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.

A new species of Puddle Frog, genus Phrynobatrachus (Amphibia: Anura: Phrynobatrachidae) from Ghana

Zootaxa ◽  
2018 ◽  
Vol 4374 (4) ◽  
pp. 565 ◽  
Author(s):  
CALEB OFORI-BOATENG ◽  
ADAM D. LEACHÉ ◽  
BRIGHT OBENG-KANKAM ◽  
N’GORAN GERMAIN KOUAMÉ ◽  
ANNIKA HILLERS ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
New Species ◽  
Sequence Data ◽  
Face Mask ◽  
West African ◽  
Rrna Gene ◽  
Forest Region ◽  
Mtdna Sequence ◽  
A New Species ◽  
The 16S Rrna Gene

We describe a new species of Phrynobatrachus from the eastern part of the Upper Guinea forest region, Ghana, West Africa. Morphologically, the new species can be distinguished from all of its congeners by the combination of a slender body, short and pointed snout, a relatively warty dorsum, a black-spotted throat in both sexes, a gular flap in males, a dark spotted chest, a white-greyish venter with occasional blackish spots, rudimentary pedal webbing, none to slightly dilated finger tips and strongly delated toe tips, presence of both inner and outer metatarsal tubercles and absence of a dark face mask, eyelid tubercles and longer dorsal ridges. We collected mitochondrial DNA (mtDNA) sequence data from the 16S rRNA gene to measure the genetic diversity of the new species, and to estimate phylogenetic relationships. The new species is a distinct and monophyletic evolutionary lineage most closely related to Phrynobatrachus gutturosus, P. fraterculus and P. maculiventris. The discovery of this new species highlights that the biodiversity of West African forests is still incompletely known and that the few remaining forests need urgent protection. 

A case of feline leprosy caused by Mycobacterium lepraemurium originating from the island of Kythira (Greece): diagnosis and treatment

2007 ◽  
Vol 9 (3) ◽  
pp. 238-241 ◽  
Author(s):  
Francois Courtin ◽  
Michel Huerre ◽  
Janet Fyfe ◽  
Paul Dumas ◽  
Maria L. Boschiroli
Keyword(s):  
Rrna Gene ◽  
Internal Transcribed Spacer Region ◽  
Leukaemia Virus ◽  
Acid Fast Bacilli ◽  
Immunodeficiency Virus ◽  
Feline Leukaemia Virus ◽  
Polymerase Chain ◽  
Mycobacterium Lepraemurium ◽  
Foamy Macrophages ◽  
The 16S Rrna Gene

A 2-year-old, 4 kg, healthy, domestic shorthair female cat presented with ulcerated subcutaneous nodules on the commissures of its mouth. The cat was negative for feline leukaemia virus and feline immunodeficiency virus. Skin mycobacteriosis was diagnosed after detection of numerous acid-fast bacilli in Ziehl Neelsen-stained smears from the ulcers. Feline leprosy was suspected following preliminary polymerase chain reaction results: positive for Mycobacterium genus but negative for Mycobacterium tuberculosis and Mycobacterium avium complexes. Mycobacterium lepraemurium was later identified following DNA sequence analysis of the 5′ end of the 16S rRNA gene and the 16S–23S internal transcribed spacer region. Microscopic lesions consisted of pyogranulomas containing mainly large foamy macrophages with 10–100 intra-cellular acid-fast bacilli per field. The cat was cured after surgery and a 14-week course of clofazimine (30 mg daily) and clarithromycin (50 mg twice daily).

scholarly journals Hematological values associated to the serological and molecular diagnostic in cats suspected of Ehrlichia canisinfection

2013 ◽  
Vol 22 (4) ◽  
pp. 470-474 ◽  
Author(s):  
ísis Assis Braga ◽  
Luana Gabriela Ferreira dos Santos ◽  
Andréia Lima Tomé Melo ◽  
Felipe Wolf Jaune ◽  
Thaysa Felfili Ziliani ◽  
...  
Keyword(s):  
Molecular Diagnostic ◽  
Rrna Gene ◽  
Igg Antibody ◽  
Mato Grosso ◽  
Pcr Products ◽  
Antibody Titers ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Veterinary Hospital ◽  
Hematological Alterations

The literature contains several studies on feline ehrlichiosis. However, information about the characteristics of Ehrlichiainfection in cats is still scanty. This study evaluated the association between Ehrlichia spp. infection and the hematologic data of 93 cats treated at the Federal University of Mato Grosso Veterinary Hospital in Cuiabá, state of Mato Grosso, Brazil. The presence of or exposure to Ehrlichia spp. infection was evaluated by Polymerase Chain Reaction (PCR) targeting the dsb and 16S rRNA gene of Ehrlichia, and by detection of anti-Ehrlichia canis IgG antibodies in Indirect Fluorescence Assay (IFA), respectively. Eight (8.6%) cats tested positive by PCR and the partial DNA sequence obtained from PCR products was a 100% match to E. canis. Forty-two (45.1%) cats showed antibody reactivity against Ehrlichia spp. Hematological alterations such as low erythrocyte count, thrombocytopenia, lymphopenia and monocytosis were observed in PCR positive cats. Among them, low erythrocyte counts were associated with IgG antibody titers of 40 to 640 and five cats also tested positive by PCR. Furthermore, PCR-positive cats showed a tendency to be lymphopenic. No correlation was found between age and sex, and no ticks were observed in any of the examined cats.

scholarly journals Genetic Diversity among Botulinum Neurotoxin-Producing Clostridial Strains

2006 ◽  
Vol 189 (3) ◽  
pp. 818-832 ◽  
Author(s):  
K. K. Hill ◽  
T. J. Smith ◽  
C. H. Helma ◽  
L. O. Ticknor ◽  
B. T. Foley ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Common Property ◽  
Animal Species ◽  
Fragment Length Polymorphism ◽  
Aflp Analysis ◽  
Rrna Gene ◽  
Botulinum Neurotoxins ◽  
The Common ◽  
Multiple Strains ◽  
The 16S Rrna Gene

ABSTRACT Clostridium botulinum is a taxonomic designation for many diverse anaerobic spore-forming rod-shaped bacteria that have the common property of producing botulinum neurotoxins (BoNTs). The BoNTs are exoneurotoxins that can cause severe paralysis and death in humans and other animal species. A collection of 174 C. botulinum strains was examined by amplified fragment length polymorphism (AFLP) analysis and by sequencing of the 16S rRNA gene and BoNT genes to examine the genetic diversity within this species. This collection contained representatives of each of the seven different serotypes of botulinum neurotoxins (BoNT/A to BoNT/G). Analysis of the16S rRNA gene sequences confirmed previous identifications of at least four distinct genomic backgrounds (groups I to IV), each of which has independently acquired one or more BoNT genes through horizontal gene transfer. AFLP analysis provided higher resolution and could be used to further subdivide the four groups into subgroups. Sequencing of the BoNT genes from multiple strains of serotypes A, B, and E confirmed significant sequence variation within each serotype. Four distinct lineages within each of the BoNT A and B serotypes and five distinct lineages of serotype E strains were identified. The nucleotide sequences of the seven toxin genes of the serotypes were compared and showed various degrees of interrelatedness and recombination, as was previously noted for the nontoxic nonhemagglutinin gene, which is linked to the BoNT gene. These analyses contribute to the understanding of the evolution and phylogeny within this species and assist in the development of improved diagnostics and therapeutics for the treatment of botulism.

scholarly journals Detection and molecular characterisation of Ehrlichia canis in naturally infected dogs in South West Nigeria

2018 ◽  
Vol 66 (1) ◽  
pp. 85-95 ◽  
Author(s):  
Olukayode Olugbenga Daramola ◽  
Michael Irewole Takeet ◽  
Ibironke Kofoworola Oyewusi ◽  
Mufutau Atanda Oyekunle ◽  
Adewale Oladele Talabi
Keyword(s):  
Ehrlichia Canis ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Rickettsial Disease ◽  
Blood Samples ◽  
Canine Ehrlichiosis ◽  
South West ◽  
Polymerase Chain ◽  
Reference Sequences ◽  
The 16S Rrna Gene

Canine ehrlichiosis is an important tick-borne rickettsial disease mainly caused by Ehrlichia canis. This study aimed to detect and characterise E. canis in dogs in Abeokuta, Nigeria by microscopy and nested PCR. Blood samples were collected from 205 dogs, thin smears were made, field-stained, and DNA was extracted from the blood samples. A partial region of the 16S rRNA gene was amplified by polymerase chain reaction (PCR) and sequenced unidirectionally. Ehrlichial morulae were detected in three dogs (1.5%). The PCR test revealed that 47 dogs (22.9%) were positive for E. canis. The lengths of the sequences obtained range from 374 bp to 376 bp with an average G-C content of 37% and 98–99% homology with the reference sequences in GenBank. The aligned autochthonous sequences were less polymorphic. The phylogenetic analysis separated sequences reported previously in Nigeria from the autochthonous sequences. The present work shows that the strain of E. canis detected in the study area is genetically different from those reported in the northern part of Nigeria and more closely related to sequences from Brazil and India.

scholarly journals Identification of Mycoplasma genitalium from clinical swabs by direct PCR

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 1993
Author(s):  
Robinson M. Irekwa ◽  
Perpetual Ndung'u ◽  
Peter Kipkemboi ◽  
Tonny Teya ◽  
Anne Wanjiru Mwangi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sex Workers ◽  
Genital Tract ◽  
Mycoplasma Genitalium ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Direct Pcr ◽  
The World ◽  
Polymerase Chain ◽  
The 16S Rrna Gene

Mycoplasma genitalium is one of the smallest self-replicating organisms. It is an obligate parasite found in the human genital tract. In men, the bacteria cause both acute and chronic non-gonococcal urethritis (NGU). In women, it has been associated with pelvic inflammatory disease and cervicitis among other related infections. Treatment of M. genitalium related infections has been effective using antibiotics such as the macrolides (e.g. azithromycin) and fluoroquinolones. However, there have been recorded cases of resistance to these antibiotics in various parts of the world as a result of a mutation in the 23SrRNA gene, although the antibiotic resistance has not been well established. The aim of this study was to detect M. genitalium in 352 swab samples collected from a clinic for sex workers in Nairobi, Kenya. DNA was extracted from the swabs and stored as a crude extract at -31°C. The swab lysates were subjected to direct polymerase chain reaction using primers that specifically target the 16S rRNA gene for M. genitalium. A total of 29 samples tested positive for M. genitalium. The data results showed a M. genitalium prevalence of 8.24% among sex workers in Nairobi, Kenya.

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.

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