Host-Adapted Cryptosporidium spp. in Canada Geese (Branta canadensis)

ABSTRACT The prevalence and distribution of Cryptosporidium spp. in the fecal droppings of the free-living waterfowl Canada geese were examined at 13 sites in Ohio and Illinois. On the basis of the analysis of the small-subunit rRNA gene by PCR, followed by restriction fragment length polymorphism analysis and DNA sequencing, 49 (23.4%) of 209 fecal specimens collected from 10 sites (76.9%) were positive for Cryptosporidium spp. The following five Cryptosporidium species and genotypes were identified: Cryptosporidium goose genotype I (in 36 specimens), Cryptosporidium goose genotype II (in 9 specimens), Cryptosporidium duck genotype (in 1 specimen), Cryptosporidium parvum (in 4 specimens), and C. hominis (in 2 specimens). Cryptosporidium goose genotype I was the most prevalent parasite and was found at all five Cryptosporidium-positive sites in Ohio and at four of five positive sites in Illinois, followed by Cryptosporidium goose genotype II, which was found at two of five positive sites in Ohio and at four of five positive sites in Illinois. Cryptosporidium goose genotype II was detected for the first time, and it is phylogenetically related to goose genotype I and the duck genotype. All three genotypes have not so far been reported in humans, and their pathogenicity in geese has not been determined. Only 10.2% of the Cryptosporidium-positive specimens had C. parvum and C. hominis. The results of this study indicate that Canada geese might only serve as accidental carriers of cryptosporidia infectious to humans and probably play a minor role in the animal-to-human transmission cycle of the pathogen.

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Study of Genetic Diversity of Eukaryotic Picoplankton in Different Oceanic Regions by Small-Subunit rRNA Gene Cloning and Sequencing

Applied and Environmental Microbiology ◽

10.1128/aem.67.7.2932-2941.2001 ◽

2001 ◽

Vol 67 (7) ◽

pp. 2932-2941 ◽

Cited By ~ 354

Author(s):

Beatriz Dı́ez ◽

Carlos PedrÃ¯Â¿Â½s-AliÃ¯Â¿Â½ ◽

Ramon Massana

Keyword(s):

Operational Taxonomic Unit ◽

Small Subunit ◽

Small Cells ◽

Rrna Genes ◽

Rrna Gene ◽

Polymorphism Analysis ◽

Small Subunit Rrna ◽

Phylogenetic Groups ◽

18S Rrna Genes ◽

A Minor

ABSTRACT Very small eukaryotic organisms (picoeukaryotes) are fundamental components of marine planktonic systems, often accounting for a significant fraction of the biomass and activity in a system. Their identity, however, has remained elusive, since the small cells lack morphological features for identification. We determined the diversity of marine picoeukaryotes by sequencing cloned 18S rRNA genes in five genetic libraries from North Atlantic, Southern Ocean, and Mediterranean Sea surface waters. Picoplankton were obtained by filter size fractionation, a step that excluded most large eukaryotes and recovered most picoeukaryotes. Genetic libraries of eukaryotic ribosomal DNA were screened by restriction fragment length polymorphism analysis, and at least one clone of each operational taxonomic unit (OTU) was partially sequenced. In general, the phylogenetic diversity in each library was rather great, and each library included many different OTUs and members of very distantly related phylogenetic groups. Of 225 eukaryotic clones, 126 were affiliated with algal classes, especially the Prasinophyceae, the Prymnesiophyceae, the Bacillariophyceae, and the Dinophyceae. A minor fraction (27 clones) was affiliated with clearly heterotrophic organisms, such as ciliates, the chrysomonad Paraphysomonas, cercomonads, and fungi. There were two relatively abundant novel lineages, novel stramenopiles (53 clones) and novel alveolates (19 clones). These lineages are very different from any organism that has been isolated, suggesting that there are previously unknown picoeukaryotes. Prasinophytes and novel stramenopile clones were very abundant in all of the libraries analyzed. These findings underscore the importance of attempts to grow the small eukaryotic plankton in pure culture.

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Cryptosporidium Species and C. parvum Subtypes in Farmed Bamboo Rats

Pathogens ◽

10.3390/pathogens9121018 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1018

Author(s):

Falei Li ◽

Wentao Zhao ◽

Chenyuan Zhang ◽

Yaqiong Guo ◽

Na Li ◽

...

Keyword(s):

Detection Rate ◽

Small Subunit ◽

Cryptosporidium Species ◽

Rrna Gene ◽

Human Pathogens ◽

Small Subunit Rrna ◽

Sequence Analyses ◽

Genotype I ◽

Cryptosporidium Spp ◽

Bamboo Rat

Bamboo rats (Rhizomys sinensis) are widely farmed in Guangdong, China, but the distribution and public health potential of Cryptosporidium spp. in them are unclear. In this study, 724 fecal specimens were collected from bamboo rats in Guangdong Province and analyzed for Cryptosporidium spp. using PCR and sequence analyses of the small subunit rRNA gene. The overall detection rate of Cryptosporidium spp. was 12.2% (88/724). By age, the detection rate in animals under 2 months (23.2% or 13/56) was significantly higher than in animals over 2 months (11.2% or 75/668; χ2 = 6.95, df = 1, p = 0.0084). By reproduction status, the detection rate of Cryptosporidium spp. in nursing animals (23.1% or 27/117) was significantly higher than in other reproduction statuses (6.8% or 4/59; χ2 = 7.18, df = 1, p = 0.0074). Five Cryptosporidium species and genotypes were detected, including Cryptosporidium bamboo rat genotype I (n = 49), C. parvum (n = 31), Cryptosporidium bamboo rat genotype III (n = 5), C. occultus (n = 2), and C. muris (n = 1). The average numbers of oocysts per gram of feces for these Cryptosporidium spp. were 14,074, 494,636, 9239, 394, and 323, respectively. The genetic uniqueness of bamboo rat genotypes I and III was confirmed by sequence analyses of the 70 kDa heat shock protein and actin genes. Subtyping C. parvum by sequence analysis of the 60 kDa glycoprotein gene identified the presence of IIoA15G1 (n = 20) and IIpA6 (n = 2) subtypes. The results of this study indicated that Cryptosporidium spp. are common in bamboo rats in Guangdong, and some of the Cryptosporidium spp. in these animals are known human pathogens.

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Cryptosporidium Genotypes in Wildlife from a New York Watershed

Applied and Environmental Microbiology ◽

10.1128/aem.01034-07 ◽

2007 ◽

Vol 73 (20) ◽

pp. 6475-6483 ◽

Cited By ~ 103

Author(s):

Yaoyu Feng ◽

Kerri A. Alderisio ◽

Wenli Yang ◽

Lisa A. Blancero ◽

William G. Kuhne ◽

...

Keyword(s):

New York ◽

Animal Species ◽

Small Subunit ◽

Storm Water ◽

Animal Origin ◽

Rrna Gene ◽

Polymorphism Analysis ◽

Small Subunit Rrna ◽

Health Significance ◽

Animal Hosts

ABSTRACT To identify the animal sources for Cryptosporidium contamination, we genotyped Cryptosporidium spp. in wildlife from the watershed of the New York City drinking water supply, using a small-subunit rRNA gene-based PCR-restriction fragment length polymorphism analysis and DNA sequencing. A total of 541 specimens from 38 species of wildlife were analyzed. One hundred and eleven (20.5%) of the wildlife specimens were PCR positive. Altogether, 21 Cryptosporidium genotypes were found in wildlife samples, 11 of which were previously found in storm runoff in the watershed, and six of these 11 were from storm water genotypes of unknown animal origin. Four new genotypes were found, and the animal hosts for four storm water genotypes were expanded. With the exception of the cervine genotype, most genotypes were found in a limited number of animal species and have no major public health significance.

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Tracking Host Sources of Cryptosporidium spp. in Raw Water for Improved Health Risk Assessment

Applied and Environmental Microbiology ◽

10.1128/aem.02788-06 ◽

2007 ◽

Vol 73 (12) ◽

pp. 3945-3957 ◽

Cited By ~ 77

Author(s):

Norma J. Ruecker ◽

Shannon L. Braithwaite ◽

Edward Topp ◽

Thomas Edge ◽

David R. Lapen ◽

...

Keyword(s):

Land Use ◽

Environmental Protection Agency ◽

Small Subunit ◽

Molecular Composition ◽

Source Tracking ◽

Molecular Evidence ◽

Rrna Gene ◽

Small Subunit Rrna ◽

Genotype I ◽

Area Source

ABSTRACT Recent molecular evidence suggests that different species and/or genotypes of Cryptosporidium display strong host specificity, altering our perceptions regarding the zoonotic potential of this parasite. Molecular forensic profiling of the small-subunit rRNA gene from oocysts enumerated on microscope slides by U.S. Environmental Protection Agency method 1623 was used to identify the range and prevalence of Cryptosporidium species and genotypes in the South Nation watershed in Ontario, Canada. Fourteen sites within the watershed were monitored weekly for 10 weeks to assess the occurrence, molecular composition, and host sources of Cryptosporidium parasites impacting water within the region. Cryptosporidium andersoni, Cryptosporidium muskrat genotype II, Cryptosporidium cervine genotype, C. baileyi, C. parvum, Cryptosporidium muskrat genotype I, the Cryptosporidium fox genotype, genotype W1, and genotype W12 were detected in the watershed. The molecular composition of the Cryptosporidium parasites, supported by general land use analysis, indicated that mature cattle were likely the main source of contamination of the watershed. Deer, muskrats, voles, birds, and other wildlife species, in addition to sewage (human or agricultural) may also potentially impact water quality within the study area. Source water protection studies that use land use analysis with molecular genotyping of Cryptosporidium parasites may provide a more robust source-tracking tool to characterize fecal impacts in a watershed. Moreover, the information is vital for assessing environmental and human health risks posed by water contaminated with zoonotic and/or anthroponotic forms of Cryptosporidium.

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Genetic Diversity of Cryptosporidium parvum in Neonatal Dairy Calves in Xinjiang, China

Pathogens ◽

10.3390/pathogens9090692 ◽

2020 ◽

Vol 9 (9) ◽

pp. 692

Author(s):

Yayun Wu ◽

Kuankuan Zhang ◽

Ying Zhang ◽

Bo Jing ◽

Yuancai Chen ◽

...

Keyword(s):

Genetic Diversity ◽

Cryptosporidium Parvum ◽

Diarrheal Disease ◽

Small Subunit ◽

Dairy Calves ◽

Rrna Gene ◽

Polymorphism Analysis ◽

Small Subunit Rrna ◽

High Genetic Diversity ◽

Glycoprotein Gene

Cryptosporidium parvum has been identified as a major cause of diarrhea and diarrhea-associated deaths in young children and neonatal calves. Infections can remain asymptomatic but may lead to malnutrition and persistent growth retardation. To assess the relationship between C. parvum genetic diversity and pathogenicity in neonatal dairy calves and determine the cause of diarrhea among these calves, 232 fecal samples from neonatal dairy calves on 12 farms in Xinjiang, China, were characterized for Cryptosporidium presence based on the small subunit rRNA gene. The Cryptosporidium prevalence was 38.4% (89/232), and three species were detected with restriction fragment length polymorphism analysis, including C. parvum (the significantly dominant species), C. ryanae, and C. bovis. Cryptosporidium prevalence was significantly higher in neonatal dairy calves with diarrhea (52.6%, 51/97) than in calves without diarrhea (28.1%, 38/135). All C. parvum-positive samples were analyzed based on the 60 KDa glycoprotein gene, and IIdA15G1, IIdA20G1, IIdA14G1, and IIdA19G1 were successfully subtyped. These data indicate that C. parvum may be a major contributor to diarrheal disease in neonatal dairy calves, and C. parvum subtypes from neonatal dairy calves in Xinjiang exhibited high genetic diversity.

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Targeted Next-Generation Sequencing and Informatics as an Effective Tool to Establish the Composition of Bovine Piroplasm Populations in Endemic Regions

Microorganisms ◽

10.3390/microorganisms9010021 ◽

2020 ◽

Vol 9 (1) ◽

pp. 21

Author(s):

Abdul Ghafar ◽

Anson V. Koehler ◽

Ross S. Hall ◽

Charles G. Gauci ◽

Robin B. Gasser ◽

...

Keyword(s):

Next Generation Sequencing ◽

Water Buffalo ◽

Hypervariable Region ◽

Small Subunit ◽

Rrna Gene ◽

Next Generation ◽

Small Subunit Rrna ◽

Tropical Theileriosis ◽

Ecological Zones ◽

Generation Sequencing

Protists of the genera Babesia and Theileria (piroplasms) cause some of the most prevalent and debilitating diseases for bovines worldwide. In this study, we established and used a next-generation sequencing-informatic approach to explore the composition of Babesia and Theileria populations in cattle and water buffalo in a country (Pakistan) endemic for these pathogens. We collected individual blood samples from cattle (n = 212) and water buffalo (n = 154), extracted genomic DNAs, PCR-amplified the V4 hypervariable region of 18S small subunit rRNA gene from piroplasms, sequenced amplicons using Illumina technology, and then analysed data using bioinformatic platforms. The results revealed piroplasms in 68.9% (252/366) samples, with overall occurrence being markedly higher in cattle (85.8%) than in water buffaloes (45.5%). Babesia (B.) occultans and Theileria (T.) lestoquardi-like species were recorded for the first time in Pakistan, and, overall, T. annulata was most commonly detected (65.8%) followed by B. bovis (7.1%), B. bigemina (4.4%), and T. orientalis (0.5%), with the genetic variability within B. bovis being pronounced. The occurrence and composition of piroplasm species varied markedly across different agro-ecological zones. The high detection of T. annulata in asymptomatic animals suggested a relatively high level of endemic stability of tropical theileriosis in the bovine population.

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Specificity of Associations between Bacteria and the Coral Pocillopora meandrina during Early Development

Applied and Environmental Microbiology ◽

10.1128/aem.01232-12 ◽

2012 ◽

Vol 78 (20) ◽

pp. 7467-7475 ◽

Cited By ~ 33

Author(s):

Amy Apprill ◽

Heather Q. Marlow ◽

Mark Q. Martindale ◽

Michael S. Rappé

Keyword(s):

Small Subunit ◽

Rrna Genes ◽

Polymorphism Analysis ◽

Roseobacter Clade ◽

Small Subunit Rrna ◽

Content Type ◽

Sterile Seawater ◽

Show Evidence ◽

Coral Health

ABSTRACTRelationships between corals and specific bacterial associates are thought to play an important role in coral health. In this study, the specificity of bacteria associating with the coralPocillopora meandrinawas investigated by exposing coral embryos to various strains of cultured marine bacteria, sterile seawater, or raw seawater and examining the identity, density, and location of incorporated cells. The isolates utilized in this experiment included members of the Roseobacter and SAR11 clades of theAlphaproteobacteria, aPseudoalteromonasspecies of theGammaproteobacteria, and aSynechococcusspecies of theCyanobacteriaphylum. Based on terminal restriction fragment length polymorphism analysis of small-subunit rRNA genes, similarities in bacterial communities associated with 170-h-old planulae were observed regardless of treatment, suggesting that bacteria may have been externally associated from the outset of the experiment. Microscopic examination ofP. meandrinaplanulae by fluorescencein situhybridization with bacterial and Roseobacter clade-specific oligonucleotide probes revealed differences in the densities and locations of planulae-associated cells. Planulae exposed to either raw seawater or strains ofPseudoalteromonasand Roseobacter harbored the highest densities of internally associated cells, of which 20 to 100% belonged to the Roseobacter clade. Planulae exposed to sterile seawater or strains of the SAR11 clade andSynechococcusdid not show evidence of prominent bacterial associations. Additional analysis of the raw-seawater-exposed planulae via electron microscopy confirmed the presence of internally associated prokaryotic cells, as well as virus-like particles. These results suggest that the availability of specific microorganisms may be an important factor in the establishment of coral-bacterial relationships.

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Differences in Microbial Activity and Microbial Populations of Peat Associated with Suppression of Damping-Off Disease Caused by Pythium sylvaticum

Applied and Environmental Microbiology ◽

10.1128/aem.00313-06 ◽

2006 ◽

Vol 72 (10) ◽

pp. 6452-6460 ◽

Cited By ~ 43

Author(s):

Paul J. Hunter ◽

Geoff M. Petch ◽

Leo A. Calvo-Bado ◽

Tim R. Pettitt ◽

Nick R. Parsons ◽

...

Keyword(s):

Microbial Activity ◽

Denaturing Gradient Gel Electrophoresis ◽

Small Subunit ◽

Disease Suppression ◽

Rrna Gene ◽

Damping Off ◽

Small Subunit Rrna ◽

Pythium Sylvaticum ◽

Gradient Gel Electrophoresis ◽

Microbial Groups

ABSTRACT The microbiological characteristics associated with disease-suppressive peats are unclear. We used a bioassay for Pythium sylvaticum-induced damping-off of cress seedlings to identify conducive and suppressive peats. Microbial activity in unconditioned peats was negatively correlated with the counts of P. sylvaticum at the end of the bioassay. Denaturing gradient gel electrophoresis (DGGE) profiling and clone library analyses of small-subunit rRNA gene sequences from two suppressive and two conducive peats differed in the bacterial profiles generated and the diversity of sequence populations. There were also significant differences between bacterial sequence populations from suppressive and conducive peats. The frequencies of a number of microbial groups, including the Rhizobium-Agrobacterium group (specifically sequences similar to those for the genera Ochrobactrum and Zoogloea) and the Acidobacteria, increased specifically in the suppressive peats, although no single bacterial group was associated with disease suppression. Fungal DGGE profiles varied little over the course of the bioassay; however, two bands associated specifically with suppressive samples were detected. Sequences from these bands corresponded to Basidiomycete yeast genera. Although the DGGE profiles were similar, fungal sequence diversity also increased during the bioassay. Sequences highly similar to those of Cryptococcus increased in relative abundance during the bioassay, particularly in the suppressive samples. This study highlights the importance of using complementary approaches to molecular profiling of complex populations and provides the first report that basidiomycetous yeasts may be associated with the suppression of Pythium-induced diseases in peats.

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Detection of Kobe-typeBabesia microtiassociated with Japanese human babesiosis in field rodents in central Taiwan and southeastern mainland China

Parasitology ◽

10.1017/s0031182008004356 ◽

2008 ◽

Vol 135 (6) ◽

pp. 691-699 ◽

Cited By ~ 12

Author(s):

A. SAITO-ITO ◽

N. TAKADA ◽

F. ISHIGURO ◽

H. FUJITA ◽

Y. YANO ◽

...

Keyword(s):

Mainland China ◽

Small Subunit ◽

Human Infection ◽

Babesia Microti ◽

Rrna Gene ◽

Microscopical Examination ◽

Small Subunit Rrna ◽

Central Taiwan ◽

Field Rodents ◽

Human Babesiosis

SUMMARYField rodent surveys forBabesiainfection were performed from 2002 to 2005 in the vicinities of human babesiosis occurrences in Taiwan and mainland China.Babesia microtiwas identified by microscopical examination and/or PCR in 1Rattus coxingaand 1Crocidura horsfieldiiin central Taiwan and in 13Niviventer confucianusand 1Apodemus agrariusin Zhejiang and Fujian Provinces of southeastern China. Of 15B. microtisamples detected by PCR, all except 1 were shown to be the Kobe-type, the aetiological small subunit rRNA gene-type of the first Japanese patient; the exception was also a Kobe-related type. The Kobe-type had been found in rodents only in a few places including the human infection occurrence place in Japan. The internal transcribed spacer 1 to 2 sequences of the Taiwanese and Chinese Kobe-types were very similar to each other but considerably different (approx. 94% pairwise identities) from that of the Japanese Kobe-type. A Taiwanese Kobe-type strain was serologically differentiated from the Kobe strain originating from the Japanese first patient. The distribution of the Kobe-type in the vicinities of human babesiosis occurrences in Taiwan and China as well as in Japan is suggestive of involvement of the Kobe-type in Asian human babesiosis.

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