Assessment and Improvement of Methods for Microbial DNA Preparation from Fecal Samples

Author(s):  
Mariko Ueno ◽  
Mami Kikuchi ◽  
Kenshiro Oshima ◽  
Seok-won Kim ◽  
Hidetoshi Morita ◽  
...  
2017 ◽  
Vol 7 (1) ◽  
pp. 1386536 ◽  
Author(s):  
Per Eriksson ◽  
Evangelos Mourkas ◽  
Daniel González-Acuna ◽  
Björn Olsen ◽  
Patrik Ellström

2020 ◽  
Vol 3 (5) ◽  
pp. 100-107
Author(s):  
Marina Camargo de Sousa ◽  
◽  
Julia Ronzani Vial ◽  
Rodrigo Hidalgo Friciello Teixeira ◽  
Andrea Cristina Higa Nakaghi ◽  
...  

Birds of the psittaciform order, composed by the Psittacidae and Loridae family have several characteristics making them more frequently kept as companion animals, promoting the increase of breeding sites in Brazil. The present study aimed to analyze the specificity and sensitivity of three different coproparasitological tests, Willis, Hoffman and Direto de feces, through statistical tests: Chi-Square and Kappa. 70 fecal samples of exotic parrots were collected from a commercial breeding site and these were submitted to the three tests, totaling 210 coproparasitological exams. Among the tests performed, 29,5% were positive for nematode eggs, cestodes and oocysts. Coproparasitological exams are inexpensive, have clinical importance, indicating the population of endoparasites and therapeutic treatments.


2011 ◽  
Vol 152 (25) ◽  
pp. 1007-1012 ◽  
Author(s):  
Gábor Reuter ◽  
Mária Új ◽  
Péter Pankovics ◽  
Tímea Kolozsi ◽  
Ilona Mihály ◽  
...  

Human parechoviruses (HPeV) belonging to the family Picornaviridae are widespread enteric pathogens and are associated with various clinical syndromes in human. At present, 16 HPeV genotypes (HPeV1–16) are known. There is no report on the detection of HPeVs in Central Europe. Aims: The aim of the retrospective study was to detect and characterize HPeVs using molecular methods in cell cultures with “enterovirus-like” cytophatic effect (CPE) archived between 1990 and 2004, in two virology laboratories, in Hungary. Materials and methods: In Laboratory I, fecal samples from children with symptoms of gastroenteritis under the age of 10 years were cultured as a previous routine diagnostic laboratory protocol for “enterovirus”. Cell cultures indicating CPE were archived between 1990 and 2000. In Laboratory II, 2 fecal samples, a liquor and a nasopharyngeal aspirate were re-tested which contained an “enterovirus-like” virus in cell cultures and were positive by HPeV1 neutralization immunosera between 2000 and 2004. Specimens were tested retrospectively for HPeV by reverse transcription–PCR (RT-PCR) method using 5’UTR conserved primers. Specific primers were designed to determine the HPeV structural region (VP0-VP3-VP1). Results: 9 of the 66 archived samples (9.1%) from Laboratory I and all the 4 samples from Laboratory II were found to be HPeV-positive. 10 samples were identified as HPeV1, 2 were HPeV4 and 1 could not be determined. 3 HPeV1 clusters were identified in Laboratory I according to the isolation date originated from years 1990/1991, 1992/1995 and 1998. HPeV1 was detected in clinical syndromes: gastroenteritis (in a 24-years-old adult), recurrent stomatitis aphtosa (in a 42-years-old adult), encephalitis and ataxia cerebellaris acuta in infants and children in Laboratory II. Conclusions: This is the first detection of HPeVs in Central Europe. Detection and genetic characterization of HPeV in available historical samples infected with previously unidentifiable agents with “enterovirus-like” cytopathogenic effect may help to understand the clinical importance and spectrum of the infections and the genetic diversity and evolution of these viruses. Orv. Hetil., 2011, 152, 1007–1012.


Author(s):  
E.V. Korneenko ◽  
◽  
А.E. Samoilov ◽  
I.V. Artyushin ◽  
M.V. Safonova ◽  
...  

In our study we analyzed viral RNA in bat fecal samples from Moscow region (Zvenigorod district) collected in 2015. To detect various virus families and genera in bat fecal samples we used PCR amplification of viral genome fragments, followed by high-throughput sequencing. Blastn search of unassembled reads revealed the presence of viruses from families Astroviridae, Coronaviridae and Herpesviridae. Assembly using SPAdes 3.14 yields contigs of length 460–530 b.p. which correspond to genome fragments of Coronaviridae and Astroviridae. The taxonomy of coronaviruses has been determined to the genus level. We also showed that one bat can be a reservoir of several virus genuses. Thus, the bats in the Moscow region were confirmed as reservoir hosts for potentially zoonotic viruses.


2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Ying Wang ◽  
Bing-Cheng Ma ◽  
Li-Ying Wang ◽  
Gongsang Quzhen ◽  
Hua-Sheng Pang

Abstract Background Echinococcosis is highly endemic in western and northern China. Tibet Autonomous Region (TAR) is the most serious prevalent area. Linzhi is located in southeastern part of TAR. Dogs are the primary infection source for the transmission of echinococcosis to humans. A control and prevention campaign based on dog management has been implemented in the past three years. This study aims to evaluate the effects of dog management on the infection rate of dogs. Methods Data of dog population, registration and de-worming of seven counties/district in Linzhi between 2017 and 2019 were obtained from the annual prevention and control report. Domestic dog fecal samples were collected from each endemic town of seven counties/district in Linzhi in 2019 to determine the infection of domestic dogs using coproantigen enzyme-linked immunosorbent assay (ELISA). Data analysis was processed using SPSS statistics to compare dog infection rate between 2016 and 2019 by chi-square test, and maps were mapped using ArcGIS. Results In Linzhi, domestic dog population has decreased from 17 407 in 2017 to 12 663 in 2019, while the registration rate has increased from 75.9% in 2017 to 98.6% in 2019. Similarly, stray dog population has decreased from 14 336 in 2017 to 11 837 in 2019, while sheltered rate has increased from 84.6% in 2017 to 96.6% in 2019. Dog de-worming frequency has increased from 4 times per annum in 2017 to 12 times in 2019, indicating that approximately every dog was dewormed monthly. A total of 2715 dog fecal samples were collected for coproantigen ELISA assay. The dog infection rate was 2.8% (77/2715) in 2019, which was significantly lower than 7.3% (45/618) in 2016 (P < 0.05). Conclusions Increased dog registration, decreased dog population, and increased dog de-worming frequency contributed to significantly decrease the dog infection rate in Linzhi. Control and prevention campaign based on dog management could significantly decrease dog infection with Echinococcus spp. in echinococcosis endemic areas.


Fermentation ◽  
2021 ◽  
Vol 7 (1) ◽  
pp. 14
Author(s):  
Nelson Mota de Carvalho ◽  
Diana Luazi Oliveira ◽  
Mayra Anton Dib Saleh ◽  
Manuela Pintado ◽  
Ana Raquel Madureira

The use of fecal inoculums for in vitro fermentation models requires a viable gut microbiota, capable of fermenting the unabsorbed nutrients. Fresh samples from human donors are used; however, the availability of fresh fecal inoculum and its inherent variability is often a problem. This study aimed to optimize a method of preserving pooled human fecal samples for in vitro fermentation studies. Different conditions and times of storage at −20 °C were tested. In vitro fermentation experiments were carried out for both fresh and frozen inoculums, and the metabolic profile compared. In comparison with the fresh, the inoculum frozen in a PBS and 30% glycerol solution, had a significantly lower (p < 0.05) bacterial count (<1 log CFU/mL). However, no significant differences (p < 0.05) were found between the metabolic profiles after 48 h. Hence, a PBS and 30% glycerol solution can be used to maintain the gut microbiota viability during storage at −20 °C for at least 3 months, without interfering with the normal course of colonic fermentation.


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