Small Subunit rRNA Research Articles

Molecular characterization of Cryptosporidium spp. and Giardia duodenalis in pet dogs in Xinjiang, China.

Cryptosporidium spp. and Giardia duodenalis are opportunistic zoonotic protozoa transmitted through several routes. In this study, a total of 604 fecal samples were collected from pet dogs in Xinjiang, China to detect the presence of Cryptosporidium spp. and G. duodenalis by PCR amplification of the small subunit (SSU) rRNA gene. The incidence rate of Cryptosporidium spp. or G. duodenalis was 5.3% (32/604). Among the collection sites, a higher number ofCryptosporidium spp. or G. duodenalis positive dogs were detectedin Hotan (9.9%, 21/213) and Shihezi (9.8%, 4/41) were higher than those in Aksu (4.5%, 1/22), Urumqi (2.1%, 4/191), and Korla (1.5%,2/137). Among the sources, dogs in pet shops (7.4%, 19/256) showed a significantly higher incidence rate than those in pet hospitals (3.0%, 4/134) and pet kennels (2.3%, 5/214). When the data were examined by age, dogs < 1year of age (6.1%, 28/459) were more likely to be infected by Cryptosporidium spp. or G. duodenalis than older (≥ 1year) dogs (2.8%, 4/145). No significant differences were observed when animals were grouped by sex (5.0%, 14/278 for males; 5.5%, 18/326 for females). Sequence analysis revealed that the Cryptosporidium spp. (n = 10) in dogs were identified as C. canis. The G. duodenalis detectedbelonged to assemblages A (n = 1), C (n = 14), and D (n = 7). Among the identified 22 G. duodenalis isolates, eight samples were subtyped according toβ-giardin (bg)and the results were consistent with the identified assemblages. To our knowledge, this is the first report of Cryptosporidium spp. and G. duodenalis infections in domesticated canines in Xinjiang, China. The C. canis and G. duodenalis assemblage A identified in pet dogs in the present study were previously associated with infections in humans, indicating the potential for zoonotic transmission.

Gross Morphology of Diseased Tissues in Nezara viridula (Hemiptera: Pentatomidae) and Molecular Characterization of an Associated Microsporidian.

Nezara viridula (L.) (Hemiptera: Pentatomidae), commonly known in the U.S. as the southern green stink bug (SGSB), is a cosmopolitan, highly polyphagous feeder that causes severe damage to a wide range of agronomically important crops such as fruit, vegetable, grain, tobacco, and cotton, throughout much of the United States, and is a global pest of considerable ecological, agricultural, and economical interest. During dissection of female Nz. viridula, conspicuous black and brown spots or lesions were observed on various internal organs. To determine the cause of these spots or lesions, tissues of fat body, spermatheca, ovaries, and ovulated eggs were collected from healthy and infected individuals. The gross morphology of the spots was characterized, and the microorganisms associated with the infection were identified by amplicon sequencing of the V4 region of the small subunit rRNA gene. The presence of a microsporidian pathogen Nosema maddoxi, Becnel, Solter, Hajek, Huang, Sanscrainte, & Estep (Microsporidia: Nosematidae) which has been observed on other species of stink bug, was evidenced for the first time. The characterization of the gross morphology of this associated microsporidian may enable more rapid determination of microsporidia infection in stink bug colonies and field populations.

The infection and molecular characterization of Cryptosporidium spp. in diarrheic pigs in southern China

Cryptosporidium spp. is recognized as an opportunistic zoonotic parasite that infects humans, wild and domestic animals, and is also a major cause of diarrhea-related disease in immunocompromised individuals, considered a global public health concern. Pig is considered as one of the reservoir hosts of Cryptosporidium spp. can transmit cryptosporidiosis to humans and other animals. However, limited studies on the distribution of Cryptosporidium spp. in diarrheic pigs have been published. Objective of the current study was to investigate the infection and species/genotypes of Cryptosporidium spp. from feces of diarrheic pigs in southern China. A total of 1254 fresh fecal samples were collected from 37 intensive pig farms in Jiangxi, Hunan and Fujian provinces, and were screened for Cryptosporidium spp. infection using a nested PCR assay targeted the small subunit rRNA (SSU rRNA) genes. The overall infection rate of Cryptosporidium spp. was 4.5% (57/1254), including 5.5% (17/307) in suckling piglets, 2.7% (8/299) in weaned piglets, 7.4% (7/95) in fattening pigs and 4.5% (25/553) in sows, respectively. In addition, two human-pathogenic species Cryptosporidium scrofarum (80.7%, 46/57) and Cryptosporidium suis (19.3%, 11/57) were identified. C. scrofarum and C. suis were observed in pigs tested in all age groups. Interestingly, a high colonization incidence of C. scrofarum (16/57) was observed in suckling piglets. This study revealed the prevalence and species of Cryptosporidium spp. in diarrheic pigs in three provinces of southern China, which suggested that diarrhea maybe not a direct factor affecting the prevalence of Cryptosporidium spp. in pigs. More prevention and control of this parasite in pigs should receive greater attention from farmers in investigated provinces.

Response of Total (DNA) and Metabolically Active (RNA) Microbial Communities in Miscanthus × Giganteus Cultivated Soil to Different Nitrogen Fertilization Rates.

ABSTRACTMiscanthus × giganteus is a promising high-yielding perennial plant to meet growing bioenergy demands; however, the degree to which the soil microbiome affects its nitrogen cycling and subsequently, biomass yield remains unclear. In this study, we hypothesize that contributions of metabolically active soil microbial membership may be underestimated with DNA-based approaches. We assessed the response of the soil microbiome to nitrogen availability in terms of both DNA and RNA soil microbial communities from the Long-term Assessment of Miscanthus Productivity and Sustainability (LAMPS) field trial. DNA and RNA were extracted from 271 samples, and 16S small subunit (SSU) rRNA amplicon sequencing was performed to characterize microbial community structure. Significant differences were observed in the resulting soil microbiomes and were best explained by the sequencing library of origin, either DNA or RNA. Similar numbers of membership were detected in DNA and RNA microbial communities, with more than 90% of membership shared. However, the profile of dominant membership within DNA and RNA differed, with varying proportions of Actinobacteria and Proteobacteria and Firmicutes and Proteobacteria. Only RNA microbial communities showed seasonal responses to nitrogen fertilization, and these differences were associated with nitrogen-cycling bacteria. The relative abundance of bacteria associated with nitrogen cycling was 7-fold higher in RNA than in DNA, and genes associated with denitrifying bacteria were significantly enriched in RNA, suggesting that these bacteria may be underestimated with DNA-only approaches. Our findings indicate that RNA-based SSU characterization can be a significant and complementing resource for understanding the role of soil microbiomes in bioenergy crop production.IMPORTANCE Miscanthus × giganteus is a promising candidate for bioeconomy cropping systems; however, it remains unclear how the soil microbiome supplies nitrogen to this low-input crop. DNA-based techniques are used to provide community characterization, but may miss important metabolically active taxa. By analyzing both DNA- and actively transcribed RNA-based microbial communities, we found that nitrogen cycling taxa in the soil microbiome may be underestimated using only DNA-based approaches. Accurately understanding the role of microbes and how they cycle nutrients is important for the development of sustainable bioenergy crops, and RNA-based approaches are recommended as a complement to DNA approaches to better understand the microbial, plant, and management interactions.

The universally conserved nucleotides of the small subunit ribosomal RNAs.

The ribosomal RNAs, along with their substrates the transfer RNAs, contain the most highly conserved nucleotides in all of biology. We have assembled a database containing structure-based alignments of sequences of the small-subunit rRNAs from organisms that span the entire phylogenetic spectrum, to identify the nucleotides that are universally conserved. In its simplest (bacterial and archaeal) forms, the small-subunit rRNA has ∼1500 nt, of which we identify 140 that are absolutely invariant among the 1961 species in our alignment. We examine the positions and detailed structural and functional interactions of these universal nucleotides in the context of a half century of biochemical and genetic studies and high-resolution structures of ribosome functional complexes. The vast majority of these nucleotides are exposed on the subunit interface surface of the small subunit, where the functional processes of the ribosome take place. However, only 40 of them have been directly implicated in specific ribosomal functions, such as contacting the tRNAs, mRNA, or translation factors. The roles of many other invariant nucleotides may serve to constrain the positions and orientations of those nucleotides that are directly involved in function. Yet others can be rationalized by participation in unusual noncanonical tertiary structures that may uniquely allow correct folding of the rRNA to form a functional ribosome. However, there remain at least 50 nt whose universal conservation is not obvious, serving as a metric for the incompleteness of our understanding of ribosome structure and function.

In-Depth Analysis of Bacillus anthracis 16S rRNA Genes and Transcripts Reveals Intra- and Intergenomic Diversity and Facilitates Anthrax Detection.

ABSTRACTAnalysis of 16S rRNA (rRNA) genes provides a central means of taxonomic classification of bacterial species. Based on presumed sequence identity among species of the Bacillus cereus sensu lato group, the 16S rRNA genes of B. anthracis have been considered unsuitable for diagnosis of the anthrax pathogen. With the recent identification of a single nucleotide polymorphism in some 16S rRNA gene copies, specific identification of B. anthracis becomes feasible. Here, we designed and evaluated a set of in situ, in vitro, and in silico assays to assess the unknown 16S state of B. anthracis from different perspectives. Using a combination of digital PCR, fluorescence in situ hybridization, long-read genome sequencing, and bioinformatics, we were able to detect and quantify a unique 16S rRNA gene allele of B. anthracis (16S-BA-allele). This allele was found in all available B. anthracis genomes and may facilitate differentiation of the pathogen from any close relative. Bioinformatics analysis of 959 B. anthracis SRA data sets inferred that abundances and genomic arrangements of the 16S-BA-allele and the entire rRNA operon copy numbers differ considerably between strains. Expression ratios of 16S-BA-alleles were proportional to the respective genomic allele copy numbers. The findings and experimental tools presented here provide detailed insights into the intra- and intergenomic diversity of 16S rRNA genes and may pave the way for improved identification of B. anthracis and other pathogens with diverse rRNA operons.IMPORTANCE For severe infectious diseases, precise pathogen detection is crucial for antibiotic therapy and patient survival. Identification of Bacillus anthracis, the causative agent of the zoonosis anthrax, can be challenging when querying specific nucleotide sequences such as in small subunit rRNA (16S rRNA) genes, which are commonly used for typing of bacteria. This study analyzed on a broad genomic scale a cryptic and hitherto underappreciated allelic variant of the bacterium’s 16S rRNA genes and their transcripts using a set of in situ, in vitro, and in silico assays and found significant intra- and intergenomic heterogeneity in the distribution of the allele and overall rRNA operon copy numbers. This allelic variation was uniquely species specific, which enabled sensitive pathogen detection on both DNA and transcript levels. The methodology used here is likely also applicable to other pathogens that are otherwise difficult to discriminate from their less harmful relatives.

Molecular-based assay for genotyping Leishmania spp. from clinically suspected cutaneous leishmaniasis lesions in the Garmian area, Kurdistan Region of Iraq

Cutaneous leishmaniasis (CL) is highly prevalent in southern Iraq and neighboring countries, but is non-endemic to the Kurdistan Region of Iraq, particularly in the Garmian area. This study aimed to investigate the causative agent of CL at the molecular level by amplifying the small subunit (18S) rRNA and internal transcribed spacer 1 (ITS1) region. The present study was conducted from December 2019 to December 2020 at Kalar General Hospital, Kalar, Kurdistan Region, Iraq. Eighty-five clinical specimens were collected selectively from patients with suspected CL lesions via fine needle aspiration. After parasitic genomic DNA was extracted from the removed fluid, PCR and DNA sequencing targeting the 18S rRNA and ITS1 region were performed for molecular detection and species identification. Additionally, for 14 samples, the target bands of amplified DNA fragments for both 18S rRNA and ITS1 were extracted and sequenced via Sanger method using both the directional primers employed in the PCR. Seventy-one (83.53%) of the 85 suspected patients had CL, based on amplification of 18S rRNA and ITS1 via PCR. The sequence analysis revealed that all samples were Leishmania major. Phylogenetic analysis based on ITS1 was also performed. Our study revealed that our molecular method was an efficient technique for detecting CL and a valuable method for identifying Leishmania species in clinical samples. Sequence analysis indicated that the causative agent of CL in the Garmian area was L. major and the disease was rural in origin.

Metahyphopichia suwanaadthiae sp. nov., an anamorphic yeast species in the order Saccharomycetales and reassignment of Candida silvanorum to the genus Metahyphopichia.

Seven yeast strains, representing a single novel anamorphic species, were isolated in Thailand. They consisted of five strains (DMKU-MRY16T, DMKU-SK18, DMKU-SK25, DMKU-SK30 and DMKU-SK32) obtained from five different mushrooms, and two strains (ST-224 and 11-14.2) derived from insect frass and soil, respectively. The pairwise sequence analysis indicated that all seven strains had identical sequences in the D1/D2 domains of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region. Metahyphopichia silvanorum was the most closely related species, but with 11.9-12.4% nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and 13.1-13.3% nucleotide substitutions in the ITS region. The phylogenetic analyses based on the concatenated sequences of the ITS region and the D1/D2 domains of the LSU rRNA gene showed that the seven strains form a well-separated subclade in a clade containing M. silvanorum and Metahyphopichia laotica with high bootstrap support. A phylogenetic analysis of a multilocus dataset including the small subunit (SSU) rRNA gene, the ITS region, the D1/D2 domains of the LSU rRNA gene, translation elongation factor 1-alpha gene, actin gene and the RNA polymerase II subunit 2 gene, confirmed the presence of the monophyletic clade that also includes M. silvanorum and M. laotica, and strongly supported the phylogenetic isolation of the seven strains from its neighbouring species. Therefore, the seven strains were assigned as a single novel species of the genus Metahyphopichia, according to their phylogenetic relationships. The name Metahyphopichia suwanaadthiae sp. nov. is proposed to accommodate the seven strains. The holotype is DMKU-MRY16T (TBRC 11775T=NBRC 114386T=PYCC 8655T). The MycoBank number of the novel species is MB 841280. In addition, Candida silvanorum is reassigned to the genus Metahyphopichia. The MycoBank number of M. silvanorum comb. nov. is MB 841279.

Testing the Presence of an Opportunistic Pathogen Nosema ceranae (Microsporidia) in Housefly (Musca domestica) using Polymerase Chain Reaction

Houseflies, being vector of serious human disease, have been extensively used in the laboratory to research on aging and sex determination. It also plays an important role in breaking down the organic matter and thus makes its contribution in food cycle. Houseflies belong to the phylum Arthropoda and serves as vector for many pathogens transmiting disease from lower to higher vertebrates. Microsporidiosis (also known as nosemosis) is a disorder of honeybee and various other pollinators belonging to the class Insecta that are affecting nation’s economy up to a great extent. Microsporidian spores have reportedly infected insects, fishes, crustaceous, human and thus causing huge threat to society. In order to establish a clear understanding for its role in the transmission of microsporidiosis, the present study was designed to detect and identify a microsporidian parasite- Nosema ceranae by amplifying 16sRNA (Small Subunit rRNA) gene using molecules technique Polymerase Chain Reaction (PCR). Results of the present study suggested no existence of Nosema ceranae spores as, in spite of performing gradient PCR, no amplification of gene occurred. The research work paves a way for other researchers to investigate the possible reasons behind the absence of Nosema ceranae infection in housefly and also on why the infection is not prevalent in this model organism.

RNA Virosphere in a Marine Zooplankton Community in the Subtropical Western North Pacific.

Zooplankton and viruses play a key role in marine ecosystems; however, their interactions have not been examined in detail. In the present study, the diversity of viruses associated with zooplankton collected using a plankton net (mesh size: 100‍ ‍μm) in the subtropical western North Pacific was investigated by fragmented and primer ligated dsRNA sequencing. We obtained 21 and 168 operational taxonomic units (OTUs) of ssRNA and dsRNA viruses, respectively, containing RNA-dependent RNA polymerase (RdRp). These OTUs presented average amino acid similarities of 43.5 and 44.0% to the RdRp genes of known viruses in ssRNA viruses and dsRNA viruses, respectively. Dominant OTUs mainly belonged to narna-like and picorna-like ssRNA viruses and chryso-like, partiti-like, picobirna-like, reo-like, and toti-like dsRNA viruses. Phylogenetic ana-lyses of the RdRp gene revealed that OTUs were phylogenetically diverse and clustered into distinct clades from known viral groups. The community structure of the same zooplankton sample was investigated using small subunit (SSU) rRNA sequences assembled from the metatranscriptome of single-stranded RNA. More than 90% of the sequence reads were derived from metazoan zooplankton; copepods comprised approximately 70% of the sequence reads. Although this ana-lysis provided no direct evidence of the host species of RNA viruses, these dominant zooplankton are expected to be associated with the RNA viruses detected in the present study. The present results indicate that zooplankton function as a reservoir of diverse RNA viruses and suggest that investigations of zooplankton viruses will provide a more detailed understanding of the role of viruses in marine ecosystems.

A Novel Gonadotropic Microsporidian Parasite (Microsporidium clinchi n. sp.) Infecting a Declining Population of Pheasantshell Mussels (Actinonaias pectorosa) (Unioinidae) from the Clinch River, USA

Freshwater mussels of the order Unionida are among the most endangered animal groups globally, but the causes of their population decline are often enigmatic, with little known about the role of disease. In 2018, we collected wild adult pheasantshell (Actinonaias pectorosa) and mucket (Actinonaias ligamentina) during an epidemiologic survey investigating an ongoing mussel mass mortality event in the Clinch River, Virginia and Tennessee, USA. Histopathology and transmission electron microscopy showed a novel microsporidian parasite primarily infecting the ovary of pheasantshell. Sequencing of the small subunit rRNA gene produced a 1333 bp sequence with the greatest similarity to Pseudonosema cristatellae (AF484694.1; 86.36%; e-value = 0), a microsporidium infecting the freshwater bryozoan (Cristatella mucedo). Microsporidia were observed in 65% (17/26) of the examined female pheasantshell (A. pectorosa) and in no (0/2) female muckets (A. ligamentina) and occurred at mortality and non-mortality sites. Our findings indicate that a novel parasite, Microsporidium clinchi n. sp., is present in pheasantshell in the Clinch River, and while likely not a cause of mass mortality, could reduce fecundity and recruitment in this declining population and threaten the success of reintroductions. Surveillance of M. clinchi n. sp. and evaluation of broodstock and their progeny for microsporidia would therefore be prudent.

Dothidea kunmingensis, a novel asexual species of Dothideaceae on Jasminum nudiflorum (winter jasmine) from Southwestern China

During a survey of saprobic microfungi in Southwest China, a coelomycetous fungus was found on dead twigs of Jasminum nudiflorum in Kunming, Yunnan Province. Based on a detailed morphological characterization coupled with multi-locus phylogenetic analyses, the fungus was identified as a new species in the genus Dothidea. Phylogenetic analyses using a combined matrix consisting of internal transcribed spacer (ITS), large subunit rRNA (LSU), small subunit rRNA (SSU), beta tubulin (tub2) and translation elongation factor-1 alpha (tef1-α) confirmed its placement in Dothideaceae and revealed a sister relationship to Dothidea eucalypti. The new species is characterized by pycnidial conidiomata, ampulliform or doliiform conidiogenous cells as well as aseptate, subglobose to ovoid, hyaline to pale-brown conidia. Comprehensive descriptions and illustrations are provided. Morphological characteristics of asexual morph taxa in Dothideaceae are also summarized and discussed.

Txikispora philomaios n. sp., n. g., a micro-eukaryotic pathogen of amphipods, reveals parasitism and hidden diversity in Class Filasterea.

This study provides a morphological, ultrastructural, and phylogenetic characterization of a novel micro-eukaryotic parasite (2.3-2.6µm) infecting amphipod genera Echinogammarus and Orchestia. Longitudinal studies across two years revealed that infection prevalence peaked in late April and May, reaching 64% in Echinogammarus sp. and 15% in Orchestia sp., but was seldom detected during the rest of the year. The parasite infected predominantly hemolymph, connective tissue, tegument, and gonad, although hepatopancreas and nervous tissue were affected in heavier infections, eliciting melanization and granuloma formation. Cell division occurred inside walled parasitic cysts, often within host hemocytes, resulting in hemolymph congestion. Small subunit (18S) rRNA gene phylogenies including related environmental sequences placed the novel parasite as a highly divergent lineage within Class Filasterea, which together with Choanoflagellatea represent the closest protistan relatives of Metazoa. We describe the new parasite as Txikispora philomaios n. sp. n. g., the first confirmed parasitic filasterean lineage, which otherwise comprises four free-living flagellates and a rarely observed endosymbiont of snails. Lineage-specific PCR probing of other hosts and surrounding environments only detected T.philomaios in the platyhelminth Procerodes sp. We expand the known diversity of Filasterea by targeted searches of metagenomic datasets, resulting in 13 previously unknown lineages from environmental samples.

Amphidinium stirisquamtum sp. nov. (Dinophyceae), a new marine sand-dwelling dinoflagellate with a novel type of body scale

Amphidinium species are amongst the most abundant benthic dinoflagellates in marine intertidal sandy ecosystems. Some of them produce a variety of bioactive compounds that have both harmful effects and pharmaceutical potential. In this study, Amphidinium cells were isolated from intertidal sand collected from the East China Sea. The two strains established were subjected to detailed examination by light, and scanning and transmission electron microscopy. The vegetative cells had a minute, irregular, and triangular-shaped epicone deflected to the left, thus fitting the description of Amphidinium sensu stricto. These strains are distinguished from other Amphidinium species by combination characteristics: (1) longitudinal flagellum inserted in the lower third of the cell; (2) icicle-shaped scales, 276 ± 17 nm in length, on the cell body surface; (3) asymmetrical hypocone with the left side longer than the right; and (4) presence of immotile cells. Therefore, they are described here as Amphidinium stirisquamtum sp. nov. The molecular tree inferred from small subunit rRNA, large subunit rRNA, and internal transcribed spacer-5.8S sequences revealed that A. stirisquamtum is grouped together with the type species of Amphidinium, A. operculatum, in a fully supported clade, but is distantly related to other Amphidinium species bearing body scale. Live A. stirisquamtum cells greatly affected the survival of rotifers and brine shrimp, their primary grazers, making them more susceptible to predation by the higher tropic level consumers in the food web. This will increase the risk of introducing toxicity, and consequently, the bioaccumulation of toxins through marine food webs.

Changes in Soil-Borne Communities of Arbuscular Mycorrhizal Fungi during Natural Regrowth of Abandoned Cattle Pastures Are Indicative of Ecosystem Restoration

Natural restoration of ecosystems includes the restoration of plant-microbial associations; however, few studies had documented those changes in tropical ecosystems. With the aim to contribute to understand soil microbial changes in a natural regrowth succession of degraded pastures that were left for natural restoration, we studied changes in arbuscular mycorrhizal (AM) fungal communities. Arbuscular mycorrhizal fungi (AMF) establish a mutualistic symbiosis with plants, improving plant nutrition. Amplification of the small subunit rRNA with specific primers and subsequent Illumina sequencing were used to search soil-borne AM fungal communities in four successional natural regrowth stages in two landscapes (hill and mountain) with soil differences, located in the Andean-Amazonian transition. Molecular results corroborated the results obtained previously by spores-dependent approaches. More abundance and virtual taxa of AMF exist in the soil of degraded pastures and early natural regrowth stages than in old-growth or mature forest soils. Although changes in AM fungal communities occurred similarly over the natural regrowth chronosequence, differences in soil texture between landscapes was an important soil feature differentiating AM fungal community composition and richness. Changes in soil-borne AM fungal communities reflect some signals of environmental restoration that had not been described before, such as the reduction of Glomus dominance and the increase of Paraglomus representativeness in the AM fungal community during the natural regrowth chronosequence.

Molecular Study of Eimeria species in Quail birds (Coturnix coturnix japonica) in Thi-Qar Province, Southern Iraq

Fecal samples were collected from 330 quail birds from different areas of Thi-Qar province, Iraq. Allsamples were examined by traditional laboratory methods. Four species of Eimeria were diagnosed inquail (Eimeria bateri, Eimeria uzura, Eimeria tsunodai and Eimeria fluminensis), with a total infectionwas 64.54% (213 out of 330). DNA was extracted from 100 positive samples confirm the small subunitrRNA (18s) gene by Polymerase chain reaction using a general primer for Eimeria species, Sevengenotypes were according to the identify of DNA sequences of Eimeria species in Japanese quail, theaccession codes to NCBI-Genbank. using A comprehensive phylogenetic tree was generated in thepresent study, this details were being analyzed. This was the first molecular study in Iraq for Eimeriain common quail.

Characterization of a Novel Hypotrich Ciliate From Heavy Metal-Contaminated Industrial Outlet in Onsan, Ulsan, South Korea.

Very few studies exist on the description of protozoan ciliates from industrially contaminated sites. In this study, we report a description of a novel hypotrich ciliate isolated from water samples collected from an industrially contaminated outlet in Onsan, Ulsan, South Korea. The oxytrichid ciliate, Histriculus tolerans n. sp., was investigated using live observation and protargol impregnation. The morphology, morphogenesis, and molecular phylogeny inferred from small-subunit (SSU) rRNA gene sequences were studied. The new species is mainly characterized by a cell size of about 70 × 40 μm in vivo, two elongate ellipsoidal macronuclear nodules and one or two micronuclei, adoral zone of about 51% of body length with 32 membranelles on average, about 34 cirri in the right and 24 cirri in the left marginal row, 18 frontoventral transverse cirri, six dorsal kineties including two dorsomarginal rows, and dorsal kinety 1 with 26 bristles. Morphogenesis is similar to that of the type species, i.e., Histriculus histrio, except that oral primordium does not contribute to anlage II of the proter. Phylogenetic analyses, based on small-subunit rRNA gene sequences, consistently place the new species within the family Oxytrichidae, clustering with H. histrio.

Arbuscular Mycorrhizal Communities in the Roots of Sago Palm in Mineral and Shallow Peat Soils

Communities of arbuscular mycorrhizal fungi (AMF) in plant roots improve host plant growth. In this study, AMF communities in the roots of the sago palm (Metroxylon sagu Rottb.) were investigated in mineral soil (MS) and shallow peat soil (SPS) in Sarawak, Malaysia. MS exhibited lower moisture content (MS, 38.1; SPS, 79.8%), higher pH (H2O) (MS, 4.6; SPS, 4.1), higher soil bulk density (MS, 1.03; SPS, 0.20 g cm−3), and higher nitrogen content (MS, 16.9; SPS, 2.7 kg m−3) than SPS at the same soil depth, while the phosphorus (P) content (Bray II) (MS, 1.6; SPS, 1.9 g P2O5 m−3) was similar. The AMF colonization rate was significantly lower in SPS (39.2 ± 12.5%) than in MS (73.2 ± 4.6%). The higher number of AMF operational taxonomic units (OTUs) was detected by amplicon sequencing of the partial small-subunit rRNA gene (MS, 78; SPS, 50). A neighbor-joining tree of obtained OTUs revealed that they belonged to Acaulosporaceae, Ambisporaceae, Claroideoglomeraceae, Gigasporaceae, and Glomeraceae. The lower abundance and diversity of AMF in SPS are possibly caused by abiotic factors, including soil physicochemical properties. Glomus and Acaulospora species detected in SPS might have strong tolerance against acidity and high soil moisture content.

Molecular Characterization of Blastocystis from Beef Cattle in Northeastern China.

Blastocystis is a common unicellular protist that lives in the intestines of humans and animals. Blastocystis infection and subtypes in cattle have been reported in several regions. However, the information of Blastocystis infection in cattle in China is still largely scant. To assess the prevalence and subtype distribution of Blastocystis in beef cattle in China, 803 fecal samples were collected from beef cattle farms in four cities of Northeast China, and were subjected to an analysis based on small subunit rRNA gene fragment. The overall prevalence of Blastocystis in beef cattle was 2.11% (17/803), with 2.15% in preweaning calves, 1.9% in postweaning calves, and 3.85% in breeding cattle, but absence in adult cattle (p > 0.05). Moreover, five Blastocystis subtypes were identified (ST10, ST21, ST23, ST25, and ST26), among which ST10 and ST26 subtypes were dominant subtypes in beef cattle. Mixed infections were detected in three specimens (ST10/ST25, ST10/ST23/ST25, and ST10/ST26). This is the first report showing Blastocystis infection in beef cattle in Northeast China. In addition, a variety of Blastocystis subtypes are reported in cattle in China for the first time. These results will benefit for better understanding the epidemiology and public health implications of Blastocystis.

Molecular detection of Babesia microti in one-humped camel (Camelus dromedarius) in Halayeb and Shalateen, Halayeb, Egypt

There is paucity in the epidemiological screening of a blood parasite of zoonotic importance, Babesia microti (B. microti) in camels in Egypt. Therefore, a total of 142 individuals of one humped camel (Camelus dromedarius) that were bred in Halayeb and Shalateen were examined for infection and subjected to phylogenetic analysis on blood parasite infection with B. microti. PCR assay focusing on the Babesia microti small subunit rRNA (ss-rRNA) gene was used for detection of its prevalence in camels. Phylogenetic analysis revealed the detection of 17 case from a total of 142 cases with an infection rate of 11.97% using nested PCR assays. Moreover, phylogenetic analyses showed that B. microti was genetically related with German strain that is isolated from voles and rats in France. In conclusion, this is the first evidence of B. microti infection in camels in Egypt with evidence of the role of one-humped camels in sustaining the babesia transmission in Egypt.