Diarrheal Stool Research Articles

Molecular studies on antibiotic resistant Bacillus cereus isolated from meat products and human in Kaliobia, Egypt

The current study was conducted on 210 random samples of meat products (beef burger, kofta, luncheon, minced meat, sausage) and diarrheic human stool of patients suffering from vomiting and diarrhoea (35 for each). The meat products were collected from different shops and hospitals at Kaliobia Governorate, Egypt, for detection of B. cereus strains, and their phenotypic characterization as well as antibiotic resistant genes. Bacteriological examination of the collected samples indicated the identification of 51 (24.3%) isolates of B. cereus from 210 samples as 11/31.4% from kofta 13/37.1% from minced meat; 9/25.7% from sausage; 7/20.0% from beef burger; 6 /17.1% from luncheon samples; and 5/14.3% from human stool specimens. Most of 51 isolated B. cereus strains had the ability for biofilm production. The antibiotic sensitivity profiles revealed that, the isolated B. cereus were highly resistant for Penicillin-G followed by methicillin; ampicillin; oxytetracycline; sulfa-trimethoprim and cefotaxime. Meanwhile, they were highly sensitive to to gentamycin and norfloxacin followed by ciprofloxacin; meropenem and florphenicol. Further, PCR declared that bla; tetA and erm genes were amplified in 9; 7; 6 out of 10 studied B. cereus isolates giving products of 680 bp.; 502 bp. and 645 bp. respectively. Therefore, one can conclude that; B. cereus especially antibiotic resistances ones are meat-borne pathogens of public health importance and they may be the causative agents in patients suffering from vomiting and diarrhoea.

Occurrence, Molecular Characteristics, and Antimicrobial Resistance of Escherichia coli O157 in Cattle, Beef, and Humans in Bishoftu Town, Central Ethiopia.

Escherichia coli O157 is a Shiga toxin-producing E. coli causing disease in humans. Cattle are the primary reservoir of the pathogen. Information regarding the contribution of cattle to diarrheal illnesses in humans through consumption of contaminated beef is scarce in Ethiopia. We collected samples from 240 cattle, 127 beef, and 216 diarrheic patients in Bishoftu town in Ethiopia to assess the occurrence and determine the virulence genes, genetic relatedness, and antimicrobial resistance of E. coli O157. E. coli O157 was detected in 7.1% of the rectal content samples from cattle in slaughterhouses, in 6.3% (n = 127) of the beef samples, and in 2.8% of the diarrheic patients' stool samples. All isolates were positive for eae gene, 24 (77%) of them were positive for stx2 gene (21 stx2c and 3 stx2a), whereas stx1 gene was not detected. Molecular typing grouped the isolates into eight pulsed-field gel electrophoresis pulsotypes with three pulsotypes containing isolates from all three sources, one pulsotype containing one isolate from human origin and one isolate from beef. The remaining four pulsotypes contained isolates unique either to beef or to humans. With the exception of 1 multidrug-resistant isolate from beef, which was resistant to 8 antimicrobial drugs, the remaining 30 isolates were susceptible to the 14 antimicrobials tested. In conclusion, the finding of genetically similar isolates in cattle, beef, and humans may indicate a potential transmission of E. coli O157 from cattle to humans through beef. However, more robust studies are required to confirm this epidemiological link.

Isolation, Identification, and Antibiotic Susceptibility Testing of Salmonella Isolated from Foodborne Outbreaks

Background: Foodborne diseases are a major problem worldwide. The epidemiological investigations in many parts of the world have shown an increase in infections caused by Salmonella serovars. Furthermore, the emergence of drug resistance among them has become a major global concern and awareness of the resistance patterns of Salmonella could be very useful in treatment of diseases. Objective: This study aimed to investigate Salmonella serotypes in foodborne outbreaks by sequencing of ITS region of 16S-23SrRNA gene and to determine their antimicrobial susceptibility pattern. Materials and Methods: A total of 614 diarrheal stool samples were collected from 173 foodborne outbreaks in different provinces of Iran during one year. Identification of Salmonella was carried out by phenotypic and molecular (16s-23srRNA gene detection) methods and antibiotic susceptibility was performed using disc diffusion method. Results: Out of 614 samples, 18 isolates were identified as Salmonella of which 16 (88.9%) isolates were Salmonella Enteritidis and 2 (11.1%) isolates as Salmonella Paratyphi A. All isolates were sensitive to ceftazidime, and high resistance was seen with nalidixic acid with 14 (77.8%) isolates. Conclusion: Increasing antibiotic resistance in many bacterial pathogens such as Salmonella has been a major threat for human health. Therefore, identifying the antibiotic resistance patterns of Salmonella serovars may help in treatment of the associated infections.

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.

Antimicrobial Resistance and Virulence-Associated Markers in Campylobacter Strains From Diarrheic and Non-diarrheic Humans in Poland.

Campylobacteriosis is one of the most common causes of bacterial gastroenteritis. However, the clinical course of the illness varies in symptoms and severity. The aim of this study was to characterize Campylobacter jejuni (34 isolates) and C. coli (9 isolates) from persons with diarrheal and non-diarrheal stools at the time of examination and fecal sampling, in Poland by using whole-genome sequencing (WGS). Multilocus sequence typing (MLST) analysis revealed a high diversity with a total of 20 sequence types (STs) among 26 Campylobacter isolates from diarrheic and 13 STs among 17 isolates from non-diarrheic persons. ST-50 and ST-257 were most common in both groups. The phenotypic resistance rate was 74.4% for ciprofloxacin, 67.4% for sulfamethoxazole/trimethoprim, 58.1% for amoxicillin, 48.8% for tetracycline, and 46.5% for ceftriaxone. Only single isolates were resistant to erythromycin, gentamicin, and amoxicillin/clavulanic acid. Overall genotypic resistance toward amoxicillin, fluoroquinolones, tetracyclines, and aminoglycosides was predicted to occur in 93.1, 67.4, 48.8, and 11.6% of the isolates, respectively. None of the isolates showed the presence of the erm(B) gene or mutation in 23S rRNA. Neither was variation found in the important target region in L4 and L22 ribosomal proteins. In regard to the CmeABC efflux pump, a set of variable mutations affecting the regulatory region was noted. All Campylobacter isolates possessed genes associated with adhesion (cadF, jlpA, porA, and pebA) and invasion (ciaB, pldA, and flaC). The type IV secretion system (T4SS) was found in isolates from both diarrheic (15.4%, CI 95%: 6.1–33.5%) and non-diarrheic (23.5%, CI 95%: 9.6–47.3%) persons. The rates of the presence of cytolethal distending toxin cdtABC gene cluster and type VI secretion system (T6SS) were higher in Campylobacter isolates obtained from persons with diarrhea (96.2%, CI 95%: 81.7–99.3% and 26.9%, CI 95%: 13.7–46.1%) compared to isolates from non-diarrheic persons (76.5%, CI 95%: 52.7–90.4% and 11.8%, CI 95%: 3.3–34.3%). The lack of statistically significant differences between two groups in tested virulence factors suggests that individual susceptibility of the host might play more determining role in the disease outcome than characteristics of the infecting strain.

Pervasiveness of diarrheagenic E. coli pathotypes and Salmonella species among gastroenteritis patients in some selected pastoral hinterlands of the Amathole district municipality, Eastern Cape, South Africa

BackgroundDiarrhea as the consequence of gastroenteritis is one of the most significant causes of infant’s deaths across the world. Over 700000 child loses occur each year as a result of gastroenteritis infection. This study aimed at elucidating potential bacterial aetiological agents of diarrhoea within the selected rural settlements of Amathole District Municipality, Eastern Cape, South Africa. Standard culture-based methods and Polymerase chain reaction techniques were employed for the detection, isolation and validation of diarrheagenic E. coli (DEC) pathotypes and Salmonella species from diarrheal stool samples. ResultsA total of 208 (64%) isolates were positively affirmed by conventional Polymerase Chain Reaction as Diarrheagenic E. coli (DEC) and were further delineated into 4 DEC pathotypes and an additional 116 (36%) isolates were confirmed as Non-diarrheagenic E. coli. Among the confirmed DEC pathotypes, Enterotoxigenic E. coli (ETEC) (51%) was the most prevalent followed by Diffusely adherent E. coli DAEC (18%), Enteroaggregative E. coli (EAEC) (16%) and Enteropathogenic E. coli (EPEC) (15%). Subsequently; 62 (23%) of 263 Salmonella phenotypic isolates were also confirmed by conventional Polymerase Chain Reaction (PCR) using genus specific primer sets. Though sought; no presumptive isolates of Campylobacter species were detected from the diarrheal stool samples obtained in the study region. ConclusionThe findings of this study elucidated bacterial pathogens co-infection of DEC and Salmonella species among diarrheal stool specimens, accentuating a significant public health concern.

Echinostoma mekongi n. sp. (Digenea: Echinostomatidae) from Riparian People along the Mekong River in Cambodia.

Echinostoma mekongi n. sp. (Digenea: Echinostomatidae) is described based on adult flukes collected from humans residing along the Mekong River in Cambodia. Total 256 flukes were collected from the diarrheic stool of 6 echinostome egg positive villagers in Kratie and Takeo Province after praziquantel treatment and purging. Adults of the new species were 9.0–13.1 (av. 11.3) mm in length and 1.3–2.5 (1.9) mm in maximum width and characterized by having a head collar armed with 37 collar spines (dorsal spines arranged in 2 alternative rows), including 5 end group spines. The eggs in feces and worm uterus were 98–132 (117) μm long and 62–90 (75) μm wide. These morphological features closely resembled those of Echinostoma revolutum, E. miyagawai, and several other 37-collar-spined Echinostoma species. However, sequencing of the nuclear ITS (ITS1-5.8S rRNA-ITS2) and 2 mitochondrial genes, cox1 and nad1, revealed unique features distinct from E. revolutum and also from other 37-collar-spined Echinostoma group available in GenBank (E. bolschewense, E. caproni, E. cinetorchis, E. deserticum, E. miyagawai, E. nasincovae, E. novaezealandense, E. paraensei, E. paraulum, E. robustum, E. trivolvis, and Echinostoma sp. IG). Thus, we assigned our flukes as a new species, E. mekongi. The new species revealed marked variation in the morphology of testes (globular or lobulated), and smaller head collar, collar spines, oral and ventral suckers, and cirrus sac compared to E. revolutum and E. miyagawai. Epidemiological studies regarding the geographical distribution and its life history, including the source of human infections, remain to be performed.

Evaluation of different laboratory techniques for diagnosis of intestinal microsporidiosis in diarrheic children

Background: Microsporidia are a group of obligate intracellular organisms that can infect all animals including man causing disease in both immunocompetent individuals and immunosuppressed patients. The most common microsporidia causing gastrointestinal infection worldwide are Enterocytozoonbieneusi and Encephalitozoonintestinalis. Objectives: The present study was designed with the aim of estimating the rate of microsporidia in diarrheic children and the evaluation of different staining techniques and nested polymerase chain reaction (PCR) for the clinical diagnosis of intestinal microsporidiosis. Subjects and Methods: One hundred and fifty diarrheic stool samples were collected from Abou El-Rich children hospital laboratory. Samples were examined by “Ryan-Blue” modified trichrome stain (MTS), modified ZiehlNeelsen (MZN), acid fast trichrome (AFT) stain, and nested polymerase chain reaction (nPCR). Results:Microsporidium spp. was detected in 12 (8%) of samples using MTS. Compared with the other detection techniques the respective sensitivities and specificities of MZN stain was 100% and 98.57%; nPCR was 80% and 100%; and AFT was 85.71% and 97.87%. Conclusion: Using specific staining techniques as MZN and AFT for diagnosis of intestinal microsporidiosis in diarrheal stool samples are nearly as efficacious as PCR, but even better in some cases with the added advantage of being an inexpensive diagnostic method compared to PCR. Another benefit is the detection of other oocyst forming parasites such as Cryptosporidium, which makes staining techniques very suitable for developing countries

Perforación duodenal por absceso retroperitoneal secundario a una apendicitis aguda perforada

Introducción: La incidencia de apendicitis perforada o gangrenosa sigue siendo alta (28-29%). Un retraso en el diagnóstico y la cirugía lleva a un aumento de los costos, la mortalidad y la morbilidad. 
 Objetivo: Describir un caso clínico relacionado con la inflamación del apéndice.
 Material y métodos: Estudio descriptivo retrospectivo, presentación de caso clínico.
 Resultados: Paciente masculino de 63 años de edad, con dolor abdominal localizado en el epigastrio y en cuadrante inferior derecho de 5 días de evolución, acompañado de deposiciones diarreicas, náuseas y vómito, fue diagnosticado con una perforación duodenal por un absceso retroperitoneal secundario a apendicitis aguda perforada, el mismo reconocido como una condición potencialmente mortal. A través de la comunicación anatómica congénita, el absceso retroperitoneal tiene el potencial de propagarse rápidamente en el espacio perinéfrico, el músculo psoas, la pared abdominal lateral y las extremidades inferiores.
 Conclusiones: La formación de grandes abscesos retroperitoneales con perforación del duodeno es una complicación grave de la apendicitis perforada aguda que debe tenerse en cuenta en todos los pacientes con cuadro clínico atípico de apendicitis aguda.

The Effectiveness of Synbiotic Preparation Containing Lactobacillus and Bifidobacterium Probiotic Strains and Short Chain Fructooligosaccharides in Patients with Diarrhea Predominant Irritable Bowel Syndrome-A Randomized Double-Blind, Placebo-Controlled Study.

The purpose of the randomized double-blind placebo-controlled trial was to assess the effectiveness of synbiotic preparation containing probiotic Lactobacillus rhamnosus FloraActive™ 19070-2, Lactobacillus acidophilus DSMZ 32418, Bifidobacterium lactis DSMZ 32269, Bifidobacterium longum DSMZ 32946, Bifidobacterium bifidum DSMZ 32403 and fructooligosaccharides in adult patients with diarrhea-dominant IBS (IBS-D). The study included eighty patients with moderate and severe IBS-D who were randomized to receive synbiotics or placebo for eight weeks. Finally, a total of sixty-eight patients finished the study. The primary endpoints included the assessment of the symptoms’ severity with IBS symptom severity scale (IBS-SSS), an improvement of IBS global symptoms with Global Improvement Scale (IBS-GIS) and adequate relief of symptoms after four and eight weeks of therapy. Secondary endpoints, which were collected by telephone interviewers three times a week included the assessment of individual IBS symptoms and adverse events. Synbiotic treatment in comparison to placebo significantly improved IBS-GIS (p = 0.043), and IBS-SSS score inducing a decrease in the total IBS-SSS (p = 0.042) and in domain-specific scores related to flatulence (p = 0.028) and bowel habit (p = 0.028) after four and eight weeks. Patients treated with synbiotics reported in weekly observations a significant amelioration in a feeling of incomplete bowel movements, flatulence, pain, stool pressure and diarrheal stools compared to those receiving placebo. There were no differences in adverse events between both groups. Concluding, the multi-strain synbiotic preparation was associated with a significant improvement in symptoms in IBS-D patients and was well-tolerated. These results suggest that the use of synbiotics offers a benefit for IBS-D patients. [Clinicaltrials.gov NCT04206410 registered 20 December 2019].

Bacteriological and Molecular Characterization of Diarrheagenic Escherichia Coli Pathotypes From Children in Duhok City, Iraq

Background: A variety of diarrheagenic E coli (DEC) are responsible for causing different types of diarrhea in children especially in developing courtiers.
 Objectives: This study was primarily aimed to isolate and bacteriological characterizing of E coli from diarrheic infant stool and to investigate their antibiotic resistance patterns and then using molecular identification of DEC pathotypes for better discrimination.
 Methods: Total of 400 fresh stools specimens were collected from children with diarrhea in Heevi Hospital in Duhok city, Iraq. The samples were cultured on selective media such as (MacConkey and MacConkey sorbitol agar). Colonies were identified through biochemical reaction and VITEK 2 system and then antibiotic susceptibility profiles were determined.
 Results: A total of 349(87.2%) samples were yielded positive for growth of E coli. Out of these, 50 phenotypically-identified E coli were then subjected to PCR assay targeting certain virulence factors (alt, eae, sxt1 and sxt2) for discrimination of pathotyes. 13/50(26%) Enterotoxigenix E. coli (ETEC) was detected, 5/50(10%) Enterohemorrhagic E coli EHEC was detected, while no Enteropathogenic E coli (EPEC) was detected. All pathotypes were more frequent in samples from male children aged between 2-3 years that were artificial feeding pattern. Moreover, all pathotypes expressed high resistant to ampicillin, cephalosporin and tetracycline while little resistance to imipenem was observed. Conclusion: The study concludes presence of diarrheagenic E coli pathotypes in our community causing diarrhea in children and emphasize on using of PCR assay for best discrimination.

Stool Samples of Acute Diarrhea Inpatients as a Reservoir of ST11 Hypervirulent KPC-2-Producing Klebsiella pneumoniae.

The emergence and spread of carbapenem-resistant hypervirulent Klebsiella pneumoniae sequence type 11 (ST11-CR-HvKP) in China are a great concern in the public health community. However, the underlying mechanism that enables its wide dissemination in China remains unclear. Here, we investigated the prevalence of carriage of carbapenemase-producing Enterobacteriaceae (CPE) among inpatients with diarrhea in a teaching hospital over 1 year to identify ST11-CR-HvKP reservoirs and to understand the genetic background and plasmid profiles of these pathogens. As assessed by stool analysis, the CPE colonization rate (12.4%) among the inpatients with diarrhea was high (12.4%). Antibiotic exposure, surgical history, and CPE positivity were correlated. Genomic investigation of 65 carbapenem-resistant K. pneumoniae isolates indicated a shared bacterial population in various wards. According to maximum likelihood phylogenetic tree analysis, these isolates were partitioned into three major clades. Analysis of the wzi locus revealed three different K types (KL105, KL47, and K64) among the ST11 isolates, indicating the genetic diversity of these isolates. Genetic and sequence mapping revealed the complexity of virulence and resistance plasmid sets harbored by the isolates. These observations indicate that the dissemination of resistant bacteria is more complex than initially anticipated and possibly involves multiple K. pneumoniae ST11 lineages and a variety of virulence plasmids. Collectively, we show for the first time that stool may be a source of ST11-CR-HvKP isolates. Furthermore, the findings reveal the silent dissemination of ST11-CR-HvKP bacteria in Zhejiang Province, China. Future investigations are warranted to determine the association between rectal colonization by ST11-CR-HvKP and clinical infections.IMPORTANCE China has been experiencing a rapid increase in the number of nosocomial infections caused by carbapenem-resistant Klebsiella pneumoniae ST11 (ST11-CRKP) for decades. The emergence of hypervirulent ST11-CRKP (ST11-CR-HvKP) strains is expected to become a serious public health issue in China, considering that carbapenem resistance and virulence have converged in an epidemic clone. K. pneumoniae strains that colonize the human intestinal tract may become a reservoir of virulence and carbapenemase-encoding genes. Here, we first characterized the genotypes and antimicrobial phenotypes of ST11-CR-HvKP strains isolated from diarrheal stool samples of inpatients in Zhejiang Province, China. Active surveillance approaches based on the findings of the present study should be implemented, particularly in intensive care units, to combat the spread of ST11-CR-HvKP and to improve treatment.

Enfeksiyöz ishallerde Campylobacter jejuni prevalansının çeşitli yöntemlerle araştırılması

In our study, stool samples that were sent to the department of Microbiology for routine bacteriologic culture and parasitological examination were used in to estimate the prevalence of Campylobacter jejuni. Samples collected for 6 months. We depend on traditional methods such as culture for identifying the C. jejuni strains and molecular methods. Fresh diarrheal stool samples were streaked on Skirrow blood agar and incubated in a microaerophilic conditions at two different temperatures (35°C, 42°C) for 48 hours. Conventional methods were used for identification of C. jejuni. Antibiotic susceptibilities of the isolates were tested by disc diffusion method and the results were evaluated according to the EUCAST guidelines. All stool specimens were re-evaluated using (BD MAX™) Enteric Bacterial Panel system by real-time PCR method targeting tuf gen regions of C. jejuni/coli. From a total of 300 stool samples tested by both culture and molecular methods, only 3 C. jejuni strains were isolated by culture; however, using molecular method a total of 5 C. jejuni/coli positive were given.In conclusion, the culture for Campylobacter jejuni should be routinely included in our laboratories because of its importance as a bacterial cause of diarrhea in children. Also molecular methods are thought to be useful in the detection.

Molecular prevalence of intestinal parasites infections in children with diarrhea in Franceville, Southeast of Gabon

BackgroundPediatric diarrhea caused by a range of pathogens, including intestinal parasites, is one of main causes of death among children under 5 years of age. The distribution of these parasitic infections overlaps in many environmental, socioeconomic and epidemiological settings. Their distribution and prevalence varies from region to region. In the current study, we assess the prevalence of intestinal parasites among pediatric patients with syndromic diarrheal disease living in Franceville, Gabon.MethodsA cross-sectional study conducted in the Amissa Bongo Regional Hospital and Chinese-Gabonese Friendship Hospital in Franceville, between November 2016 and August 2017, enrolled a total of 100 diarrheic children between 0 and 180 months of age. Parasite detection in stool samples was performed using molecular diagnostic by PCR. Difference in means were tested by Student’s t test and ANOVA while principal component analysis was used to determine the correlation between parasite distributions and age groups.ResultsThe overall prevalence of intestinal parasite infection was 61% (61/100). Hymenolepis sp and Cryptosporidium hominis/parvum were the most common parasites (31 and 19%, respectively), followed by Encephalitozoon intestinalis (15%), Trichuris trichiura (4%), Dientamoeba fragilis (4%), and Enterocytozoon bieneusi (2%). The polyparasitism rate was 19.7%, with 83.3% double and 16.7% triple infections. Protozoan infections (66.7%) were more prevalent than helminths infections (33.3%). Seasonal association of the circulation of intestinal parasite was statistically significant (p = 0.03). Correlations between different parasites was also observed.ConclusionThe prevalence of intestinal parasitic infections is highest in diarrheic pediatric children. The prevalence of parasitic infections indicates that protozoa and helminths are the most common parasites in the Franceville environment. This study reinforces the importance of routine examination of diarrheic stool samples for the diagnostic of intestinal parasites. Further analyses are required to better understand the local epidemiology and risk factors associated with the transmission of intestinal parasites in Franceville, Gabon.Keyswordsdiarrhea, children, intestinal parasitic infections, molecular diagnostic, Franceville, Gabon.

Antimicrobial susceptibility of Clostridioides difficile isolated from diarrhoeal stool specimens of Canadian patients: summary of results from the Canadian Clostridioides difficile (CAN-DIFF) surveillance study from 2013 to 2017.

To summarize data generated by the Canadian Clostridioides difficile (CAN-DIFF) surveillance study from 2013 to 2017. Isolates of C. difficile (n = 2158) were cultured from toxin-positive diarrhoeal stool specimens submitted by eight hospital laboratories to a coordinating laboratory. Antimicrobial susceptibility testing was performed according to the CLSI agar dilution method (M11, 2018). Isolate ribotypes were determined using an international, standardized, high-resolution capillary gel-based electrophoresis protocol. Of the 2158 isolates of C. difficile, 2133 (98.8%) had vancomycin MICs ≤2 mg/L [i.e. were vancomycin susceptible (EUCAST breakpoint tables, v 9.0, 2019) or WT (CLSI M100, 29th edition, 2019)]. Fidaxomicin MICs were lower than those of all other agents tested (MIC90, 0.5 mg/L); however, one isolate with a fidaxomicin MIC of >8 mg/L was identified. Metronidazole MICs ranged from 0.12 to 4 mg/L; all isolates were metronidazole susceptible by the CLSI breakpoint (≤8 mg/L) compared with 96.8% susceptible by the EUCAST breakpoint (≤2 mg/L). In total, 182 different ribotypes were identified from 2013 to 2017. The most common ribotypes identified were 027 (19.3% of isolates) and 106 (8.2%). Ribotype 027 isolates were frequently moxifloxacin resistant (87.3% of isolates) and MDR (48.6%), associated with vancomycin (10/25, 40.0%) and metronidazole (58/69, 84.1%) resistance and from patients aged ≥80 years. The prevalence of ribotype 027 decreased significantly (P < 0.0001) from 2013 (27.5%) to 2017 (9.0%) and was replaced by increases in ribotype 106 (P = 0.0003) and multiple less common ribotypes. Periodic surveillance is required to monitor clinical isolates of C. difficile for changes to in vitro susceptibility testing profiles and ribotype evolution.

Relationship between faecal metronidazole and lactoferrin concentrations to clinical response of patients with Clostridioides difficile.

We investigated patients with Clostridioides difficile-associated diarrhoea to see if clinical resolution correlated with faecal concentrations of metronidazole or markers of inflammation. Faecal metronidazole, lactoferrin and serum CRP were measured daily. These were then compared with clinical progress. Metronidazole concentration correlated with lactoferrin (ρ = 0.17, p = 0.015), CRP (ρ = 0.23, p < 0.001) and number of diarrhoeal stools per day (ρ = 0.29, p < 0.001). Lactoferrin correlated with CRP (ρ = 0.57, p < 0.001) and the number of diarrhoeal stools per day (ρ = 0.52, p < 0.001) as did CRP (ρ = 0.52, p < 0.001). We found no association between cessation of diarrhoea and metronidazole or lactoferrin concentrations. There was a relationship between metronidazole concentrations and markers of inflammation and stool frequency.

Clostridium difficile in patients with nosocomial diarrhea, Northwest of Iran.

Background: Clostridium difficile is known as a prevalent pathogen leading to infections ranging from mild diarrhea to severe disease and death. The aim of the present study was to evaluate the incidence of C. difficile from inpatients with nosocomial diarrhea hospitalized in different wards in the northwest region of Iran. Methods: In this cross-sectional study, 485 diarrheal stool samples were collected from 384 patients referred from different wards of Imam Reza, Sina and Pediatric hospitals, Tabriz and transferred to the laboratory from 25 March 2015 till 1 March 2018. Immuno-chromatographicassay for detection of toxins A and B of C. difficile was used for identification. Results: Clostridium difficile was isolated from 24 (4.7%) out of 485 samples. Fifteen patients(62.5%) were males and 9 were females (37.5%). Twelve positive patients were from the gastrointestinal ward (50%), 5 patients (20.8%) from surgery ward, 3 patients from infectious disease ward (12.5%), 3 patients from rheumatology ward (12.5%) and 1 patient (4.1%) were collected from neurology ward. 95.3% of diarrhea samples had no signs from toxin A and B. Conclusion: These results indicate most of infected patients were from the gastrointestinaland surgery wards which show a different pattern of infection compared to previous studies.The neurology department had the lowest rate of infection. C. difficile is a health threat afterantibiotic consumption and for health promotion, developing strategies for less antibioticconsumption and preventing these emerging infections is critical. The low rate of this infection shows improvement in knowledge and effect of stewardships in physicians.

Antibiotic sensitivity pattern of diarrhoeal pathogens in under five children

Background: Different bacterial enteropathogens are responsible for diarrhoea in children less than 5 years of age in Bangladesh. This study aimed to determine the antimicrobial sensitivity pattern of diarrhoeal pathogens that is necessary to know for the specific management of acute bacterial diarrhoea.&#x0D; Objectives: To find out the bacterial enteropathogens causing diarrhoea and their antimicrobial sensitivity pattern in under 5 children.&#x0D; Methodology: This was an observational cross sectional study carried out at Microbiology Department of Sir Salimullah Medical College, Dhaka. A total of 217 diarrhoeal patients 0-5 years age, attending Pediatric unit of Sir Salimullah Medical College and Mitford Hospital. Dhaka and Dhaka Shishu Hospital, Dhaka during the period of July, 2015 to June, 2016, were enrolled in the study. For isolation of the common enteric bacteria, diarrhoeal stools were cultured on MacConkey agar media and for Campylobacter spp, diarrhoeal stools were cultured on selective Modified Campy BAP media and nonselective Blood agar media with membrane filter. Positive culture was confirmed by Gram stain morphology and standard biochemical tests. Antimicrobial susceptibility test were used to identify the bacterial sensitivity pattern in the isolates.&#x0D; Results: Out of 217 diarrhoeal stools, bacteria was isolated from 97 (44.70%) patients by culture. Among 97 culture positive cases Eschericha coli (51- 52.58%), Campylobacter spp. (23-23.71%), Shigella spp. (15-15.46%) and Salmonella spp. (8-8.25%) were isolated. Mecillinam is the most sensitive drug for Escherichia coli, Campylobacter spp., Shigella spp. and Salmonella spp. Among the other antimicrobials, Campylabacter spp. were sensitive to azithromycin, erythromycin and nitrofurantoin. Escherichia coli were sensitive to nitrofurantoin, gentamicin and azithromycin. Shigella spp. were sensitive to ceftriaxone, ciprofloxacin and nitrofurantoin and Salmonella spp. were sensitive to ceftriaxone, cotrimoxazole and gentamicin.&#x0D; Conclusion: Mecillinam and nitrofurantoin were highly effective against bacteria causing diarrhoea in children.&#x0D; Bang Med J (Khulna) 2019; 52 : 35-39

Unveiling Pharmacological Responses and Potential Targets Insights of Identified Bioactive Constituents of Cuscuta reflexa Roxb. Leaves through In Vivo and In Silico Approaches.

Cuscuta reflexa Roxb. is traditionally used by the indigenous communities of Bangladesh to treat different diseases, such as pain, edema, tumor, jaundice, and skin infections. This study tested neuro-pharmacological, anti-nociceptive, and antidiarrheal activities by in vivo and in silico experiments for the metabolites extracted (methanol) from the leaves of Cuscuta reflexa (MECR). During the anxiolytic evaluation analyzed by elevated plus maze and hole board tests, MECR (200 and 400 mg/kg) exhibited a significant dose-dependent reduction of anxiety-like behavior in mice. Similarly, mice treated with MECR demonstrated a dose-dependent decrease in the time of immobility in both forced swimming and tail suspension tests. In addition, anti-nociceptive activity was assessed by the chemical-induced (acetic acid and formalin) pain models. In both cases, 400 mg/kg was found to be most effective and significantly (p < 0.001) inhibited acetic acid stimulated writhing and formalin-induced licking (pain response) in mice. Furthermore, antidiarrheal efficacy determined by the castor-oil induced diarrheal model manifested an evident inhibition of diarrheal stool frequency. In parallel, previously isolated bioactive compounds were documented based on the biological activities and subjected to in silico studies to correlate with the current pharmacological outcomes. The selected isolated compounds (15) displayed favorable binding affinities to potassium channels, human serotonin receptor, COX-1, COX-2, M3 muscarinic acetylcholine receptor, and 5-HT3 receptor proteins. Additionally, the ADME/T and toxicological properties were justified to unveil their drug-like properties and toxicity level. Overall, Cuscuta reflexa is bioactive and could be a potential source for the development of alternative medicine.

Etiologic features of diarrheagenic microbes in stool specimens from patients with acute diarrhea in Thailand

Many microbial species have been recognized as enteropathogens for humans. Here, we predicted the causative agents of acute diarrhea using data from multiplex quantitative PCR (qPCR) assays targeting 19 enteropathogens. For this, a case-control study was conducted at eight hospitals in Thailand. Stool samples and clinical data were collected from 370 hospitalized patients with acute diarrhea and 370 non-diarrheal controls. Multiple enteropathogens were detected in 75.7% and 13.0% of diarrheal stool samples using multiplex qPCR and bacterial culture methods, respectively. Asymptomatic carriers of enteropathogens were found among 87.8% and 45.7% of individuals by qPCR and culture methods, respectively. These results suggested the complexity of identifying causative agents of diarrhea. An analysis using the quantification cut-off values for clinical relevance drastically reduced pathogen-positive stool samples in control subjects from 87.8% to 0.5%, whereas 48.9% of the diarrheal stool samples were positive for any of the 11 pathogens. Among others, rotavirus, norovirus GII, Shigella/EIEC, and Campylobacter were strongly associated with acute diarrhea (P-value < 0.001). Characteristic clinical symptoms, epidemic periods, and age-related susceptibility to infection were observed for some enteropathogens. Investigations based on qPCR approaches covering a broad array of enteropathogens might thus improve our understanding of diarrheal disease etiology and epidemiological trends.