Molecular characterization and sequence analysis of human astroviruses circulating in Hungary

Human astroviruses (HAstVs) are major pathogens in viral gastroenteritis worldwide. Twenty-five HAstV strains were detected from stool specimens of children hospitalized for acute gastroenteritis in Budapest, Hungary, between 1995 and 1999. Sequence analysis was performed at the 3' end of the capsid gene to determine genotypic diversity of HAstVs circulating in Hungary. Five different genotypes of HAstVs were identified: HAstV-1 was predominant, followed by types 5, 8, 3 and 4. Two different subtypes of HAstV-1 were detected, but only one at a time in the community. This is the first report on the genetic diversity of HAstVs in Hungary and Central/Eastern Europe.     

Enteroaggregative and cell-detaching<Emphasis Type="Italic">Escherichia coli</Emphasis> strains among polish children with and without diarrhea

To determine the association of enteroaggregative (EAEC) and cell-detaching (CDEC)Escherichia coli with diarrhea of unknown origin among children from Wroc?aw (Poland),E. coli strains isolated from stool specimens of children with diarrhea were examined for mannose-resistant adherence to HEp-2 cells. EAEC were isolated from 10 of 39 (26%) children examined with diarrhea and 4 of 20 (20%) age-matched controls. CDEC were present in 14 (36%) cases of diarrhea and 7 (35%) healthy subjects. Cell-detaching activity was distinctly associated with hemolysin production. Among hemolytic CDEC strains cytotoxic necrotizing factor 1 (CNF1) synthesis prevailed among isolates obtained from cases of diarrhea (57%) in comparison with isolates obtained from healthy controls (14.3%). Although neither EAEC nor CDECE. coli strains were associated with diarrhea of children in this setting, there were differences among EAEC and CDEC strains isolated from children with and without diarrhea.     

Prevalence of enterotoxigenic Bacteroides fragilis in patients with diarrhea: a controlled study

In this age matched controlled study performed in Malatya, a city in east region of Turkey, enterotoxigenic Bacteroides fragilis (ETBF) was investigated in stool specimens obtained from children and adults with and without diarrhea. A nested polymerase chain reaction (PCR) method was used to detect the enterotoxin gene of B. fragilis in a total of 418 stool samples, including 221 samples from 117 children (aged 0-16 years) and 104 adults (aged >16 years) with diarrhea, and 197 samples from 102 children and 95 adults as control group that was the same age group with those having diarrhea. ETBF was detected in 13 of 117 diarrheal children (11.1%) and 8 of 102 control children (7.8%) (P>0.05). In children aged 1-5 years, the rate of ETBF was significantly higher in patients than in controls (25% versus 9.5%, respectively; P<0.05). On the other hand ETBF was detected similar rates (2.2% and 2.4%, respectively) in children younger than 1 year in both patients and controls. ETBF positivity was not significantly difference between patient and control groups who were older than 5 years of age and adults. The frequency of ETBF in the controls was slightly higher in older persons than in younger ones; however, it was not significant. The rate of ETBF as the only enteropathogen in the patients with ETBF was significantly higher than in controls with ETBF (88% versus 39%, respectively; P<0.02). We found that in east region of Turkey, the prevalence of ETBF was higher in the childhood diarrhea, particularly in aged 1-5. As the only enteropathogen, ETBF may play an important role in diarrheal diseases. Persons after 6 years old can be carrier for ETBF regardless diarrhea.     

Etiology and Epidemiology of Diarrhea in Hospitalized Children from Low Income Country: A Matched Case-Control Study in Central African Republic

Background

In Sub-Saharan Africa, infectious diarrhea is a major cause of morbidity and mortality. A case-control study was conducted to identify the etiology of diarrhea and to describe its main epidemiologic risk factors among hospitalized children under five years old in Bangui, Central African Republic.

Methods

All consecutive children under five years old hospitalized for diarrhea in the Pediatric Complex of Bangui for whom a parent’s written consent was provided were included. Controls matched by age, sex and neighborhood of residence of each case were included. For both cases and controls, demographic, socio-economic and anthropometric data were recorded. Stool samples were collected to identify enteropathogens at enrollment. Clinical examination data and blood samples were collected only for cases.

Results

A total of 333 cases and 333 controls was recruited between December 2011 and November 2013. The mean age of cases was 12.9 months, and 56% were male. The mean delay between the onset of first symptoms and hospital admission was 3.7 days. Blood was detected in 5% of stool samples from cases. Cases were significantly more severely or moderately malnourished than controls. One of the sought-for pathogens was identified in 78% and 40% of cases and controls, respectively. Most attributable cases of hospitalized diarrhea were due to rotavirus, with an attributable fraction of 39%. Four other pathogens were associated with hospitalized diarrhea: Shigella/EIEC, Cryptosporidium parvum/hominis, astrovirus and norovirus with attributable fraction of 9%, 10%, 7% and 7% respectively. Giardia intestinalis was found in more controls than cases, with a protective fraction of 6%.

Conclusions

Rotavirus, norovirus, astrovirus, Shigella/EIEC, Cryptosporidium parvum/hominis were found to be positively associated with severe diarrhea: while Giardia intestinalis was found negatively associated. Most attributable episodes of severe diarrhea were associated with rotavirus, highlighting the urgent need to introduce the rotavirus vaccine within the CAR’s Expanded Program on Immunization. The development of new medicines, vaccines and rapid diagnostic tests that can be conducted at the bedside should be high priority for low-resource countries.     

枯草杆菌二联活菌肠溶胶囊预防艾滋病相关性腹泻的临床研究

目的:观察枯草杆菌二联活菌肠溶胶囊对艾滋病(AIDS)相关性腹泻的防治作用。方法:选择2013年7月至2014年5月在我科门诊进行高效抗逆转录病毒(HAART)治疗的AIDS患者80例,随机分成对照组40例和观察组40例。对照组给予单用HAART治疗,观察组在对照组的基础上联合使用枯草杆菌二联活菌肠溶胶囊,两组疗程均为24周。治疗结束后观察两组的HAART疗效、AIDS相关性腹泻的发生率,以及大便培养真菌感染情况。结果:观察组和对照组HAART治疗有效率分别为75%和70%,两组比较差异无统计学意义(P0.05)。对照组腹泻发生率明显高于观察组,比较差异有统计学意义(P0.05),且对照组大便培养真菌感染率明显高于观察组,比较差异有统计学意义(P0.05)。结论:枯草杆菌二联活菌肠溶胶囊能有效预防AIDS相关性腹泻的发生,并减少继发肠道真菌感染的发生。     

Evaluation of the prevalence of enterotoxigenic Bacteroides fragilis and the distribution bft gene subtypes in patients with diarrhea

The aim of this study was to determine the prevalence of enterotoxigenic Bacteroides fragilis (ETBF) in the patients with diarrhea in our region and to assess the association between diarrhea and bft gene subtypes. The presence of ETBF and bft gene subtypes were investigated in 200 stool samples from patients with diarrhea, diagnosed as gastroenteritis, which were sent to Clinical Microbiology Laboratory at Zonguldak Karaelmas University, Training and Research Hospital and in 200 stool samples from age-matched healthy subjects between April 14, 2009 and October 28, 2009. Nested – polymerase chain reaction was used to detect the presence of bft gene directly from stool samples. The bft gene subtypes were determined by PCR in case of ETBF detection. The presence of bft gene was detected in 29 (15%) of patients and 27 (14%) of control group. bft-1 and bft-2 were found in 24 and five stool samples from 29 diarrheic patients with ETBF, respectively. Among 27 control patients with ETBF, bft-1 and bft-2 were found in 24 and three samples, respectively. No bft-3 subtypes were identified in our study. ETBF was found as a single pathogen in 9% of the patients with diarrhea, while there was an accompanying pathogen in 6% of the patients. The proportion of coinfection with another pathogen among ETBF positive patients was 38%. Cooccurance with ETBF was present in nine of 18 patients with Rotavirus and two of five patients with Entamoeba histolytica. In conclusion; there was no statistically significant difference between the prevalence of ETBF in diarrheal patients and that of the control group. When the patients and controls were compared for each age group, no statistically significant difference in ETBF rates was found. There was no significant difference between groups with respect to bft subtypes; bft-1 was identified as the most common subtype. The rate of coinfection of ETBF and Rotavirus was high.     

An imported human case of hookworm infection with worms in the rectum]

An imported case of rectal hookworm infection was diagnosed by stool examination and recovery of adult worms from the rectal mucosa by sigmoidoscopy. The chief complaints of a patient were diarrhea, abdominal pain and weight loss for about 1 month after returning from his travel abroad to the Southeast Asia. Leukocytosis(16,750/microliters) and peripheral eosinophilia(33.7%) were noticed without anemia. Typical hookworm eggs were detected by stool examination, and 3 worms were collected by sigmoidoscopy from rectal mucosa of this patient. Those worms were confirmed as adult worms of Ancylostoma duodenale(male:1, female:2) based on their morphological characteristics. The symptoms were relieved after treatment with anthelmintics. This case was considered as one of the imported parasitic infections in Korea, and a rare case of hookworm infection on human rectal mucosa.     

Household Transmission of Rotavirus in a Community with Rotavirus Vaccination in Quininde,Ecuador

Background

We studied the transmission of rotavirus infection in households in peri-urban Ecuador in the vaccination era.

Methods

Stool samples were collected from household contacts of child rotavirus cases, diarrhea controls and healthy controls following presentation of the index child to health facilities. Rotavirus infection status of contacts was determined by RT-qPCR. We examined factors associated with transmissibility (index-case characteristics) and susceptibility (household-contact characteristics).

Results

Amongst cases, diarrhea controls and healthy control household contacts, infection attack rates (iAR) were 55%, 8% and 2%, (n = 137, 130, 137) respectively. iARs were higher from index cases with vomiting, and amongst siblings. Disease ARs were higher when the index child was <18 months and had vomiting, with household contact <10 years and those sharing a room with the index case being more susceptible. We found no evidence of asymptomatic infections leading to disease transmission.

Conclusion

Transmission rates of rotavirus are high in households with an infected child, while background infections are rare. We have identified factors associated with transmission (vomiting/young age of index case) and susceptibility (young age/sharing a room/being a sibling of the index case). Vaccination may lead to indirect benefits by averting episodes or reducing symptoms in vaccinees.     

Prevalence of Clostridium spp. and Clostridium difficile in children with acute diarrhea in São Paulo city,Brazil

Species of Clostridium are widely distributed in the environment, inhabiting both human and animal gastrointestinal tracts. Clostridium difficile is an important pathogen associated with outbreaks of pseudomembranous colitis and other intestinal disorders, such as diarrhea. In this study, the prevalence of Clostridium spp. and C. difficile, from hospitalized children with acute diarrhea, was examined. These children were admitted to 3 different hospitals for over 12 months. Eighteen (20%) and 19 (21%) stool specimens from children with (90) and without (91) diarrhea respectively, were positive to clostridia. Only 10 C. difficile strains were detected in 5.5% of the stool samples of children with diarrhea. None healthy children (without diarrhea) harbored C. difficile. From these 10 C. difficile, 9 were considered as toxigenic and genotyped as tcdA+/tcdB+ or tcdA-/tcdB+, and 1 strain as nontoxigenic (tcdA-/tdcB-). They were detected by the citotoxicity on VERO cells and by the multiplex-polymerase chain reaction. Thirty clinical fecal extracts produced minor alterations on VERO cells. The presence of C. difficile as a probable agent of acute diarrhea is suggested in several countries, but in this study, the presence of these organisms was not significant. More studies will be necessary to evaluate the role of clostridia or C. difficile in diarrhoeal processes in children.     

Gastrointestinal myiasis by larvae of Sarcophaga sp. and Oestrus sp. in Egypt: report of cases, and endoscopical and morphological studies

Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and us-ing a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.     

双歧杆菌三联活菌散治疗儿童抗生素相关性腹泻的疗效及安全性观察

目的探讨双歧杆菌三联活菌散治疗儿童抗生素相关性腹泻的疗效及安全性观察。方法选取抗生素相关性腹泻的患儿86例,采用随机数字表将纳入患者分为观察组（n=43例）和对照组（n=43例）。两组患儿均予以调整饮食、口服或静脉补液及口服蒙脱石散等常规治疗。观察组患儿加用双歧杆菌三联活菌散1.0 g/次,3次/d,溶解于温水中口服。对照组患儿除不使用双歧杆菌三联活菌散外余治疗同观察组。观察并记录两组患儿治疗后大便次数恢复时间和性状改变时间,并比较其临床疗效及不良反应。结果观察组患儿大便次数恢复时间和性状改变时间明显短于对照组（P〈0.05）;治疗72 h后,观察组患儿的临床总有效率为明显优于对照组（95.35%vs.81.40%）（χ^2=4.07,P〈0.05）,两组患儿治疗中均未出现明显的不良反应。结论双歧杆菌三联活菌散治疗抗生素相关性腹泻具有较好效果及安全性,能明显减少患儿腹泻次数及程度,改善患儿大便性状,较快缓解患儿的病情。     

婴幼儿腹泻病原学及其快速诊断

目的探讨小儿腹泻标本病原学特点及流行病学特征,寻求小儿感染性腹泻的快速诊断方法。方法对我院2003年10月至2005年9月门诊及住院的1160例2个月-5岁腹泻患儿的腹泻标本进行细菌学检查及pH检测。结果(1)1160例患儿腹泻标本病原学阳性605例(52.16%),检出病原20种621株,外源性肠道病原菌、内源性肠道条件致病菌以及轮状病毒3种病原构成比依次为35.75%、32.69%、31.56%。主要病原:(1)轮状病毒196株(31.56%),志贺菌118株(19.00%),克雷伯菌86株(13.8%),枸橼酸杆菌67株(10.79%),致泻性大肠埃希菌60株(9.66%)。(2)1160例患儿腹泻标本6个月-2岁患儿占881例(69.91%)。轮状病毒腹泻多发于秋冬季、大便为水样、多见于2岁以内婴幼儿,志贺菌以夏秋季与脓血便检出率较高。(3)病原菌分离阳性腹泻标本pH>7占明显优势,轮状病毒阳性标本、病原检查阴性标本、健康体检对照标本pH<6占明显优势,前者pH与后三者pH统计学比较,差异有高度显著性(P<0.01)。结论引起小儿腹泻的病原种类繁多,轮状病毒居首位,内源性肠道条件致病菌在腹泻标本中的构成比已接近外源性肠道致病菌而成为腹泻的重要病原菌,大便pH可作为急性腹泻病早期简单、快速、准确、价廉的初步诊断及合理应用抗生素的重要指标。     

Enteropathogens associated with diarrheal disease in infants of poor urban areas of Porto Velho, Rondônia: a preliminary study

One hundred and thirty cases of diarrhea and 43 age-matched controls, 0 to 5 years old, were studied in a pediatric outpatient unit from a poor peri urban area of Porto Velho, Rond?nia. Eighty percent of diarrheal cases were observed in the groups under 2 years of age. Rotavirus (19.2%) was the most frequent enteropathogen associated with diarrhea, followed by Shigella flexneri (6.15%) and S. sonnei (1.5%) and Salmonella sp. (6.9%). Four cases of E. coli enterotoxigenic infections (3.1%), E. coli enteropathogenic (EPEC)(2.3%) one case of E. coli enteroinvasive infection (0.8%) and one case of Yersinia enterocolitica (0.8%) were also identified. Mixed infections were frequent, associating rotavirus, EPEC and Salmonella sp. with Entamoeba histolytica and Giardia lamblia.     

Analysis of chromosomal structural variation in patients with congenital left‐sided cardiac lesions

Background: We sought to characterize the landscape of structural variation associated with the subset of congenital cardiac defects characterized by left‐sided obstruction. Methods: Cases with left‐sided cardiac defects (LSCD) and pediatric controls were uniformly genotyped and assessed for copy number variant (CNV) calls. Significance testing was performed to ascertain differences in overall CNV incidence, and for CNV enrichment of specific genes and gene functions in LSCD cases relative to controls. Results: A total of 257 cases of European descent and 962 ethnically matched, disease‐free pediatric controls were included. Although there was no difference in CNV rate between cases and controls, a significant enrichment in rare LSCD CNVs was detected overall (p = 7.30 × 10^?3, case/control ratio = 1.26) and when restricted either to deletions (p = 7.58 × 10^?3, case/control ratio = 1.20) or duplications (3.02 × 10^?3, case/control ratio = 1.43). Neither gene‐based, functional nor knowledge‐based analyses identified genes, loci or pathways that were significantly enriched in cases as compared to controls when appropriate corrections for multiple tests were applied. However, several genes of interest were identified by virtue of their association with cardiac development, known human conditions, or reported disruption by CNVs in other patient cohorts. Conclusion: This study examines the largest cohort to date with LSCD for structural variation. These data suggest that CNVs play a role in disease risk and identify numerous genes disrupted by CNVs of potential disease relevance. These findings further highlight the genetic heterogeneity and complexity of these disorders. Birth Defects Research (Part A) 100:951–964, 2014. © 2014 Wiley Periodicals, Inc.     

Vibrio mimicus diarrhea following ingestion of raw turtle eggs.

Clinical and epidemiological characteristics of diarrhea associated with Vibrio mimicus were identified in 33 hospitalized patients referred to the Costa Rican National Diagnostic Laboratory Network between 1991 and 1994. The relevant symptoms presented by patients included abundant watery diarrhea, vomiting, and severe dehydration that required intravenous Dhaka solution in 83% of patients but not fever. Seroconversion against V. mimicus was demonstrated in four patients, from whom acute- and convalescent-phase sera were obtained. Those sera did not show cross-reaction when tested against Vibrio cholerae O1 strain VC-12. All the V. mimicus isolates from these cases produced cholera toxin (CT) and were susceptible to commonly used antibiotics. Attempts to isolate this bacterium from stool samples of 127 healthy persons were not successful. Consumption of raw turtle eggs was recalled by 11 of the 19 (58%) individuals interviewed. All but two V. mimicus diarrheal cases were sporadic. These two had a history of a common source of turtle (Lepidochelys olivacea) eggs for consumption, and V. mimicus was isolated from eggs from the same source (a local market). Among the strains, variations in the antimicrobial susceptibility pattern were observed. None of the strains recovered from market turtle eggs nor the four isolates from river water showed CT production. Further efforts to demonstrate the presence of CT-producing V. mimicus strains in turtle eggs were made. Successful results were obtained when nest eggs were tested. In this case, it was possible to isolate CT- and non-CT-producing strains, even from the same egg. For CT detection we used PCR, enzyme-linked immunosorbent assay (ELISA), and Y-1 cell assay, obtaining a 100% correlation between ELISA and PCR results. Primers Col-1 and Col-2, originally described as specific for the V. cholerae O1 ctxA gene, also amplified a 302-bp segment with an identical restriction map from V. mimicus. These results have important implications for epidemiological surveillance in tropical countries where turtle eggs are used for human consumption, serving as potential sources of cholera-like diarrhea.     

Identification of MW Polyomavirus, a Novel Polyomavirus in Human Stool

We have discovered a novel polyomavirus present in multiple human stool samples. The virus was initially identified by shotgun pyrosequencing of DNA purified from virus-like particles isolated from a stool sample collected from a healthy child from Malawi. We subsequently sequenced the virus' 4,927-bp genome, which has been provisionally named MW polyomavirus (MWPyV). The virus has genomic features characteristic of the family Polyomaviridae but is highly divergent from other members of this family. It is predicted to encode the large T antigen and small T antigen early proteins and the VP1, VP2, and VP3 structural proteins. A real-time PCR assay was designed and used to screen 514 stool samples from children with diarrhea in St. Louis, MO; 12 specimens were positive for MWPyV. Comparison of the whole-genome sequences of the index Malawi case and one St. Louis case demonstrated that the two strains of MWPyV varied by 5.3% at the nucleotide level. The number of polyomaviruses found in the human body continues to grow, raising the question of how many more species have yet to be identified and what roles they play in humans with and without manifest disease.     

320例腹泻患者病原菌分离及流行病学分析

目的了解广州市2009年至2010年肠道门诊就诊的细菌性腹泻患者病原谱的分布情况,为制定针对重点人群肠道传染病防治策略提供依据。方法收集2009年5月至2010年5月暨南大学附属第一医院腹泻患者的粪便标本,使用卡-布运送培养基,增菌培养后,采用生化反应和氧化酶试验进行菌株鉴定,并用梅里埃API生化板条进行验证,用病原菌诊断血清进行细菌分型。结果从320份粪便标本中分离到38株菌株,其中沙门菌15株,产毒大肠埃希菌12株,致病大肠埃希菌6株,志贺菌2株,出血性大肠埃希菌、霍乱弧菌、气单胞菌各1株。0—20岁年龄段高发,以1岁以内婴幼儿为主;7—10月为发病高峰期。结论来该院肠道门诊就诊的细菌性腹泻患者,其病原体以沙门菌为主,其次为产毒大肠埃希菌。因此．广州地区细菌件腹泻的预防．廊右针对件的面向雷占人群和重占病厢莴．