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The near-complete genomes of two picobirnaviruses (PBVs) in diarrheal stool samples, human picobirnaviruses D and E (HuPBV-D and -E), were genetically characterized. Their RNA-dependent RNA polymerase (RdRp) protein sequences had <66% identities to known PBVs. Due to a single nucleotide insertion, the open reading frame 2 (ORF2) in segment 1 of HuPBV-D was interrupted by a stop codon. A small stem-loop structure overlying the stop codon may result in translational readthrough into the rest of ORF2.
To determine the source of Streptobacillus notomytis bacteremia in a woman in Japan with signs of rat-bite fever, we examined rat feces from her home. After culture and PCR failed to identify the causative organism in the feces, next-generation sequencing detected Streptobacillus spp., illustrating this procedure's value for identifying causative environmental organisms.
Lactic acid produced by intestinal bacteria is fermented by lactate-utilizing bacteria. In this study, we developed a selective culture medium (KMI medium) for Megasphaera elsdenii, a lactate-utilizing bacterium that is abundant in pig intestines. Supplementation of the medium with lactate and beef extract powder was necessary for the preferential growth of M. elsdenii. In addition, we designed a species-specific primer set to detect M. elsdenii. When pig fecal samples were plated on KMI agar medium, approximately 60-100% of the resulting colonies tested positive using the M. elsdenii-specific PCR primers. In fact, nearly all of the large, yellow-white colonies that grew on the KMI agar medium tested positive by PCR with this primer set. The 16S rRNA gene sequences of three representative PCR-positive strains showed strong similarities to that of M. elsdenii ATCC 25940T (98.9-99.2% identity). These three strains were approximately 1.5 μm sized cocci that were primarily arranged in pairs, as was observed for M. elsdenii JCM 1772T. The selective KMI medium and species-specific primer set developed in this study are useful for the isolation and detection of M. elsdenii and will be useful in research aimed at increasing our understanding of intestinal short-chain fatty acid metabolism in pigs.
Sonoran felids are threatened by drought and habitat fragmentation. Vector range expansion and anthropogenic factors such as habitat encroachment and climate change are altering viral evolutionary dynamics and exposure. However, little is known about the diversity of viruses present in these populations. Small felid populations with lower genetic diversity are likely to be most threatened with extinction by emerging diseases, as with other selective pressures, due to having less adaptive potential. We used a metagenomic approach to identify novel circoviruses, which may have a negative impact on the population viability, from confirmed bobcat (Lynx rufus) and puma (Puma concolor) scats collected in Sonora, Mexico. Given some circoviruses are known to cause disease in their hosts, such as porcine and avian circoviruses, we took a non-invasive approach using scat to identify circoviruses in free-roaming bobcats and puma. Three circovirus genomes were determined, and, based on the current species demarcation, they represent two novel species. Phylogenetic analyses reveal that one circovirus species is more closely related to rodent associated circoviruses and the other to bat associated circoviruses, sharing highest genome-wide pairwise identity of approximately 70% and 63%, respectively. At this time, it is unknown whether these scat-derived circoviruses infect felids, their prey, or another organism that might have had contact with the scat in the environment. Further studies should be conducted to elucidate the host of these viruses and assess health impacts in felids.
Using a metagenomic approach and molecular cloning methods, we identified, cloned, and sequenced the complete genome of a novel circular DNA virus, porcine stool-associated virus (PoSCV4), from pig feces. Phylogenetic analysis of the deduced replication initiator protein showed that PoSCV4 is most related to a fur seal feces-associated circular DNA virus.
CrAssphages are a diverse group of related phages detected in human feces where they are the most prevalent and abundant prokaryotic virus. CrAssphages' cellular host has been identified as the anaerobic Bacteroides intestinalis. CrAssphage has also been reported in non-human primates and environmental samples and has been proposed as a marker of human fecal contamination. Here we describe crAssphage DNA in a feline fecal sample. 95% of the ~ 100 Kb genome could be assembled and classified in genus 1 of the recently proposed Alphacrassvirinae subfamily. The cat origin of the fecal sample was confirmed by partial mitochondrial DNA sequencing. High levels of Bacteroides intestinalis DNA could also be detected in this cat's feces. Fecal samples longitudinally collected over a 4-week period showed the continuous shedding of crAssphage DNA. We therefore report the first genome sequence-confirmed detection of crAssphage in fecal samples of a non-primate mammal.
Staphylococcus epidermidis has emerged as the leading agent causing neonatal late-onset sepsis in preterm neonates; although the severity of the episodes caused by this species is often underestimated, it might exert relevant short- and long-term detrimental effects on neonatal outcomes. In this context, the objective of this study was to characterize a collection of S. epidermidis strains obtained from meconium and feces of preterm infants, and to assess the potential role of the enteral feeding tubes as potential reservoirs for this microorganism. A total of 26 preterm infants were enrolled in the study. Meconium and fecal samples were collected weekly during their first month of life (n = 92). Feeding samples were collected after their pass through the enteral feeding tubes (n = 84). S. epidermidis was present in the fecal samples of all the infants in, at least, one sampling time at concentrations ranging from 6.5 to 7.8 log10 CFU/g. Initially, 344 isolates were obtained and pulsed-field gel electrophoresis (PFGE) profiling allowed the reduction of the collection to 101 strains. Among them, multilocus sequence typing (MLST) profiling showed the presence of 32 different sequence types (ST). Globally, most of the STs to hospital-adapted high-risk clones and belonged to clonal complexes (CC) associated to the hospital environment, such as CC2. The virulence gene most commonly detected among the strains was altE. High resistance rates to macrolides and aminoglycosides were detected and 64% of the strains harboured the mecA gene, which was codified in SCCmec types. Our results indicates the existence of a complex and genetically diverse S. epidermidis population in the NICU environment. A better knowledge of S. epidermidis strains may help to devise strategies to avoid their conversion from symbiont to pathobiont microorganisms in the NICUs.
A divergent rotavirus I was detected using viral metagenomics in the feces of a cat with diarrhea. The eleven segments of rotavirus I strain Felis catus encoded non-structural and structural proteins with amino acid identities ranging from 25 to 79% to the only two currently sequenced members of that viral species both derived from canine feces. No other eukaryotic viral sequences nor bacterial and protozoan pathogens were detected in this fecal sample suggesting the involvement of rotavirus I in feline diarrhea.
We present proteome data from the microbiota (feces) after a diet shift from a natural diverse to a monocultural meadow with Dactylis glomerata. The abundant grasshopper species, Chorthippus dorsatus, was taken from the wild and kept in captivity and were fed with Dactylis glomerata for five days. For phytophagous insects, the efficiency of utilization of hemicellulose and cellulose depends on the gut microbiota. Shifts in environmental and management conditions alter the presence and abundance of plant species which may induce adaptations in the diversity of gut microbiota. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD005126.
In the present research, the 20 lactobacilli isolated from children feces aged 4-15 years old were investigated for their capabilities to survive at pH 2.0, 2.5, 3.0 and in the presence of 0.25, 0.50 and 0.75% bile salts, their effect on the growth of pathogens, in addition to their sensitivity against 13 selected antibiotics. All the lactobacilli strains were able to survive in low pH and bile salt conditions at pH 2.0 and 0.25% bile salt for 2 h. Moreover, all lactobacilli strains exhibited inhibitory activity against Escherichia coli ATCC 11229, Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213. In addition, all lactobacilli strains indicated resistance to teicoplanin, vancomycin, and bacitracin. The amount of exopolysaccharide (EPS) produced by the strains was 70 and 290 mg/L. The capabilities to autoaggregation and coaggregate with E. coli ATCC 11229 of the strains were also evaluated. High EPS-producing strains indicated significant autoaggregation and coaggregation capability with test bacteria (p < 0.01). The maximum cholesterol removal (76.5%) was observed by strain Lactobacillus pentosus T3, producing a high amount of exopolysaccharide, in 0.3%oxgall concentration (p < 0.05). Our results demonstrate that the capability to EPS production, acid-bile tolerance, antimicrobial activity, antibiotic resistance, aggregation and cholesterol removal of lactobacilli could be utilized for preliminary screening in order to identify potentially probiotic bacteria suitable for human.
The Guide for the Care and Use of Laboratory Animals (the Guide) recommends that terrestrial mammals be provided space free of urine and feces in which to rest. To evaluate the feasibility of meeting this recommendation, the author examined the availability of feces-free resting areas in standard rodent cages over time. Adult rodents (C57BL/6J mice and Wistar rats) were housed singly, in pairs or in trios in shoebox cages at densities that met the space recommendations of the Guide. As housing density increased, the availability of unsoiled resting space declined. For C57BL/6J mice housed singly, in pairs or in trios, most cages lacked unsoiled resting area within 3-6 days (depending on cage size), 2 days or 1 day, respectively. Similarly, for Wistar rats housed singly, in pairs or in trios, most cages lacked adequate unsoiled resting space within 3 days, 2 days or 1 day, respectively. Because most cages lacked adequate unsoiled resting space within 3 days of housing animals, the author concludes that standard cage change frequencies of once a week for adult C57BL/6J mice and twice a week for adult Wistar rats may be inadequate to provide unsoiled resting areas for rodents.
Enterococcus is ubiquitous in nature and is a commensal of both the bovine and human gastrointestinal (GI) tract. It is also associated with clinical infections in humans. Subtherapeutic administration of antibiotics to cattle selects for antibiotic resistant enterococci in the bovine GI tract. Antibiotic resistance genes (ARGs) may be present in enterococci following antibiotic use in cattle. If located on mobile genetic elements (MGEs) their dissemination between Enterococcus species and to pathogenic bacteria may be promoted, reducing the efficacy of antibiotics.
Canine parvovirus (CPV) is a contagious disease in dogs that has high morbidity and mortality. In cases of infection, the pups tend to have a higher mortality and more severe clinical symptoms than the adult dogs because the dehydration is difficult for pups to bear. Following the natural infection, there is a rapid antibody response neutralizing the extracellular virus. As a result, virus titers in tissue and feces become markedly reduced. Hence, it is important to have an effective symptomatic therapy of supporting animals to survive in the early stages of CPV infection. Furthermore, the co-infection with bacteria could increase the severity of lesions and clinical signs as well. In this paper, we obtained the bacterial diversity in feces of CPV infected dogs with the enrichment of five bacteria genera (Shigella, Peptoclostridium, Peptostreptococcus, Streptococcus, Fusobacterium). These microorganisms may partly result in the intestinal pathology of the infection. In summary, the discussion of the bacterial biodiversity in feces of CPV infected dogs provides further insights into the pathology of CPV disease and the targets of developing more effective treatment strategies.
Most human microbiota studies focus on bacteria inhabiting body surfaces, but these surfaces also are home to large populations of viruses. Many are bacteriophages, and their role in driving bacterial diversity is difficult to decipher without the use of in vitro ecosystems that can reproduce human microbial communities.
Bird feces are commonly used as a proxy for measuring dietary metal exposure levels in wild populations. Our study aims to improve the reliability and repeatability of fecal metal measurements and gives some recommendations for sampling. First, we studied levels of variation in metallic element (arsenic, calcium, cadmium, cobalt, copper, nickel, lead) concentrations: temporal variation within an individual, among siblings in a brood and among-brood/spatial variation. Second, we explored the variation caused by dual composition (urate vs. feces) of bird droppings. Two sets of fresh fecal samples were collected from pied flycatcher (Ficedula hypoleuca) nestlings living in a metal polluted area in summers 2017 (dataset 1) and 2018 (dataset 2). We found a great deal of temporal intra-individual variation in metal levels, suggesting that dietary exposure varied markedly in a short time scale (within a day). A sample from only one nestling per brood did not well describe the brood mean value, and we recommend that at least four siblings should be sampled. Brood level samples give relatively good temporal repeatability for most metals. For all the metals, the levels in the fecal portion were more than double to those in the urate portion. Since the mass proportion of urate in the bird droppings varied a great deal among samples, standardizing sampling, e.g., by collecting only the fecal part, would markedly reduce the variation due to composition. Alternatively, urate portion could be used for biomonitoring of internally circulated bioavailable metal.
Lactobacillus plantarum FH185 was isolated from the feces of healthy adults. In our previous study, L. plantarum FH185 was demonstrated that it has anti-obesity effect in the in vitro and in vivo test. In order to determine its potential for use as a probiotic, we investigated the physiological characteristics of L. plantarum FH185. The optimum growth temperature of L. plantarum FH185 was 40℃. L. plantarum FH185 showed higher sensitivity to novobiocin in a comparison of fifteen different antibiotics and showed higher resistance to polymyxin B and vancomycin. It also showed higher β-galactosidase and N-acetyl-β-glucosaminidase activities. Moreover, it was comparatively tolerant to bile juice and acid, and inhibited the growths of Salmonella Typhimurium and Staphylococcus aureus with rates of 44.76% and 53.88%, respectively. It also showed high adhesion activity to HT-29 cells compared to L. rhamnosus GG.
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