scholarly journals Detection of Anaplasma sp. phylogenetically related to A. phagocytophilum in a free-living bird in Brazil

2017 ◽  
Vol 26 (4) ◽  
pp. 505-510 ◽  
Author(s):  
Anna Claudia Baumel Mongruel ◽  
Jyan Lucas Benevenute ◽  
Priscila Ikeda ◽  
Marcos Rogério André ◽  
Rosangela Zacarias Machado ◽  
...  
Keyword(s):  
Wild Bird ◽  
Molecular Detection ◽  
Bird Species ◽  
Rrna Gene ◽  
Wild Animals ◽  
Domestic Cats ◽  
Free Living ◽  
Likelihood Analysis ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

Abstract Wild animals play an important role in carrying vectors that may potentially transmit pathogens. Several reports highlighted the participation of wild animals on the Anaplasma phagocytophilum cycle, including as hosts of the agent. The aim of this study was to report the molecular detection of an agent phylogenetically related to A. phagocytophilum isolated from a wild bird in the Midwest of the state of Paraná, Brazil. Fifteen blood samples were collected from eleven different bird species in the Guarapuava region. One sample collected from a Penelope obscura bird was positive in nested PCR targeting the 16S rRNA gene of Anaplasma spp. The phylogenetic tree based on the Maximum Likelihood analysis showed that the sequence obtained was placed in the same clade with A. phagocytophilum isolated from domestic cats in Brazil. The present study reports the first molecular detection of a phylogenetically related A. phagocytophilum bacterium in a bird from Paraná State.

scholarly journals Chlamydiales and hemotropic mycoplasma in captive and free-living bats

2020 ◽  
Author(s):  
Janine Fritschi ◽  
Hanna Marti ◽  
Helena M.B. Seth-Smith ◽  
Sébastien Aeby ◽  
Gilbert Greub ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Rrna Gene ◽  
Free Living ◽  
Hemotropic Mycoplasma ◽  
Total Prevalence ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

Abstract Background: Bats are hosts for a variety of microorganisms, however, little is known about the presence of Chlamydiales and hemotropic mycoplasma. This study investigated 475 free-living and captive bats from Switzerland, Germany and Costa Rica for the occurrence of Chlamydiales and hemotropic mycoplasma.Results: Screening for Chlamydiales was performed using a Chlamydiaceae-specific real-time PCR targeting the 23S rRNA gene and a pan-Chlamydiales PCR targeting the 16S rRNA gene resulting in a total prevalence of 31.4%. For sequencing, a PCR with the specifically designed inner primers panFseq and panRseq was performed, and criteria published by Pillonel et al. were used to classify the 19 obtained sequences, resulting in the formation of two groups. Groups one and two shared sequence identities to Chlamydiaceae and to Chlamydia-like organisms, including Rhabdochlamydiaceae and unclassified Chlamydiales from environmental samples, respectively.Analysis for the presence of hemotropic mycoplasma was performed using a universal SYBR Green hemoplasma screening real-time PCR targeting the 16S rRNA gene, real-time PCRs specific for M. haemofelis-like and 'Candidatus M. haemominutum'-like organisms and two conventional PCRs targeting an 871-bp and 1030-bp region of the 16S rRNA gene resulting in a total prevalence of 0.7%. Sequencing and phylogenetic analysis of the 871-bp and 1030-bp region of the 16S rRNA gene were used to classify positive specimens and infer their phylogenetic relationships. Three sequences with identities to other unidentified mycoplasma found in vampire bats and Chilean bats were obtained.Conclusions: Bats can harbor Chlamydiales and hemotropic mycoplasma and the newly described sequences in this study indicate that the diversity of these bacteria in bats is much larger than thought before. Both, Chlamydiales and hemotropic mycoplasmas are not restricted to certain bat species or countries and free-living as well as captive bats can be colonized. In conclusion, bats represent another potential host or vector for novel, previously unidentified, Chlamydiales and hemotropic mycoplasma.

scholarly journals Molecular Detection of Novel Borrelia Species, Candidatus Borrelia javanense, in Amblyomma javanense Ticks from Pangolins

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 728
Author(s):  
Bao-Gui Jiang ◽  
Ai-Qiong Wu ◽  
Jia-Fu Jiang ◽  
Ting-Ting Yuan ◽  
Qiang Xu ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Molecular Detection ◽  
Southern China ◽  
Natural Circulation ◽  
Rrna Gene ◽  
The Novel ◽  
Flagellin Gene ◽  
Manis Javanica ◽  
The 16S Rrna Gene ◽  
Novel Agent

A novel Borrelia species, Candidatus Borrelia javanense, was found in ectoparasite ticks, Amblyomma javanense, from Manis javanica pangolins seized in anti-smuggling operations in southern China. Overall, 12 tick samples in 227 (overall prevalence 5.3%) were positive for Candidatus B. javanense, 9 (5.1%) in 176 males, and 3 (5.9%) in 51 females. The phylogenetic analysis, based on the 16S rRNA gene and the flagellin gene sequences of the Borrelia sp., exhibited strong evidence that Candidatus B. javanense did not belong to the Lyme disease Borrelia group and the relapsing fever Borrelia group but another lineage of Borrelia. The discovery of the novel Borrelia species suggests that A. javanense may be the transmit vector, and the M. javanica pangolins should be considered a possible origin reservoir in the natural circulation of these new pathogens. To our knowledge, this is the first identification of a novel Borrelia species agent in A. javanense from pangolins. Whether the novel agent is pathogenic to humans is unknown and needs further research.

scholarly journals Genetic diversity of Ehrlichia canisstrains from naturally infected dogs in Rio de Janeiro, Brazil

2014 ◽  
Vol 23 (3) ◽  
pp. 301-308 ◽  
Author(s):  
Renata Fernandes Ferreira ◽  
Aloysio de Mello Figueiredo Cerqueira ◽  
Tatiana Xavier de Castro ◽  
Eliane de Oliveira Ferreira ◽  
Felipe Piedade Gonçalves Neves ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Rio De Janeiro ◽  
Complete Sequence ◽  
Rrna Gene ◽  
Hybrid Strain ◽  
The Us ◽  
Polymerase Chain ◽  
Hematological Manifestations ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of 1the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.

scholarly journals Molecular Detection of Borrelia Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity

Diagnostics ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 2088
Author(s):  
Malin Lager ◽  
Peter Wilhelmsson ◽  
Andreas Matussek ◽  
Per-Eric Lindgren ◽  
Anna J. Henningsson
Keyword(s):  
Cerebrospinal Fluid ◽  
16S Rrna ◽  
Clinical Diagnostics ◽  
Systematic Evaluation ◽  
Clinical Samples ◽  
Rrna Gene ◽  
Analytical Sensitivity ◽  
Gene 16S Rrna ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

The main tools for clinical diagnostics of Lyme neuroborreliosis (LNB) are based on serology, i.e., detection of antibodies in cerebrospinal fluid (CSF). In some cases, PCR may be used as a supplement, e.g., on CSF from patients with early LNB. Standardisation of the molecular methods and systematic evaluation of the pre-analytical handling is lacking. To increase the analytical sensitivity for detection of Borrelia bacteria in CSF by PCR targeting the 16S rRNA gene, parameters were systematically evaluated on CSF samples spiked with a known amount of cultured Borrelia bacteria. The results showed that the parameters such as centrifugation time and speed, the use of complementary DNA as a template (in combination with primers and a probe aiming at target gene 16S rRNA), and the absence of inhibitors (e.g., erythrocytes) had the highest impact on the analytical sensitivity. Based on these results, a protocol for optimised handling of CSF samples before molecular analysis was proposed. However, no clinical evaluation of the proposed protocol has been done so far, and further investigations of the diagnostic sensitivity need to be performed on well-characterised clinical samples from patients with LNB.

scholarly journals Genotype diversity of Mycoplasma Hyopneumoniae in Chinese swine herds based on multilocus sequence typing

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Hui Zhang ◽  
Yuanyuan Wang ◽  
Lu Gao ◽  
Yan Wang ◽  
Rong Wei
Keyword(s):  
Multilocus Sequence Typing ◽  
Clinical Signs ◽  
Mycoplasma Hyopneumoniae ◽  
Rrna Gene ◽  
Genotype Diversity ◽  
Swine Diseases ◽  
Swine Herds ◽  
Pcr Targeting ◽  
Sequence Types ◽  
The 16S Rrna Gene

Abstract Background Between 2018 and 2020, 989 clinical specimens from pigs showing clinical signs of a variety of swine diseases in 27 provinces in China were sampled and submitted for further testing. Nested PCR targeting the 16S rRNA gene of Mycoplasma hyopneumoniae and subsequent sequencing were used to analyse these specimens. Mycoplasma hyopneumoniae-positive samples were assayed by multilocus sequence typing (MLST). The aim of the study was to reveal the distribution of M. hyopneumoniae and determine the genotypes of M. hyopneumoniae in pig herds in China based on MLST. Results Among these 989 samples, 199 samples were M. hyopneumoniae-positive. The M. hyopneumoniae positivity rate was 7.2% (35/494) in 2018, 18.4% (38/207) in 2019, and 43.8% (126/288) in 2020. In total, 47 samples were successfully assayed by MLST. Sixteen new M. hyopneumoniae sequence types from 9 provinces were recorded in the present study. Conclusions This is the first report on sample positivity rates and molecular typing results for M. hyopneumoniae in swine herds in China. MLST has revealed high genotype diversity among M. hyopneumoniae from different provinces of China.

Detection of Mycoplasma galliseptium infection in chickens from Tamil Nadu State of India

2018 ◽  
Author(s):  
K. Manimaran ◽  
Adarsh Mishra ◽  
S. Hemalatha ◽  
K. Karthik ◽  
P.I. Ganesan
Keyword(s):  
Tamil Nadu ◽  
Molecular Detection ◽  
Age Groups ◽  
Chronic Respiratory Disease ◽  
Rrna Gene ◽  
Histopathological Changes ◽  
Tissue Samples ◽  
Air Sacs ◽  
Tamil Nadu State ◽  
Pcr Targeting

Mycoplasma gallisepticum (MG) is one of the major respiratory tract pathogens affecting chickens. It causes Chronic Respiratory Disease (CRD) among chickens of various age groups. The present study describes the isolation, molecular detection and histopathological changes associated with CRD in chickens. A total of 790 samples viz., trachea, lungs and air sacs were collected from chickens showing the symptoms of CRD from different parts of Tamil Nadu state. All the samples were processed for isolation and molecular detection of MG. A total of 91 samples were found positive by isolation and 105 samples were found positive through MG specific PCR targeting 16S rRNA gene. The histopathological changes in tissue samples of trachea, sinuses, air sacs and lungs collected from naturally infected M. gallisepticum infection were suggestive of subacute to chronic nature of infection. Though isolation is considered to be a gold standard, still PCR is a rapid, sensitive and cheap method for early diagnosis of MG which can help poultry farmers to avoid severe economic loss.

scholarly journals Molecular detection of Anaplasmataceae agents in Dasyprocta azarae in northeastern Brazil

2018 ◽  
Vol 27 (1) ◽  
pp. 98-104 ◽  
Author(s):  
Maria do Socorro Costa de Oliveira Braga ◽  
José Gomes Pereira ◽  
Simone de Jesus Fernandes ◽  
Ingrid Carolinne Lopes Marques ◽  
Renata Passos de Jesus ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Detection ◽  
Evolutionary Model ◽  
Northeastern Brazil ◽  
Likelihood Method ◽  
Rrna Gene ◽  
Nested Pcr Assays ◽  
Tick Vectors ◽  
The 16S Rrna Gene

Abstract Recently, the importance of wild-living rodents for maintenance of pathogens of the family Anaplasmataceae in the environment was investigated. These mammals play a role as reservoirs for these pathogens and act as hosts for the immature stages of tick vectors. The aim of the present study was to investigate the prevalence of Ehrlichia sp. and Anaplasma sp. in 24 specimens of Azara’s agouti (Dasyprocta azarae) that had been trapped in the Itapiracó Environmental Reserve, in São Luís, Maranhão, northeastern Brazil, using molecular methods. Four animals (16.7%) were positive for Ehrlichia spp. in nested PCR assays based on the 16S rRNA gene. In a phylogenetic analysis based on the 16S rRNA gene, using the maximum likelihood method and the GTRGAMMA+I evolutionary model, Ehrlichia sp. genotypes detected in Azara’s agoutis were found to be closely related to E. canis and to genotypes relating to E. canis that had previously been detected in free-living animals in Brazil. The present work showed the first molecular detection of Ehrlichia sp. in Azara’s agoutis in Brazil.

scholarly journals Antimicrobial susceptibility profiles of Staphylococcus spp. from domestic and wild animals

2016 ◽  
Vol 46 (12) ◽  
pp. 2148-2151 ◽  
Author(s):  
Isabela de Godoy ◽  
Danny Franciele da Silva Dias Moraes ◽  
Letícia Camara Pitchenin ◽  
Janaina Marcela Assunção Rosa ◽  
Francielle Cristina Kagueyama ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Rrna Gene ◽  
Wild Animals ◽  
Species Determination ◽  
Disk Diffusion Test ◽  
Susceptibility Profiles ◽  
Staphylococcus Spp ◽  
The 16S Rrna Gene

ABSTRACT: The aim of this study was to determine the prevalence and diversity of veterinary clinical isolates of Staphylococcus and analyze their antimicrobial susceptibility. One hundred Staphylococcus spp. clinical isolates from domestic and wild animals were subjected to partial sequencing of the 16S rRNA gene to species determination. Antimicrobial susceptibility was obtained by a disk diffusion test against six antibiotics: amoxicillin (AMX), cephalexin (LEX), ciprofloxacin (CIP), erythromycin (ERY), gentamicin (GEN) and trimethoprim-sulfamethoxazole (SXT). The most common specie was S. pseudintermedius (61%, 61/100) and resistance to ERY (57%, 57/100), SXT (50%, 50/100) and AMX (46%, 46/100) was detected most frequently. In total, 40% (40/100) of Staphylococcus spp. exhibited a multidrug-resistant (MDR) phenotype. Results of this study emphasize that animals are reservoir of MDR Staphylococcus spp.

scholarly journals Accuracy of Conventional PCR Targeting the 16S rRNA Gene with the Ot-16sRF1 and Ot-16sRR1 Primers for Diagnosis of Scrub Typhus: a Case-Control Study: TABLE 1

2015 ◽  
Vol 54 (1) ◽  
pp. 178-179 ◽  
Author(s):  
Choon-Mee Kim ◽  
Min Keun Cho ◽  
Dong-Min Kim ◽  
Na-Ra Yun ◽  
Seok Won Kim ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Scrub Typhus ◽  
Rrna Gene ◽  
Conventional Pcr ◽  
Content Type ◽  
Increased Sensitivity ◽  
Good Diagnostic Accuracy ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

We retrospectively evaluated the accuracy of conventional PCR targeting the 16S rRNA gene (16S C-PCR) using the Ot-16sRF1/Ot-16sRR1 primers for diagnosing scrub typhus. The diagnosis ofOrientia tsutsugamushiinfection by 16S C-PCR presented an increased sensitivity of 87.0% and specificity of 100% compared with those obtained with other targets and is thus a simple and clinically useful method with good diagnostic accuracy.

scholarly journals The hierarchical system of the ‘Alphaproteobacteria’: description of Hyphomonadaceae fam. nov., Xanthobacteraceae fam. nov. and Erythrobacteraceae fam. nov.

2005 ◽  
Vol 55 (5) ◽  
pp. 1907-1919 ◽  
Author(s):  
Kyung-Bum Lee ◽  
Chi-Te Liu ◽  
Yojiro Anzai ◽  
Hongik Kim ◽  
Toshihiro Aono ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Phylogenetic Tree ◽  
Small Subunit ◽  
Rrna Gene ◽  
Small Subunit Rrna ◽  
Gene Sequences ◽  
16S Rrna Gene Sequences ◽  
Likelihood Analysis ◽  
The 16S Rrna Gene

Phylogenetic analysis of the class ‘Alphaproteobacteria’, including physiologically diverse species, was conducted by using small-subunit rRNA gene sequences. The 16S rRNA gene sequences of 261 species in the class ‘Alphaproteobacteria’ were obtained from GenBank/EMBL/DDBJ for constructing a phylogenetic tree by using maximum-likelihood analysis. In the resulting tree, members of the class ‘Alphaproteobacteria’ were subdivided into five major clusters, which were compared with the taxonomic outline of Bergey's Manual of Systematic Biology and the arb tree. Based on this phylogenetic tree, three novel families are proposed: Hyphomonadaceae fam. nov. to accommodate the bacterial genera Hyphomonas, Hirschia, Maricaulis and Oceanicaulis, Xanthobacteraceae fam. nov. to include the genera Xanthobacter, Azorhizobium, Ancylobacter, Labrys and Starkeya, and Erythrobacteraceae fam. nov. to accommodate the genera Erythrobacter, Porphyrobacter and Erythromicrobium. The phylogenetic tree of 16S rRNA gene sequences established in this study may provide a sound basis for future taxonomic reconstruction of the class ‘Alphaproteobacteria’.

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