scholarly journals COMPARISON OF DIAGNOSTIC EFFECTIVENESS OF METHODS OF DETECTION OF COXIELLA BURNETII IN BLOOD OF PATIENTS WITH Q FEVER BASED ON AMPLIFICATION OF 16S rRNA GENE FRAGMENTS (STANDARD PCR) AND groEL GENE (REALTIME PCR)

2016 ◽  
pp. 70-74
Author(s):  
Yu. A. Panferova ◽  
O. A. Freilikhman ◽  
N. K. Tokarevich ◽  
S. F. Karpenko ◽  
Kh. M. Galimzyanov
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Clinical Material ◽  
Rrna Gene ◽  
Gene Fragment ◽  
Rt Pcr ◽  
Pcr Marker ◽  
Infectious Process

Aim. Comparison of diagnostic capabilities of 2 variants of PCR for detection of Coxiella burnetii persistence in dynamics of infectious process in patients with Q fever. Materials and methods. 110 samples of clinical material, obtained from patients with Q fever in an endemic region for this infection (Astrakhan region), were studied. The samples were studied in a standard PCR (marker - 16S rRNA gene fragment) and in real-time PCR (RT-PCR) (marker - groEL gene fragment). Results. Both markers were established to be perspective for detection of C. burnetii DNA in clinical material, and RT-PCR detects positive result including late stages of the disease (illness day 21 - 31). Conclusion. This study is the first Russian publication on comparison on different PCR variants for detection of C. burnetii in blood of Q fever patients in dynamics of the infectious process.

scholarly journals First isolation and molecular phylogenetic analysis of Coxiella burnetii in lactating cows

2021 ◽  
Vol 24 (4) ◽  
pp. 508-519
Author(s):  
H. A. J. Gharban ◽  
A. A. Yousif
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Q Fever ◽  
Age Groups ◽  
Sampling Period ◽  
Rrna Gene ◽  
Pcr Assay ◽  
Milk Samples ◽  
Lactating Cows ◽  
Specific Pathogen

Q fever is an infectious disease of animals and humans, caused by globally distributed C. burnetii. In Iraq, there are no previous studies associated with the detection of the organism in cattle. An overall of 130 lactating cows were submitted to direct collection of milk samples. Initially, the samples of milk were tested using the molecular polymerase chain reaction (PCR) assay targeting three genes (16S rRNA, IS1111a transposase, and htpB). However, positive results (18.46%; 24/130) were detected only with the 16s rRNA gene. Concerning risk factors, the highest prevalence of C. burnetii was showed in the district of Badra (42.86%), whereas the lowest - in Al-Numaniyah and Al-Suwaira districts (P=0.025). There was no significant variation in positivity between the months of sampling period (P=0.082) and between age groups (P=0.076). Crossbred cows (20.69%) showed a higher positivity than local and pure breeds (P=0.043). Milk of positive samples (n=24) was used for cultivation of C. burnetii into specific pathogen free-embryonated chicken eggs (SPF-ECEs). After three passages into SPF-ECEs, contents of yolk sac were collected, subjected for DNA extraction, and re-tested by PCR assay using the primer of 16s rRNA gene only. Of 24 cultivated milk samples, 12.5% (3/24) were positive for C. burnetii. Finally, the positive local isolates were analysed phylogenetically and reported in NCBI-Genbank under the accession numbers of MN121700.1, MN121701.1, and MN121702.1. In conclusion, this is a unique study as it detected C. burnetii in Iraqi lactating cows, and confirmed that organism was shed actively through milk, suggesting that these animals can play a role as a reservoir for organism with potential risk for transmission of infection from these animals to humans as well as to other animal species.

scholarly journals Phylogenetic position of Odorrana macrotympana (Yang, 2008) (Anura, Ranidae) and extension of its geographical distribution

Herpetozoa ◽  
2022 ◽  
Vol 35 ◽  
pp. 9-19
Author(s):  
Shuo Liu ◽  
Dingqi Rao ◽  
Dongru Zhang ◽  
Ye Htet Lwin ◽  
Mingzhong Mo ◽  
...  
Keyword(s):  
16S Rrna ◽  
Molecular Phylogeny ◽  
16S Rrna Gene ◽  
Geographical Distribution ◽  
New Records ◽  
First Record ◽  
Phylogenetic Position ◽  
Rrna Gene ◽  
Gene Fragment ◽  
Wildlife Sanctuary

Based on a 16S rRNA gene fragment, a molecular phylogeny for the genus Odorrana Fei, Ye & Huang, 1990 was reconstructed, the validity of the poorly-known ranid species O. macrotympana (Yang, 2008) was confirmed and its phylogenetic position was evaluated. In addition, we report the first country record of O. macrotympana from Myanmar, based on our new records from Htamanthi Wildlife Sanctuary, Sagaing Division and present a supplementary description of this species. This report also constitutes the first record of O. macrotympana from outside of China.

Detection of Psychrophilic Clostridium spp. in Fecal Samples from Cattle of Different Ages Sampled at the Slaughterhouse Level

2020 ◽  
Vol 84 (1) ◽  
pp. 58-62
Author(s):  
JOSEPH WAMBUI ◽  
GIOVANNI GHIELMETTI ◽  
MARINA MORACH ◽  
MIRJAM HOCHREUTENER ◽  
ROGER STEPHAN
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Rrna Gene ◽  
Rt Pcr ◽  
Defibrinated Sheep Blood ◽  
Fecal Samples ◽  
Spoilage Bacteria ◽  
Different Ages ◽  
Young Cattle ◽  
Clostridium Spp

ABSTRACT Clostridium estertheticum and C. estertheticum–like spp. are obligate anaerobic psychrophiles causing “blown pack” spoilage of chilled vacuum-packed meat. The present study aimed at detecting and isolating these spoilage bacteria in fecal samples of cattle of different ages at the slaughterhouse level. One hundred two swab fecal samples were obtained and enriched anaerobically in prereduced peptone-yeast-glucose-starch (PYGS) medium for 3 weeks at 4°C and then screened for C. estertheticum and C. estertheticum–like spp. by using a 16S rRNA gene–based real-time PCR (RT-PCR) assay. The RT-PCR–positive samples were further enriched for 3 weeks in prereduced PYGS medium and then subjected to an ethanol (50%, v/v) and lysozyme (4 mg/mL) treatment. Isolation was carried out anaerobically on Columbia agar with 5% defibrinated sheep blood at 4°C for 3 weeks. Isolated strains were identified morphologically and by the 16S rRNA gene. Forty (39%) of 102 samples were RT-PCR positive. The frequency of positive samples was the following: 9 (45%) of 20 in calves (aged ≤160 days), 23 (43%) of 54 in young cattle (aged 161 to 1,000 days), and 8 (29%) of 28 in cows or bulls (aged >1,000 days). Six strains were isolated from 6 of 40 RT-PCR–positive samples. Of these, five were from the calves (n = 1) and young cattle (n = 4). The six isolates were identified as C. estertheticum (n = 1), Clostridium frigoriphilum (n = 1), and C. estertheticum–like spp. (n = 4). The present findings confirm that feces of cattle are an important source of psychrophilic Clostridium spp. The fecal carriage among livestock animals at slaughter is strongly correlated with the risk of carcass contamination. Therefore, the maintenance of slaughter hygiene is of central importance. HIGHLIGHTS

scholarly journals Systemic mycobacteriosis along with concurrent fibroma in Goldfish, Carassius auratus from India

2021 ◽  
Vol 42 (4) ◽  
pp. 1019-1025
Author(s):  
T.R. Swaminathan ◽  
◽  
N. Sood ◽  
P.K. Pradhan ◽  
M.K. Yadav ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Carassius Auratus ◽  
Ornamental Fish ◽  
Rrna Gene ◽  
Gene Fragment ◽  
Internal Organs ◽  
Goldfish Carassius Auratus ◽  
The Family ◽  
Maximum Similarity

Aim: Goldfish (Carassius auratus L.), a freshwater fish belonging to the family Cyprinidae, is an important ornamental fish species in the world. Herewith, we report a case of systemic mycobacteriosis along with concurrent fibroma in goldfish. Methodology: The protruding mass and internal organs exhibiting lesions were collected for histopathology and molecular diagnosis. The sections of cutaneous mass were stained with haematoxylin and eosin (H&E) and Masson trichrome, whereas sections from spleen as well as kidney were stained with H&E and Ziehl-Neelsen. For identifying the etiological agent, 16S rRNA gene fragment was amplified and sequenced, using genus-specific primers for Mycobacterium sp. Results: In histopathology, multiple granulomas could be observed in kidney and spleen sections. Importantly, typical acid-fast bacilli were demonstrated in granulomas in spleen. The sequence of 16S rRNA gene fragment amplified from spleen and kidney showed maximum similarity to Mycobacterium stomatepiae. Besides, the histopathological characteristics of the protruding mass were consistent with cutaneous fibroma. Interpretation: These findings assume significance as several Mycobacterium species, including the current one from ornamental fish are reported to be of zoonotic significance.

scholarly journals Phylogenetic inference of Coxiella burnetii by 16S rRNA gene sequencing

PLoS ONE ◽  
2017 ◽  
Vol 12 (12) ◽  
pp. e0189910 ◽  
Author(s):  
Heather P. McLaughlin ◽  
Blake Cherney ◽  
Janetta R. Hakovirta ◽  
Rachael A. Priestley ◽  
Andrew Conley ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Coxiella Burnetii ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Phylogenetic Inference ◽  
Rrna Gene ◽  
Rrna Gene Sequencing

scholarly journals Hemotropic mycoplasmas in bats from forest fragments, state of Paraná, southern Brazil

2022 ◽  
Vol 43 (1) ◽  
pp. 431-440
Author(s):  
Flávia Carolina Meira Collere ◽  
◽  
Larissa Dantas Roeder Ferrari ◽  
Ricardo Nascimento Drozino ◽  
Jéssica Damiana Marinho Valente ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
23S Rrna ◽  
Rrna Gene ◽  
Forest Fragments ◽  
Gene Fragment ◽  
Southern Brazil ◽  
Pcr Assay ◽  
Hemotropic Mycoplasma ◽  
The 16S Rrna Gene

The order Chiroptera is the second largest group of mammals with bats being identified as reservoir of several viral zoonoses, although, little is known about their role in other groups of pathogens, including hemotropic Mycoplasma spp. To date, hemoplasma species have been found infecting several species of bats with high genetic diversity between 16S rRNA gene sequences. On this study, we aimed to identify the occurrence and characterize 16S and 23S rRNA genes of hemoplasma species in four bats species (Artibeus lituratus, Carollia perspicillata, Sturnira lilium and Sturnira tildae) from forest fragments in Paraná State, southern Brazil, using PCR-based assays. Spleen tissue samples were collected, DNA extracted and further screened by a pan‑hemoplasma PCR assay. All samples consistently amplified the mammal endogenous gapdh gene. One out of 15 (6.66%; 95% CI: 0.2-31%) bats tested positive for hemotropic Mycoplasma sp. by the PCR assay targeting the 16S rRNA gene. Sequencing of the 16S rRNA gene fragment from the hemoplasma-positive bat showed 99.14% identity with hemotropic Mycoplasma sp. detected in Sturnira parvidens from Belize. Sequencing of the 23S rRNA gene fragment from the hemoplasma-positive bat showed 86.17% identity with ‘Candidatus Mycoplasma haemosphiggurus’ detected in orange-spined hairy dwarf porcupines (Sphiggurus villosus) from Southern Brazil.

scholarly journals Survey of Tick-borne-disease from Ornithodoros spp. in Uninhabited Islands of Korea.

2019 ◽  
Vol 11 (1) ◽  
Author(s):  
Sun-Woo Han ◽  
Yong-Sun Jo ◽  
Jeong Byoung Chae ◽  
Yoon Kyoung Cho ◽  
Nam-Shik Shin ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Anaplasma Phagocytophilum ◽  
Economic Losses ◽  
Rrna Gene ◽  
Gene Fragment ◽  
Sequence Identification ◽  
Tick Borne Disease ◽  
Anaplasma Bovis ◽  
Breeding Place

ObjectiveThe aim of this study is to survey data of ticks distribution of Korean islands and to investigate pathogens in Argasid ticks.IntroductionTicks and tick-borne diseases have been thought global important issues, because it's affect to animal and human health and are the cause of significant economic losses. The genus Ornithodoros spp., which is included in Family Argasidae, is usually associated with wild animals including seabirds and it was difficult to investigate because seabirds’ nests are found in inaccessible uninhabited islands. However, Ornithodoros spp. has been known for the vector of many diseases including African swine fever.MethodsIn this study, nest with soil and litter of seabirds were collected, to investigate Ornithodoros species from 9 uninhabited islands, Nan-do, Chilsan-do, Chilbal-do, Sogukhol-do, Googul-do, Gaerin-do, Sasu-do, Hong-do (Hallyeohaesang) and Dok-do located western and southern part of the Korea from July, 2017 to September in 2018. The islands are known for breeding places of migratory and resident birds. Maximum ten nests with soil and litter of seabirds were collected from one uninhabited island for the conservation of the islands environment. Ticks were collected from nest with soil and litter of seabirds using Tullgren funnel and were assayed for tick identification by PCR using 16S rRNA gene and tick-borne pathogens including Rickettia spp., Borrelia spp., Bartonella spp., Ehrlichia chaffensis, Ehrlichia canis, Anaplasma phagocytophilum and Anaplasma bovis by nested PCR.ResultsTotal 65 Ornithodoros species ticks from 338 seabird's (black-tailed gull, Larus crassirotris; streaked shearwater, Calonectris leucomelas and Swinhoe's storm petrel, Oceanodroma monorhis) nesting soil with litter in 9 uninhabited islands. In the sequence identification of 16S rRNA gene fragment of Ornithodoros species, O. capensis and O. sawaii were 37 and 28, respectively. In the analyses of tick-borne pathogens, Borrelia spp. (n=5) was detected in O. sawaii from Gaerin-do and Googul-do. The total detection rate of Borrelia sp. from Ornithodoros spp. was 7.69% (5/65).ConclusionsIn this research, we discovered that O. sawaii are habitat in west and southern part of uninhabited islands to breeding place of black-tailed gulls and streaked shearwater and O. capensis are habitat in uninhabited islands related to breeding place of streaked shearwater. This is first report of the Borrelia spp. from Ornithodoros sawaii in Korea. 

scholarly journals Species Diversity of Bifidobacteria in the Intestinal Microbiota Studied Using MALDI-TOF Mass-Spectrometry

2015 ◽  
Vol 70 (4) ◽  
pp. 435-440 ◽  
Author(s):  
A. V. Chaplin ◽  
A. G. Brzhozovskii ◽  
T. V. Parfenova ◽  
L. I. Kafarskaia ◽  
N. N. Volodin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Intestinal Microbiota ◽  
Rrna Gene ◽  
Growing Up ◽  
Gene Fragment ◽  
Bifidobacterium Adolescentis ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof

Background: The members of genus Bifidobacterium represent a significant part of intestinal microbiota in adults and predominate in infants. Species repertoire of the intestinal bifidobacteria is known to be subjected to major changes with age; however, many details of this process are still to be elucidated.Objective: Our aim was to study the diversity of intestinal bifidobacteria and changes of their qualitative and quantitative composition characteristics during the process of growing up using MALDI-TOF mass-spectrometric analysis of pure bacterial cultures.Methods: A cross-sectional study of bifidobacteria in the intestinal microbiota was performed in 93 healthy people of the ages from 1 month to 57 years. Strains were identified using Microflex LT MALDI-TOF MS, the confirmation was performed by 16S rRNA gene fragment sequencing.Results: 93% of isolated bifidobacterial strains were successfully identified using MALDI-TOF mass-spectrometry. At least two of the strains from each species were additionally identified by 16S rRNA gene fragment sequencing, in all of the cases the results were the same. It was shown that the total concentration of bifidobacteria decreases with age (p 0.001) as well as the frequency of isolation of Bifidobacterium bifidum (p =0.020) and Bifidobacterium breve (p 0.001), and the frequency of isolation of Bifidobacterium adolescentis, increases (p 0.001), representing the continuous process of transformation of microbiota.Conclusion: The method of MALDI-TOF mass spectrometry demonstrated the ability to perform rapid and reliable identification of bifidobacteria that allowed the study of changes in the quantitative and qualitative characteristics of human microbiota in the process of growing up.

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