scholarly journals Molecular survey of Cryptosporidium spp. in calves from the state of Mato Grosso, Brazil

2020 ◽  
Vol 41 (5supl1) ◽  
pp. 2437-2444
Author(s):  
Thábata dos Anjos Pacheco ◽  
Felippe Danyel Cardoso Martins ◽  
Sayanne Luns Hatum de Almeida ◽  
Thiago Borges Fernandes Semedo ◽  
Michelle Igarashi Watanabe ◽  
...  
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
The State ◽  
Severe Illness ◽  
Rrna Gene ◽  
Mato Grosso ◽  
Fecal Samples ◽  
Cryptosporidium Spp ◽  
Gp60 Gene ◽  
Wide Range

Cryptosporidium spp. is a protozoan that infects a wide range of vertebrate hosts; it has been reported to be the cause of severe illness or death in livestock worldwide, which leads to decreased performance and production losses, especially in young animals. This study investigated the presence of Cryptosporidium in calves from beef farms in the state of Mato Grosso, midwestern Brazil. For this purpose, fecal samples from 237 animals aged ? 45 days, raised in 20 rural properties were subjected to DNA extraction and nested polymerase chain reaction (nPCR) targeting 18S ribosomal RNA (18S rRNA) gene followed by sequencing. Additionally, positive samples, previously identified as Cryptosporidium parvum by sequencing and phylogenetic analyses based on 18S rRNA gene, were subsequently analyzed focusing the amplification and sequencing using nPCR of a fragment of the 60 kDa glycoprotein (gp60) gene. Of the 237 fecal samples analyzed by PCR (18S rRNA), 50 (21.1%) fecal samples were positive for Cryptosporidium spp., while 14 (70%) of the 20 properties had at least one positive animal. The following Cryptosporidium species were detected: C. bovis, C. parvum, and C. ryanae. Thereafter, two potentially zoonotic subtypes (IIaA15G2R1 and IIaA16G3R1) of C. parvum were identified based on gp60 gene sequences. This study resulted in the detection of subtype IIaA16G3R1 for the first time in South America and showed a wide distribution of the protozoan in beef farms in the studied area of the State.

scholarly journals High-resolution molecular identification of smalltooth sawfish prey

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Taylor L. Hancock ◽  
Gregg R. Poulakis ◽  
Rachel M. Scharer ◽  
S. Gregory Tolley ◽  
Hidetoshi Urakawa
Keyword(s):  
18S Rrna ◽  
Stomach Content Analysis ◽  
18S Rrna Gene ◽  
Taxonomic Resolution ◽  
Molecular Technique ◽  
Rrna Gene ◽  
Fecal Samples ◽  
Predator Prey ◽  
Critically Endangered Species ◽  
Wide Range

AbstractThe foundation of food web analysis is a solid understanding of predator-prey associations. Traditional dietary studies of fishes have been by stomach content analysis. However, these methods are not applicable to Critically Endangered species such as the smalltooth sawfish (Pristis pectinata). Previous research using the combination of stable isotope signatures from fin clips and 18S rRNA gene sequencing of fecal samples identified the smalltooth sawfish as piscivorous at low taxonomic resolution. Here, we present a high taxonomic resolution molecular technique for identification of prey using opportunistically acquired fecal samples. To assess potential biases, primer sets of two mitochondrial genes, 12S and 16S rRNA, were used alongside 18S rRNA, which targets a wider spectrum of taxa. In total, 19 fish taxa from 7 orders and 11 families native to the Gulf of Mexico were successfully identified. The sawfish prey comprised diverse taxa, indicating that this species is a generalist piscivore. These findings and the molecular approach used will aid recovery planning for the smalltooth sawfish and have the potential to reveal previously unknown predator-prey associations from a wide range of taxa, especially rare and hard to sample species.

scholarly journals Emergence of Babesia Conradae Infection in Coyote-hunting Greyhounds in Oklahoma, USA

2021 ◽  
Author(s):  
Erin M Stayton ◽  
Megan Lineberry ◽  
Jennifer Thomas ◽  
Tina Bass ◽  
Kelly Allen ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
18S Rrna Gene ◽  
Post Treatment ◽  
Rrna Gene ◽  
Babesia Gibsoni ◽  
Wide Range ◽  
Pcr Products ◽  
Life Threatening ◽  
Tick Vectors

Abstract Background: Babesia species are intraerythrocytic Apicomplexan parasites that infect a wide range of vertebrate hosts. These pathogens are typically transmitted either by tick vectors or by direct blood-to-blood contact, and may cause life-threatening clinical disease such as thrombocytopenia, hemolytic anemia, and acute renal failure in canine hosts. While Babesia vogeli and Babesia gibsoni infections have both been reported in Oklahoma, reports of B. conradae infections have been limited to California. Methods: Whole blood samples were collected in EDTA tubes from all dogs in four separate kennels in Oklahoma. DNA was extracted from each blood sample and a nested PCR was performed using general Apicomplexan primers for the partial 18S rRNA gene. PCR products were electrophoresed in agarose matrix and appropriately sized amplicons were sequenced. Sequences were compared to reference 18S rRNA sequences available in GenBank, and samples with >98% homology to B. conradae (GenBank MK256976) were considered positive. B. conradae positive dogs were then treated with atovaquone (13.5 mg/kg TID) and azithromycin (10 mg/kg SID) for 10 days and retested at 30 and 60 days post treatment by PCR. Results: Fifteen of 40 dogs tested positive for B. conradae with 98–100% sequence homology to B. conradae from California. All positive cases were coyote-hunting Greyhounds. Treatment of clinically ill dogs with atovaquone and azithromycin resulted in complete clinical recovery in clinically ill dogs and all treated dogs had negative follow-up PCR at 30 and 60 days post treatment. Conclusions: Collectively, this study (i) documents the occurrence of B. conradae in Oklahoma, (ii) highlights this pathogen as a differential to be considered when clinical signs are present, and (iii) supports the use of atovaquone and azithromycin as effective treatment in these cases.

scholarly journals Cryptosporidium spp. in Domestic Dogs: the “Dog” Genotype

2000 ◽  
Vol 66 (5) ◽  
pp. 2220-2223 ◽  
Author(s):  
Una M. Morgan ◽  
Lihua Xiao ◽  
Paul Monis ◽  
Abbie Fall ◽  
Peter J. Irwin ◽  
...  
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Domestic Dogs ◽  
Heat Shock Gene ◽  
Valid Species ◽  
Rrna Gene ◽  
Phylogenetic Characterization ◽  
Cryptosporidium Spp ◽  
Shock Gene

ABSTRACT Genetic and phylogenetic characterization ofCryptosporidium isolates at two loci (18S rRNA gene and heat shock gene) from both Australian and United States dogs demonstrated that dog-derived Cryptosporidium isolates had a distinct genotype which is conserved across geographic areas. Phylogenetic analysis provided support for the idea that the “dog” genotype is, in fact, a valid species.

Molecular identification ofSarcocystisspp. helped to define the origin of green pythons (Morelia viridis) confiscated in Germany

Parasitology ◽  
2013 ◽  
Vol 141 (5) ◽  
pp. 646-651 ◽  
Author(s):  
GASTÓN MORÉ ◽  
NIKOLA PANTCHEV ◽  
DALAND C. HERRMANN ◽  
MAJDA GLOBOKAR VRHOVEC ◽  
SABINE ÖFNER ◽  
...  
Keyword(s):  
18S Rrna ◽  
Sequence Data ◽  
18S Rrna Gene ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Intermediate Hosts ◽  
Apicomplexan Parasites ◽  
Large Numbers ◽  
Wide Range ◽  
18S Rrna Genes

SUMMARYSarcocystisspp. represent apicomplexan parasites. They usually have a heteroxenous life cycle. Around 200 species have been described, affecting a wide range of animals worldwide, including reptiles. In recent years, large numbers of reptiles have been imported into Europe as pets and, as a consequence, animal welfare and species protection issues emerged. A sample of pooled feces from four confiscated green pythons (Morelia viridis) containingSarcocystisspp. sporocysts was investigated. These snakes were imported for the pet trade and declared as being captive-bred. Full length 18S rRNA genes were amplified, cloned into plasmids and sequenced. Two differentSarcocystisspp. sequences were identified and registered asSarcocystissp. fromM. viridisin GenBank. Both showed a 95–97% sequence identity with the 18S rRNA gene ofSarcocystis singaporensis.Phylogenetic analysis positioned these sequences together with otherSarcocystisspp. from snakes and rodents as definitive and intermediate hosts (IH), respectively. Sequence data and also the results of clinical and parasitological examinations suggest that the snakes were definitive hosts forSarcocystisspp. that circulate in wild IH. Thus, it seems unlikely that the infected snakes had been legally bred. Our research shows that information on the infection of snakes withSarcocystisspp. may be used to assess compliance with regulations on the trade with wildlife species.

scholarly journals Molecular characterisation of Cryptosporidium spp. in lambs in the South Central region of the State of São Paulo

2015 ◽  
Vol 67 (2) ◽  
pp. 441-446 ◽  
Author(s):  
A.S. Zucatto ◽  
M.C.C. Aquino ◽  
S.V. Inácio ◽  
R.N. Figueiredo ◽  
J.C. Pierucci ◽  
...  
Keyword(s):  
Central Region ◽  
18S Rrna Gene ◽  
Sao Paulo ◽  
The State ◽  
Rrna Gene ◽  
São Paulo ◽  
The South ◽  
Cryptosporidium Spp ◽  
South Central ◽  
One Year

Considering the proximity of sheep farmers to animals that are possibly diseased or releasing fecal oocysts into the environment and the marked pathogenicity in lambs, the aim of this study was to determine the occurrence and to molecularly characterize the infection by Cryptosporidium spp. in lambs in the South Central region of the state of São Paulo, Brazil. A total of 193 fecal samples were collected from sheep of several breeds, males and females, aged up to one year. Polymerase chain reaction (nested-PCR) was used to amplify DNA fragments from the subunit 18S rRNA gene and indicated 15% positivity; sequencing of amplified fragments was possible for 19 samples. Analysis of the obtained sequences showed that the identified species were Cryptosporidium xiaoi for 15 samples, constituting thus the first molecular characterization study of this Cryptosporidium species in Brazil. Cryptosporidium ubiquitum was identified for three samples and Cryptosporidium meleagridis for one sample; the latter two are considered zoonotic species.

A new set of primers directed to 18S rRNA gene for molecular identification of Cryptosporidium spp. and their performance in the detection and differentiation of oocysts shed by synanthropic rodents

2013 ◽  
Vol 135 (3) ◽  
pp. 551-557 ◽  
Author(s):  
Sheila O.S. Silva ◽  
Leonardo J. Richtzenhain ◽  
Iracema N. Barros ◽  
Alessandra M.M. C. Gomes ◽  
Aristeu V. Silva ◽  
...  
Keyword(s):  
Molecular Identification ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Cryptosporidium Spp ◽  
Synanthropic Rodents

scholarly journals pr2-primers: an 18S rRNA primer database for protists

2021 ◽  
Author(s):  
Daniel Vaulot ◽  
Stefan Geisen ◽  
Frédéric Mahé ◽  
David Bass
Keyword(s):  
Web Application ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Taxonomic Distribution ◽  
System A ◽  
Microbial Eukaryotes ◽  
Wide Range ◽  
Amplicon Size ◽  
On Line

AbstractMetabarcoding of microbial eukaryotes (collectively known as protists) has developed tremendously in the last decade, almost uniquely relying on the 18S rRNA gene. As microbial eukaryotes are extremely diverse, many primers and primer pairs have been developed. To cover a relevant and representative fraction of the protist community in a given study system, a wise primer choice is needed as no primer pair can target all protists equally well. As such, a smart primer choice is very difficult even for experts and there are very few on-line resources available to list existing primers. We built a database listing 179 primers and 76 primer pairs that have been used for eukaryotic 18S rRNA metabarcoding. In silico performance of primer pairs was tested against two sequence databases: PR2 for eukaryotes and a subset of Silva for prokaryotes. This allowed to determine the taxonomic specificity of primer pairs, the location of mismatches as well as amplicon size. We developed a R-based web application that allows to browse the database, visualize the taxonomic distribution of the amplified sequences with the number of mismatches, and to test any user-defined primer set (https://app.pr2-primers.org). This tool will provide the basis for guided primer choices that will help a wide range of ecologists to implement protists as part of their investigations.

scholarly journals The first study of molecular prevalence and species characterization of Cryptosporidium in free-range chicken (Gallus gallus domesticus) from Brazil

2017 ◽  
Vol 26 (4) ◽  
pp. 472-478 ◽  
Author(s):  
Maria Paula de Carvalho Ewald ◽  
Felippe Danyel Cardoso Martins ◽  
Eloiza Teles Caldart ◽  
Fernando Emmanuel Gonçalves Vieira ◽  
Milton Hissashi Yamamura ◽  
...  
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Gallus Gallus Domesticus ◽  
Free Range ◽  
Cryptosporidium Spp ◽  
High Prevalence ◽  
Cryptosporidium Oocysts ◽  
Genotype C

Abstract Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

scholarly journals Molecular detection of genotypes and subtypes of Cryptosporidium infection in diarrheic calves, lambs, and goat kids from Turkey

2020 ◽  
Author(s):  
Mohammad Hazzaz Bin Kabir ◽  
Onur Ceylan ◽  
Ceylan Ceylan ◽  
Ayman Ahmed Shehata ◽  
Hironori Bando ◽  
...  
Keyword(s):  
Small Subunit ◽  
Rrna Gene ◽  
Ssu Rrna ◽  
Cryptosporidium Infection ◽  
Cryptosporidium Spp ◽  
Gp60 Gene ◽  
Wide Range ◽  
Cryptosporidium Oocysts ◽  
Few Data ◽  
Almost All

Abstract Background: Cryptosporidium spp. are enteric protozoan parasites that infect a wide range of animals as well as humans. The studies on Cryptosporidium infections of animals in Turkey are mostly rely on microscopic observation. Few data are available regarding the distribution of Cryptosporidium genotypes and subtypes infection. The aim of this study is to analyse the zoonotic potential of Cryptosporidium oocysts shed from young ruminant livestock. Methods: A total of 415 diarrheic fecal specimens from 333 calves, 67 lambs, and 15 goat kids were examined for the presence of Cryptosporidium oocysts by microscopy. Microscopic positive specimens were then analyzed for Cryptosporidium genotypes and subtypes detection by use of nested PCR of the small subunit ribosomal RNA (SSU rRNA) gene and the highly polymorphic 60 kDa glycoprotein (gp60) gene followed by sequence analyses. Results: The results of this study revealed that 25.6% (106 of 415) of the specimens were positive for Cryptosporidium spp. infection by microscopic examination and molecular analysis. We identified 27.4% (91/333), 19.4% (13/67), and 13.4% (2/15) of positivity in calves, lambs and goat kids, respectively. Genotyping of the SSU rRNA indicated that almost all positive specimens were of C . parvum , except for one calf which was of C. bovis . Sequence analysis of the gp60 gene revealed the most common zoonotic subtypes (IIa and IId) of C . parvum. We detected 11 subtypes (IIaA11G2R1, IIaA11G3R1, IIaA12G3R1, IIaA13G2R1, IIaA13G4R1, IIaA14G1R1, IIaA14G3R1, IIaA15G2R1, IIdA16G1, IIdA18G1, IIdA22G1); three of them (IIaA12G3R1, IIaA11G3R1 and IIaA13G4R1) was novel subtypes found in calves and lambs. Additionally, three subtypes (IIaA11G2R1, IIaA14G3R1, and IIdA16G1) were detected in calves, lambs, and goat kid for the first time in Turkey. Conclusions: The findings illustrate the high occurrence of Cryptosporidium infection in Turkey and suggest that calves, lambs, and goat kids are likely a major reservoir of C. parvum and a potential source of zoonotic transmission, which may have public health implications. Keywords: Calves, C. bovis, C. parvum , Cryptosporidium , Diarrhea, Goat kids, Lambs, Subtypes, Turkey.

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