Non-diarrheic Children Research Articles

Enteroaggregative Escherichia coli (EAEC) isolates obtained from non-diarrheic children carry virulence factor-encoding genes from Extraintestinal Pathogenic E. Coli (ExPEC).

Enteroaggregative E. coli (EAEC) is one of the most frequent pathogens isolated from diarrheal patients as well as from healthy individuals in Brazil and has recently also been implicated as an extraintestinal pathogenic E. coli (ExPEC) associated with bloodstream and urinary tract infections. In this study, 37 EAEC isolates, obtained from fecal samples of non-diarrheic children, were molecularly and phenotypically characterized to access the pathogenic features of these isolates. The EAEC isolates were assigned into the phylogroups A (54.1%), D (29.7%), B1 (13.5%) and B2 (2.7%); and harbored genes responsible for encoding the major pilin subunit of the aggregative adherence fimbriae (AAFs) or aggregate-forming pili (AFP) adhesins as follows: aggA (24.3%), agg3A (5.4%), agg4A (27.0%), agg5A (32.4%) and afpA (10.8%). The most frequent O:H serotypes were O15:H2 (8.1%), O38:H25 (5.4%) and O86:H2 (5.4%). Twenty-one isolates (56.8%) produce the aggregative adherence (AA) pattern on HeLa cells, and biofilm formation was more efficient among EAEC isolates harboring the aggA and agg5A genes. PFGE analysis showed that 31 (83.8%) of the isolates were classified into 10 distinct clusters, which reinforces the high diversity found among the isolates studied. Of note, 40.5% (15/37) of the EAEC isolates have a genetic profile compatible with E. coli isolates with intrinsic potential to cause extraintestinal infections in healthy individuals, and therefore, classified as EAEC/ExPEC hybrids. In conclusion, we showed the presence of EAEC/ExPEC hybrids in the intestinal microbiota of non-diarrheic children, possibly representing the source of some endogenous extraintestinal infections.

Prevalence of Cryptosporidium Infection among Children Attending Maryam Abacha Women and Children Hospital, Sokoto State, Nigeria

Study's Excerpt/Novelty This study investigates the prevalence of Cryptosporidium among children attending Maryam Abacha Women and Children Hospital, Sokoto, using Formol ether concentration and Modified Ziehl Neelsen staining methods. It identifies a high prevalence of (71.1%) and shows a strong correlation between Cryptosporidium infection, diarrhea, and source of drinking water. These findings underscore the necessity for routine diagnostic testing of Cryptosporidium among children for improved detection, treatment, and overall patient outcomes in the region. Full Abstract Cryptosporidium infection is ranked as the second most common cause of diarrheal disease and fatalities among children in developing nations. Nonetheless, due to the fact that many healthcare providers do not consistently request Cryptosporidium diagnostic tests, the prevalence of cryptosporidiosis is probably underestimated. The current study is therefore aimed at investigating the prevalence of cryptosporidiosis among children attending Maryam Abacha Women and Children Hospital (MAWCH), Sokoto. During the timeframe of April to June 2021, stool samples were collected from 114 diarrheic and non-diarrheic children and analyzed using Formol ether concentration and Modified Ziehl Neelsen staining methods. Out of 114 samples analyzed, 81(71.1%) were positive for Cryptosporidium oocysts. A higher prevalence of Cryptosporidiosis was observed in males 45(39.5%) compared to females 36(31.6%) participants and higher 41(36.0%) in younger children aged 0-1 years than the older children. Similarly, a greater prevalence of Cryptosporidiosis was observed in diarrheic children (42.1%) than in non-diarrheic children (28.9%). A statistically significant relationship was observed between the occurrence of Cryptosporidium and diarrhea (P = 0.002) and the source of drinking water (P = 0.001). Routine diagnosis of Cryptosporidiosis among children is vital and can enhance effective diagnosis, treatment, and consequently, improvement in the patients' well-being.

Infectious etiology and indicators of malabsorption or intestinal injury in childhood diarrhea.

The multifactorial etiology of gastroenteritis emphasizes the need for different laboratory methods to identify or exclude infectious agents and evaluate the severity of diarrheal disease. To diagnose the infectious etiology in diarrheic children and to evaluate some fecal markers associated with intestinal integrity. The study group comprised 45 children with diarrheal disease, tested for enteropathogens and malabsorption markers, and 76 children whose feces were used for fat evaluation by the traditional and acid steatocrit tests. We observed acute diarrhea in 80% of the children and persistent diarrhea in 20%. Of the diarrheic individuals analyzed, 40% were positive for enteropathogens, with rotavirus (13.3%) and Giardia duodenalis (11.1%) the most frequently diagnosed. Among the infected patients, occult blood was more evident in those carrying pathogenic bacteria (40%) and enteroviruses (40%), while steatorrhea was observed in infections by the protozoa G. duodenalis (35.7%). Children with diarrhea excreted significantly more lipids in feces than non-diarrheic children, as determined by the traditional (p<0.0003) and acid steatocrit (p<0.0001) methods. Moreover, the acid steatocrit method detected 16.7% more fecal fat than the traditional method. Childhood diarrhea can lead to increasingly severe nutrient deficiencies. Steatorrhea is the hallmark of malabsorption, and a stool test, such as the acid steatocrit, can be routinely used as a laboratory tool for the semi-quantitative evaluation of fat malabsorption in diarrheic children.

Detection of Clostridium difficile among diarrheic children using cultural and polymerase chain reaction technique

Introduction: Clostridium difficile is the most common cause of antibiotic-associated diarrhea and colitis. Several methods are available for the detection of C. difficile in stool samples. This study aimed to use glutamate dehydrogenase (GDH), toxin detection, culture and polymerase chain reaction (PCR) techniques for the diagnosis of this pathogen. Methodology: A total of 300 stool samples were collected from children with hospital acquired diarrhea (HA-D), community acquired diarrhea (CA-D), and hospitalized non-diarrheic children as control with ages ranging from 6 months to 6 years (mean 3.7 ± 1.7). Each stool sample was divided into two parts; one part was tested for the enzyme GDH, toxin A and B and then cultured on selective media; and the other part for direct DNA extraction. Results: From a total of 300 stool samples, 9 (3.0%) were positive for C. difficile by the PCR technique, 7 (7%) samples of which were from HA-D cases and 2 (2.0%) from CA-D cases; the control group samples were negative. The enzyme GDH was detected in 12 (12%) samples and toxins A and B in 8 (8%) samples from HA-D cases compared to 5 (5%) and 2 (2%), respectively from CA-D cases. Both GDH and the toxins were negative in control samples. Only 19 (19.0%) samples from HA-D cases gave suspected growth and all of these were negative by PCR. Conclusions: Based on the results of this study, we conclude that the PCR technique is the only reliable method for the diagnosis of this pathogen.

Molecular epidemiology and antimicrobial susceptibility of diarrheagenic Escherichia coli isolated from children under age five with and without diarrhea in Central Ethiopia.

Diarrhea is a serious health problem in children, with the highest mortality rate in sub-Saharan Africa. Diarrheagenic Escherichia coli (DEC) is among the major bacterial causes of diarrhea in children under age five. The present study aims to determine molecular epidemiology and antimicrobial resistance profiles of DEC and identify contributing factors for acquisition among children under age five in Central Ethiopia. A health facility-centered cross-sectional study was conducted in Addis Ababa and Debre Berhan, Ethiopia, from December 2020 to August 2021. A total of 476 specimens, 391 from diarrheic and 85 from non-diarrheic children under age five were collected. Bacterial isolation and identification, antimicrobial susceptibility, and pathotype determination using polymerase chain reaction (PCR) were done. Of the 476 specimens analyzed, 89.9% (428/476) were positive for E. coli, of which 183 were positive for one or more genes coding DEC pathotypes. The overall prevalence of the DEC pathotype was 38.2% (183/476). The predominant DEC pathotype was enteroaggregative E. coli (EAEC) (41.5%, 76/183), followed by enterotoxigenic E. coli (21.3%, 39/183), enteropathogenic E. coli (15.3%, 28/183), enteroinvasive E. coli (12.6%, 23/183), hybrid strains (7.1%, 13/183), Shiga toxin-producing E. coli (1.6%, 3/183), and diffusely-adherent E. coli (0.6%, 1/183). DEC was detected in 40.7% (159/391) of diarrheic and 28.2% (24/85) in non-diarrheic children (p = 0.020). The majority of the DEC pathotypes were resistant to ampicillin (95.1%, 174/183) and tetracycline (91.3%, 167/183). A higher rate of resistance to trimethoprim-sulfamethoxazole (58%, 44/76), ciprofloxacin (22%, 17/76), ceftazidime and cefotaxime (20%, 15/76) was seen among EAEC pathotypes. Multidrug resistance (MDR) was detected in 43.2% (79/183) of the pathotypes, whereas extended spectrum ß-lactamase and carbapenemase producers were 16.4% (30/183) and 2.2% (4/183), respectively. All six common DEC pathotypes that have the potential to cause severe diarrheal outbreaks were found in children in the study area; the dominant one being EAEC with a high rate of MDR.

Epidemiologic and Clinical Characteristics of Human Bocavirus Infection in Children with or without Acute Gastroenteritis in Acre, Northern Brazil.

Human bocavirus (HBoV) is an emerging virus detected around the world that may be associated with cases of acute gastroenteritis (AGE). However, its contribution to AGE has not been elucidated. This study aimed to describe the frequency, clinical features, and HBoV species circulation in children up to 5 years with or without AGE symptoms in Acre, Northern Brazil. A total of 480 stool samples were collected between January and December 2012. Fecal samples were used for extraction, nested PCR amplification, and sequencing for genotyping. Statistical analysis was applied to verify the association between epidemiological and clinical characteristics. Overall, HBoV-positivity was 10% (48/480), with HBoV-positive rates of 8.4% (19/226) and 11.4% (29/254) recorded in diarrheic and non-diarrheic children, respectively. The most affected children were in the age group ranging between 7 and 24 months (50%). HBoV infection was more frequent in children who live in urban areas (85.4%), use water from public networks (56.2%), and live with adequate sewage facilities (50%). Co-detection with other enteric viruses was 16.7% (8/48) and the most prevalent coinfection was RVA+ HBoV (50%, 4/8). HBoV-1 was the most frequent species detected in diarrheic and non-diarrheic children, responsible for 43.8% (21/48) of cases, followed by HBoV-3 (29.2%, 14/48) and HBoV-2 (25%, 12/48). In this study, HBoV infection was not always associated with AGE, as most HBoV cases belonged to the non-diarrheal group. Future studies are warranted in order to determine the role of HBoV in causing acute diarrhea disease.

Antimicrobial Susceptibility and Genetic Basis of Resistance of &amp;lt;i&amp;gt;Klebsiella spp&amp;lt;/i&amp;gt; Isolated from Diarrheic and Non-Diarrheic Children at Health Facilities in Mukuru Informal Settlement, Nairobi, Kenya

Antimicrobial resistance (AMR) is a global threat to public health and particularly to children. This study aimed to determine the prevalence of multidrug resistance of fecal Klebsiella spp on selected beta lactam (3rd generation cephalosporins and carbapenems) and fluoroquinolone classes of drugs in four health facilities serving the slum communities of Nairobi city in Kenya. Additionally, determine the genetic basis for the multidrug resistance observed. A cross sectional laboratory based study was undertaken where a total of 1171 children below 16 years were selected, from whom stool samples were collected, tested and analyzed. 395 (33.73%) Klebsiella spp were isolated, consisting of 365 (92.4%) Klebsiella pneumoniae and 30 (7.6%) Klebsiella oxytoca were isolated. The proportion of multi-drug resistance (MDR) K. pneumoniae and MDR K. oxytoca was 64.1% (234/365) and 96.67% (29/30) respectively. Third generation cephalosporins, cefotaxime ceftriaxone and ceftazidime showed the highest resistance of 30.7%, 29.9% and 27.4% respectively, whereas carbapenems including imipenem and meropenem had the least resistance of 1.6%, each, to K. pneumoniae. A significant association was observed in diarrheic children (OR = 1.88; p = 0.01) and those below 50 months (OR = 0.43; p = 0.002) and carrying K. pneumoniae resistance to one or more third generation cephalosporins. Genes associated with resistance included bla TEM 100%, bla CTX-M 95.2%, bla SHV 57.1%, bla OXA-1 66.7%, qnrS 54.1%, qnrB 47.6% and bla NDM 7.1%. In conclusion, there’s need for more effective infection control measures, antimicrobial stewardship to reduce emergence of antimicrobial resistance, improved drinking water, sanitation and hygiene (WASH) practices.

Helicobacter pylori and enteric parasites co-infection among diarrheic and non-diarrheic Egyptian children: seasonality, estimated risks, and predictive factors

Helicobacter pylori (H. pylori) and intestinal parasites are known for their high prevalence in children. Both of them infect the gastrointestinal tract with overlapping clinical pictures. This study was conducted to determine H. pylori prevalence and its association with intestinal parasites in children, moreover to estimate risk and predictive factors for their detection in stool samples. Single fecal samples were collected from 226 Egyptian pediatric patients (125 diarrheic and 101 non-diarrheic) attending gastroenterology outpatients' clinics, from February 2016 to June 2017. All stool specimens were microscopically examined to search for ova and parasites. Copro-DNAs detection of H. pylori and Cryptosporidium were performed using nested-PCR assays. H. pylori was detected molecularly in 36.8% of the total study population, with a higher prevalence in diarrheic than in non-diarrheic children. Intestinal parasites were detected in 27.4% of the total study populations, of these, 43.9% had co-existence with H. pylori colonized patients and was significantly associated with Cryptosporidium spp. and G. intestinalis. Estimated risk of the presence of H. pylori was in January. Our data provide a better understanding of the epidemiology of H. pylori infection when associated with intestinal parasites. H. pylori co-existence with G. intestinals and Cryptosporidium may suggest the association of H. pylori infection with markers of fecal exposure. Whether H. pylori provides favorable conditions for intestinal parasitosis or vice versa, still further investigations are needed with an emphasis upon determining correlation with gut microbiomes.

Enteric pathogens and associated risk factors among under-five children with and without diarrhea in Wegera District, Northwestern Ethiopia.

IntroductionChildhood diarrhea is highly prevalent in slums in developing countries, but it remains understudied. The objectives of this study were to explore the prevalence of Giardia, rotavirus and bacterial enteropathogens among diarrheic and non-diarrheic children and investigate socio-environmental determinants of diarrhea in two Ethiopian towns.MethodsA cross-sectional study was conducted from June to October 2016. Prevalence of childhood diarrhea was established using information gathered during interviews with mothers/guardians. Saline wet mounts of fresh stool samples were used to test for the presence of Giardia. Stool samples were cultured on MacConkey agar and suspected colonies were characterized using biochemical tests. Susceptibility testing was done by the disk diffusion method. ELISA was used to screen for rotavirus.ResultsA total of 225 children were included in this study. Four enteropathogens (Giardia, rotavirus, Shigella and Salmonella) were identified from 31% (35/112) diarrheic and 12% (14/113) from non-diarrheic children (p < 0.001). The prevalence of rotavirus infection was 18.0% among diarrheic children and 3.3% among non-diarrheic children unvaccinated against rotavirus (p < 0.01). The prevalence of Giardia was 21.0% among diarrheic and 8.0% among non-diarrheic children (p < 0.01). Diarrheic children had significantly higher rates of bloody stool (p < 0.02), vomiting, fever and breastfeeding for children beyond 23 months of age (p < 0.001). Giardia and rotavirus were identified in more diarrheic than non-diarrheic children.ConclusionThe high prevalence of Giardia and rotavirus in the study area indicates the need for coordinated healthcare activities in the two communities. Vaccination against rotavirus infections and educational interventions are recommended.

Comprehensive Analysis of Prevalence, Epidemiologic Characteristics, and Clinical Characteristics of Monoinfection and Coinfection in Diarrheal Diseases in Children in Tanzania.

The role of interactions between intestinal pathogens in diarrheal disease is uncertain. From August 2010 to July 2011, we collected stool samples from 723 children admitted with diarrhea (cases) to 3 major hospitals in Dar es Salaam, Tanzania, and from 564 nondiarrheic children (controls). We analyzed the samples for 17 pathogens and assessed interactions between coinfections in additive and multiplicative models. At least one pathogen was detected in 86.9% of the cases and 62.8%, of the controls. Prevalence of coinfections was 58.1% in cases and 40.4% in controls. Rotavirus, norovirus genogroup II, Cryptosporidium, and Shigella species/enteroinvasive Escherichia coli were significantly associated with diarrhea both as monoinfections and as coinfections. In the multiplicative interaction model, we found 2 significant positive interactions: rotavirus + Giardia (odds ratio (OR) = 23.91, 95% confidence interval (CI): 1.21, 470.14) and norovirus GII + enteroaggregative E. coli (OR = 3.06, 95% CI: 1.17, 7.98). One significant negative interaction was found between norovirus GII + typical enteropathogenic E. coli (OR = 0.09, 95% CI: 0.01, 0.95). In multivariate analysis, risk factors for death were presence of blood in stool and severe dehydration. In conclusion, coinfections are frequent, and the pathogenicity of each organism appears to be enhanced by some coinfections and weakened by others. Severity of diarrhea was not affected by coinfections.

Cryptosporidium genotypes and associated risk factors in a cohort of Egyptian children

Cryptosporidium is one of the most common, worldwide diarrheal diseases caused by parasites. Due to absence of an effective treatment, determining the prevailing species of Cryptosporidium is a key in identifying its transmission dynamics and a necessary precursor required for the planning and implementation of effective preventive and control strategies. This PCR-RFLP study was done to determine the prevalence of Cryptosporidium species in the stool of a cohort of Egyptian children and evaluate/assess associated risk factors for susceptibility to cryptosporidiosis, due to the lack of existent studies addressing Cryptosporidium transmission dynamics in humans and assessed risk factors in Egypt. Stool samples were collected from 431 children; 331 diarrheic and 100 apparently healthy non-diarrheic children; their data were recorded. Samples were processed for Copro-nPCR targeting Hsp90 gene and PCR-RFLP analysis for species identification. Variables which showed statistical significance for Cryptosporidium were included in a logistic regression analysis to identify the estimated risk. Out of 84 (19.5%) Cryptosporidium-positive samples (78 diarrheic and 6 non-diarreic), 75 (89.3%) were Cryptosporidium hominis, 6 (7.1%) were Cryptosporidium parvum, and 3 (3.6%) were non-typed. There was a significant association between Cryptosporidium detection in stool and the estimated risk factors: diarrhea, soft stool, and drinking from tap water. Cryptosporidium is an indigenous, prevailing intestinal parasite among children in Cairo that physicians must consider, especially in diarrheic, preschool-aged children, who drink from tap water. The finding of a predominance of C. hominis indicates anthroponotic rather than zoonotic transmission.

High prevalence of G12P[8] rotavirus strains in Rio Branco, Acre, Western Amazon, in the post-rotavirus vaccine introduction period.

The present study aimed to provide a molecular characterization of circulating rotavirus (RVA) strains in Rio Branco, Acre, in the post-rotavirus vaccination period, particularly with regard to the emerging, increasingly prevalent G12P[8] genotype. A total of 488 fecal specimens from diarrheic and non-diarrheic children were obtained between January and December 2012. RVA detection was initially performed using enzyme-linked immunosorbent assay (ELISA) method, followed by reverse-transcription polymerase chain reaction (RT-PCR) using specific primers. RVA was detected in 18.3% (44/241) of the children with acute diarrhea and in 1.2% (3/247) of the non-diarrheic children (P < 0.001), with overall RVA-positivity of 9.6% (47/488). The most common genotype was G2P[4] with 43.2% (19/44) of the diarrheic cases, followed by G12P[8] (27.3%, 12/44), G3P[6] (18.2%, 8/44), G3P[8] (4.5%, 2/44), and G12P[6] (2.3%, 1/44). G12 samples belonged to lineage III and were from children aged 4-52 months. All of these children had acute diarrhea associated with fever (83.3%, 10/12) and vomiting (66.7%, 8/12). Most of the cases occurred in August (58.3%, 7/12), 75% (9/12) of which having received the full vaccination scheme with Rotarix™. For the first time G12 was reported at relative high prevalence in Brazil. Our findings warrant further monitoring studies on the molecular characterization of circulating RVA strains after rotavirus vaccine introduction in Brazil and elsewhere, since the occurrence of either unusual our emerging genotypes may pose a challenge to vaccination strategies.

Laboratory utility of coproscopy, copro immunoassays and copro nPCR assay targeting Hsp90 gene for detection of Cryptosporidium in children, Cairo, Egypt.

Cryptosporidium is a significant cause of diarrhea worldwide especially in children. Infection may end fatally in immunocompromised patients. Multi-attribute analysis was used to determine the lab utility of 4 diagnostics; coproscopy of AF stained fecal smear, fecal immunoassays by ICT and ELISA and copro-nPCR assay targeting Hsp90 gene, for detection of Cryptosporidium in stool of 250 Egyptian children (150 diarrheic and 100 non-diarrhaeic children). Also, to determine Cryptosporidium molecular prevalence. Cryptosporidium was an important enteric pathogen among both diarrheic and non-diarrheic study children with a clearly high prevalence of 16.4% (n=41). Conventional methods had perfect specificity (100%) but couldn`t be used as a consistent single detection method due to their lowered sensitivities. Multi-attribute analysis ranked nPCR the highest test for lab use. Being the test with the best diagnostic yield, nPCR is a reliable diagnostic test and is going to replace conventional methods for reliable detection of Cryptosporidium.

Phenotypic and genotypic characteristics associated with biofilm formation in clinical isolates of atypical enteropathogenic Escherichia coli (aEPEC) strains.

BackgroundBiofilm formation by enteropathogenic Escherichia coli (EPEC) have been recently described in the prototype typical EPEC E2348/69 strain and in an atypical EPEC O55:H7 strain. In this study, we sought to evaluate biofilm formation in a collection of 126 atypical EPEC strains isolated from 92 diarrheic and 34 nondiarrheic children, belonging to different serotypes. The association of biofilm formation and adhesin-related genes were also investigated.ResultsBiofilm formation occurred in 37 (29%) strains of different serotypes, when the assays were performed at 26°C and 37°C for 24 h. Among these, four strains (A79, A87, A88, and A111) formed a stronger biofilm than did the others. The frequency of biofilm producers was higher among isolates from patients compared with isolates from controls (34.8% vs 14.7%; P = 0.029). An association was found between biofilm formation and expression of type 1 fimbriae and curli (P < 0.05). Unlike the previously described aEPEC O55:H7, one aEPEC O119:HND strain (A111) formed a strong biofilm and pellicle at the air-liquid interface, but did not express curli. Transposon mutagenesis was used to identify biofilm-deficient mutants. Transposon insertion sequences of six mutants revealed similarity with type 1 fimbriae (fimC, fimD, and fimH), diguanylate cyclase, ATP synthase F1, beta subunit (atpD), and the uncharacterized YjiC protein. All these mutants were deficient in biofilm formation ability.ConclusionThis study showed that the ability to adhere to abiotic surfaces and form biofilm is present in an array of aEPEC strains. Moreover, it seems that the ability to form biofilms is associated with the presence of type 1 fimbriae and diguanylate cyclase. Characterization of additional biofilm formation mutants may reveal other mechanisms involved in biofilm formation and bring new insights into aEPEC adhesion and pathogenesis.

Detection and genetic analysis of the enteroaggregative Escherichia coli heat-stable enterotoxin (EAST1) gene in clinical isolates of enteropathogenic Escherichia coli (EPEC) strains.

BackgroundThe enteroaggregative E. coli heat-stable enterotoxin 1 (EAST1) encoded by astA gene has been found in enteropathogenic E. coli (EPEC) strains. However, it is not sufficient to simply probe strains with an astA gene probe due to the existence of astA mutants (type 1 and type 2 SHEAST) and EAST1 variants (EAST1 v1-4). In this study, 222 EPEC (70 typical and 152 atypical) isolates were tested for the presence of the astA gene sequence by PCR and sequencing.ResultsThe astA gene was amplified from 54 strains, 11 typical and 43 atypical. Sequence analysis of the PCR products showed that 25 strains, 7 typical and 18 atypical, had an intact astA gene. A subgroup of 7 atypical strains had a variant type of the astA gene sequence, with four non-synonymous nucleotide substitutions. The remaining 22 strains had mutated astA gene with nucleotide deletions or substitutions in the first 8 codons. The RT-PCR results showed that the astA gene was transcribed only by the strains carrying either the intact or the variant type of the astA gene sequence. Southern blot analysis indicated that astA is located in EAF plasmid in typical strains, and in plasmids of similar size in atypical strains. Strains carrying intact astA genes were more frequently found in diarrheic children than in non-diarrheic children (p < 0.05).ConclusionIn conclusion, our data suggest that the presence of an intact astA gene may represent an additional virulence determinant in both EPEC groups.

Enteroaggregative Escherichia coli in diarrheic children in Egypt: molecular characterization and antimicrobial susceptibility.

Little information is available regarding the significance of enteroaggregative Escherichia coli (EAEC) in pediatric diarrhea in Egypt. Escherichia coli was isolated from stool samples of 62 diarrheic and 43 non-diarrheic (control) Egyptian children. Samples were screened for genes specific for enteroaggregative E. coli (EAEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), Shiga toxin-producing E. coli (STEC), and enteroinvasive E. coli (EIEC) using polymerase chain reaction (PCR). Diarrheagenic E. coli were grouped phylogenetically using PCR and tested for their susceptibility to antibiotics using the disk diffusion method. Isolates designated as EAEC were examined for eight virulence factors (VFs) using PCR. EAEC was detected in 19 (30.7%) and 4 (9.3%), EPEC in 2 (3.2%) and 1 (2.3%), and ETEC in 2 (3.2%) and 0 (0.0%) diarrheic and control children, respectively; STEC and EIEC were not detected. Only EAEC was significantly isolated from diarrheic children compared with controls (p < 0.01, OR = 4.31).Three or more VFs (multivirulent isolates) were found in 52.6% and 50% of EAEC isolated from diarrheic children and controls, respectively. More than 73% (17/23) of EAEC isolates were identified as belonging to phylogenetic group D. Multiple-antibiotic resistance (resistance to three or more drugs) was observed in more than 91% of EAEC. Multivirulent EAEC is a significant causative agent of pediatric diarrhea in Egypt, with the majority of isolated EAEC belong to phylogenetic group D. Multiple-antibiotic resistance among EAEC has the potential to be a serious public health problem for the country.

Molecular Characterization of Noroviruses and HBGA from Infected Quilombola Children in Espirito Santo State, Brazil

Noroviruses (NoV) are the main etiological agents of gastroenteritis outbreaks worldwide and susceptibility to NoV infection has been related to the histo-blood group antigen (HBGA). This study aimed to determine the prevalence of NoV strains and to evaluate the HBGA phenotype and genotype of children from semi-isolated Quilombola communities, descendents of black slaves in Brazil. A total of 397 children up to eleven years old, with and without diarrhea, from Quilombola Communities in the Espirito Santo State, Brazil, were investigated for the presence of NoV from August 2007 to September 2009. Feces were collected from all the children, and blood from the NoV positive children. NoV was screened by reverse transcription-PCR with primers for the RNA-dependent RNA polymerase region; genogroup was determined by PCR with primers for the C and D regions and genotyped by sequencing. HBGA phenotype was performed by gel-spinning and FUT2 and FUT3 were analyzed by PCR or sequencing analysis. NoV were detected in 9.2% (12/131) of diarrheic and 1.5% (4/266) of non-diarrheic children (p<0.05, Fisher’s exact test). GI and GII genogroups were present in 12.5% and 87.5% of the samples, respectively. The following genotypes were characterized: GII.4 (25%), GII.12 (25%), GII.6 (12.5%) and GI.1 (6.3%), GI.3 (12.5%) and GI.4 (6.3%). Children infected with NoV showed the A (n = 6), O (n = 6), and B (n = 2) HBGA phenotypes, and 13 of them were classified as secretors (Se) and one as a non secretor (se). Mutations of Se 40, 171,216,357,428,739,960 were found for the FUT2 gene and mutations of Le 59, 202, 314 for the FUT3 gene. The only se child was infected by NoV GI, whereas the Se children were indiscriminately infected by GI or GII. This study showed rates of NoV infection in symptomatic and asymptomatic Quilombola children consistent with other studies. However, children under 12 months were seven times more affected than those between 1 and 5 years old. GII.12 was as frequent as GII.4 and GI.1 and GI.4 were described for the first time in Brazil. Owing to the small number of cases studied, no clear pattern of susceptibility and/or HBGA resistance could be inferred.

The prevalence of group A rotavirus infection and some risk factors in pediatric diarrhea in Zaria, North central Nigeria

Group A rotavirus cause infectious, severe and dehydrating diarrhea which have great impact on childhood morbidity and mortality worldwide. This study was designed to investigate the disease burden and risk factors of rotavirus infection among children 1 – 5 years in Zaria, Nigeria. Stool samples were collected from 666 diarrheic children and 170 matched non-diarrheic controls and screened for rotavirus antigen by ELISA. Their socio-demographic information and clinical presentations were also noted. There was a 15.6 and 7.6% prevalence of rotavirus infections among the diarrheic and non-diarrheic children, respectively, with the peak of infection occurring between 1 - 6 months of age. The most common clinical features included dehydration (59.1%), fever and vomiting (41.3%), vomiting (34.6%) and fever (19.2%). Age, feeding regimen, and the presence of another person in the house with diarrhea were found to be risk factors for rotavirus diarrhea while gender and socioeconomic status were not. In view of the peak of infection in infants in the study area, rotavirus vaccine will be best administered in early infancy. Key words: Rotavirus, prevalence, risk factors, pediatric, diarrhea.

Estimation of Interleukin-8 Level in Diarrheic Children Infected with Rotavirus

Background and objective: Rotavirus is the most important cause of severe diarrhea in children worldwide. We have investigated that certain cytokines especially interleukin- 8 (IL-8) as a chemokine, may play an important role in the pathogenesis of and the protection against rotavirus disease in children. The purpose of this study was to reveal the association between IL-8 and child diarrhea with rotavirus. Methods: Stool samples were collected from 150 children under 5 years of age who developed diarrhea and 45 from non-diarrheic children of equivalent age. All samples were detected for rotavirus and serum IL-8 level. The study was undertaken from May to December of 2006. Enzyme linked immunosorbant assay (ELISA) was applied for detection rotavirus and IL-8. Results: Among the children, female patients with breast feeding were less infected with rotavirus compared to males with artificial and mixed feeding. The IL-8 concentration was highly significantly between the study groups (P&lt; 0.01). Regarding the gender, serum IL-8 level was significantly higher in males than females (P&lt;0.05). However, no significance prevalence in IL-8 level was recorded between gender in healthy control group (P &gt;0.05). Conclusion: Rotavirus infection induces the expression of chemokines as IL-8. These data support the hypothesis that chemokine secretion may play a role in the initiation and modulation of the immune response to rotavirus infection.

Symptomatic and Asymptomatic Cryptosporidiosis in Young Children in Iran

The present study was conducted during the period May 2003 to October 2003 in 616 children less than three years of age with and without diarrheal disease presenting at the pediatric clinic of teaching hospitals in Kermanshah, Iran. Single stool specimens were collected from 515 diarrheic and 99 non-diarrheic children. Two smears were made from each stool samples and were stained by a modified Ziehl Neelsen technique. Cryptosporidium parvum (C. parvum) were detected in 10.4% of children. The C. parvum infection rate was significantly higher in diarrheic children (11.6%) than in non-diarrheic children (4.0%). C. parvum was observed more frequently in stool samples of children who lived in rural areas (15.2%) than those who lived in urban areas (7.2%). In regard to the presence of animals, the infection rate was 18.5% among children who lived in association with animals in comparison with 8.2% among those who lived in compounds with no animals. The majority of C. parvum cases occurred in children between the ages of 0-12 months (11.9%), followed by in children between the ages of 13-24 months (9.2%) and in children between the ages of 25-36 months (3.0%). The data suggest that C. parvum is relatively endemic in young children and that Cryptosporidium may be an important pathogen associated with diarrhea in young children.