Pathogens In Fecal Samples Research Articles

Microbiological pathogens in fecal samples of foals during the first year of life.

The objective of the study was to investigate the age dependent occurrence of different infectious agents in foals with diarrhea. Fecal samples, which were submitted to a commercial laboratory for a PCR-profile "Foal Diarrhea Pathogens" from 01.01.2021 up to 31.12.2022 (n=144), were examined for Equine Coronavirus (ECoV), Lawsonia (L.) intracellularis, Rhodococcus (R.) hoagii (=R. equi), Clostridium (Cl.) perfringens toxin-encoding genes cpa, cpe, cpb, etx and netF, Clostridioides (C.) difficile toxin-encoding genes tcdA and tcdB, as well as Rotavirus A via PCR. Pathogens could be detected in a high proportion (42.9%) of the samples. Rotavirus A was the most prevalent pathogen in the current study, followed by clostridial species. Especially in foals younger than one month, netF-producing Cl. perfringens was detected frequently. In this age group, netF-producing Cl. perfringens was as prevalent as Rotavirus A. In comparison, R. hoagii, L. intracellularis and ECoV were detected rarely. Cl. perfringens toxin-encoding genes cpb and etx were not present in the examined samples. In general, the previously known age dependency of the investigated pathogens could be confirmed. Nevertheless, Rotavirus A and netF-positive Cl. perfringens could also be detected outside of the most susceptible age group. Coinfections with the examined pathogens had a low prevalence in the current study. In general, the examined pathogens showed an age dependent occurrence, but infections in foals outside of the common age group could not be ruled out with certainty due to small sample numbers in some of the age groups. Although Rotavirus A was the most prevalent pathogen in this study, netF-producing Cl. perfringens is an important differential diagnosis, especially in newborn foals. The diagnostic approach in diarrheic foals should contain a broad spectrum of pathogens. This is not only important to detect coinfections, but also to detect shedders, in order to protect other horses in the stable.

Analysis of Cultured Gut Microbiota Using MALDI-TOF MS in COVID-19 Patients from Serbia during the Predominance of the SARS-CoV-2 Omicron Variant.

The currently dominant SARS-CoV-2 omicron variant, while causing mild respiratory symptoms, exhibits high transmissibility, drug resistance, and immune evasion. We investigated whether the presence of the SARS-CoV-2 affected the dynamics of fecal microbial composition isolated in culture in moderate COVID-19 patients. Blood, stool, and medical records were collected from 50 patients with confirmed SARS-CoV-2 infection. Two samples were taken per patient, at disease onset (within 5 days) and after symptom resolution (30-35 days). The part of the gut microbiota identifiable using MALDI-TOF MS was analyzed, and inflammatory cytokines and blood markers were measured in serum. The analysis identified 566 isolates at the species level, including 83 bacterial and 9 fungal species. Our findings indicate a change in the gut microbiota composition isolated in culture during the initial phase of infection, characterized by the proliferation of opportunistic bacteria such as Enterococcus spp. and Citrobacter spp., at the expense of beneficial commensal bacteria from the genus Bacillus and Lactobacillus. Additionally, the enrichment of fungal pathogens in fecal samples collected 30 days after the cessation of disease symptoms might suggest a prolonged disruption of the gut microbiota even after the resolution of COVID-19 symptoms. This study contributes to a growing body of evidence on the systemic effects of SARS-CoV-2 and highlights the importance of considering gastrointestinal involvement in the management and treatment of COVID-19.

Microbiome composition recovery after liver transplantation correlates with initial liver disease severity and antibiotics treatment

Liver transplantation (LT) is crucial for end-stage liver disease, but it is linked to infection risks. Pathobionts, microorganisms potentially harmful under specific conditions, can cause complications posttransplant. Monitoring such pathogens in fecal samples can be challenging and therefore remains underexplored post-LT. This study aimed to analyze the gut microbiome before and after LT, tracking pathobionts and correlating clinical data. The study involved 17 liver transplant recipients, 17 healthy relatives (spouses), and 13 donors. Gut samples collected pretranplantation and posttransplantation underwent bacterial and fungal profiling through DNA sequencing. Quantitative polymerase chain reaction was used to assess microbial load. Statistical analyses included alpha and beta diversity measures, differential abundance analysis, and correlation tests between microbiome and clinical parameters. Microbiome analysis revealed dynamic changes in diversity posttransplant. Notably, high-severity patients showed persistent and greater dysbiosis during the first months post-LT compared with low-severity patients, partly due to an antibiotic treatment pre-LT. The analysis identified a higher proportion of pathogens such as Escherichia coli/Shigella flexneri in high-severity cases posttransplant. Furthermore, butyrate producers including Roseburia intestinalis, Anaerostipes hadrus, and Eubacterium coprostanoligenes were positively correlated with levels of albumin. This study offers valuable insights into post-LT microbiome changes, shedding light on the need for tailored prophylactic treatment post-LT.

Reduction of Pathogens in Feces and Lymph Nodes Collected from Beef Cattle Fed Lactobacillus salivarius (L28), Lactobacillus acidophilus (NP51) and Propionibacterium freudenreichii (NP28), Commercially Available Direct-Fed Microbials

The purpose of the study was to evaluate the prevalence and concentration of foodborne pathogens in the feces and peripheral lymph nodes (PLNs) of beef cattle when supplemented with direct-fed microbials (DFMs) in feedlots. Fecal samples were collected from the pen floors over a 5-month period at three different feedlots in a similar geographical location in Nebraska, where each feed yard represented a treatment group: (i.) control: no supplement, (ii.) Bovamine Defend: supplemented with NP51 and NP24 at a target dose of 9 log10CFU/g/head/day, and (iii.) Probicon: supplemented with L28 at a target dose of 6 log10CFU/g/head/day. Each fecal sample was tested for the prevalence of E. coli O157:H7 and Salmonella, and concentration of E. coli O157:H7, Enterobacteriaceae and Clostridium perfringens. Cattle were harvested and PLNs were collected on the harvest floor. Real-time Salmonella PCR assays were performed for each PLN sample to determine Salmonella presence. The cattle supplemented with both DFMs had reduced foodborne pathogens in fecal samples, but feces collected from the pens housing the cattle supplemented with Probicon consistently had significantly less E. coli O157:H7 and Salmonella prevalence as well as a lower C. perfringens concentration. While DFMs do not eliminate foodborne pathogens in fecal shedding and PLNs, the use of DFMs as a pre-harvest intervention allows for an effective way to target multiple pathogens reducing the public health risks and environmental dissemination from cattle.

Norovirus Is the Most Frequent Cause of Diarrhea in Hospitalized Patients in Monterrey, Mexico.

Little information is available regarding the pathogens that cause diarrhea in hospitalized patients who also have various clinical problems. The purpose of this study was to determine the presence of pathogens in fecal samples of hospitalized patients all suffering diarrhea in addition to other problems in Mexico. Diarrheic stools from 240 patients were obtained in a third-level hospital in Monterrey, Mexico. PCR was used for the detection of Salmonella spp., Shigella spp., Campylobacter spp., Yersinia spp., Aeromonas spp., Clostridioides difficile, and norovirus GI and GII. The presence of trophozoites, cysts of protozoa, eggs, and/or helminth larvae was determined by microscopic observation. Of the 240 patients analyzed, 40.4% presented at least one of the pathogens analyzed. Norovirus was the pathogen most frequently found (28.6%), followed by bacteria (11.7%), and parasites (8.3%). The majority of co-infections were parasites + norovirus, and bacteria + norovirus. Norovirus was detected mainly in children aged 0 to 10 years (9/15, 60%). Patients aged 0–20 years did not present co-infections. Entamoeba coli and Entamoeba histolytica were the most common parasites, (8/240), and Salmonella was the most prevalent bacteria (10/240). This information can help design specific strategies useful for hospitalized people with a compromised status.

Epidemiology and genotype diversity of norovirus infections among children aged

BackgroundFollowing rotavirus vaccine introduction, norovirus has emerged as a significant pathogen associated with acute gastroenteritis among children in some high- and middle-income countries. In a case-control study following rotavirus vaccination in Malawi, we used PCR to test for multiple enteric pathogens in fecal samples from children aged <5 years hospitalized with diarrhea, and from asymptomatic community controls (Iturriza-Gómara et al. 2019). ObjectivesTo describe the epidemiology and genotype diversity of norovirus infections among infants and young children in Blantyre, Malawi, following rotavirus vaccine introduction in 2012. Study DesignWe analysed data from the case-control study to assess annual and agespecific norovirus prevalence and the presence of co-infection. Norovirus-containing specimens were amplified by PCR and sequenced to determined genotype. ResultsNorovirus prevalence in cases was similar for each complete year of study (11.4% in 2013, 9.3% in 2014 and 11.2% in 2015). Prevalence of norovirus among children aged <6 months, 6-11 months, 12-23 months and 24+ months was 15.3% (11/72), 13.3% (44/331), 11.0% (24/219) and 6.6% (4/61) respectively in cases and 6.7% (2/30), 13.1% (30/229), 4.2% (8/192) and 7.1 (5/70) in controls. Co-pathogens were commonly detected in norovirus positive cases (77/83) and controls (44/45). Norovirus GII.4 was the most commonly identified genotype, comprising 48% and 41% of genotyped strains among cases and controls, respectively. ConclusionsNorovirus disease prevalence was unchanged during the study period, and was greatest amongst infants. Frequent co-infection and asymptomatic shedding suggests intense community transmission of norovirus and other enteric pathogens in this low-income, African setting.

Marker genes of fecal indicator bacteria and potential pathogens in animal feces in subtropical catchments

We investigated the abundance of marker genes of two fecal indicator bacteria (FIB) and eight potential pathogens in fecal samples of humans (n = 14) and 10 domestic and native wild animals (n = 134). For each target animal, between 10 and 14 individual fecal samples were collected (n = 148 individual fecal samples in total). The abundance of FIB and potential pathogens within each sample was determined using quantitative PCR (qPCR) assays. All animals tested were positive for Escherichia coli (EC) and the concentrations ranged from 6.13 (flying fox) to 8.87 (chicken) log10 GC/g of feces. These values for Enterococcus spp. (ENT) were 5.25 log10 GC/g for flying fox and 8.12 log10 GC/g of feces for chicken. Moderate correlations were observed between EC with P. aeruginosa, EC O157 and Cryptosporidium parvum, whereas weak correlations were observed between EC and Salmonella spp. and Giardia lamblia, Mycobacterium avium complex (MAC) and Campylobacter spp. The prevalence of MAC and P. aeruginosa were low in dog (14.3% each) and moderate (57.2%, MAC; 42.9% P. aeruginosa) in Eastern grey kangaroo fecal samples. Cryptosporidium parvum was detected in one cattle and one human fecal sample, while G. lamblia was detected in one dog, one flying fox, and one pig fecal samples. Among the eight potential pathogens tested, five pathogens were detected in chicken and dog fecal samples. The remaining animal species contained up to three potential pathogens in their feces. The data generated in this study may aid in the calculation of pathogen loads in the environment, and hence to assess the risks from human and animal fecal contamination of source waters.

Application of real-time PCR for detection of Lawsonia intracellularis and Brachyspira hyodysenteriae in fecal samples from pigs

The aim of the study was to develop and validate real-time PCR method for the quantification of Lawsonia intracellularis and Brachyspira hyodysenteriae in porcine feces. Before the optimization process was performed two different extraction methods were compared to select the more efficient one. Based on the results achieved at this stage the boiling procedure was rejected and a commercially available silica-membrane based method was chosen for further analysis. The primers and the Taqman probe for B. hyodysenteriae and L. intracellularis were based on the sequence of NADH oxidase gene and 16S rDNA gene, respectively. The detection limit of the real-time PCR for suspension of feces inoculated with B. hyodysenteriae and L. intracellularis was determined to be 1.5x10(3) CFU/ml and 6.5x10(1) CFU/ml, respectively. The results of this study demonstrate that our real-time PCR is able to detect low number of B. hyodysenteriae and L. intracellularis cells which is satisfying in routine diagnosis of swine dysentery and proliferative enteropathy. Therefore, it is possible to identify both subclinically infected pigs and those representing an acute form of mentioned diseases. In summary, the quantitative real-time PCR is useful for routine diagnosis of L. intracellularis and B. hyodysenteriae. Compared to conventional PCR, the new validated quantification method based on real-time PCR is fast and with reduced risk of laboratory contamination. The novel technique is specific and even more sensitive than the previously used one. Furthermore, the new real-time PCR enables quick detection and quantification of both pathogens in fecal samples, which helps to estimate the health status of a pig herd.

Diarrheagenic Pathogens in Polymicrobial Infections

During systematic active surveillance of the causes of diarrhea in patients admitted to the Infectious Diseases and Beliaghata General Hospital in Kolkata, India, we looked for 26 known gastrointestinal pathogens in fecal samples from 2,748 patients. Samples from about one-third (29%) of the patients contained multiple pathogens. Polymicrobial infections frequently contained Vibrio cholerae O1 and rotavirus. When these agents were present, some co-infecting agents were found significantly less often (p = 10–5 to 10–33), some were detected significantly more often (p = 10–5 to 10–26), and others were detected equally as often as when V. cholerae O1 or rotavirus was absent. When data were stratified by patient age and season, many nonrandom associations remained statistically significant. The causes and effects of these nonrandom associations remain unknown.

Diarrheagenic Pathogens in Polymicrobial Infections

During systematic active surveillance of the causes of diarrhea in patients admitted to the Infectious Diseases and Beliaghata General Hospital in Kolkata, India, we looked for 26 known gastrointestinal pathogens in fecal samples from 2,748 patients. Samples from about one-third (29%) of the patients contained multiple pathogens. Polymicrobial infections frequently contained Vibrio cholerae O1 and rotavirus. When these agents were present, some co-infecting agents were found significantly less often (p = 10 (-5) to 10 (-33), some were detected significantly more often (p = 10 (-5) to 10 (-26), and others were detected equally as often as when V. cholerae O1 or rotavirus was absent. When data were stratified by patient age and season, many nonrandom associations remained statistically significant. The causes and effects of these nonrandom associations remain unknown.

Comparison of the QIAsymphony automated nucleic acid extraction and PCR setup platforms with NucliSens easyMAG and Corbett CAS-1200 liquid handling station for the detection of enteric pathogens in fecal samples

In this study we compared the QIAsymphony Sample Preparation and Assay Setup modules (Qiagen), which provide automated nucleic acid extraction and PCR setup, to the NucliSens easyMAG (bioMérieux) and CAS-1200 liquid handling station 1 1 Corbett, a QIAGEN company, currently vendors the CAS-1200 liquid handling station under the name QIAgility. (Corbett) for a molecular screening approach of enteric pathogens in fecal samples using multiplex real-time PCR. The relative DNA recovery of both platforms, within- and between-run reproducibility and a prospective study, including 510 clinical fecal samples, were performed. The results demonstrated that the QIAsymphony Sample Preparation and Assay Setup modules were highly reproducible and achieve equal performance, quantitative and qualitative, when compared with the NucliSens easyMAG and CAS-1200 systems for the molecular screening analysis of enteric pathogens in fecal samples.

Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method

To develop a new, rapid and accurate reverse dot blot (RDB) method for the detection of intestinal pathogens in fecal samples. The 12 intestinal pathogens tested were Salmonella spp., Brucella spp., Escherichia coli O157:H7, Clostridium botulinum, Bacillus cereus, Clostridium perfringens, Vibrio parahaemolyticus, Shigella spp., Yersinia enterocolitica, Vibrio cholerae, Listeria monocytogenes and Staphylococcus aureus. The two universal primers were designed to amplify two variable regions of bacterial 16S and 23S rDNA genes from all of the 12 bacterial species tested. Five hundred and forty fecal samples from the diarrhea patients were detected using the improved RDB assay. The methods could identify the 12 intestinal pathogens specifically, and the detection limit was as low as 103 CFUs. The consistent detection rate of the improved RDB assay compared with the traditional culture method was up to 88.75%. The hybridization results indicated that the improved RDB assay developed was a reliable method for the detection of intestinal pathogen in fecal samples.

Deaths from Norovirus among the Elderly, England and Wales

The number of deaths in England and Wales associated with gastrointestinal pathogens, norovirus in particular, in persons >or=65 years was estimated for 2001-2006. Regression analysis was used to model monthly counts of gastrointestinal pathogens in fecal samples from infected patients against monthly counts of deaths from infectious and noninfectious intestinal diseases. Data came from the Office of National Statistics (death registrations from local registrars) and from the Health Protection Agency (laboratory results). Model results suggest that 20% (13.3%-26.8%) of deaths in persons >or=65 years of age caused by infectious intestinal disease other than Clostridium difficile were associated with norovirus infection in this period and that 13% (7.5%-18.5%) of deaths caused by noninfectious intestinal disease were associated with norovirus. An estimated 80 deaths each year in this age group may be associated with norovirus infection.

下痢, 乳量減少および食欲不振が認められた搾乳牛の牛B群ロタウイルス感染

An epizootic outbreak of diarrhea, milk drop and inappetence in lactating cows occurred on a dairy farm in Fukui Prefecture, Japan. Rotavirus particles were observed in fecal samples under electron microscopy. On polyacrylamide gel electrophoresis, the dsRNA migration pattern from fecal samples was characteristic of bovine group B rotavirus (GBR). Fecal samples were positive for GBR by reverstranscriptase PCR. No diarrheal pathogens in fecal samples were found on bacteriological or parasitological testing. Antibody testing for GBR by ELISA showed seroconversion in most lactating cows on the farm concerned. We subsequently investigated 130 serum samples from 26 herds to determine the prevalence of infection with GBR in Fukui Prefecture. Results were positive in 21 of 26 herds (81%) and in 65 of 130 samples (50%). These results suggest that there is some doubt that occurrences of diarrhea, milk drop and inappetence in lactating cows are related toinfection with bovine GBR or that GBR infection is prevalent in Fukui Prefecture.

Detection of Viral, Bacterial, and Parasitological RNA or DNA of Nine Intestinal Pathogens in Fecal Samples Archived as Part of the English Infectious Intestinal Disease Study

Fecal samples were collected from cases and controls as part of the Infectious Intestinal Disease (IID) study in England and were stored as frozen suspensions for 8 to 12 years. The purpose of this study was to apply PCR-based procedures to assess the stability of pathogen-specific nucleic acid sequences present in this archive. Samples from which Cryptosporidium, Giardia, Salmonella, Campylobacter, enteroaggregative Escherichia coli (EAggEC), enterotoxigenic Clostridium perfringens, rotaviruses, noroviruses, or sapoviruses had been previously detected during the IID study using conventional methods were selected from the archive. A generic nucleic acid extraction method to recover RNA or DNA was used. Complementary DNA was generated from RNA by reverse transcription with random priming. Block-based and real-time PCR assays were used to amplify and detect gene fragments from each of these pathogens. The percentage reconfirmation of target was as follows: Giardia duodenalis 68%, Cryptosporidium 96%, Campylobacter 98%, Salmonella 98%, enterotoxigenic C perfringens 34%, EAggEC 93.3%, rotavirus 95%, norovirus 73%, and sapovirus 85%. This study has shown that nucleic acid can be extracted and specific sequences amplified and detected from archived fecal samples. The IID archive therefore represents a valuable resource for further studies, especially the investigation of the samples from which no pathogens had previously been detected.