scholarly journals Detection of Pneumocystis in lungs of bats from Brazil by PCR amplification

2009 ◽  
Vol 29 (6) ◽  
pp. 469-473 ◽  
Author(s):  
Edna Maria Cavallini Sanches ◽  
Susi M. Pacheco ◽  
Alison S. Cericatto ◽  
Rosane M. Melo ◽  
Edson Molleta Colodel ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Urban Areas ◽  
Pcr Amplification ◽  
Small Subunit ◽  
Rrna Gene ◽  
Wild Animals ◽  
Tadarida Brasiliensis ◽  
Desmodus Rotundus ◽  
Mato Grosso ◽  
Wide Range

Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3% = 5/19), Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis (2/3, 66.7%)and Diphylla ecaudata (1/2, 50%). PCR products which could indicate the presence of Pneumocystis (21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the first report of detection of Pneumocystis in bats from Brazil.

scholarly journals Genetic characterization of Blastocystis from wild animals in Sichuan Wolong National Natural Reserve, southwestern of China-Zoonotic potential

2021 ◽  
Author(s):  
Shanyu Chen ◽  
Wangyu Meng ◽  
Ziyao Zhou ◽  
Lei Deng ◽  
Xiaogang Shi ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Human Infection ◽  
Rrna Gene ◽  
Zoonotic Potential ◽  
Wild Animals ◽  
Genetic Characteristics ◽  
Natural Reserve ◽  
Faecal Samples ◽  
Wide Range

Abstract Background Blastocystis, a highly prevalent eukaryotic parasite, has been identified in a wide range of hosts, including humans, domestic and wild animals. Many animals are potential sources of Blastocystis infection for humans, while few information about the prevalence of Blastocystis in wild animals have being documented. Therefore, the present study was designed to investigate the prevalence and subtypes of Blastocystis in wild animals of Sichuan Wolong National Natural Reserve, southwestern of China, so as to assess the zoonotic potential of these animals. Methods A total of 300 faecal samples were collected from 27 wildlife species in three areas of Sichuan Wolong National Natural Reserve in southwestern China. The subtype (ST) genetic characteristics and prevalence of Blastocystis were determined by PCR amplification of the barcode region (a fragment of ∼600 bp) of the SSU rRNA gene, and phylogenetic analysis were further performed to determine the genetic characteristics of Blastocystis subtypes. Results 30 of 300 faecal samples (10.0%) were Blastocystis-positive. The highest prevalence of Blastocystis was found in Yinchanggou (18.3%), which was significantly higher than that in Niutoushan (7.5%), and Genda (5.5%) (P < 0.05). Specifically, the highest prevalence of Blastocystis was found in primates (20.0%, 1/5), followed by rodentia 14.3% (1/7), artiodactyla 13.1% (26/198), carnivora 2.3% (2/87), galliformes 0% (0/3). Sequence analysis showed 5 subtypes (ST1, ST3, ST5, ST13, and ST14), with ST13 and ST14 as the predominant subtype (33.3%, 10/30), followed by ST1 (20.0%, 6/30). Conclusions To the best of our knowledge, this is the first molecular investigation on Blastocystis infection in wild animals in southwestern of China. ST1, ST3, and ST5 were identified in both humans and wild animals, suggesting that these wild animals may be potential reservoirs of Blastocystis for human infection.

scholarly journals Prevalence and Genetic Characterization of Cryptosporidium Infection in Java Sparrows (Lonchura oryzivora) in Northern China

2017 ◽  
Vol 2017 ◽  
pp. 1-4 ◽  
Author(s):  
Qiu-Xia Yao ◽  
Xiao-Xuan Zhang ◽  
Kai Chen ◽  
Jian-Gang Ma ◽  
Wen-Bin Zheng ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Northern China ◽  
Small Subunit ◽  
Rrna Gene ◽  
Ssu Rrna ◽  
Ssu Rrna Gene ◽  
Wide Range ◽  
Baseline Information ◽  
And Control

Cryptosporidiosis is a cosmopolitan parasitosis that affects a wide range of hosts including birds. As information concerning Cryptosporidium in birds is limited, the present study examined the prevalence and genotypes of Cryptosporidium in Java sparrows in Beijing and Shangqiu, northern China. Three hundred and fifty fecal samples were collected from Java sparrows (Lonchura oryzivora, 225 white Java sparrows and 125 gray Java sparrows) in Beijing and Shangqiu in October 2015, and the samples were examined by PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene. The overall Cryptosporidium prevalence is 13.42% (47/350), with 16.44% (37/225) in white Java sparrows and 8.00% (10/125) in gray Java sparrows. Cryptosporidium prevalence was 9.82% (16/163) in Java sparrows from Beijing and 16.58% (31/187) in Java sparrows from Shangqiu. The prevalence of Cryptosporidium in females and males was 40.63% (26/64) and 7.34% (21/286), respectively. The Cryptosporidium prevalence in Java sparrows of different ages varied from 10.47% to 16.33%. Sequence analysis of the SSU rRNA gene revealed that all the samples represented C. baileyi. This is the first report of Cryptosporidium in gray Java sparrows in China, which extend the host range for C. baileyi. These results provide baseline information for further studies of molecular epidemiology and control of Cryptosporidium infection in poultry in China.

scholarly journals Genetic characterization of Blastocystis from wild animals in Sichuan Wolong National Natural Reserve, southwestern of China-Zoonotic potential

2021 ◽  
Author(s):  
Shanyu Chen ◽  
Wangyu Meng ◽  
Ziyao Zhou ◽  
Lei Deng ◽  
Xiaogang Shi ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Human Infection ◽  
Rrna Gene ◽  
Zoonotic Potential ◽  
Wild Animals ◽  
Genetic Characteristics ◽  
Natural Reserve ◽  
Faecal Samples ◽  
Wide Range

Abstract Background: Blastocystis, a highly prevalent eukaryotic parasite, has been identified in a wide range of hosts, including humans, domestic and wild animals. Many animals are potential sources of Blastocystis infection for humans, while few information about the prevalence of Blastocystis in wild animals have being documented. Therefore, the present study was designed to investigate the prevalence and subtypes of Blastocystis in wild animals of Sichuan Wolong National Natural Reserve, southwestern of China, so as to assess the zoonotic potential of these animals.Methods: A total of 300 faecal samples were collected from 27 wildlife species in three areas of Sichuan Wolong National Natural Reserve in southwestern China. The subtype (ST) genetic characteristics and prevalence of Blastocystis were determined by PCR amplification of the barcode region (a fragment of ∼600 bp) of the SSU rRNA gene, and phylogenetic analysis were further performed to determine the genetic characteristics of Blastocystis subtypes.Results: 30 of 300 faecal samples (10.0%) were Blastocystis-positive. The highest prevalence of Blastocystis was found in Yinchanggou (18.3%), which was significantly higher than that in Niutoushan (7.5%), and Genda (5.5%) (P < 0.05). Specifically, the highest prevalence of Blastocystis was found in primates (20.0%, 1/5), followed by rodentia 14.3% (1/7), artiodactyla 13.1% (26/198), carnivora 2.3% (2/87), galliformes 0% (0/3). Sequence analysis showed 5 subtypes (ST1, ST3, ST5, ST13, and ST14), with ST13 and ST14 as the predominant subtype (33.3%, 10/30), followed by ST1 (20.0%, 6/30).Conclusions: To the best of our knowledge, this is the first molecular investigation on Blastocystis infection in wild animals in southwestern of China. ST1, ST3, and ST5 were identified in both humans and wild animals, suggesting that these wild animals may be potential reservoirs of Blastocystis for human infection.

scholarly journals Genetic characterization and zoonotic potential of Blastocystis from wild animals in Sichuan Wolong National Natural Reserve, Southwest China

Parasite ◽  
2021 ◽  
Vol 28 ◽  
pp. 73
Author(s):  
Shanyu Chen ◽  
Wanyu Meng ◽  
Ziyao Zhou ◽  
Lei Deng ◽  
Xiaogang Shi ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Rrna Gene ◽  
Zoonotic Potential ◽  
Wild Animals ◽  
Genetic Characteristics ◽  
Fecal Samples ◽  
Wildlife Species ◽  
Natural Reserve ◽  
Wide Range ◽  
In The Wild

Blastocystis is a prevalent eukaryotic parasite that has been identified in a wide range of hosts. Several species are considered potential sources of Blastocystis infection in humans, but little is known about the prevalence of Blastocystis in wild animals. In this study, the prevalence and subtypes of Blastocystis were investigated to assess the zoonotic potential of wild animals in Sichuan Wolong National Natural Reserve. A total of 300 fecal samples were collected from 27 wildlife species in three areas of the Reserve. The subtype (ST), genetic characteristics, and prevalence of Blastocystis were determined by PCR amplification of part (~600 bp) of the SSU rRNA gene. Thirty fecal samples (10.0%) were Blastocystis-positive. The highest prevalence of Blastocystis was found in Yinchanggou (18.3%), with significantly less found in Niutoushan (7.5%) and Genda (5.5%) (p < 0.05). No significant differences were associated with different orders of animals in prevalence, which may be because of the small number of positive samples obtained. Sequence analysis showed five subtypes (ST1, ST3, ST5, ST13, and ST14), with ST13 and ST14 being predominant (33% each), followed by ST1 (20%). This is the first molecular investigation of Blastocystis infection in the wild animals of southwestern China. Subtypes ST1, ST3, ST5, and ST14 have previously been identified in humans, suggesting that wild animals may be potential reservoirs of Blastocystis for humans.

scholarly journals Molecular Subtyping of Blastocystis sp. Isolated from Farmed Animals in Southern Italy

2021 ◽  
Vol 9 (8) ◽  
pp. 1656
Author(s):  
Simona Gabrielli ◽  
Marialetizia Palomba ◽  
Federica Furzi ◽  
Emanuele Brianti ◽  
Gabriella Gaglio ◽  
...  
Keyword(s):  
Current Knowledge ◽  
Pcr Amplification ◽  
Epidemiological Studies ◽  
Fallow Deer ◽  
Small Subunit ◽  
Zoonotic Transmission ◽  
Ssu Rrna ◽  
Molecular Approaches ◽  
Wide Range ◽  
Blastocystis Sp

Blastocystis is a common intestinal protist distributed worldwide, infecting humans and a wide range of domestic and wild animals. It exhibits an extensive genetic diversity and, so far, 25 distinct small subunit ribosomal RNA (SSU rRNA) lineages termed subtypes (STs)) have been characterized; among them, 12 have thus far been reported in humans. The aims of the present study were to detect and genetically characterize Blastocystis sp. in synantropic animals to improve our current knowledge on the distribution and zoonotic transmission of Blastocystis STs in Italy. Samples were collected from N = 193 farmed animals and submitted to DNA extraction and PCR amplification of the SSU rRNA. Blastocystis was detected in 60 samples (31.08%) and successfully subtyped. Phylogenetic analysis evidenced that the isolates from fallow deer, goats, and pigs (N = 9) clustered within the ST5; those from pheasants (N = 2) in the ST6; those from chickens (N = 8) in the ST7; those from sheep (N = 6) in the ST10; and those from water buffaloes (N = 9) in the ST14 clade. The comparison between the present isolates from animals and those previously detected in humans in Italy suggested the animal-to-human spillover for ST6 and ST7. The present study represents the widest Blastocystis survey performed thus far in farmed animals in Italy. Further epidemiological studies using molecular approaches are required to determine the occurrence and distribution of Blastocystis STs in other potential animal reservoirs in Italy and to define the pathways of zoonotic transmission.

scholarly journals Quantitative Measure of Small-Subunit rRNA Gene Sequences of the Kingdom Korarchaeota

1998 ◽  
Vol 64 (12) ◽  
pp. 5064-5066 ◽  
Author(s):  
Clifford F. Brunk ◽  
Nicole Eis
Keyword(s):  
Internal Standard ◽  
Pcr Amplification ◽  
Quantitative Measure ◽  
Pcr Primers ◽  
Small Subunit ◽  
Ssu Rdna ◽  
Rdna Sequence ◽  
Rrna Gene ◽  
Small Subunit Rrna ◽  
Rdna Sequences

ABSTRACT Comparative PCR amplification of small-subunit (SSU) rRNA gene (rDNA) sequences indicates substantial preferential PCR amplification of pJP27 sequences with korarchaeote-specific PCR primers. The coamplification of a modified SSU rDNA sequence can be used as an internal standard to determine the amount of a specific SSU rDNA sequence.

scholarly journals Molecular identification and subtyping of Blastocystis sp. in laboratory rats in China

Parasite ◽  
2020 ◽  
Vol 27 ◽  
pp. 35
Author(s):  
Junqiang Li ◽  
Yueyue Yuan ◽  
Yuxi Jiang ◽  
Wen Wang ◽  
Liqin Chao ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Small Subunit ◽  
Laboratory Animals ◽  
Rrna Gene ◽  
Sprague Dawley ◽  
Laboratory Rearing ◽  
Spontaneously Hypertensive ◽  
Laboratory Rats ◽  
Oral Transmission ◽  
Blastocystis Sp

Blastocystis sp. is a ubiquitous protist that has been frequently reported in humans and animals worldwide. A total of 355 fecal samples of experimental rats were collected from four laboratory rearing facilities in China, and Blastocystis sp. was detected by PCR amplification of the partial small subunit ribosomal (SSU) rRNA gene. Twenty-nine (8.2%, 29/355) samples were positive for Blastocystis sp., with the highest infection rate (20.7%, 24/116) in rats of the Zhengzhou1, followed by that in the Zhengzhou2 (5.0%, 2/40), Shenyang (3.0%, 3/100) and Wuhan (0) rearing facilities. Among the three rat strains, Sprague–Dawley (SD) rats had higher infection rates (11.3%, 17/151) compared to Wistar rats (8.7%, 9/104) and spontaneously hypertensive (SH) rats (3.0%, 3/100). Two Blastocystis sp. subtypes (ST4 and ST7) were identified. ST4 was the predominant subtype detected in 26 samples (89.7%). A phylogenetic analysis demonstrated that the sequences of ST4 and ST7 obtained in this study were clustered with their reference subtypes. To our knowledge, this is the first report of Blastocystis sp. in experimental rats in China. Pathogen infections in laboratory animals need to be monitored due to fecal-oral transmission.

scholarly journals Cryptosporidium species and subtypes in diarrheal children and HIV-infected persons in Ebonyi and Nsukka, Nigeria

2017 ◽  
Vol 11 (02) ◽  
pp. 173-179 ◽  
Author(s):  
Boniface Nwofoke Ukwah ◽  
Ifeoma Maureen Ezeonu ◽  
Chinonyelum Thecla Ezeonu ◽  
Dawn Roellig ◽  
Lihua Xiao
Keyword(s):  
Species Distribution ◽  
Highly Active Antiretroviral Therapy ◽  
Urban Areas ◽  
Age Groups ◽  
Small Subunit ◽  
Cryptosporidium Species ◽  
Common Disease ◽  
Rrna Gene ◽  
Highly Active ◽  
Cell Counts

Introduction: Cryptosporidiosis is a common disease of children and immune-compromised persons. This study evaluated the diversity and distribution of Cryptosporidium species in diarrheal children and HIV-infected persons on highly active antiretroviral therapy (HAART) and those not on HAART. Methodology: A total of 394 fecal specimens were collected from patients attending clinics in Nsukka and Ebonyi, Nigeria. Detection and identification of Cryptosporidium species were conducted by PCR-RFLP of the small subunit (SSU) rRNA gene, whereas subtyping was done by sequence analysis of the 60 kDa glycoprotein (gp60) gene. Results: Twenty-five (6.3%) specimens yielded four Cryptosporidium species, including C. hominis, C. parvum, C. felis, and C. viatorum. C. hominis was the most dominant species with 48.0% occurrence and three identified subtype families: Ia (six specimens), Ib (three specimens), Ie (two specimens), and one un-subtyped species. C. parvum had 44.0% occurrence and two subtype families: IIc (eight specimens) and IIe (three specimens), while C. felis and C. viatorum each had 4.0% occurrence. There were significant differences in Cryptosporidium species distribution between age groups in children and HIV-infected persons, between suburban and urban areas, and between low and high CD4+ cell counts in HIV-infected patients. There were no significant differences in infection rate and species distribution between HIV-infected patients on HAART and those not on HAART. Conclusions: The results from this study show that there is a high diversity of Cryptosporidium spp. in humans in Ebonyi and Nsukka, Nigeria, and that all the C. parvum subtypes identified are most likely anthroponotic in origin.

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.

Detection of filamentous genus Gordonia in foam samples using genus-specific primers combined with PCR – denaturing gradient gel electrophoresis analysis

2007 ◽  
Vol 53 (6) ◽  
pp. 768-774 ◽  
Author(s):  
Fo-Ting Shen ◽  
Hsuan-Ru Huang ◽  
A.B. Arun ◽  
Hui-Ling Lu ◽  
Ta-Chen Lin ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Nested Pcr ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Wastewater Treatment Plants ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Gradient Gel Electrophoresis

A nested-PCR amplification combined with denaturing gradient gel electrophoresis (PCR–DGGE) approach was used to detect and identify Gordonia populations from wastewater treatment plant foam samples. The PCR-amplified region (position 722–1119) by specifically designed primers G699F and G1096R covered the hypervariable region of the Gordonia 16S rRNA gene sequence. This approach successfully distinguished Gordonia species to the interspecies level. The differential ability of PCR–DGGE analysis was effectively used to separate 12 Gordonia species belonging to different 16S rRNA gene-based phylogenetic lineages into 8 groups. Based on this method, the minimum limit of Gordonia detection was 5 × 104CFU·g–1in the seeded soil samples. The PCR–DGGE bands obtained were excised and identified by sequence analysis. Gordonia polyisoprenivorans , Gordonia amicalis , DGGE type II Gordonia species, and an uncertain Gordonia species dominated the activated sludge foam samples. Results of this study indicate that the detection and analyses of genus Gordonia within a complex microbial community could be accomplished using the PCR–DGGE approach to a larger extent, with certain limitations. Detection of diverse Gordonia populations in foam samples from wastewater treatment plants revealed the significant role of Gordonia in biological foaming during wastewater treatment. The nested-PCR amplification and DGGE can be used as a diagnostic tool for the early detection of foaming incidents in wastewater treatment plants using Gordonia as indicator organism.

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