Animal Stool Samples Research Articles

A Double Lysis Method for Animal Rotavirus RNA Extraction From Stool Samples.

Globally, porcine rotavirus is a leading cause of gastroenteritis in nursing and post-weaning piglets, as well as adult pigs. Between February 2015 and June 2016, 156 fecal samples were collected from pigs in the Northeastern part of Accra, Ghana, and screened for Group A rotavirus using the ProflowTM Kit. Here, we describe different extraction methods that were employed to recover high-quality RNA for downstream analysis, with emphasis on a novel hybrid extraction method. The hybrid approach with a kit and manual extraction method led to a 10-fold greater RNA yield versus the kit-based method alone. The new extraction method gave an average purity ratio (A260/A280) of 1.8, which was also significantly higher than that obtained solely from the manual or kit-based extraction methods. Our novel hybrid approach will be useful in the extraction of rotavirus from animal fecal samples, thus improving the yield of RNA for downstream analysis. © 2024 Wiley Periodicals LLC. Basic Protocol: Hybrid 2: A double lysis method for RNA extraction from animal stool samples Support Protocol 1: The GenElute extraction method Support Protocol 2: Hybrid 1 extraction method.

Molecular Detection of Cryptosporidium spp. and Microsporidia in Human and Animal Stool Samples.

Cryptosporidium spp. and Microsporidia are opportunistic microorganisms with remarkable zoonotic transmission potential due to their capacity to infect humans and animals. The aim of this study was to evaluate the prevalence of these microorganisms in stool samples of animal and human origin. In total, 369 stool samples (205 from human patients with diarrhea and 164 of animal origin) were included in the study. Cryptosporidium spp. and Microsporidia presence were determined by using multiplex nested PCR. Positive results were analyzed by using Sanger sequencing of the amplicon, utilizing BLASTN and ClustalX software to confirm identification. Cryptosporidium spp. were found in 0.97% and 4.26% of human and animal samples, respectively. Enterocytozoon bieneusi was detected in human and animal stools in 6.82% and 3.05% of the samples, respectively. No associations were found when analyzing the presence of Cryptosporidium spp. and E. bieneusi and the demographic and clinical variables of patients and animals. This study demonstrates the presence of these microorganisms in human and animal samples from different species, and the most interesting findings are the detection of Cryptosporidium spp. in pets (e.g., rodents) that are not usually included in this type of study, and the identification of E. bieneusi in patients with diarrhea without underlying disease.

Genomic and phylogenetic features of the Picobirnaviridae suggest microbial rather than animal hosts.

The RNA virus family Picobirnaviridae has traditionally been associated with the gastrointestinal systems of terrestrial mammals and birds, with the majority of viruses detected in animal stool samples. Metatranscriptomic studies of vertebrates, invertebrates, microbial communities, and environmental samples have resulted in an enormous expansion of the genomic and phylogenetic diversity of this family. Yet picobirnaviruses remain poorly classified, with only one genus and three species formally ratified by the International Committee of Virus Taxonomy. Additionally, an inability to culture picobirnaviruses in a laboratory setting or isolate them in animal tissue samples, combined with the presence of bacterial genetic motifs in their genomes, suggests that these viruses may represent RNA bacteriophage rather than being associated with animal infection. Utilising a data set of 2,286 picobirnaviruses sourced from mammals, birds, reptiles, fish, invertebrates, microbial communities, and environmental samples, we identified seven consistent phylogenetic clusters likely representing Picobirnavirus genera that we tentatively name 'Alpha-', 'Beta-', 'Gamma-', 'Delta-', 'Epsilon-', 'Zeta-', and 'Etapicobirnavirus'. A statistical analysis of topological congruence between virus-host phylogenies revealed more frequent cross-species transmission than any other RNA virus family. In addition, bacterial ribosomal binding site motifs were more enriched in Picobirnavirus genomes than in the two groups of established RNA bacteriophage-the Leviviricetes and Cystoviridae. Overall, our findings support the hypothesis that the Picobirnaviridae have bacterial hosts and provide a lower-level taxonomic classification for this highly diverse and ubiquitous family of RNA viruses.

On the nature of picobirnaviruses.

The picobirnaviruses (Picobirnaviridae, Picobirnavirus, PBVs) are currently thought to be animal viruses, as they are usually found in animal stool samples. However, no animal model or cell culture for their propagation has yet been found. In 2018, a hypothetical assumption about PBVs belonging to prokaryotic viruses was put forward and experimentally substantiated. This hypothesis is based on the presence of Shine-Dalgarno sequences in the genome of all PBVs before three reading frames (ORF) at the ribosomal binding site, with which the prokaryotic genome is saturated, while in the eukaryotic genome such regions occur with low frequency. The genome saturation with the Shine-Dalgarno sequences, as well as the preservation of this saturation in the progeny, according to scientists, allows us to attribute PBVs to prokaryotic viruses. On the other hand, there is a possibility that PBVs belong to viruses of eukaryotic hosts - fungi or invertebrates, since PBV-like sequences similar to the genome of fungal viruses from the families of mitoviruses and partitiviruses have been identified. In this regard, the idea arose that, in terms of reproduction mode, PBVs resemble fungal viruses. The divergence of views on the true PBV host(s) has sparked discussions among scientists and required further research to elucidate their nature. The review highlights the results of the search for a PBV host. The reasons for the occurrence of atypical sequences among the PBV genome sequences that use an alternative mitochondrial code of lower eukaryotes (fungi and invertebrates) for the translation of viral RNA-dependent RNA polymerase (RdRp) instead of the standard genetic code are analyzed. The purpose of the review was to collect arguments in support of the hypothesis about the phage nature of PBVs and to find the most realistic explanation of the reasons for identifying non-standard genomic sequences for PBVs. Based on the hypothesis about the genealogical relationship of PBVs with RNA viruses from other families with similar segmented genomes, such as Reoviridae, Cystoviridae, Totiviridae and Partitiviridae, virologists support the assumption of a decisive role in the origin of atypical PBV-like reassortment strains between PBVs and viruses of the listed families. The collected arguments given in this review indicate a high probability of a phage nature of PBVs. The data presented in the review show that the belonging of PBV-like progeny to prokaryotic or eukaryotic viruses is determined not only by its genome saturation level with a prokaryotic motif, standard or mitochondrial genetic code. The primary structure of the gene encoding the viral capsid protein responsible for the presence or absence of specific proteolytic properties of the virus that determine its ability for independent horizontal transmission into new cells may also be a decisive factor.

Ocurrence of enteroparasites with zoonotic potential in animals of the rural area of San Andres, Chimborazo, Ecuador.

The aim of this research was the identification of the enteroparasites harbored by the animals of the San Andrés community, to evaluate their role as susceptible hosts and sources of infection for other animals, humans (zoonoses), as well as parasite forms spreaders to the environment in this rural area, located in the province of Chimborazo, Ecuadorian Andean region. The study was carried out combining 3 coproparasitological techniques: direct examination, Ritchie and Ziehl-Neelsen in 300 animal stool samples RESULTS: Blastocystis sp., Entamoeba spp., Giardia spp., Balantidium spp., Cryptosporidium spp., Ascaris spp., Toxocara spp., Ancylostoma spp., Strongylida, Hymenolepis nana and Echinococcus spp., were detected. Infection by protozoa (87.3%) was higher than helminths (31.0%). All cattle, sheep and guinea pigs were found parasitized, and the presence of Blastocystis sp., Entamoeba spp. and Cryptosporidium spp. by all groups of animals stands out. It is also remarkable the presence of Giardia spp. in swine (19.2%), big herbivores-livestock (11.5%), leporids (8.3%) and carnivores (5.9%); Balantidium spp. in swine (19.2%), big herbivores-livestock (5.8%) and carnivores (1.2%); Hymenolepis nana in guinea pigs (2.1%); and Toxocara spp. (15.7%), Echinococcus spp. (9.6%) and Ancylostoma spp. (6.0%) in dogs. Animals from San Andrés have a wide spectrum of intestinal parasitic forms in their feces, being a source of infection to other animals and humans, and a source of contamination of the environment, posing a risk factor and reinforcing the idea of the need for more effective treatments and hygienic measures to improve livestock production and cutting its transmission.

Molecular epidemiology of Cryptosporidium spp. in an agricultural area of northern Vietnam: A community survey.

The purpose of this study was to investigate the occurrence of Cryptosporidium infection and the potential for transmission of Cryptosporidium spp. between animals and humans in northern Vietnam. A total of 2715 samples (2120 human diarrheal samples, 471 human non-diarrheal samples, and 124 animal stool samples) were collected through our community survey in an agricultural area. All samples were tested for Cryptosporidium spp. by direct immunofluorescence assay (DFA) using a fluorescent microscope. DNA extraction, PCR amplification of three genes (COWP, SSU-rRNA, and GP60), and sequencing analysis were performed to identify Cryptosporidium spp. Of 2715 samples, 15 samples (10 diarrheal samples, 2 non-diarrheal samples, and 3 animal stool samples) tested positive by PCR for the COWP gene. Three species of Cryptosporidium spp. were identified as C. canis (from six human diarrheal samples, two human non-diarrheal samples, and one dog sample), C. hominis (from four human diarrheal samples), and C. suis (from two pig samples) by sequencing the amplified COWP and/or SSU-rRNA genes. In terms of C. hominis, the GP60 subtype IeA12G3T3 was detected in all four human diarrheal samples. Although the number of positive samples was very small, our epidemiological data showed that the emerging pattern of each of the three species (C. canis, C. hominis, and C. suis) was different at this study site. While C. hominis and C. suis were only detected in human and pig samples, respectively, C. canis was detected in samples from both dogs and humans. We suspect that C. canis infections in humans at this study site may be due to environmental contamination with animal and human feces.

Seasonal prevalence, risk factors, and One Health intervention for prevention of intestinal parasitic infection in underprivileged communities on the Thai-Myanmar border

BackgroundTha Song Yang District, located on the Thai-Myanmar border, contributes to the second highest cases of amoebic dysentery due to intestinal parasitic infections (IPI). However, there were limited disease prevalence data, specific surveillance systems, and interventions available. ObjectiveThis study aimed to explore the epidemiological features of the IPIs and apply the One Health (OH) approach to solve IPI-related problems. MethodsPrevalence of asymptomatic infections in human and animals, yearly symptomatic cases, and associated risk factors were investigated. The OH intervention included improving the knowledge, attitude, and practice (KAP) of the community, microscopic diagnosis training, and stakeholder engagement for IPI prevention designs. ResultsThe prevalence of asymptomatic cases was much higher than that of the symptomatic cases. Infective stages of the intestinal parasites were discovered in animal stool and water samples, indicating possible transmission routes. One year after the intervention, there were significant declines in asymptomatic IPIs and symptomatic cases of amoebic dysentery. Significant improvements in KAP and awareness regarding water and manure-waste management of the community were observed. ConclusionWe reported the successful application of the OH intervention in reducing the IPI prevalence and mitigating disease-related risks. The intervention might be applied to address other infectious diseases in the future.

Molecular epidemiology of Giardia spp. in northern Vietnam: Potential transmission between animals and humans

Giardia spp. is detected frequently in humans and animals. Although many studies have been conducted on the epidemiology of giardiasis, there is a scarcity of information on the genetic diversity and the dynamics of transmission of Giardia spp. in Vietnam. The zoonotic potential of Giardia spp. remains elusive. The objective of this study was to determine the genetic diversity of Giardia spp. in both humans and livestock to assess the existence of a route of infection between livestock and humans. Our goal was to assess the role animals play in the epidemiology of human infection in northern Vietnam. In Hien Khanh commune in northern Vietnam, 311 households with 1508 residents were randomly selected for a diarrheal cohort study. Of these, 2120 human diarrheal samples were collected from 1508 residents in 2014 and 2017. Of these, non-diarrheal samples were cross-sectionally collected from 471 residents. At the same site, livestock samples from buffalo, dairy and beef cattle, pigs, and dogs were collected. All stool samples were examined for Giardia spp. by Direct Immunofluorescence Assay (DFA) using fluorescent microscope. DNA extraction, PCR analysis of the 3 genes (bg, gdh, tpi), and sequencing analysis were continuously carried out. A total of 23 animal stool samples, 8 human non-diarrheal samples, and 36 human diarrheal samples were Giardia spp. were positive by PCR using the bg and gdh genes. Giardia spp. assemblage AII and E were detected in both animal samples and human samples in this study site. The detection of assemblage E in human stool samples suggests the first human case report in Vietnam. We assume that the unexpected human infection of all Giardia assemblages including A, B, and E may be due to an environment contaminated with animal and human feces in this village.

The molecular characters and antibiotic resistance of Clostridioides difficile from economic animals in China

BackgroundIt has been performed worldwidely to explore the potential of animals that might be a reservoir for community associated human infections of Clostridioides difficile. Several genetically undistinguished PCR ribotypes of C. difficile from animals and human have been reported, illustrating potential transmission of C. difficile between them. Pig and calf were considered as the main origins of C. difficile with predominant RT078 and RT033, respectively. As more investigations involved, great diversity of molecular types from pig and calf were reported in Europe, North American and Australia. However, there were quite limited research on C. difficile isolates from meat animals in China, leading to non-comprehensive understanding of molecular epidemiology of C. difficile in China.ResultsA total of 55 C. difficile were isolated from 953 animal stool samples, within which 51 strains were from newborn dairy calf less than 7 days in Shandong Province. These isolates were divided into 3 STs and 6 RTs, of which ST11/RT126 was predominant type, and responsible for majority antibiotic resistance isolates. All the isolates were resistant to at least one tested antibiotics, however, only two multidrug resistant (MDR) isolates were identified. Furthermore, erythromycin (ERY) and clindamycin (CLI) were the two main resistant antibiotics. None of the isolates were resistant to vancomycin (VAN), metronidazole (MTZ), tetracycline (TET), and rifampin (RIF).ConclusionsIn this study, we analyzed the prevalence, molecular characters and antibiotic resistance of C. difficile from calf, sheep, chicken, and pig in China. Some unique features were found here: first, RT126 not RT078 were the dominant type from baby calf, and none isolates were got from pig; second, on the whole, isolates from animals display relative lower resistant rate to these 11 tested antibiotics, compared with isolates from human in China in our previous report. Our study helps to deep understanding the situation of C. difficile from economic animals in China, and to further study the potential transmission of C. difficile between meat animals and human.

Humoral responses to Schistosoma japonicum soluble egg antigens in domestic animals in Lindu Subdistrict, Central Sulawesi Province, Indonesia

Background and Aim: Schistosomiasis japonica, a disease caused by Schistosoma japonicum, is a public health problem in the Philippines, the Republic of Indonesia, and the People's Republic of China. The disease is known as zoonotic, meaning other than humans, animals are involved as the reservoirs. In Indonesia, schistosomiasis surveillance in animals is not continuous. Thus, the study to determine the prevalence of the disease in animals is needed. The study was aimed to determine the seroprevalence of S. japonicum infection among four species of domestic animals in the Lindu Sub-district, Central Sulawesi Province of Indonesia. Materials and Methods: Blood samples of domestic animals were collected and analyzed for the presence of anti-S. japonicum immunoglobulin G antibodies against S. japonicum soluble egg antigens using the indirect hemagglutination assay. Animal stool samples were collected, and the miracidia-hatching assay was used for the detection of S. japonicum infection. Additional data concerning the animal identity and the management practices were obtained through a questionnaire used in surveys and interviews. Results: A total of 146 sera from 13 cattle, 24 buffaloes, 54 pigs, and 55 dogs were collected. The overall schistosomiasis seroprevalence was 64.4%. The serology prevalence in cattle, buffalo, pig, and dog was 100.0%, 41.7%, 74.1%, and 56.4%, respectively. Domestic animals in all of five villages have previous exposure with S. japonicum as seropositive animals detected in every village. A total of 104 animal stool samples from 146 animals sampled were obtained. The overall schistosomiasis prevalence determined by the miracidia hatching assay was 16.35%. The sensitivity and specificity of indirect hemagglutination assay (IHA) in the current study were 88.24% and 41.37%, respectively, with miracidia hatching assay as the gold-standard method. Conclusion: This study has shown a high seroprevalence of schistosomiasis japonica among domestic animals in the Lindu Subdistrict. IHA can be used as the screening method for the detection of S. japonicum infection in domestic animals. Chemotherapy and animal livestock grazing management programs to reduce the parasite burden and Schistosoma egg contamination in the environment must be implemented as part of one health approaches, in addition to other control measures.

An exploration of the gut and environmental resistome in a community in northern Vietnam in relation to antibiotic use

BackgroundAntibiotic resistance is a major global public health threat. Antibiotic use can directly impact the antibiotic resistant genes (ARGs) profile of the human intestinal microbiome and consequently the environment through shedding.MethodsWe determined the resistome of human feces, animal stools, human food and environmental (rain, well, and irrigative water) samples (n = 304) in 40 households within a community cohort and related the data to antibiotic consumption. Metagenomic DNA was isolated and qPCR was used to determine presence of mobile colistin resistance (mcr) genes, genes encoding extended-spectrum β-lactamases (ESBL), carbapenemases and quinolone resistance genes.ResultsNearly 40 % (39.5%, 120/304) of samples contained ESBL genes (most frequent were CTX-M-9 (23.7% [72/304]), CTX-M-1 (18.8% [57/304]). Quinolone resistance genes (qnrS) were detected in all human and 91% (41/45) of animal stool samples. Mcr-1 and mcr-3 were predominantly detected in human feces at 88% (82/93) and 55% (51/93) and animal feces at 93% (42/45) and 51% (23/45), respectively. Mcr-2, mrc-4 and mcr-5 were not detected in human feces, and only sporadically (< 6%) in other samples. Carbapenemase-encoding genes were most common in water (15% [14/91]) and cooked food (13% [10/75]) samples, while their prevalence in human and animal stools was lower at 4% in both human (4/93) and animal (2/45) samples. We did not find an association between recent antibiotic consumption and ARGs in human stools. Principal component analysis showed that the resistome differs between ecosystems with a strong separation of ARGs profiles of human and animal stools on the one hand versus cooked food and water samples on the other.ConclusionsOur study indicated that ARGs were abundant in human and animal stools in a rural Vietnamese community, including ARGs targeting last resort antibiotics. The resistomes of animal and human stools were similar as opposed to the resistomes from water and food sources. No association between antibiotic use and ARG profiles was found in a setting of high background rates of AMR.

Antibiotic resistance, ESBL genes, integrons, phylogenetic groups and MLVA profiles of Escherichia coli pathotypes isolated from patients with diarrhea and farm animals in south-east of Iran.

The aims of this study were to investigate the prevalence, antibiotic resistance, presence of class 1 and 2 integrons, Extended Spectrum β-Lactamases (ESBL) genes, phylogenetic group and epidemiological relationships of EPEC, ETEC and EHEC pathotypes isolated from patients with diarrhea and farm animals in south east region of Iran. A total of 671 diarrheagenic E. coli (DEC) were collected from stool samples of 395 patients with diarrhea and 276 farm cattles and goats. Presence of EPEC, ETEC and EHEC were identified using multiplex-PCR employing primers targeted the shiga toxin (stx), intimin (eae), bundle forming pili (bfp), and enterotoxins (lt and st) genes. The highest proportion of the patients (64%) were children under age 1-15 year (p ≤ 0.05). Among the isolates, atypical EPEC was detected in 26 patients and 14 animal stool samples, while typical EPEC was found in 2 cattles. ETEC isolates were detected in stools of 13 patients and 4 EHEC was identified in 3 goats and one cattle. The isolates were checked for susceptibility to 14 antibiotics. 50% (n = 13) of EPEC and 61.5% (n =8) of ETEC showed multi-drug resistance (MDR) profiles and one EPEC was found to be extensive drug resistant (XDR). In contrast, EHEC isolates were susceptible to the majority of antimicrobial agents. The MDR isolates were positive for blaTEM and blaCTX-M ESBL genes and carried class 1 integrons. Further study on the biofilm formation indicated that, 3 out of 4 EHEC isolates showed strong biofilm, while other pathotypes had either moderate, weak or no biofilm activity. Majority of EPEC isolates were belonged to phylogenetic group B1, all except one ETEC were classified as phylogenetic group A and two EHEC were belonged to phylogroup D, respectively. A multilocus variable tandem repeat analysis (MLVA) exhibited 22 distinct patterns. In conclusion, MLVA data showed high clonal diversity. Presence of EHEC in animal origins pose public health concern in this region.

Extensive conservation of prokaryotic ribosomal binding sites in known and novel picobirnaviruses

Currently, the Leviviridae and Cystoviridae are the only two recognized families of prokaryotic RNA viruses. Picobirnaviruses, which are bisegmented double-stranded RNA viruses commonly found in animal stool samples, are currently thought to be animal viruses, but have not been propagated in cell culture or in an animal model. We hypothesize that picobirnaviruses are prokaryotic RNA viruses. We identified and analyzed the genomes of 38 novel picobirnaviruses and determined that a classical bacterial sequence motif, the ribosomal binding site (RBS), is present in the 5′ untranslated regions (5′ UTRs) of all of the novel as well as all previously published picobirnavirus sequences. Among all viruses, enrichment of the RBS motif is only observed in viral families that infect prokaryotes and not in eukaryotic infecting viral families. These results will enable future studies to more accurately understand the biology of picobirnaviruses.

BLASTOCYSTIS HOMINIS: A MYSTERIOUS AND COMMONLY DISREGARDED PARASITE

Blastocystis hominis (B. hominis) is an anaerobic, single-cell protozoan, commonly present in human and animal stool samples. It can be found in healthy people as well and it still has not been elucidated whether it is a commensal organism or a pathogen. Blastocystosis is a disease caused by the protozoan in humans. The prevalence of the parasitosis varies both between the countries, and between certain population groups within individual countries. Due to poor hygienic conditions, common exposure to animals and intake of contaminated water and food, people in the developing countries have got a higher prevalence of blastocystosis, but economically developed countries have not been spared either. The taxonomy of B. hominis is still a matter of debates. For the reasons of genetic diversity, it has been suggested that the name B. hominis should be replaced with „Blastocystis species‟. Seventeen subtypes of the species have been so far identified, and a definitive characterization of Blastocystis spp. is possible at the molecular level only. The parasite is transferred by the fecal-oral route. A variety of hosts have been identified, and animal-to-human and vice versa transfers have been documented. The most common manifestations of the infection with the organism are diarrhea, abdominal pain, nausea, and bloating. This infection has also been associated with the irritable bowel syndrome (IBS), non-specific colitis, chronic inflammatory bowel disease (CIBD), and urticaria. The diagnosis can be made using the methods of conventional microscopy (CVM), phase-contrast and electron microscopy, cultivation, serodiagnosis, and by using molecular methods. The infection caused by the parasite does not always require treatment. In symptomatic patients, the first line medical treatment is metronidazole. Further studies are required to resolve all dilemmas regarding the parasite.

Serotype identification and antibiotic susceptibility of Shiga toxin-producing Escherichia coli in the Weishan area in Shandong Province, China

Objective: To determine the serotypes and drug resistance profiles of Shiga toxin-producing Escherichia coli (STEC) in animal stools from the Weishan area in Shandong Province, China. To provide the basis for further study. Methods: Five hundred animal stool samples (from pigs, cattle, sheep, dogs and birds) were collected from the Weishan area and STEC strains were isolated from these samples. Strains were serotyped by a serum agglutination test, and their drug resistance profiles were determined through antimicrobial sensitivity experiments. In this study, PCR was used to detect tetracycline resistance genes (tetA, tetB, tetC, tetD) and beta-lactam resistance genes (blaSHV-1, blaCTX-M, blaTEM). Results: Sixteen strains of STEC were isolated from animal stool samples. Thirteen strains were isolated from pig stool samples, two from bovine stool samples and one from a sheep stool sample. Two of the strains were identified as E. coli O157:H7, and other 14 strains were non-O157 STEC of different serotypes. Antimicrobial sensitivity experiments showed that 15 of the strains were multidrug resistant. The rates of resistance were as follows: nalidixic acid (12/16 strains), sulfisoxazole (11/16), trimethoprim and sulphame-thoxazole (11/16), doxycycline (9/16), azithromycin (9/16), tetracycline (9/16), chloramphenicol (8/16) and streptomycin (8/16). Therefore, nalidixic acid showed the highest rate of resistance among the strains, followed by trimethoprim and sulphame-thoxazole, and sulfisoxazole. Resistance to cefepime or imipenem was not detected. In total, three types of drug resistance genes (tetA, tetB and tetC) were detected among the 16 strains. Conclusion: The results showed that STEC strains isolated from animals in the Weishan area were of a range of serotypes. The 16 strains of STEC isolated from animal stools in this area were resistant to a number of antibiotics, with many strains displaying multidrug resistance.

Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) Genotyping of Escherichia coli Strains Isolated from Different Animal Stool Specimens

Escherichia coli is a commensal-pathogenic organism, which includes a wide range of strains. Despite several advanced molecular-genomic technologies for detecting and identifying different strains ofE. coli, Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) technique is a quick, sharp and cost effective fingerprint method. The major purpose of the present study was to determine the distribution of ERICs within E. colistrains isolated from different healthy animal stool specimens including hens, sheep, and cows, as an appropriate and quick molecular-genomic tool. The animal stool samples were obtained during 1 year (October 2012 to October 2013), from animal husbandries around Tehran and Alborz provinces, Iran. After screening processes, theE. colibacteria were isolated and cultured via standard microbiological methods. The DNA molecules ofE. coli bacteria were harvested and Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) was applied for bacterial molecular genotyping. The ERIC-PCR products were run on 1% gel electrophoresis. The final images regarding gel electrophoresis banding patterns were used for dendrogram generation via the GelClust software. Of 120 isolated samples, 115 different strains were recognized asE. coli. The fingerprint patterns involved 380 to 3280 bp bands. The predominant bands included 2900 bp, 1200 bp, and 1200 bp in stool samples of hens, sheep, and cows, respectively. The highest frequencies and diversities were seen amongE. colistrains isolated from hens and sheep stool samples. The DNA profiles were clearly detectable via specific fingerprint patterns. The ERIC-PCR seemed to be a good approach for molecular typing ofE. colistrains isolated from different animal sources.

Identification of Blastocystis subtypes in clinical stool samples from Sao Paulo City, Brazil

SUMMARY Blastocystis sp. is a protozoan commonly found in human and animal stool samples. Several pathogenic and zoonotic aspects of this organism are still unknown. The aim of the present study was to investigate Blastocystis subtypes (STs) in samples from patients of the Hospital das Clínicas of the Faculdade de Medicina at the Universidade de São Paulo (HC-FMUSP), Brazil. Blastocystis sp.-positive stool samples diagnosed at the Section of Parasitology of the Central Laboratory (HC-FMUSP) were used for DNA isolation. Polymerase chain reaction (PCR) was performed using specific primers targeting the small-subunit rRNA gene. Direct DNA sequencing of the PCR products was performed and the DNA sequences were then aligned and compared with other sequences obtained from the GenBank database. Phylogenetic analysis was used to identify STs and determine the phylogenetic relationships between the sequences. Four STs were identified: ST1 (22·5%), ST2 (12·5%), ST3 (60%) and ST6 (5%). In conclusion, ST3 was the most prevalent ST among the human isolates followed by ST1. The present study is one of the few providing STs data from the human population in South America. Determining ST prevalence in human samples may contribute to the monitoring of Blastocystis sp. infection transmission in endemic regions.

A Cross-Sectional Study of Helicobacter Infection Among Laboratory Animals and Animal Research Workers

A Cross-Sectional Study of Helicobacter Infection Among Laboratory Animals and Animal Research Workers

Detection of Shiga toxin variants among Shiga toxin-formingEscherichia coliisolates from animal stool, meat and human stool samples in India

To study the prevalence and distribution of various variants in the stx gene of Shiga toxin-producing Escherichia coli (STEC) isolated from diverse environmental sources (animal stool, meat) and human illness, from a large geographic area in India, and to understand the association between variants, serotype distribution and human disease. A surveillance for STEC was conducted in the semi-urban and rural areas of Punjab, Himachal, Haryana and Chandigarh. Shiga toxin-producing Escherichia coli isolates (80 animal stool, 39 meat, 21 human stool from diarrhoea and HUS cases) were characterized for stx variants by PCR. Shiga-like toxin (Stx) was detected using Ridascreen-EIA assay. Variant stx2c was the most common (25·1%), followed by stx1d (13%), stx1c (10·7%) and stx2d (9·2%), whereas stx2e, stx2f and stx2g were absent. Only 8/21 (38%) human isolates harboured stx variants, of which stx2c and stx2d were found in 2 and 1 isolates, respectively. The low frequency of carriage of these potentially more pathogenic variants may explain the low severity of human illness seen in India. Shiga-like toxin was detected in only 42 of the isolates positive for the stx genes probably due to the low levels of toxins produced. Serogroup distribution was found to be diverse, suggesting the lack of any predominant circulating type. The presence of stx variants 1c, 1d, 2c and stx2d in diverse environmental and human sources in India was demonstrated. The prevalence of the most common subtype stx2c found in this study in animal isolates may pose a threat to the public health. We report the subtyping of human STEC isolates and report the presence of stx1d subtype for the first time from India. We demonstrated the presence of potentially pathogenic subtypes in the environmental specimens which may act as a reservoir for human infections. Serogroups new to India were also reported.

Carbapenem resistance and Acinetobacter baumannii in Senegal: the paradigm of a common phenomenon in natural reservoirs.

Incidence of carbapenem-resistant Acinetobacter baumannii is rising in several parts of the world. In Africa, data concerning this species and its resistance to carbapenems are limited. The objective of the present study was to identify the presence of A. baumannii carbapenem-resistant encoding genes in natural reservoirs in Senegal, where antibiotic pressure is believed to be low. From October 2010 to January 2011, 354 human head lice, 717 human fecal samples and 118 animal fecal samples were screened for the presence of A. baumannii by real time PCR targeting bla OXA51-like gene. For all samples positive for A. baumannii, the carbapenemase-hydrolysing oxacillinases blaOXA23-like and blaOXA24-like were searched for and sequenced, and the isolates harbouring an oxacillinase were genotyped using PCR amplification and sequencing of recA gene. The presence of A. baumannii was detected in 4.0% of the head lice, in 5.4% of the human stool samples and in 5.1% of the animal stool samples tested. No bla OXA24 gene was detected but six fecal samples and three lice were positive for bla OXA23-like gene. The bla OXA23-like gene isolated in lice was likely a new oxacillinase sequence. Finally, the A. baumannii detected in stools were all of recA genotype 3 and those detected in lice, of recA genotype 4. This study shows for the first time a reservoir of bla OXA23-like-positive gene in human head lice and stool samples in Senegal.