Here we summarize the results from 10 cruises in the Chukchi Sea, in August, each year from 2011 to 2020. Samples for the qualitative analysis of the microzooplankton were obtained from stations located across the Chukchi Sea using a 20μm plankton net. Conditions encountered, in terms of sea ice coverage and chlorophyll concentrations, varied widely from year to year without any obvious relationship with the composition of the microzooplankton assemblage. Examining a total of 242 samples gathered, we found a total of 44 tintinnid species (morphologically distinct forms). Plotting cumulative number of tintinnid species encountered vs cumulative number of samplings gave a typical species accumulation curve showing no sign of saturation suggesting that continued sampling in the Chukchi Sea will likely yield increases in the tintinnid species catalogue. The tintinnid species found ranged widely in lorica opening diameters (LOD) from about 11 μm to 80 μm in diameter. However, the median size of the LOD of the tintinnid assemblages varied little from year to year ranging only from about 30 μm to 40 μm. Most of the forms encountered were found in samples from only 1 or 2 cruises. Very few forms were found every year throughout the 10 years of sampling. These were 5 species of tintinnids (Acanthostomella norvegica, Leprotintinnus pellucidus, Pytchocylis obtusa, Salpingella acuminata, Salpingella faurei) and the nasselarian radiolarian Amphimelissa setosa. Examples of the morphological variability observed among individuals of Acanthostomella norvegica and Pytchocylis obtusa within single samples are shown with some individuals easily confused with forms described as other species are shown. To our knowledge, our data are the most extensive data set on Chukchi Sea microplankton. We provide all of the data recorded, which may serve as a baseline from which to assess changes projected in Arctic Sea systems, in a supplementary data file.

Cyclospora is an intracellular, gastrointestinal parasite found in birds and mammals worldwide. Limited accessibility of the protozoan for experimental use, scarcity, genome heterogeneity of the isolates and narrow panel of molecular markers hamper zoonotic investigations. One of the significant limitation in zoonotic studies is the lack of precise molecular tools that would be useful in linking animal vectors as a source of human infection. Strong and convincing evidence of zoonotic features will be achieved through proper typing of Cyclospora spp. taxonomic units (e.g. species or genotypes) in animal reservoirs. The most promising method that can be employ for zoonotic surveys is next-generation sequencing.

Selenoproteins have been described in all three domains of life and their function has been mainly associated with oxidative stress defense. Canonical elements required for selenoprotein production have been identified in members of the kinetoplastid group supporting the existence of a complete selenocysteine synthesis pathway in these organisms. Currently, nothing is known regarding the selenocysteine pathway in Trypanosoma evansi. In this study, we identified the expression of the elements selB, selC, selD, PSTK and selTRYP at the mRNA level in T. evansi. All translated proteins (selD, PSTK, selTRYP and selB) have the domains predicted and higher identity with Trypanosoma brucei. gambiense. The selenophosphate synthetase protein was localized in the cytoplasm. Our results support the existence of an active selenocysteine pathway in T. evansi.

The Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae) is one of the most important stored product pests. Fumigation plays a significant role in the management of insect pests in stored-products. However, the use of fumigants is problematic because of their effects on the environment and high costs. Entomopathogenic organisms are environmentally friendly control agents and suppress pest populations under natural conditions. In this study, distribution and occurrence of a microsporidian pathogen, Vairimorpha plodiae (Opisthokonta: Microspora) in the populations of P. interpunctella from 12 localities representing Turkey between 2019 and 2020 are presented for the first time by confirming its effectiveness on natural populations. The presence of the microsporidian pathogen was found in 11 of 12 (91.7%) populations. In total, 863 of 3,044 samples were infected by the pathogen. Infection mean was 28.4% for all populations. Our results showed that V. plodiae infection reached to a considerably high prevalence (88.77%) in P. interpunctella populations and varied from 5.1 to 88.7% between the populations. In addition, microsporidia infections have been identified throughout Turkey. We found that V. plodiae can infect all life stages of P. interpunctella. Totally, 623 (28.5%) of 2187 larvae, 14 (37.8%) of 37 pupae, 226 (27%) of 820 adults were found to be infected by the pathogen. There were considerable differences between the dead and living larvae. The microsporidian infection was found in 26 (11.6%) of 225 living larvae, whereas it was found in 595 (30.5%) of 1,952 dead larvae. These results confirm that the microsporidia pathogen has a high spreading potential in P. interpunctella populations and can be a natural biological suppression factor on pest populations.

Naegleria belongs to the free-living amoeba family and is well-known as a human pathogen. It is recognized as etiological agent of primary amoebic meningoencephalitis involving central nervous system which always leads to death. To date, there is not a single report demonstrating Naegleria isolation and identification from environmental sources of Rawlakot, Azad Jammu and Kashmir Pakistan, and thus the aim of this study. Naegleria was isolated on non-nutrient agar plates seeded with heat killed E. coli and confirmed by morphological properties of the both stages of cyst or trophozoites. Furthermore, PCR was conducted along with direct sequencing of the PCR product for molecular identification. PCR and sequencing data verified the amplification of Naegleria sp. (07) and Vahlkampfia sp. (01) from both water and soil samples. Interestingly two species were successfully isolated and cultured on both 30 and 45°C. To the best of our knowledge this is the first report demonstrating the Naegleria isolation and molecular characterization from environmental sources of Rawlakot, Azad Jammu and Kashmir, Pakistan. The author is anxious for further evaluation of the pathogenic potential of the identified species and explores drinking water across Pakistan to investigate its quality and frequency of FLA, which might be a possible human hazard in future.

A dinoflagellate identified as Prorocentrum dentatum, P. shikokuense or P. donghaiense is responsible of massive harmful events. Blooms of a species identified as P. shikokuense have been recently reported in the Mediterranean Sea, and an exotic origin, tentatively introduced by ballast waters from Asia, has been hypothesized. The molecular data based on the small-, large subunit, and internal transcriber spacers ribosomal RNA gene (SSU-, LSU-, ITS rRNA) sequences confirmed P. shikokuense in the Mediterranean Sea. The Mediterranean ribotype is identical to a subtropical North Atlantic ribotype, and with slight divergence from the numerous sequences from the Pacific Ocean. To revisit the relationship between P. shikokuense (=P. donghaiense) and P. obtusidens, we provide the first micrographs of P. dentatum and P. obtusidens, the latter collected from the type locality. Our observations indicate that P. dentatum, P. obtusidens, and P. shikokuense are three different species. Their diagnostic morphological characters are: Prorocentrum dentatum is 44–60 μm long, leaf-shaped, pointed and central posterior end, conspicuous anterior shoulder; P. obtusidens is 33–41 μm long, irregular parallelepiped, almost paralel valve margins, eccentric and pointed posterior end, moderate anterior shoulder; and P. shikokuense is 25 μm long, sunflower seed-shaped, round and centric posterior end, inconspicuous anterior shoulder.

Microbial diversity is known to be huge but remains only partly documented. Testate amoebae are a relatively well studied group or free-living protists that build morphologically characteristic shells on which the taxonomy of this group is mainly based. Among testate amoebae, the Hyalospheniformes are especially well documented and are a model group for microbial biogeography. Here we describe a new species of genus Certesella from Sphagnum mosses and forest litter samples collected in the Dominican Republic and Chile. We name this species Certesella larai to honour our esteemed colleague Enrique Lara. This species bears two large pores connected by tube, which are characteristic of the genera Porosia and Certesella. The new species fits best in the latter genus given the presence of a distinct, sometimes denticulated, neck with a bulge that is absent in genus Porosia but occurs in Certesella. Its elongated shape (length/breadth ratio 2.03–2.58) only overlaps with C. certesi, which is distinguishable by the presence of a distinct longitudinal groove in the neck region that is absent from the new species. Interestingly most specimens lacked the internal teeth which are usually present in members of genus Certesella. As such, while the morphology of C. larai strongly suggests a belonging to genus Certesella, molecular analyses are still required to confirm its phylogenetic position, clarify the relationships among all species in genera Porosia and Certesella and the definition and validity of these two genera. The presence of this new species in two rather distant regions – Caribbean and central Chile – suggests it is likely widespread, but possibly rare. However, this perception may be due to a sampling bias in favour of Sphagnum mosses and wet and organic-rich habitats. This illustrates the need for a more systematic sampling effort to document the full and mostly unknown taxonomic diversity of soil protists.

Here we summarize the results from 10 cruises in the Chukchi Sea, in August, each year from 2011 to 2020. Samples for the qualitative analysis of the microzooplankton were obtained from stations located across the Chukchi Sea using a 20μm plankton net. Conditions encountered, in terms of sea ice coverage and chlorophyll concentrations, varied widely from year to year without any obvious relationship with the composition of the microzooplankton assemblage. Examining a total of 242 samples gathered, we found a total of 44 tintinnid species (morphologically distinct forms). Plotting cumulative number of tintinnid species encountered vs cumulative number of samplings gave a typical species accumulation curve showing no sign of saturation suggesting that continued sampling in the Chukchi Sea will likely yield increases in the tintinnid species catalogue. The tintinnid species found ranged widely in lorica opening diameters (LOD) from about 11 μm to 80 μm in diameter. However, the median size of the LOD of the tintinnid assemblages varied little from year to year ranging only from about 30 μm to 40 μm. Most of the forms encountered were found in samples from only 1 or 2 cruises. Very few forms were found every year throughout the 10 years of sampling. These were 5 species of tintinnids (Acanthostomella norvegica, Leprotintinnus pellucidus, Pytchocylis obtusa, Salpingella acuminata, Salpingella faurei) and the nasselarian radiolarian Amphimelissa setosa. Examples of the morphological variability observed among individuals of Acanthostomella norvegica and Pytchocylis obtusa within single samples are shown with some individuals easily confused with forms described as other species are shown. To our knowledge, our data are the most extensive data set on Chukchi Sea microplankton. We provide all of the data recorded, which may serve as a baseline from which to assess changes projected in Arctic Sea systems, in a supplementary data file.

Cyclospora is an intracellular, gastrointestinal parasite found in birds and mammals worldwide. Limited accessibility of the protozoan for experimental use, scarcity, genome heterogeneity of the isolates and narrow panel of molecular markers hamper zoonotic investigations. One of the significant limitation in zoonotic studies is the lack of precise molecular tools that would be useful in linking animal vectors as a source of human infection. Strong and convincing evidence of zoonotic features will be achieved through proper typing of Cyclospora spp. taxonomic units (e.g. species or genotypes) in animal reservoirs. The most promising method that can be employ for zoonotic surveys is next-generation sequencing.

Selenoproteins have been described in all three domains of life and their function has been mainly associated with oxidative stress defense. Canonical elements required for selenoprotein production have been identified in members of the kinetoplastid group supporting the existence of a complete selenocysteine synthesis pathway in these organisms. Currently, nothing is known regarding the selenocysteine pathway in Trypanosoma evansi. In this study, we identified the expression of the elements selB, selC, selD, PSTK and selTRYP at the mRNA level in T. evansi. All translated proteins (selD, PSTK, selTRYP and selB) have the domains predicted and higher identity with Trypanosoma brucei. gambiense. The selenophosphate synthetase protein was localized in the cytoplasm. Our results support the existence of an active selenocysteine pathway in T. evansi.

The Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae) is one of the most important stored product pests. Fumigation plays a significant role in the management of insect pests in stored-products. However, the use of fumigants is problematic because of their effects on the environment and high costs. Entomopathogenic organisms are environmentally friendly control agents and suppress pest populations under natural conditions. In this study, distribution and occurrence of a microsporidian pathogen, Vairimorpha plodiae (Opisthokonta: Microspora) in the populations of P. interpunctella from 12 localities representing Turkey between 2019 and 2020 are presented for the first time by confirming its effectiveness on natural populations. The presence of the microsporidian pathogen was found in 11 of 12 (91.7%) populations. In total, 863 of 3,044 samples were infected by the pathogen. Infection mean was 28.4% for all populations. Our results showed that V. plodiae infection reached to a considerably high prevalence (88.77%) in P. interpunctella populations and varied from 5.1 to 88.7% between the populations. In addition, microsporidia infections have been identified throughout Turkey. We found that V. plodiae can infect all life stages of P. interpunctella. Totally, 623 (28.5%) of 2187 larvae, 14 (37.8%) of 37 pupae, 226 (27%) of 820 adults were found to be infected by the pathogen. There were considerable differences between the dead and living larvae. The microsporidian infection was found in 26 (11.6%) of 225 living larvae, whereas it was found in 595 (30.5%) of 1,952 dead larvae. These results confirm that the microsporidia pathogen has a high spreading potential in P. interpunctella populations and can be a natural biological suppression factor on pest populations.

Naegleria belongs to the free-living amoeba family and is well-known as a human pathogen. It is recognized as etiological agent of primary amoebic meningoencephalitis involving central nervous system which always leads to death. To date, there is not a single report demonstrating Naegleria isolation and identification from environmental sources of Rawlakot, Azad Jammu and Kashmir Pakistan, and thus the aim of this study. Naegleria was isolated on non-nutrient agar plates seeded with heat killed E. coli and confirmed by morphological properties of the both stages of cyst or trophozoites. Furthermore, PCR was conducted along with direct sequencing of the PCR product for molecular identification. PCR and sequencing data verified the amplification of Naegleria sp. (07) and Vahlkampfia sp. (01) from both water and soil samples. Interestingly two species were successfully isolated and cultured on both 30 and 45°C. To the best of our knowledge this is the first report demonstrating the Naegleria isolation and molecular characterization from environmental sources of Rawlakot, Azad Jammu and Kashmir, Pakistan. The author is anxious for further evaluation of the pathogenic potential of the identified species and explores drinking water across Pakistan to investigate its quality and frequency of FLA, which might be a possible human hazard in future.

A dinoflagellate identified as Prorocentrum dentatum, P. shikokuense or P. donghaiense is responsible of massive harmful events. Blooms of a species identified as P. shikokuense have been recently reported in the Mediterranean Sea, and an exotic origin, tentatively introduced by ballast waters from Asia, has been hypothesized. The molecular data based on the small-, large subunit, and internal transcriber spacers ribosomal RNA gene (SSU-, LSU-, ITS rRNA) sequences confirmed P. shikokuense in the Mediterranean Sea. The Mediterranean ribotype is identical to a subtropical North Atlantic ribotype, and with slight divergence from the numerous sequences from the Pacific Ocean. To revisit the relationship between P. shikokuense (=P. donghaiense) and P. obtusidens, we provide the first micrographs of P. dentatum and P. obtusidens, the latter collected from the type locality. Our observations indicate that P. dentatum, P. obtusidens, and P. shikokuense are three different species. Their diagnostic morphological characters are: Prorocentrum dentatum is 44–60 μm long, leaf-shaped, pointed and central posterior end, conspicuous anterior shoulder; P. obtusidens is 33–41 μm long, irregular parallelepiped, almost paralel valve margins, eccentric and pointed posterior end, moderate anterior shoulder; and P. shikokuense is 25 μm long, sunflower seed-shaped, round and centric posterior end, inconspicuous anterior shoulder.

Microbial diversity is known to be huge but remains only partly documented. Testate amoebae are a relatively well studied group or free-living protists that build morphologically characteristic shells on which the taxonomy of this group is mainly based. Among testate amoebae, the Hyalospheniformes are especially well documented and are a model group for microbial biogeography. Here we describe a new species of genus Certesella from Sphagnum mosses and forest litter samples collected in the Dominican Republic and Chile. We name this species Certesella larai to honour our esteemed colleague Enrique Lara. This species bears two large pores connected by tube, which are characteristic of the genera Porosia and Certesella. The new species fits best in the latter genus given the presence of a distinct, sometimes denticulated, neck with a bulge that is absent in genus Porosia but occurs in Certesella. Its elongated shape (length/breadth ratio 2.03–2.58) only overlaps with C. certesi, which is distinguishable by the presence of a distinct longitudinal groove in the neck region that is absent from the new species. Interestingly most specimens lacked the internal teeth which are usually present in members of genus Certesella. As such, while the morphology of C. larai strongly suggests a belonging to genus Certesella, molecular analyses are still required to confirm its phylogenetic position, clarify the relationships among all species in genera Porosia and Certesella and the definition and validity of these two genera. The presence of this new species in two rather distant regions – Caribbean and central Chile – suggests it is likely widespread, but possibly rare. However, this perception may be due to a sampling bias in favour of Sphagnum mosses and wet and organic-rich habitats. This illustrates the need for a more systematic sampling effort to document the full and mostly unknown taxonomic diversity of soil protists.