 Log10 for E. coli and 0.6 Log10 for the MS2 bacteriophages caused mainly by predation. The 18S sequencing analysis revealed high operational taxonomic units (OTUs) of free-living protozoa genera Rhogostoma and Telotrochidium concerning the whole eukaryotic community. Attached ciliates propagated after the addition of the E. coli, an active contribution of the genera Epistylis, Vorticella, and Pseudovorticella was found when the reactor reached stability. In contrast, no significant growth of predators occurred when spiking the system with MS2 bacteriophages, indicating a low contribution of protozoa on the phage removal. Settling did not contribute to the removal of the studied bacterial and viral surrogates. M.L. Barrios-Hernández acknowledges the Technological Institute of Costa Rica for providing the fellowship (Grant Number 007-2014-M) to pursue her PhD programme (2016-2020) at IHE-Delft, the Netherlands. K. Mora-Cabrera acknowledges the Generalitat Valenciana (GRISOLIAP/2017/173) and the European Social Funds (BEFPI/2019/065) for their financial support.

An important functional trait of organisms is their trophic mode. It determines their position within food webs, as well as their function within an ecosystem. For the better part of the 20th century, aquatic protist communities were thought to consist mainly of producers (phytoplankton) and consumers (protozooplankton). Phytoplankton cover their energy requirements through photosynthesis (phototrophy), while protozooplankton graze on prey and organic particles (phagotrophy). However, over the past decades, it was shown that another trophic group (mixoplankton) comprise a notable part of aquatic protist communities. Mixoplankton employ a third trophic mode by combining phototrophy and phagotrophy (mixotrophy). Due to the historical dichotomy, it is not straightforward to gain adequate and correct information on the trophic mode of aquatic protists. Long hours of literature research or expert knowledge are needed to correctly assign trophic modes. Additionally, aquatic protists also have a long history of undergoing taxonomic changes which make it difficult to compare past and present literature. While WoRMS, the World Register of Marine Species, keeps track of the taxonomic changes and assigns each species a unique AphiaID that can be linked to its various historic and present taxonomic hierarchy, there is currently no machine-readable database to query aquatic protists for their trophic modes. This paper describes a dataset that was submitted to WoRMS and links aquatic protist taxa, with a focus on marine taxa, to their AphiaID and their trophic mode. The bulk of the data used for this dataset stems from (routine) monitoring stations in the North Sea and the Baltic Sea. The data were augmented and checked against state-of-the-art knowledge on mixoplankton taxa by consulting literature and experts. Thus, this dataset provides a first attempt to make the trophic mode of aquatic protists easily accessible in both a human- and machine-readable format.

This paper describes a phytoplankton data series generated through systematic observations in the Belgian Part of the North Sea (BPNS). Phytoplankton samples were collected during multidisciplinary sampling campaigns, visiting nine nearshore stations with monthly frequency and an additional eight offshore stations on a seasonal basis. The data series contain taxon-specific phytoplankton densities determined by analysis with the Flow Cytometer And Microscope (FlowCAM®) and associated image-based classification. The classification is performed by two separate semi-automated classification systems, followed by manual validation by taxonomic experts. To date, 637,819 biological particles have been collected and identified, yielding a large dataset of validated phytoplankton images. The collection and processing of the 2017–2018 dataset are described, along with its data curation, quality control and data storage. In addition, the classification of images using image classification algorithms, based on convolutional neural networks (CNN) from 2019 onwards, is also described. Data are published in a standardised format together with environmental parameters, accompanied by extensive metadata descriptions and finally labelled with digital identifiers for traceability. The data are published under a CC‐BY 4.0 licence, allowing the use of the data under the condition of providing the reference to the source.

We investigated changes in protozoa and metazoa community in relation to process parameters in activated sludge from four wastewater treatment plants (WWTPs) throughout the period of 1 year. Principal component analysis (PCA) showed that activated sludge from investigated treatment plants had different dominating species representatives and community composition mainly depends on individual features of the treatment plants. Redundancy analysis (RDA) showed that the temperature in bioreactors was the most relevant factor explaining changes in the microorganism community, whereas reduction rate of chemical oxygen demand (COD), biological oxygen demand (BOD5), suspended solids (SS), and total nitrogen (TN) did not sufficiently explain the variation in protozoa and metazoan community composition. The results indicate that in stable working WWTP it is difficult to find a pronounced link between activated sludge species composition, process parameters, and plant configuration. Applied multivariate analysis can be a valuable tool for the exploration of the relations between community composition and WWTP process parameters. Electronic supplementary material The online version of this article (10.1007/s11356-020-10684-5) contains supplementary material, which is available to authorized users.

Background: Testate amoebae are a polyphyletic group of protists living preferentially in soils, freshwaters and wetlands. These Protozoa have a worldwide distribution, but their presence and diversity in the Azores (a remote oceanic archipelago) is poorly known, with only twelve taxa recorded so far. The published information reflects occasional collections from sporadic field visits from naturalists to São Miguel Island, mainly in the nineteenth century. To overcome this limitation, a standardised survey was carried out on the Island, sampling different types of habitats from several localities to provide the distribution and information on species ecology of testate amoebae. New information: In this study, 43 species of testate amoebae were recorded (within a total of 499 occurrences), belonging to two orders of Protista (26 Arcellinida and 17 Euglyphida). The most frequently occurring testate amoebae were Euglypha strigosa, Trinema lineare, Euglypha rotunda, Assulina muscorum and Cyclopyxis eurystoma. The most diverse genus was Euglypha (six species). A total of 38 species are new records for the Azores Archipelago. These data help to improve knowledge of the geographical distribution of testate amoebae in the northern hemisphere and their diversity in the Azores Archipelago This work was funded by FCT – Foundation for Science and Technology, the European Union, QREN, FEDER, COMPETE programmes (PMR - DL57/2016/ICETA/EEC2018/25; MSS - ICETA/EEC2018/25; DiscoverAzores Project - PTDC/CTA-AMB/28511/2017; UID/BIA/50027/2020 and POCI-01-0145-FEDER-006821), AZORESBIOPORTAL – PORBIOTA (ACORES-01-0145-FEDER-000072) and Consolidation and Structuring Project 2018 GRC-ED431C 2018/32 of Xunta de Galicia government SI

Ciliates represent a morphologically and genetically distinct group of single-celled eukaryotes that segregate germline and somatic functions into two types of nuclei and exhibit complex cytogenetic events during the sexual process of conjugation, which is under the control of the so-called “mating type systems”. Studying conjugation in ciliates may provide insight into our understanding of the origins and evolution of sex and fertilization. In the present work, we studied in detail the sexual process of conjugation using the model species Euplotes vannus, and compared these nuclear events with those occurring in other ciliates. Our results indicate that in E. vannus: 1) conjugation requires about 75 hours to complete: the longest step is the development of the new macronucleus (ca. 64h), followed by the nuclear division of meiosis I (5h); the mitotic divisions usually take only 2h; 2) there are three prezygotic divisions (mitosis and meiosis I and II), and two of the eight resulting nuclei become pronuclei; 3) after the exchange and fusion of the pronuclei, two postzygotic divisions occur; two of the four products differentiate into the new micronucleus and macronucleus, respectively, and the parental macronucleus degenerates completely; 4) comparison of the nuclear events during conjugation in different ciliates reveals that there are generally three prezygotic divisions while the number of postzygotic divisions is highly variable. These results can serve as reference to investigate the mating type system operating in this species and to analyze genes involved in the different steps of the sexual process.

Cancer recurrence has remained a significant challenge, despite advances in therapeutic approaches. In part, this is due to our incomplete understanding of the biology of cancer stem cells and the underlying molecular mechanisms. The phenomenon of differentiation and dedifferentiation (phenotypic switching) is not only unique to stem cells but it is also observed in several other organisms, as well as evolutionary-related microbes. Here, we propose the use of a primitive eukaryotic unicellular organism, Acanthamoeba castellanii, as a model to study the molecular mechanisms of cellular differentiation and dedifferentiation.

Acanthamoeba castellanii (Ac) are ubiquitously distributed in nature, and by contaminating medical devices such as heart valves and contact lenses, they cause a broad range of clinical presentations to humans. Although several molecules have been described to play a role in Ac pathogenesis, including parasite host-tissue invasion and escaping of host-defense, little information is available on their mechanisms of secretion. Herein, we describe the molecular components secreted by Ac, under different protein availability conditions to simulate host niches. Ac extracellular vesicles (EVs) were morphologically and biochemically characterized. Dynamic light scattering analysis of Ac EVs identified polydisperse populations, which correlated to electron microscopy measurements. High-performance thin liquid chromatography of Ac EVs identified phospholipids, steryl-esters, sterol and free-fatty acid, the last two also characterized by GC-MS. Secretome composition (EVs and EVs-free supernatants) was also determined and proteins biological functions classified. In peptone-yeast-glucose (PYG) medium, a total of 179 proteins were identified (21 common proteins, 89 exclusive of EVs and 69 in EVs-free supernatant). In glucose alone, 205 proteins were identified (134 in EVs, 14 common and 57 proteins in EVs-free supernatant). From those, stress response, oxidative and protein and amino acid metabolism proteins prevailed. Qualitative differences were observed on carbohydrate metabolism enzymes from Krebs cycle and pentose phosphate shunt. Serine proteases and metalloproteinases predominated. Analysis of the cytotoxicity of Ac EVs (upon uptake) and EVs-free supernatant to epithelial and glioblastoma cells revealed a dose-dependent effect. Therefore, the Ac secretome differs depending on nutrient conditions, and is also likely to vary during infection.

The inhibition of α-, β-, γ-, and δ-class carbonic anhydrases (CAs, EC 4.2.1.1) from bacteria (Vibrio cholerae and Porphyromonas gingivalis) and diatoms (Thalassiosira weissflogii) with a panel of N’-aryl-N-hydroxy-ureas is reported. The α-/β-CAs from V. cholerae (VchCAα and VchCAβ) were effectively inhibited by some of these derivatives, with KIs in the range of 97.5 nM – 7.26 µM and 52.5 nM – 1.81 µM, respectively, whereas the γ-class enzyme VchCAγ was less sensitive to inhibition (KIs of 4.75 – 8.87 µM). The β-CA from the pathogenic bacterium Porphyromonas gingivalis (PgiCAβ) was not inhibited by these compounds (KIs > 10 µM) whereas the corresponding γ-class enzyme (PgiCAγ) was effectively inhibited (KIs of 59.8 nM – 6.42 µM). The δ-CA from the diatom Thalassiosira weissflogii (TweCAδ) showed effective inhibition with these derivatives (KIs of 33.3 nM – 8.74 µM). As most of these N-hydroxyureas are also ineffective as inhibitors of the human (h) widespread isoforms hCA I and II (KIs > 10 µM), this class of derivatives may lead to the development of CA inhibitors selective for bacterial/diatom enzymes over their human counterparts and thus to anti-infectives or agents with environmental applications.

Background: In developing countries, diarrhoea is the most common cause of death for children under five years of age, with Giardia lamblia, Cryptosporidium and Entamoeba histolytica as the most frequent pathogenic parasites. Traditional microscopy for stool parasites has poor sensitivity and specificity, while new molecular methods may provide more accurate diagnostics. In poor regions with sample storage hampered by uncertain electricity supply, research would benefit from a method capable of analysing dried stools. Methods: A real-time multiplex PCR method with internal inhibition control was developed for detecting Giardia lamblia, Cryptosporidium hominis/parvum and Entamoeba histolytica directly from stool specimens. Applicability to dried samples was checked by comparing with fresh ones in a small test material. Finally, the assay was applied to dried specimens collected from Guinea-Bissauan children with diarrhoea. Results: The PCR's analytical sensitivity limit was 0.1 ng/ml for G. lamblia DNA, 0.01 ng/ml for E. histolytica DNA and 0.1 ng/ml for Cryptosporidium sp. In the test material, the assay performed similarly with fresh and dried stools. Of the 52 Guinea-Bissauan samples, local microscopy revealed a parasite in 15%, while PCR detected 62% positive for at least one parasite: 44% of the dried samples had Giardia, 23% Cryptosporidium and 0% E. histolytica. Conclusions: Our new multiplex real-time PCR for protozoa presents a sensitive method applicable to dried samples. As proof of concept, it worked well on stools collected from Guinea-Bissauan children with diarrhoea. It provides an epidemiological tool for analysing dried specimens from regions poor in resources.

Motivation Antimicrobial peptides (AMPs) are innate immune molecules that exhibit activities against a range of microbes, including bacteria, fungi, viruses and protozoa. Recent increases in microbial resistance against current drugs has led to a concomitant increase in the need for novel antimicrobial agents. Over the last decade, a number of AMP prediction tools have been designed and made freely available online. These AMP prediction tools show potential to discriminate AMPs from non-AMPs, but the relative quality of the predictions produced by the various tools is difficult to quantify. Results We compiled two sets of AMP and non-AMP peptides, separated into three categories—antimicrobial, antibacterial and bacteriocins. Using these benchmark data sets, we carried out a systematic evaluation of ten publicly available AMP prediction methods. Among the six general AMP prediction tools—ADAM, CAMPR3(RF), CAMPR3(SVM), MLAMP, DBAASP and MLAMP—we find that CAMPR3(RF) provides a statistically significant improvement in performance, as measured by the area under the receiver operating characteristic (ROC) curve, relative to the other five methods. Surprisingly, for antibacterial prediction, the original AntiBP method significantly outperforms its successor, AntiBP2 based on one benchmark dataset. The two bacteriocin prediction tools, BAGEL3 and BACTIBASE, both provide very good performance and BAGEL3 outperforms its predecessor, BACTIBASE, on the larger of the two benchmarks. Supplementary information Supplementary data are available at Bioinformatics online.

The co-treatment of two synthetic faecal sludges (FS-1 and FS-2) with municipal synthetic wastewater (WW) was evaluated in an aerobic granular sludge (AGS) reactor. After characterisation, FS-1 showed the following concentrations, representative for medium-strength FS: 12,180 mg TSS L−1, 24,300 mg total COD L−1, 93.8 mg PO3-P L−1, and 325 mg NH4-N L−1. The NO3-N concentration was relatively high (300 mg L−1). For FS-2, the main difference with FS-1 was a lower nitrate concentration (18 mg L−1). The recipes were added consecutively, together with the WW, to an AGS reactor. In the case of FS-1, the system was fed with 7.2 kg total COD m−3d−1 and 0.5 kg Nitrogen m−3d−1. Undesired denitrification occurred during feeding and settling resulting in floating sludge and wash-out. In the case of FS-2, the system was fed with 8.0 kg total COD m−3d−1 and 0.3 kg Nitrogen m−3d−1. The lower NO3-N concentration in FS-2 resulted in less floating sludge, a more stabilised granular bed and better effluent concentrations. To enhance the hydrolysis of the slowly biodegradable particulates from the synthetic FS, an anaerobic stand-by period was added and the aeration period was increased. Overall, when compared to a control AGS reactor, a lower COD consumption (from 87 to 35 mg g−1 VSS h−1), P-uptake rates (from 6.0 to 2.0 mg P g VSS−1 h−1) and NH4-N removal (from 2.5 to 1.4 mg NH4-N g VSS−1 h−1) were registered after introducing the synthetic FS. Approximately 40% of the granular bed became flocculent at the end of the study, and a reduction of the granular size accompanied by higher solids accumulation in the reactor was observed. A considerable protozoa Vorticella spp. bloom attached to the granules and the accumulated particles occurred; potentially contributing to the removal of the suspended solids which were part of the FS recipe.

International audience; Background - The Biological Field Station of Paimpont (Station Biologique de Paimpont, SBP), owned by the University of Rennes and located in the Brocéliande Forest of Brittany (France), has been hosting student scientific research and field trips during the last 60 years. The study area of the SBP is a landscape mosaic of 17 ha composed of gorse moors, forests, prairies, ponds and creeks. Land use has evolved over time. Historical surveys by students and researchers focused on insects and birds. With this study, we aimed to increase the range of taxa observations, document changes in species composition and landscape and provide a basis for interdisciplinary research perspectives. We gathered historical data, implemented an all-taxon biodiversity inventory (ATBI) in different habitats of the SBP study area, measured abiotic factors in the air, water and soil and performed a photographical landscape observation during the BioBlitz held in July 2017. New information - During the 24 h BioBlitz, organised by the SBP and the EcoBio lab from the University of Rennes and the French National Center of Scientific Research (CNRS), different habitats were individually sampled. Seventy-seven experts, accompanied by 120 citizens and 12 young people participating in the European Volunteer Service, observed, identified and databased 660 species covering 5 kingdoms, 8 phyla, 21 classes, 90 orders and 247 families. In total, there were 1819 occurrences including records identified to higher taxon ranks, thereby adding one more kingdom and four more phyla. Historical data collection resulted in 1176 species and 4270 occurrences databased. We also recorded 13 climatic parameters, 10 soil parameters and 18 water parameters during the BioBlitz. Current habitats were mapped and socio-ecological landscape changes were assessed with a diachronic approach using 32 historical photographs and historical maps. The coupling of historical biodiversity data with new biotic and abiotic data and a photographic comparison of landscape changes allows an integrative understanding of how the SBP changed from agriculturally-used land to a managed natural area within the last 60 years. Hence, this BioBlitz represents an important holistic sampling of biodiversity for studies on trophic webs or on trophic interactions or on very diverse, but connected, habitats. The integration of social, biotic and abiotic data opens innovative research opportunities on the evolution of socio-ecosystems and landscapes.

Effluents from textile dyeing industry are so toxic that it threatens the environment in various ways. Hence, it should be treated before discharge. Biotreatment is the most promising approach for the remediation of toxic industrial effluent where microorganisms breakdown the dye molecule into its nontoxic basic elements. In effluent treatment plants (ETP), protozoa accelerate dye degradation process by stimulating the growth of acting bacteria and also play a key role in clarification of the secondary effluent through engulfment of excess bacteria. However, only a few types of protozoa can survive and thrive on this toxic environment. This study deals with the identification and survey of the protozoa present in the effluent treatment plant of a dyeing industry. Collected samples were stained with Leishman reagent and observed under microscope. A total of 10 protozoa were observed and 6 of them were identified up to genus viz. Paramecium, Nassula, Opercularium, Stylonychia, Telotrochidium, Trachelomonas on the basis of their morphological and structural arrangements observed under microscope. Hence, this study reveals that some protozoa can thrive in toxic dyeing effluent and play an important role in maintaining the sustainability of an effluent treatment plant (ETP).Bangladesh J Microbiol, Volume 30, Number 1-2,June-Dec 2013, pp 39-42

The ongoing and increasing worldwide demand for fish has caused a steady increase in aquaculture production during the last decades. This emphasizes the importance of farming systems with a low ecological footprint, like recirculating aquaculture systems (RAS), which are an alternative to traditional open systems. Furthermore, implementing microalgae treatments in RAS, sustainable water management and low discharge of concentrated wastewater could be achieved, allowing its reuse in the system. The influence of three factors on microalgae treatment efficiency in RAS water were studied: i) microalgae species (Chlorella vulgaris, Tetradesmus obliquus), ii) water pre-treatment (sterile filtration), and iii) sampling location within the RAS (e.g. from fish tank, after UV-disinfection, etc.). To this end, fully factorial, replicated cultivations were carried out in 100-ml flasks, and nutrient removal, microalgae growth, and density of bacteria and protozoa were measured for up to 18 days. Results show that both species are able to grow in RAS water and effectively remove nutrients in it, yet their performance depended greatly on water quality. In sterile RAS water, growth and nutrient removal efficiency of C. vulgaris surpassed that of T. obliquus. In non-sterile RAS water, the pattern reversed because of grazing protozoa. The location of sampling within the RAS had no discernible effect on microalgae growth or nutrient removal efficiency. The results confirm that a microalgae-based technology to treat and valorise RAS water is technically feasible, yet caution that inferences made can be reversed depending on the choice of the species and the pretreatment of the RAS water prior to cultivation.

There is a scarcity of recent epidemiological data on intestinal parasitic infections in France. We conducted a prospective study aimed at estimating the prevalence of 10 enteric parasites in Marseille, France, using real-time polymerase chain reaction (PCR)-based diagnosis. A total of 643 faeces from 488 patients referred to the Parasitology-Mycology Laboratory of the University Hospital of Marseille over a 6 months period were included. DNA was extracted using a semi-automated method. Parasites of interest were detected using singleplex quantitative PCRs (qPCRs). For positive samples, theBlastocystissubtype was determined by sequence analysis. During the study, the overall prevalence of enteric parasites was 17%.Blastocystis sp.was the most frequent species (10.5%), followed byDientamoeba fragilis(2.3%) andGiardia intestinalis(2.3%). The prevalence of other parasites was Blastocystissubtype was predominant (43.6%) and the other subtypes identified were ST1, ST2, ST4 and ST6. This is the first time that a qPCR-based diagnosis has been used to survey the prevalence of 10 enteric parasites in a French University Hospital. This study confirms that fast, specific, sensitive and simultaneous detection in a single stool sample by qPCR clearly outperforms conventional microscopy-based diagnosis. Furthermore, qPCR is particularly well suited to surveying gastroenteritis agents.

Six myxomycete species new to the Åland Islands are presented: Comatricha elegans, Cribraria intricata, Didymium minus, Hemitrichia clavata, Licea variabilis and Trichia favoginea. The record of Cribraria intricata is the third in Finland. Specimens were collected in September 2014. Altogether the number of myxomycete species found from the Åland Islands is now 55.

Understanding how closely related wildlife species and their domesticated counterparts exchange or share parasites, or replace each other in parasite life cycles, is of great interest to veterinary and human public health, and wildlife ecology. Grey wolves (Canis lupus) host and spread endoparasites that can either directly infect canid conspecifics or their prey serving as intermediate hosts of indirectly transmitted species. The wolf recolonization of Central Europe represents an opportunity to study parasite transmission dynamics between wildlife and domestic species for cases when a definitive host returns after local extinction – a situation equivalent to a ‘removal experiment’. Here we investigate whether the re–appearance of wolves has increased parasite pressure on hunting dogs – a group of companion animals of particular interest as they have a similar diet to wolves and flush wolf habitats when hunting. We compared prevalence (P) and species richness (SR) of helminths and the protozoan Sarcocystis to determine whether they were higher in hunting dogs from wolf areas (ndogs = 49) than a control area (ndogs = 29) without wolves. Of particular interest were S. grueneri and S. taeniata, known as ‘wolf specialists’. Five helminth and 11 Sarcocystis species were identified, of which all helminths and eight Sarcocystis species were shared between dogs and wolves. Overall prevalence and species richness of helminths (P:38.5% vs. 24.1%; SRmean:0.4 vs. 0.3 species) and Sarcocystis (P:63.3% vs. 65.5%, SRmean:2.1 vs. 1.8 species) did not differ between study sites. However, hunting dogs were significantly more likely to be infected with S. grueneri in wolf areas (P:45.2% vs. 10.5%; p = 0.035). The findings suggest that wolves indirectly increase S. grueneri infection risk for hunting dogs since cervids are intermediate hosts and occasionally fed to dogs. Furthermore, a periodic anthelminthic treatment of hunting dogs may be an effective measure to control helminth infections regardless of wolf presence., Graphical abstract Image 1, Highlights • General parasite burden in hunting dogs is not increased by wolves. • General Sarcocystis burden in hunting dogs is high due to raw feeding. • ‘Wolf specialist’ parasite S. grueneri more prevalent in hunting dogs from wolf areas.

Cryptosporidium and Giardia protozoa are zoonotic parasites that cause human gastroenteritis and can be transmitted to human through the fecal-oral route and water or food. Several species belong to these genera and their resistant forms occur in water, but only some of them are infectious to human. Health risk depends on the occurrence of infectious Cryptosporidium and Giardia species and genotypes in water, and only molecular techniques allow detecting them, as well as enable to identify the contamination source. In this work, genotyping and phylogenetic analysis have been performed on the basis of 18S rDNA and s-giardin genes sequences of Cryptosporidium and Giardia, respectively, in order to provide the molecular characterization of these parasites detected earlier in five natural water bodies in Poland and to track possible sources of their (oo)cysts in water. Genotyping revealed a high similarity (over 99 up to 100 %) of analyzed sequences to cattle genotype of C. parvum isolated from cattle and human and to G. intestinalis assemblage B isolated from human. The sequences obtained by others originated from patients with clinical symptoms of cryptosporidiosis or giardiasis and/or with the infection confirmed by different methods. The contamination of three examined lakes is probably human-originated, while the sources of contamination of two remaining lakes are wild and domestic animals. Obtained phylogenetic trees support suggestions of other authors that the bovine genotype of C. parvum should be a separate species, as well as A and B assemblages of G. intestinalis.

Piggery slaughterhouse wastewater poses serious issues in terms of disposal feasibility and environmental impact, due to its huge organic load and variability. It is commonly treated by means of activated sludge processes, whose performance, in case of municipal wastewater, can be monitored by means of specific analyses, such as Sludge Biotic Index (SBI), Sludge Index (SI) and floc and filamentous bacteria observation. Therefore, this paper was aimed at assessing the applicability of these techniques to piggery slaughterhouse sewage. A plant located in Northern Italy was monitored for 1 year. Physical, chemical and operation parameters were measured; the activated sludge community (ciliates, flagellates, amoebae and small metazoa) was analysed for calculating SBI and SI. Floc and filamentous bacteria were examined and described accordingly with internationally adopted criteria. The results showed the full applicability of the studied techniques for optimizing the operation of a piggery slaughterhouse wastewater treatment plant.

In aquatic ecosystems, bacteria are controlled by several organisms in the food chain, such as protozoa, that use them as food source. This study aimed to quantify the ingestion and clearance rates of bacteria by ciliates and heterotrophic nanoflagellates (HNF) in a subtropical freshwater reservoir (Monjolinho reservoir - São Carlos - Brazil) during one year period, in order to verify their importance as consumers and controllers of bacteria in two seasons, a dry/cold and a rainy/warm one. For this purpose, in situ bacterivory experiments were carried out bimonthly using fluorescently labeled bacteria with 5-(4,6 diclorotriazin-2yl) aminofluorescein (DTAF). Although ciliates have shown the highest individual ingestion and clearance rates, bacterivory was dominated by HNF, who showed higher population ingestion rates (mean of 9,140 bacteria h(-1) mL(-1)) when compared to ciliates (mean of 492 bacteria h(-1) mL(-1)). The greater predation impact on bacterial communities was caused mainly by the small HNF (5 μm) population, especially in the rainy season, probably due to the abundances of these organisms, the precipitation, trophic index state and water temperature that were higher in this period. Thus, the protozoan densities together with environmental variables were extremely relevant in determining the seasonal pattern of bacterivory in Monjolinho reservoir.

Analyzing molecular determinants of Plasmodium parasite cell death is a promising approach for exploring new avenues in the fight against malaria. Three major forms of cell death (apoptosis, necrosis and autophagic cell death) have been described in multicellular organisms but which cell death processes exist in protozoa is still a matter of debate. Here we suggest that all three types of cell death occur in Plasmodium liver-stage parasites. Whereas typical molecular markers for apoptosis and necrosis have not been found in the genome of Plasmodium parasites, we identified genes coding for putative autophagy-marker proteins and thus concentrated on autophagic cell death. We characterized the Plasmodium berghei homolog of the prominent autophagy marker protein Atg8/LC3 and found that it localized to the apicoplast. A relocalization of PbAtg8 to autophagosome-like vesicles or vacuoles that appear in dying parasites was not, however, observed. This strongly suggests that the function of this protein in liver-stage parasites is restricted to apicoplast biology.