Prevalence of Blastocystis and Dientamoeba fragilis in diarrheal patients in Corum, Türkiye.

To investigate the prevalence of Blastocystis and Dientamoeba fragilis in diarrhea patients and healthy individuals in Corum, Türkiye, fecal samples from 92 diarrhea patients and 50 healthy individuals were collected and evaluated using direct microscopy and molecular methods to screen for bacteria, protozoa, and viruses. The prevalence of Blastocystis was 24.6% in total and more frequent in the healthy group (30.0%). The commonly detected STs (subtypes) were ST3 (40.0%) and ST2 (34.2%). The distribution of Blastocystis STs in the healthy and diarrheal groups did not show any difference in sex and age, but ST3 was detected more frequently in patients aged from 40 to 59 years (p < 0.05). Alleles 4 (8/12) and 2 (4/12) were present in ST1; 9 (3/5) and 12 (2/5) in ST2; 34 (9/14), 36 (3/14), and 38 (2/14) in ST3; and only allele 42 (2/2) in ST4. D. fragilis was present in 8.4% of the population. However, there was no statistically significant difference between the healthy and diarrheic groups (12.0% and 6.5%, respectively), neither with respect to age nor sex. Co-infection was 58.3% and was more frequent in healthy individuals (33.3%) than in diarrhea patients (25.0%). Blastocystis ST3 was the most common subtype detected, with D. fragilis at 33.3%. Salmonella, Shigella, or helminth eggs were not observed in all groups, but Entamoeba histolytica, Giardia intestinalis, Cryptosporidium, Rotavirus, Adenovirus, and Clostridium difficile toxin were found only in diarrhea patients. These findings support the hypothesis that Blastocystis and D. fragilis may be part of the healthy human gut microbiome.

Enterocytozoon bieneusi in patients with diarrhea and in animals in the northeastern Chinese city of Yichun: genotyping and assessment of potential zoonotic transmission.

Enterocytozoon bieneusi is a common microsporidia species in humans and animals. Due to lack of effective vaccines and drugs, understanding of its epidemiological status and characteristics in different hosts is an important step in controlling the infection. The present study aimed at determining the prevalence of E. bieneusi in humans with diarrhea and animals in Yichun, in northeastern China, and assessing the epidemiological role of animals in the transmission of microsporidiosis. A total of 540 fecal samples were collected from diarrheal patients (n = 222) and 11 animal species (n = 318). Enterocytozoon bieneusi was identified and genotyped by polymerase chain reaction (PCR) and sequencing of the internal transcribed spacer (ITS) region of the rRNA gene. Enterocytozoon bieneusi was detected in 1.4% (3/222) of diarrheal patients, and genotype D and novel genotypes YCHH1 and YCHH2 were identified. Enterocytozoon bieneusi was detected in wild boars (7.7%), sika deer (8.2%), dogs (3.2%), and ostriches (10.7%), and genotypes D, Type IV, Peru6, BEB6 and novel genotypes YCHA1, YCHA2 and YCHA3 were identified. Genotypes YCHH1, YCHH2 and YCHA1 were phylogenetically assigned to group 1, while YCHA2 and YCHA3 to groups 2 and 11, respectively. The finding of genotype D in humans and animals, and the identification of zoonotic genotypes Peru6, Type IV, BEB6 in animal-derived E. bieneusi isolates indicate the potential of zoonotic transmission of microsporidiosis in the investigated area. The observation of the three novel genotypes in group 1 indicates their zoonotic potential.

Title: Enterocytozoon bieneusi chez des patients souffrant de diarrhée et des animaux dans la ville de Yichun, au nord-est de la Chine: génotypage et évaluation de la transmission zoonotique potentielle. Abstract: Enterocytozoon bieneusi est une espèce de microsporidie commune chez les humains et les animaux. En raison du manque de vaccins et de médicaments efficaces, la compréhension de son statut épidémiologique et de ses caractéristiques chez différents hôtes est une étape importante dans le contrôle de l'infection. La présente étude visait à déterminer la prévalence d'E. bieneusi chez les humains souffrant de diarrhée et les animaux à Yichun, dans le nord-est de la Chine, et à évaluer le rôle épidémiologique des animaux dans la transmission de la microsporidiose. Cinq cent quarante échantillons fécaux ont été prélevés chez des patients diarrhéiques (n = 222) et 11 espèces animales (n = 318). Enterocytozoon bieneusi a été identifié et génotypé par PCR et séquençage de la région de l'espaceur interne transcrit (ITS) du gène de l'ARNr. Enterocytozoon bieneusi a été détecté chez 1,4 % (3/222) des patients souffrant de diarrhée, et le génotype D et les nouveaux génotypes YCHH1 et YCHH2 ont été identifiés. Enterocytozoon bieneusi a été détecté chez des sangliers (7,7 %), des cerfs sika (8,2 %), des chiens (3,2 %) et des autruches (10,7 %), et les génotypes D, Type IV, Peru6, BEB6 et les nouveaux génotypes YCHA1, YCHA2 et YCHA3 ont été identifiés. Les génotypes YCHH1, YCHH2 et YCHA1 ont été phylogénétiquement assignés au groupe 1, et YCHA2 et YCHA3 respectivement aux groupes 2 et 11. La découverte du génotype D chez les humains et les animaux et l'identification des génotypes zoonotiques Peru6, Type IV, BEB6 dans les isolats d'E. bieneusi d'origine animale indiquent le potentiel de transmission zoonotique de la microsporidiose dans la zone étudiée. L'observation des trois nouveaux génotypes dans le groupe 1 indique leur potentiel zoonotique.

Giardia duodenalis in patients with diarrhea and various animals in northeastern China: prevalence and multilocus genetic characterization.

BACKGROUND: Giardia duodenalis is a common parasitic diarrheal agent in humans, especially in developing countries. The aim of this study was to investigate the prevalence and multilocus genetic characterization of G. duodenalis in patients with diarrhea and animals in northeastern China, and to assess the epidemiological role of animals in the transmission of human giardiasis. METHODS: A total of 1739 fecal specimens from 413 diarrheal patients and 1326 animals comprising 16 mammal species were collected in Heilongjiang Province of China and screened for G. duodenalis by PCR and sequencing of the SSU rRNA gene. All G. duodenalis-positive specimens were subtyped by PCR and sequencing of the bg, tpi, and gdh genes. To detect additional mixed infections of different assemblages, assemblage A/B/E-specific PCRs were performed to amplify the tpi gene. RESULTS: Sequence analysis of the SSU rRNA gene determined the prevalence of G. duodenalis (5.81%, 24/413) in diarrheal patients, with a peak in minors aged 5-17 years, and identified assemblages A and B. MLG-AII and MLG-B1 were obtained based on concatenated nucleotide sequences of the bg, tpi, and gdh genes, with MLG-AII being identical to a cat-derived isolate reported previously. By sequence analysis of the SSU rRNA gene, G. duodenalis was detected in 214 (16.14%) animals belonging to 11 mammal species, with the prevalence ranging from 1.69 to 53.85%, and assemblages A to G were identified. Sequence analysis of the bg, tpi, and gdh genes from 46 specimens produced 31 MLGs, including MLG-AI (n = 1), MLG-B2-B8 (n = 18), and MLG-E1-E23 (n = 27). CONCLUSIONS: The finding of G. duodenalis in diarrheal patients enhances consciousness of detecting G. duodenalis in clinical practice and emphasizes the importance of health education in local inhabitants, especially in the age group of 5-17 years. The identification of seven assemblages (A to G) and 33 MLGs reveals genetic heterogeneity of G. duodenalis in the investigated areas. Due to insufficient homology data on the zoonotic transmission of G. duodenalis, the precise epidemiological role that animals play in the transmission of human giardiasis needs to be assessed by more large-scale molecular epidemiological investigations of local humans and animals.

Prevalence of Antimicrobial Resistant of Vibrio parahaemolyticus Isolated from Diarrheal Patients - Six PLADs, China, 2016-2020.

WHAT IS ALREADY KNOWN ON THIS TOPIC?: Vibrio parahaemolyticus (V. parahaemolyticus) is frequently resistant to common antimicrobials such as ampicillin and generally highly susceptible to most clinically used antimicrobials. WHAT IS ADDED BY THIS REPORT?: V. parahaemolyticus were highly resistant to cefazolin and ampicillin: 94.4% and 37.0%, respectively. However, it was below 3% resistance to all 10 other antimicrobials including clinically relevant agents and even imipenem. The overall levels of antimicrobial resistance and multidrug resistance were 95.1% and 3.3%, respectively. The distribution of antimicrobial resistance and the multidrug resistance had regional, temporal, sexual, and isolated source strain variation. WHAT ARE THE IMPLICATIONS FOR PUBLIC HEALTH PRACTICE?: This study provides data on drug resistance of V. parahaemolyticus in Chinese clinical settings, which will help develop a public health strategy.

Investigation of the effect of chitosan and silver nanoparticles on the antibiotic resistance of Escherichia coliO157:H7 isolated from some milk products and diarrheal patients in Sohag city, Egypt.

BACKGROUND AND AIM: Antimicrobial-resistant Escherichia coli O157:H7 causes serious diseases in humans, especially when circulated in their food. This study was designed to detect the presence of E. coli O157:H7 using the fliC H7 gene in some milk products as kareish cheese, labena, and yoghurt sold in Sohag city, Egypt, and among diarrheal patients admitted to governmental hospitals in Sohag and also to highlight the risk factors associated with their infection. In addition, the antimicrobial resistance and the effect of chitosan nanoparticles (CNP) and silver nanoparticles (SNP) on E. coli O157:H7 isolates obtained from both milk products and patients were investigated. MATERIALS AND METHODS: Microbiological culture methods and polymerase chain reaction were used for detecting E. coli O157:H7 in 150 milk products and 150 stool samples. Resistance against some antimicrobials that were used in the treatment of animals and humans was investigated using the disk diffusion technique. CNP and SNP at two concentrations (30 and 60 µg/mL) and average sizes of 25.1 and 26.5 nm, respectively, were identified by transmission electron microscopy. Their effect on E. coli O157:H7 isolates was examined using the well diffusion method. Risk factors for infection were investigated using statistical analysis. RESULTS: There were 11.3% and 14.7% of milk products and stool samples positive for E. coli O157:H7, respectively. These isolates exhibited high antimicrobial resistance to ampicillin, tetracycline, and gentamycin. CNP and SNP demonstrated inhibitory effects on E. coli O157:H7 growth, which significantly increased at high concentrations (60 µg/mL), with mean inhibition zones of 31.941±3.749 and 30.681±3.871 mm for CNP in milk products and patient isolates, respectively. The respective values for SNP were 33.588±3.675 mm and 32.500±2.444 mm, indicating a higher bactericidal effect than that of CNP. Regarding risk factors for infection, both young and elderly subjects and those in contact with infected persons and/or having chronic diseases were infected. CONCLUSION: CNP and SNP are suitable for both medical and agricultural applications for disease control and enhancement of food quality.

Comparison of the Filtration Culture and Multiple Real-Time PCR Examination for Campylobacter spp. From Stool Specimens in Diarrheal Patients.

Campylobacter is one of the most common pathogens leading to the bacterial diarrheal illness. In order to set up one effective culture independent assay for the screen of the Campylobacter infection in the diarrheal patients, the quadruple real-time PCR method comparing to the culture based on the enriched filtration method which was recognized as the most effective isolation method was assessed for 190 stool samples from the diarrheal patients collected during the Foodborne Diseases Active Surveillance Network in Beijing. This multiple real-time PCR was designed to identify the Campylobacter genus, C. jejuni, C. coli, and C. lari simultaneously. With the enrichment culture method, 23 (12.1%, 23/190) Campylobacter isolates were obtained (20 C. jejuni and 3 C. coli), however, 31 samples (16.3%, 31/190) were detected positively with the real-time PCR (21 C. jejuni, 8 C. coli, and 2 Campylobacter genus only). With the comparison, the real-time-PCR method is more sensitive than the enrichment filtration method (16.3 vs. 12.1%, p = 0.021). Among the culture-positive samples, 95.7% (22/23) were detected positively by PCR which indicate the specificity of this method was higher. These two methods were consistent well (Kappa = 0.785, p < 0.05). Comparing to the culture methods, the result of the multiple real-time PCR method is sensitive, reliable and rapid. The present study indicated this multiple real-time PCR can be used both for the surveillance network and the preceding screen for bacteria isolation. This is first comparative study between the culture and multiple real-time PCR method for Campylobacter identification in stool specimens from the diarrheal patients.

Genetic and Antibiotic Resistance Characteristics of Campylobacter jejuni Isolated from Diarrheal Patients, Poultry and Cattle in Shenzhen.

OBJECTIVE: To investigate genetic and antibiotic resistance characteristics of Campylobacter jejuni (C. jejuni) isolated from Shenzhen. METHODS: Multilocs sequence typing and agar dilution methods were used to define the genotype and antibiotic resistance of C. jejuni, respectively. RESULTS: In total, 126 C. jejuni strains were isolated. The prevalence of C. jejuni was 5.3% in diarrheal patients. The prevalence in poultry meat (36.5%) was higher than that in cattle meat (1.1%). However, the prevalence in poultry cloacal swabs (27.0%) was lower than that in cattle stool (57.3%). Sixty-two sequence types were obtained, among which 27 of the STs and 10 alleles were previously unreported. The most frequently observed clonal complexes were ST 21 (11.9%), ST-22 (10.3%), and ST-403 (7.1%). ST-21, ST-45, ST-354, ST-403, and ST-443 complexes overlapped between isolates from patients and cattle, whereas ST-45 and ST-574 complexes overlapped between isolates from patients and poultry. All C. jejuni were resistant to at least one antibiotic. The highest resistance rate was toward ciprofloxacin (89.7%), followed by tetracycline (74.6%), and nalidixic acid (69.0%). CONCLUSION: This is the first report of the genotypes and antibiotic resistance of C. jejuni in Shenzhen. Overlapping clonal complexes were found between isolates from patients and cattle, and between patients and poultry.

Prevalence and Molecular Characterization of Campylobacter spp. Isolated from Patients with Diarrhea in Shunyi, Beijing.

Bacterial pathogens have been confirmed as the major cause of acute diarrhea among outpatients in China. In this study, 370 stool samples from the patients aged from 15 to 87 years old with diarrhea were collected over 12 months (from May 2016 to April 2017) in two hospitals in Shunyi, Beijing. Bacterial isolation was performed for the common enteric pathogens: Campylobacter, Salmonella, Shigella, Diarrheagenic Escherichia coli, and Vibrio parahaemolyticus for 370 samples. The filtration method was used for the Campylobacter isolation in this study. The prevalence and molecular characterization of the Campylobacter were investigated. The isolation ratio for Campylobacter, Salmonella, Shigella, Diarrheagenic E. coli, and V. parahaemolyticus was 7.0% (26/370), 6.2% (23/370), 0.3% (1/370), 7.3% (27/370), and 10.3% (38/370), respectively. Based on the isolation result, Campylobacter positive cases presented in almost every month of the whole year and the isolation ratio was the highest among the tested pathogens during October to March. There was no significant difference between genders of Campylobacter positive cases. More Campylobacter positive cases presented dehydration compared with those who were positive for Salmonella. Twenty-six Campylobacter isolates were obtained in this study and 24 of these were Campylobacter jejuni. The antibiotic susceptibility tests indicated that 83.3% (20/24) of the isolates exhibited resistance to three or more types of antibiotic. Twenty STs were identified for the 26 Campylobacter isolates and four novel STs were identified in this study. No clonal cluster was found among these isolates. This is the first study for Campylobacter isolated using the filtration method in China which indicated the Campylobacter infection might be seriously under-ascertained in the diarrheal patients in China.

Prevalence of binary-toxin genes (cdtA and cdtB) among clinical strains of Clostridium difficile isolated from diarrheal patients in Iran.

AIM: In this study we investigated the prevalence of binary toxin genes, cdtA and cdtB, in clinical isolates of C. difficile from hospitalized patients with diarrhea. BACKGROUND: C. difficile binary toxin (CDT) is an action-specific ADP-ribosyltransferase that is produced by some strains of C. difficile. Co-expression of this toxin with tcdA and tcdB can lead to more severe disease in CDI patients. METHODS: Totally, 930 patients suspected of having CDI was included in this study. All samples were treated with methanol and cultured on selective C. difficile agar plates. The C. difficile isolates were further identified by PCR. Presence of tcdA, tcdB, cdtA, and cdtB genes among the strains were examined by PCR. RESULTS: Analysis of the PCR results showed a prevalence of 85.2% (144/169) for toxigenic C. diffidile. Toxin genotyping of the strains for tcdA and tcdB genes revealed the toxin profiles of A+B+, A+B-, A-B+ accounting for 86.1% (124/144), 7.6% (11/144), 6.2% (9/144) among the strains, respectively. Totally, 12.4% (21/169) of the C. difficile strains were binary toxin-positive. cdtA-B+, cdtA+ B + and cdtA+B- were detected in 43% (9/21), 38% (8/21) and 19% (4/21) of the strains, respectively. Interestingly, 12% (3/25) of nontoxigenic C. difficile strains (tcdA-B-) had either cdtA+ B + or cdtA-B+ profiles. CONCLUSION: This is the first report for the prevalence of binary toxin genes in C. difficile strains isolated from Iran. Further studies are required to investigate the exact role of binary toxins in the pathogenesis of C. difficile particularly in patients with chronic diarrhea among Iranian populations.

Prevalence of Antibiotic Resistance in Escherichia coli Fecal Isolates From Healthy Persons and Patients With Diarrhea.

OBJECTIVES: This study aimed to investigate the prevalence of antibiotic resistance in fecal Escherichia coli isolates from healthy persons and patients with diarrhea. METHODS: E. coli isolates (n = 428) were obtained from fecal samples of apparently healthy volunteers and hospitalized patients with diarrhea. Susceptibility patterns of isolates to 16 antimicrobial agents were determined by agar disc diffusion. RESULTS: Most E. coli isolates exhibited less than 10% resistance against imipenem, cefotetan, aztreonam, cefepime, cefoxitin, amikacin and netilamicin, although greater than 65% were resistant to ampicillin and tetracycline. No significant difference in resistance rates for all tested antibiotics was found between isolates from the healthy-and diarrheal-patient groups, including for multi-drug resistance (p = 0.22). The highest number of resistant antibiotics was 12 antibiotics. No significant differences in antibiotic resistance were found among the sex and age strata for isolates from healthy individuals. However, antibiotic resistance rates to cefoxitin, cefotaxime, amikacin, and netilamicin were significantly higher in the isolates of men than those of women (p < 0.05) in isolates from patients with diarrhea. Furthermore, isolates from patients with diarrhea older than 40-years of age showed higher resistance to cefepime and aztreonam (p < 0.05). CONCLUSION: High resistance to the antibiotics most frequently prescribed for diarrhea was found in isolates from patients with diarrhea and apparently healthy individuals without any significant difference.