scholarly journals Seasonal monitoring of Cryptosporidium species and genetic diversity in neonatal dairy calves on two large-scale farms in Xinjiang, China

2020 ◽  
Author(s):  
Yayun Wu ◽  
Kuankuan Zhang ◽  
Bo Jing ◽  
Chunyan Xu ◽  
Yuancai Chen ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Large Scale ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Small Subunit ◽  
Dairy Calves ◽  
Rrna Gene ◽  
Glycoprotein Gene ◽  
Zoonotic Infections ◽  
Seasonal Monitoring

Abstract Background: Neonatal dairy calves infected with Cryptosporidium can possess a significant source of zoonotic infections and disease. To assess seasonal variations in the prevalence and genetic diversity of Cryptosporidium in neonatal dairy calves, 380 fecal samples from neonatal dairy calves on two large-scale farms in Xinjiang (Alaer and Wensu) were screened for the Cryptosporidium small subunit (SSU) rRNA gene. Results: The overall prevalence of Cryptosporidium was 48.7% (185/380): 48.6% (108/222) in Alaer and 48.7% (77/158) in Wensu. Cryptosporidium was most frequent in summer (56.8%, 54/95), followed by spring (50.0%, 44/88), winter (46.8%, 44/94), and autumn (41.7%, 43/103) (P > 0.05). Cryptosporidium was significantly more prevalent in calves with diarrhea (72.4%, 113/156) than in those without (32.1%, 72/224) (P < 0.01). Based on a restriction fragment length polymorphism (RFLP) analysis, C. parvum (n = 173), C. bovis (n = 7), C. ryanae (n = 3), and co-infections of the three species (n = 2) were identified. Most (172/175) C. parvum samples were successfully sequenced at the 60-kDa glycoprotein gene (gp60), revealing two zoonotic subtypes: IIdA14G1 (n = 94) and IIdA15G1 (n = 7) in Alaer and IIdA15G1 (n = 71) in Wensu.Conclusions: These results showed that neonatal dairy calves were commonly infected with Cryptosporidium throughout the year, and there was a significant association between the occurrence of diarrhea and Cryptosporidium infection. Presence of IIdA14G1 and IIdA15G1 indicated neonatal dairy calves may be a source of zoonotic C. parvum subtypes.

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

Pathogens ◽  
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.

scholarly journals Genetic Diversity and Phylogeny of Rhizobia That Nodulate Acacia spp. in Morocco Assessed by Analysis of rRNA Genes

1998 ◽  
Vol 64 (12) ◽  
pp. 4912-4917 ◽  
Author(s):  
Bouchaib Khbaya ◽  
Marc Neyra ◽  
Philippe Normand ◽  
Karim Zerhari ◽  
Abdelkarim Filali-Maltouf
Keyword(s):  
Genetic Diversity ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Rrna Genes ◽  
23S Rrna ◽  
Rrna Gene ◽  
The Third ◽  
The 16S Rrna Gene ◽  
Rdna Analysis ◽  
Restriction Fragment Length

ABSTRACT Forty rhizobia nodulating four Acacia species (A. gummifera, A. raddiana, A. cyanophylla, and A. horrida) were isolated from different sites in Morocco. These rhizobia were compared by analyzing both the 16S rRNA gene (rDNA) and the 16S-23S rRNA spacer by PCR with restriction fragment length polymorphism (RFLP) analysis. Analysis of the length of 16S-23S spacer showed a considerable diversity within these microsymbionts, but RFLP analysis of the amplified spacer revealed no additional heterogeneity. Three clusters were identified when 16S rDNA analysis was carried out. Two of these clusters include some isolates which nodulate, nonspecifically, the four Acacia species. These clusters, A and B, fit within the Sinorhizobiumlineage and are closely related to S. meliloti and S. fredii, respectively. The third cluster appeared to belong to theAgrobacterium-Rhizobium galegae phylum and is more closely related to the Agrobacterium tumefaciens species. These relations were confirmed by sequencing a representative strain from each cluster.

scholarly journals Genetic Diversity within Cryptosporidium parvum and Related Cryptosporidium Species

1999 ◽  
Vol 65 (8) ◽  
pp. 3386-3391 ◽  
Author(s):  
Lihua Xiao ◽  
Una M. Morgan ◽  
Josef Limor ◽  
Ananias Escalante ◽  
Michael Arrowood ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Cryptosporidium Parvum ◽  
Fragment Length Polymorphism ◽  
Distinct Species ◽  
Small Subunit ◽  
Rrna Gene ◽  
Ssu Rrna ◽  
Specific Host ◽  
Pcr Product ◽  
Rrna Sequences

ABSTRACT To assess the genetic diversity in Cryptosporidium parvum, we have sequenced the small subunit (SSU) rRNA gene of seven Cryptosporidium spp., various isolates of C. parvum from eight hosts, and a Cryptosporidiumisolate from a desert monitor. Phylogenetic analysis of the SSU rRNA sequences confirmed the multispecies nature of the genusCryptosporidium, with at least four distinct species (C. parvum, C. baileyi, C. muris, and C. serpentis). Other species previously defined by biologic characteristics, including C. wrairi, C. meleagridis, and C. felis, and the desert monitor isolate, clustered together or within C. parvum. Extensive genetic diversities were present among C. parvum isolates from humans, calves, pigs, dogs, mice, ferrets, marsupials, and a monkey. In general, specific genotypes were associated with specific host species. A PCR-restriction fragment length polymorphism technique previously developed by us could differentiate mostCryptosporidium spp. and C. parvum genotypes, but sequence analysis of the PCR product was needed to differentiateC. wrairi and C. meleagridis from some of theC. parvum genotypes. These results indicate a need for revision in the taxonomy and assessment of the zoonotic potential of some animal C. parvum isolates.

scholarly journals Genetic Diversity of Cryptosporidium spp. in Captive Reptiles

2004 ◽  
Vol 70 (2) ◽  
pp. 891-899 ◽  
Author(s):  
Lihua Xiao ◽  
Una M. Ryan ◽  
Thaddeus K. Graczyk ◽  
Josef Limor ◽  
Lixia Li ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Cryptosporidium Parvum ◽  
Fragment Length Polymorphism ◽  
Small Subunit ◽  
Rrna Gene ◽  
Small Subunit Rrna ◽  
Cryptosporidium Spp ◽  
Small Subunit Rrna Gene ◽  
Different Types ◽  
Restriction Fragment Length

ABSTRACT The genetic diversity of Cryptosporidium in reptiles was analyzed by PCR-restriction fragment length polymorphism and sequence analysis of the small subunit rRNA gene. A total of 123 samples were analyzed, of which 48 snake samples, 24 lizard samples, and 3 tortoise samples were positive for Cryptosporidium. Nine different types of Cryptosporidium were found, including Cryptosporidium serpentis, Cryptosporidium desert monitor genotype, Cryptosporidium muris, Cryptosporidium parvum bovine and mouse genotypes, one C. serpentis-like parasite in a lizard, two new Cryptosporidium spp. in snakes, and one new Cryptosporidium sp. in tortoises. C. serpentis and the desert monitor genotype were the most common parasites and were found in both snakes and lizards, whereas the C. muris and C. parvum parasites detected were probably the result of ingestion of infected rodents. Sequence and biologic characterizations indicated that the desert monitor genotype was Cryptosporidium saurophilum. Two host-adapted C. serpentis genotypes were found in snakes and lizards.

scholarly journals Genetic Characterization of Cryptosporidium cuniculus from Rabbits in Egypt

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 775
Author(s):  
Doaa Naguib ◽  
Dawn M. Roellig ◽  
Nagah Arafat ◽  
Lihua Xiao
Keyword(s):  
Rflp Analysis ◽  
Small Subunit ◽  
Rrna Gene ◽  
Small Subunit Rrna ◽  
Genetic Characteristics ◽  
Glycoprotein Gene ◽  
Cryptosporidium Spp ◽  
Small Subunit Rrna Gene ◽  
Pcr Rflp

Rabbits are increasingly farmed in Egypt for meat. They are, however, known reservoirs of infectious pathogens. Currently, no information is available on the genetic characteristics of Cryptosporidium spp. in rabbits in Egypt. To understand the prevalence and genetic identity of Cryptosporidium spp. in these animals, 235 fecal samples were collected from rabbits of different ages on nine farms in El-Dakahlia, El-Gharbia, and Damietta Provinces, Egypt during the period from July 2015 to April 2016. PCR-RFLP analysis of the small subunit rRNA gene was used to detect and genotype Cryptosporidium spp. The overall detection rate was 11.9% (28/235). All 28 samples were identified as Cryptosporidium cuniculus. The 16 samples successfully subtyped by the sequence analysis of the partial 60 kDa glycoprotein gene belonged to two subtypes, VbA19 (n = 1) and VbA33 (n = 15). As C. cuniculus is increasingly recognized as a cause of human cryptosporidiosis, Cryptosporidium spp. in rabbits from Egypt have zoonotic potential.

scholarly journals Genetic Diversity of Cryptosporidium in Bactrian Camels (Camelus bactrianus) in Xinjiang, Northwestern China

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 946
Author(s):  
Yangwenna Cao ◽  
Zhaohui Cui ◽  
Qiang Zhou ◽  
Bo Jing ◽  
Chunyan Xu ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Sequence Analysis ◽  
Small Subunit ◽  
Cryptosporidium Species ◽  
Northwestern China ◽  
Rrna Gene ◽  
Cryptosporidium Spp ◽  
Gene Sequence Analysis ◽  
Bactrian Camels ◽  
Ssu Rrna Gene Sequence

Cryptosporidium species are ubiquitous enteric protozoan pathogens of vertebrates distributed worldwide. The purpose of this study was to gain insight into the zoonotic potential and genetic diversity of Cryptosporidium spp. in Bactrian camels in Xinjiang, northwestern China. A total of 476 fecal samples were collected from 16 collection sites in Xinjiang and screened for Cryptosporidium by PCR. The prevalence of Cryptosporidium was 7.6% (36/476). Six Cryptosporidium species, C. andersoni (n = 24), C. parvum (n = 6), C. occultus (n = 2), C. ubiquitum (n = 2), C. hominis (n = 1), and C. bovis (n = 1), were identified based on sequence analysis of the small subunit (SSU) rRNA gene. Sequence analysis of the gp60 gene identified six C. parvum isolates as subtypes, such as If-like-A15G2 (n = 5) and IIdA15G1 (n = 1), two C. ubiquitum isolates, such as subtype XIIa (n = 2), and one C. hominis isolate, such as Ixias IkA19G1 (n = 1). This is the first report of C. parvum, C. hominis, C. ubiquitum, and C. occultus in Bactrian camels in China. These results indicated that the Bactrian camel may be an important reservoir for zoonotic Cryptosporidium spp. and these infections may be a public health threat in this region.

scholarly journals Identification of Trichomonadid Protozoa from the Bovine Preputial Cavity by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism Typing

2003 ◽  
Vol 15 (4) ◽  
pp. 390-394 ◽  
Author(s):  
Dawn C. Hayes ◽  
Rebecca R. Anderson ◽  
Richard L. Walker
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Rrna Gene ◽  
5.8S Rrna Gene ◽  
5.8S Rrna ◽  
Restriction Fragment Length

Accurate identification of the bovine pathogen Tritrichomonas foetus is sometimes complicated by the presence of other trichomonadid protozoa in clinical samples. A highly specific and reproducible approach for differentiating 3 common types of bovine trichomonadid protozoa found in the bovine preputial cavity, T. foetus, Pentatrichomonas hominis, and a Tetratrichomonas species, was developed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. Universal trichomonadid protozoa primers, TFR1 and TFR2, were used to amplify the 5.8S rRNA gene and internal transcribed spacer regions (ITSRs), and the products were digested with the restriction enzyme HpyCH4IV. Restriction fragment length polymorphism analysis was performed on 55 trichomonad isolates from bovine preputial washing and scraping samples. The RFLP results correlated 100% with 5.8S rRNA gene and ITSR sequence results and PCR results with primers specific for T. foetus. The results of this study demonstrate that PCR and RFLP analysis can be used in lieu of DNA sequencing to identify the specific trichomonadid protozoa isolated from the bovine preputial cavity.

scholarly journals Use of Oxford Nanopore MinION to generate full-length sequences of the Blastocystis small subunit (SSU) rRNA gene

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Jenny G. Maloney ◽  
Aleksey Molokin ◽  
Monica Santin
Keyword(s):  
Genetic Diversity ◽  
Small Subunit ◽  
Full Length ◽  
Reference Sequence ◽  
Rrna Gene ◽  
Gene Sequences ◽  
Oxford Nanopore ◽  
Long Read ◽  
Main Gene ◽  
Enteric Parasites

Abstract Background Blastocystis sp. is one of the most common enteric parasites of humans and animals worldwide. It is well recognized that this ubiquitous protist displays a remarkable degree of genetic diversity in the SSU rRNA gene, which is currently the main gene used for defining Blastocystis subtypes. Yet, full-length reference sequences of this gene are available for only 16 subtypes of Blastocystis in part because of the technical difficulties associated with obtaining these sequences from complex samples. Methods We have developed a method using Oxford Nanopore MinION long-read sequencing and universal eukaryotic primers to produce full-length (> 1800 bp) SSU rRNA gene sequences for Blastocystis. Seven Blastocystis specimens representing five subtypes (ST1, ST4, ST10, ST11, and ST14) obtained both from cultures and feces were used for validation. Results We demonstrate that this method can be used to produce highly accurate full-length sequences from both cultured and fecal DNA isolates. Full-length sequences were successfully obtained from all five subtypes including ST11 for which no full-length reference sequence currently exists and for an isolate that contained mixed ST10/ST14. Conclusions The suitability of the use of MinION long-read sequencing technology to successfully generate full-length Blastocystis SSU rRNA gene sequences was demonstrated. The ability to produce full-length SSU rRNA gene sequences is key in understanding the role of genetic diversity in important aspects of Blastocystis biology such as transmission, host specificity, and pathogenicity.

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