Diarrhea In Young Children Research Articles

Performance and operational feasibility of two diagnostic tests for cryptosporidiosis in children (CRYPTO-POC): a clinical, prospective, diagnostic accuracy study

SummaryBackgroundCryptosporidiosis is a common cause of diarrhoea in young children (aged younger than 24 months) in low-resource settings but is currently challenging to diagnose. Light-emitting diode fluorescence microscopy with auramine-phenol staining (LED-AP), recommended for tuberculosis testing, can also detect Cryptosporidium species. A lateral-flow test not requiring refrigerator storage (by contrast with most immunochromatographic lateral-flow assays) has also recently been developed for Cryptosporidium spp detection. We aimed to evaluate the diagnostic accuracy and operational feasibility of LED-AP and the lateral-flow test strip for cryptosporidiosis in children.MethodsWe did a prospective diagnostic accuracy study in two health-care facilities in Ethiopia, in a consecutive series of children younger than 5 years of age with diarrhoea (three or more loose stools within the previous 24 h) or dysentery (at least one loose stool with stains of blood within the previous 24 h). Stool samples were tested for Cryptosporidium spp by LED-AP and the lateral-flow test strip; accuracy of each test was estimated by independent and blind comparison with a composite reference standard comprising quantitative immunofluorescent antibody test (qIFAT), ELISA, and quantitative PCR (qPCR). Quantitative cutoff values for diarrhoea-associated infection were established in an embedded case-control substudy, with cases of cryptosporidiosis coming from the 15 districts in and around Jimma and the eight districts surrounding Serbo, and community controls without diarrhoea in the previous 48 h recruited by weekly frequency matching by geographical district of the household, age group, and enrolment week.FindingsStool samples from 912 children with diarrhoea or dysentery and 706 controls from the case-control substudy were tested between Dec 22, 2016, and July 6, 2018. Estimated reference-standard cutoff values for cryptosporidiosis positivity were 2·3 × 105 DNA copies per g of wet stool for qPCR, and 725 oocysts per g for qIFAT. LED-AP had a sensitivity for cryptosporidiosis of 88% (95% CI 79–94; 66 of 75 samples) and a specificity of 99% (98–99; 717 of 726 samples); the lateral-flow test strip had a sensitivity of 89% (79–94; 63 of 71 samples) and a specificity of 99% (97–99; 626 of 635 samples).InterpretationLED-AP has high sensitivity and specificity for cryptosporidiosis and should be considered as a dual-use technology that can be easily integrated with existing laboratory infrastructures in low-resource settings. The lateral-flow test strip has similar sensitivity and specificity and provides an alternative that does not require microscopy, although purchase cost of the test strip is unknown as it is not yet available on the market.FundingNorwegian Research Council GLOBVAC fund, The Bill & Melinda Gates Foundation, Norwegian Society for Medical Microbiology, University of Bergen, and Vestfold Hospital Trust.

Prevalence of Rotaviruses in the Etiology of Acute Diarrhea in Young Children, Clinical forms, Extraintestinal Manifestations and Complications.

Rotavirus is highly contagious factor with dominant feces-oral transmission. Because it is stable in external environment, transmission clusters are possible by close contact, ingestion of contaminated water or food or contact with contaminated surfaces. It survives within hours and days on hands and contaminated surfaces. This makes it the most common enteric and nosocomial pathogen in the world, especially in early childhood. In addition to the rapid dehydration with pronounced electrolyte disturbances, numerous extraintestinal possibilities have been recorded in the clinical picture, which emphasizes the need for prevention of this disease.In the period from 1.02.2018 to 31.01.2020 at the Clinic for Infectious diseases were treated 1060 patients with diarrheal disease, of which 502 children (47.36%). Rotavirus etiology was confirmed in 23.30% of the children. According to the protocols, laboratory and biochemical investigations were done to all 117 children, with tracking parameters and their dynamics of admission and discharge from the hospital. Most of the children, 84 (82.0 6%) are from urban areas, with a more confirmed epidemiological survey of 59 (42.00%). The average age of the children was 8 months, with a small percentage of children on maternal food (breastfed 25, i.e. 21.37%), with high febrile admission in 99% of children with an average temperature of 38.5oC and an average febrile duration of 4 days, with an average of 7 (+ 2.49) of stools and 5 (+ 2.12) of vomiting. There was a significant difference in hematocrit, leukocyte, electrolyte, glycaemia, and CRP values on admission and discharge. There was predominant isonatremic dehydration, and the compensatory mechanisms followed by the values of the electrolytes ABS, Ph, BE showed a tendency to maintain within the physiological limits. The clinical picture of extraintestinal manifestations included bronchitis, mesenteric lymphadenitis, upper respiratory infections and rash.Rotavirus infection is a serious health and economic problem in our country, so it needs continuous prevention and monitoring in order to reduce the incidence, and thus the need for hospitalization and cure of rotavirus disease.

Health sequelae of human cryptosporidiosis in industrialised countries: a systematic review

BackgroundCryptosporidium is a protozoan parasite which is a common cause of gastroenteritis worldwide. In developing countries, it is one of the most important causes of moderate to severe diarrhoea in young children; in industrialised countries it is a cause of outbreaks of gastroenteritis associated with drinking water, swimming pools and other environmental sources and a particular concern in certain immunocompromised patient groups, where it can cause severe disease. However, over recent years, longer-term sequelae of infection have been recognised and a number of studies have been published on this topic. The purpose of this systematic review was to examine the literature in order to better understand the medium- to long-term impact of cryptosporidiosis.MethodsThis was a systematic review of studies in PubMed, ProQuest and Web of Science databases, with no limitations on publication year or language. Studies from any country were included in qualitative synthesis, but only those in industrialised countries were included in quantitative analysis.ResultsFifteen studies were identified for qualitative analysis which included 3670 Cryptosporidium cases; eight studies conducted in Europe between 2004–2019 were suitable for quantitative analysis, including five case-control studies. The most common reported long-term sequelae were diarrhoea (25%), abdominal pain (25%), nausea (24%), fatigue (24%) and headache (21%). Overall, long-term sequelae were more prevalent following infection with Cryptosporidium hominis, with only weight loss and blood in stool being more prevalent following infection with Cryptosporidium parvum. Analysis of the case-control studies found that individuals were 6 times more likely to report chronic diarrhoea and weight loss up to 28 months after a Cryptosporidium infection than were controls. Long-term abdominal pain, loss of appetite, fatigue, vomiting, joint pain, headache and eye pain were also between 2–3 times more likely following a Cryptosporidium infection.ConclusionsThis is the first systematic review of the long-term sequelae of cryptosporidiosis. A better understanding of long-term outcomes of cryptosporidiosis is valuable to inform the expectations of clinicians and their patients, and public health policy-makers regarding the control and prevention of this infection.Systematic review registration PROSPERO Registration number CRD42019141311

Genotypic determination of human group A rotaviruses from Goa and Meghalaya states, India

IntroductionRotavirus is the leading cause of diarrhoea in young children in India, responsible for an estimated 21357 mean numbers of deaths in 2010. Various genotypes of rotaviruses evolved due to mutational changes have been recognized. In this study, we determined the genotypes of rotaviruses involved in diarrhea in Goa and Meghalaya states of India. MethodsThe dsRNA of rotaviruses was extracted from stool samples and detected by Ribonucleic Acid-Polyacrylamide gel electrophoresis (RNA PAGE) and Reverse transcription-polymerase Chain Reaction (RT-PCR) targeting the partial VP7 gene. The full length VP7 and partial VP4 genes of rotavirus strains were amplified by RT-PCR followed by nucleotide sequencing. The RotaC classification tool was used to determine the genotypes. ResultsThe positivity of rotavirus by PAGE and RT-PCR was observed to be 43.10% and 39.65% in Goa and 38% and 36% in Meghalaya, respectively. Though long electrophoretic profile was appeared to be the most predominant rotavirus type in circulation in these two states, 96% of long and 84.61% short electropherotype profiles could be detected by RT-PCR. The dsRNA of rotavirus extracted from 36 samples could be transcribed and amplified by beg9end9 primers for G genotyping, while, 41 by con3con2 primers for P genotyping. G1P[8] and G1P[6] genotypes were commonly circulated in Goa and G1P[8] and G1P[4] genotypes in Meghalaya. On nucleotide analysis, 6 samples from Goa showed G1 genotype specificity, while, 3 showed P[8] specificity indicating the G1P[8] rotavirus circulating in Goa. In Meghalaya state, 3 strains showed P[8] and 2 showed P[4] genotype specificity. The majority of the G and P genotypes were closely related to each other and G1 genotypes appeared in two separate clusters, while, P[8] and P[4] appeared in the respective clusters. ConclusionThe circulation of G1P[8], G1P[6] genotypes in Goa and the presence of G1P[8] and G1P[4] genotypes in Meghalaya was observed.

An Association Between HBoV and Acute Gastroenteritis in a 2-Month-Old Infant: A Case Report

Introduction: The Human bocavirus (HBoV) was first identified from the nasopharyngeal aspirate specimen in 2005, which includes four subtypes (HBoV1-4). The HBoV-1 is a major subtype in acute respiratory infections of children, and others (HBoV2-4) present in the stool specimens. The pathogenic role of HBoV2-4 in acute gastroenteritis has not confirmed yet, therefore, it has been considered widely. Case Presentation: In this report, we presented a 2-month-old boy with acute gastroenteritis admitted to the Shahid Beheshti Hospital of Kashan, Iran. The stool sample of the patient was tested for HBoV by polymerase chain reaction (PCR) of the NP-1 gene. The other major gastrointestinal pathogens of Salmonella spp., Shigella spp., Giardia lamblia, and Entamoeba histolytica were confirmed by specialized microbiological procedures and viral pathogen of Rotavirus by the enzyme-linked immunosorbent assay. This case was confirmed by NP-1 plasmid cloned as a positive control. All clinical manifestations were analyzed by a pediatric nurse through hospital admission. Conclusions: This case was found HBoV-positive for the NP-1 gene of 354 bp by PCR. The major signs were diarrhea, fever, dehydration, and abdominal pain. This case was charged after supportive therapies for dehydration. We showed that HBoV could be a gastrointestinal pathogen in pediatric patients and causing diarrhea in young children. However, more studies are needed to confirm.

Dietary iron modulates fecal microbiota and severity of enterotoxigenic E. coli infection in a weanling pig model

BackgroundEnterotoxigenic E. coli (ETEC) infection is among the leading causes of diarrhea in young children who live in resource‐limited countries, wherein iron deficiency anemia is usually more prevalent. It is debated whether iron supplementation increases the risk of enteric infection.Objectivethe study used a weanling piglet model to investigate the effect of dietary iron on severity of ETEC infection and host microbiota.MethodsThirty‐two weaned piglets were blocked by body weight and randomized to the treatments. Pigs were fed 1) an iron‐replete diet (150 mg iron/kg) and received sham inoculation (CON), 2) a low‐iron diet (54 mg iron/kg) and orally inoculated with ETEC (LOI), 3) the iron‐replete diet and orally inoculated with ETEC (COI), or 4) a high‐iron diet (700 mg elemental iron /kg) and orally inoculated with ETEC (HII). Pigs were individually housed with free access to water and diets. After 10 days adaptation to diet, pigs were orally inoculated with a 3‐mL dose of F18+ ETEC (1010 cfu) or sterile saline solution for 3 days. Alertness and fecal consistency were scored daily since day 0 post‐inoculation (d0PI). Fecal samples were collected for 16S RNA sequencing and examining the presence of pathogen. Data was analyzed using PROC MIXED of SAS (v.9.4) with repeated measures for variables measured over time.ResultsPathogen was positive in fecal samples from inoculated pigs on d3PI, but was absent in most pigs on d6PI. Pigs in LOI had lower hemoglobin and hematocrit compared to those in HII before ETEC infection (P < 0.05), but dietary iron content did not affect growth before infection. Enterotoxigenic E. coli infection reduced hemoglobin and hematocrits compared to CON, but only pigs in LOI had significantly retarded growth after infection (P < 0.05). There were more days of diarrhea (fecal score > 2) in LOI than in CON and COI (P < 0.05). Treatment did not affect alpha‐diversity on d0 or d7PI. Relative abundance of Actinobacteria and Firmicutes were affected by treatment on d7PI. Within the phylum of Actinobacteria, Bifidobacteriaceae was more abundant in LOI (37.3%) than that in CON (2.0%) and HII (5.0%), while the opposite pattern was observed for Coriobacteriaceae. Within the phylum of Firmicutes, Lactobacillaceae had the greatest abundance in LOI (28.8% and 31.7%) in comparison with all the other groups (7.8% – 17.7%) on d0 and d7PI (P < 0.05). Same pattern was observed for Veillonellaceae, while Clostridiaceae had the highest abundance in HII (22.8%) and the lowest in LOI (5.7%) on d7PI (P = 0.07).ConclusionIron deficiency predisposed host to increased vulnerability to ETEC infection in weanling pigs. Both iron‐deficient and iron‐excess diet alters fecal microbiota with more prominent effect post‐ETEC infection.Support or Funding InformationNIFA hatch project

Predominance of Human Bocavirus Genotype 1 and 3 in Outpatient Children with Diarrhea from Rural Communities in South Africa, 2017-2018.

Human bocavirus (HBoV) is an emerging virus globally associated with diarrhea in young children. This study aims to investigate the prevalence of HBoV genotypes in children (≤5 years) from rural communities in South Africa (SA) suffering from acute gastroenteritis (AGE). A total of 141 fecal samples of children ≤5 years with acute gastroenteritis (AGE) were collected from rural primary health care facilities in the Vhembe district of SA between June 2017 and July 2018. Clinical symptoms and demographic data were also recorded. A total of 102 (72%) were outpatients, and 39 (28%) were hospitalized patients. Human bocavirus (HBoV) genotypes were determined using real-time multiplex PCR. DNA extracts of positive samples were confirmed by conventional PCR targeting the NS1 gene. Co-infection with other enteric viruses were determined in HBoV-positive samples using real-time PCR. HBoV was detected in eight (5.7%) children with AGE, of which three (37.5%) were HBoV1, three (37.5%) were HBoV3, and two (25%) were HBoV2. The majority of positive cases were identified in outpatients (62%) between the ages of 1 and 24 months. Co-infection in HBoV-positive samples with other enteric viruses included rotavirus (37.5%), adenovirus (37.5%), norovirus (25%), and astrovirus (12.5%). HBoV infections could be seen as a potential emerging diarrheal pathogen in South Africa. However, more studies are needed to understand the role of HBoV infections in children with AGE.

Recombinant hexosaminidases conjugated to magnetite nanoparticles: Alternative therapeutic treatment routes in GM2 fibroblasts

Rotaviruses are icosahedral, nonenveloped viruses with a genome consisting of 11 segments of double-stranded RNA. They are the leading cause of severe diarrhea in young children in countries without vaccination programs, and accounted for the death of approximately 200,000 children less than 5 years of age worldwide in 2013, primarily in developing countries. Two main RV vaccines have been approved for use in more than 100 of countries, where they have effectively reduced the disease burden and death rate due to this pathogen.

Acute Infective Bloody Diarrhea in Children Below Five Years Admitted to Children Welfare Hospital in Medical City - Baghdad During 2015

Bloody diarrhea in young children is usually a sign of invasive enteric infection that carry a substantial risk of serious morbidity and death. Objectives : to identify the proportion of infective bloody diarrhea out of total number of gastro-enteritis cases ,also to identify the prevalence of different etiological agents and to find the relation between the main infective etiological types of bloody diarrhea and some sociodemographic factors of this study. Retrospective study was conducted in Children Welfare Teaching Hospital / Baghdad during period extending from 1 st of January to 30 th of June 2016. Information was collected by reviewing the records of patients with acute infective bloody diarrhea under 5 years of age admitted during 2015. The study revealed that the proportion of infective bloody diarrhea was 10. 9% out of the total gastroenteritis (788) cases admitted during 2015, and Entamoeba Histolytica was the most common isolated pathogen in the study42 (48%) patients. followed by E.coli 15 cases (17.4%), shigella, salmonella 7(8%) and 4(4.7%) respectively . The most vulnerable age was those below 2 years (58%) and males were affected more than females (1.3:1), (37%) were bottle fed. The majority had chlorinated tap water supply (82.6%).

Occurrence of Diarrhea and Feeding Practices among Children below Two Years of Age in Southwestern Saudi Arabia

Growing evidence suggests that feeding practices in early childhood play a major role in the occurrence of childhood diarrhea. However, there is a lack of information regarding feeding practices and its relationship with occurrences of diarrhea in young children from Saudi Arabia. The present study is aimed to measure the prevalence of diarrhea and assess its relationship with feeding practices among children between two months and two years of age in Saudi Arabia. A cross-sectional study was carried out in two large cities in the Aseer region in southwest Saudi Arabia. A total of 302 mothers attending well-baby clinics across six primary health centers were included. A structured questionnaire was used to collect data. Factors associated with diarrheal disease were identified by multivariable logistic regression analysis. The prevalence of diarrhea among children during the study period was 56.3% (95% CI: 50.7%–61.8%). Only 15.9% of children in our study were exclusively breastfed. The occurrence of diarrhea was significantly associated with age 7–12 months (aOR = 2.64, 95% CI: 1.42–4.91). We found that diarrhea was prevalent among children between two months and two years of age, and that exclusive breastfeeding was not a common practice in this region. Health education programs should be directed towards mothers to improve rates of breastfeeding, weaning practices, food hygiene, and childcare. Special attention and support should be provided for working mothers.

Economic Analysis of Applying Biomass Stove to Produce Hot Water Serving Child Development Center: The Case Study Of Omkoi District, Chiangmai, Thailand


 
 
 
 This research aims to perform the techno-economic feasibility of the applied hybrid biomass stove to produce hot water for serving Child Development Center (CDC) in Omkoi District, Chiang Mai, Thailand. The hot water generated from the stove could help to decrease LPG usage and enhance household economics. Many activities in CDC (i.e., children showering during the winter season, cloth washing, and utensils cleaning among others) require hot water to prevent the contamination of harmful germs which could cause many diseases such as diarrhea in young children. Presently, Asian Development College for Community Economy and Technology, Chiang Mai Rajabhat University (adiCET) has installed the hybrid stove to the in Omkoi district. The biomass stove could generate 90 °C hot water using plant/agriculture waste as the fuel. However, the economical parameters (i.e., Payback Period, Net Present Value and Internal Rate of Return) could strengthen the technology. The results from this study could be used as one of the guidelines for policymakers to design the appropriate programs for enhancing the CDC using the waste-to-energy concept.
 
 
 

Heterogeneity in enterotoxigenic Escherichia coli and shigella infections in children under 5 years of age from 11 African countries: a subnational approach quantifying risk, mortality, morbidity, and stunting

SummaryBackgroundDiarrhoea, a global cause of child mortality and morbidity, is linked to adverse consequences including childhood stunting and death from other diseases. Few studies explore how diarrhoeal mortality varies subnationally, especially by cause, which is important for targeting investments. Even fewer examine indirect effects of diarrhoeal morbidity on child mortality. We estimated the subnational distribution of mortality, morbidity, and childhood stunting attributable to enterotoxigenic Escherichia coli (ETEC) and shigella infection in children younger than 5 years from 11 eastern and central African countries. These pathogens are leading causes of diarrhoea in young children and have been linked to increased childhood stunting.MethodsWe combined proxy indicators of morbidity and mortality risk from the most recent Demographic and Health Surveys with published relative risks to estimate the potential distribution of diarrhoeal disease risk. To estimate subnational burden, we used country-specific or WHO region-specific morbidity and mortality estimates and distributed them subnationally by three indices that integrate relevant individual characteristics (ie, underweight, probability of receiving oral rehydration treatment of diarrhoea, and receiving vitamin A supplementation) and household characteristics (ie, type of drinking water and sanitation facilities).FindingsCharacterising ETEC and shigella subnational estimates of indirect morbidity (infection-attributable stunting) and indirect mortality (stunting-related deaths from other infectious diseases) identified high-risk areas that could be missed by traditional metrics. Burundi and Democratic Republic of the Congo had the highest ETEC-associated and shigella-associated mortality and stunting rates. Mozambique, Democratic Republic of the Congo, and Zimbabwe had the greatest subnational heterogeneity in most ETEC and shigella mortality measures. Inclusion of indirect ETEC and shigella mortality in burden estimates resulted in a 20–30% increase in total ETEC and shigella mortality rates in some subnational areas.InterpretationUnderstanding the indirect mortality and morbidity of diarrhoeal pathogens on a subnational level will strengthen disease control strategies and could have important implications for the relative impact and cost-effectiveness of new enteric vaccines. Because our methods rely on publicly available data, they could be employed for national planning.FundingBill & Melinda Gates Foundation.

Adenovirus infection is associated with altered gut microbial communities in a non-human primate

Adenovirus (AdV) infections are one of the main causes of diarrhea in young children. Enteric AdVs probably disrupt gut microbial defences, which can result in diarrhea. To understand the role of the gut microbiome in AdV-induced pathologies, we investigated the gut microbiome of a naturally AdV-infected non-human primate species, the Malagasy mouse lemur (Microcebus griseorufus), which represents an important model in understanding the evolution of diseases. We observed that AdV infection is associated with disruption of the gut microbial community composition. In AdV+ lemurs, several commensal taxa essential for a healthy gut microbiome decreased, whereas genera containing potential pathogens, such as Neisseria, increased in abundance. Microbial co-occurrence networks revealed a loss of important microbial community interactions in AdV+ lemurs and an overrepresentation of Prevotellaceae. The observation of enteric virus-associated loss of commensal bacteria and associated shifts towards pathobionts may represent the missing link for a better understanding of AdV-induced effects in humans, and also for their potential as drivers of co-infections, an area of research that has been largely neglected so far.

Review of Newly Identified Functions Associated With the Heat-Labile Toxin of Enterotoxigenic Escherichia coli.

Heat-labile toxin (LT) is a well-characterized powerful enterotoxin produced by enterotoxigenic Escherichia coli (ETEC). This toxin is known to contribute to diarrhea in young children in developing countries, international travelers, as well as many different species of young animals. Interestingly, it has also been revealed that LT is involved in other activities in addition to its role in enterotoxicity. Recent studies have indicated that LT toxin enhances enteric pathogen adherence and subsequent intestinal colonization. LT has also been shown to act as a powerful adjuvant capable of upregulating vaccine antigenicity; it also serves as a protein or antigenic peptide display platform for new vaccine development, and can be used as a naturally derived cell targeting and protein delivery tool. This review summarizes the epidemiology, secretion, delivery, and mechanisms of action of LT, while also highlighting new functions revealed by recent studies.

Prevalence of Enterobacteriaceae Isolated from Childhood Diarrhoea in Mukuru Slums, Nairobi- Kenya

Diarrhoea in young children continues to be a major public health concern in developing countries, including Kenya. Poor sanitation among other factors can predispose a child to diarrhoea. Therefore, the present study sought to determine the prevalence of enterobacteriaceae isolated from childhood diarrhoea in Mukuru Slums, Nairobi. It employed a cross-sectional design targeting children below 5 years of age. Stool specimens were obtained aseptically and cultured on MacConkey agar and Salmonella-Shigella agar. Biochemical tests were used to identify the isolated bacteria to genus and species using biochemical characterization scheme and the Analytic Profile Index 20E. Drugs sensitivity Antimicrobial susceptibility tests (AST) were done using standard techniques. Escherichia coli ATCC 25922 was included as a control strain. Analysis of gender verses diarrhoea revealed that there was no statistical significant association between the gender (male and female) and area of residence in relation to diarrhoea in this study (p = 0.146 > 0.05). There was no statistical significant difference between the participant’s characteristics and their area of residence (p= 0.144). Age of the participants had significant association with the prevalence of diarrhoea (p=0.00). The E. coli bacteria showed the highest percentage of enteric pathogens isolated (35.2%) from female children at Mukuru kwa Njenga and 29.4% from male children, Salmonella spp being second (4.9%) from female at Mukruru kwa Reuben and the least was Shigella sonnei (3.2%) from female children at Sinai. Emphasis should therefore be placed on primary preventive measures such as ensuring good sewage management and safe supply of drinking water in the study area and Kenya at large especially in the slums.

Identification of a potent benzoxaborole drug candidate for treating cryptosporidiosis

Cryptosporidiosis is a leading cause of life-threatening diarrhea in young children and causes chronic diarrhea in AIDS patients, but the only approved treatment is ineffective in malnourished children and immunocompromised people. We here use a drug repositioning strategy and identify a promising anticryptosporidial drug candidate. Screening a library of benzoxaboroles comprised of analogs to four antiprotozoal chemical scaffolds under pre-clinical development for neglected tropical diseases for Cryptosporidium growth inhibitors identifies the 6-carboxamide benzoxaborole AN7973. AN7973 blocks intracellular parasite development, appears to be parasiticidal, and potently inhibits the two Cryptosporidium species most relevant to human health, C. parvum and C. hominis. It is efficacious in murine models of both acute and established infection, and in a neonatal dairy calf model of cryptosporidiosis. AN7973 also possesses favorable safety, stability, and PK parameters, and therefore, is an exciting drug candidate for treating cryptosporidiosis.

Establishment and application of isothermal multiple-self-matching-initiated amplification (IMSA) in detecting Type II heat-labile enterotoxin of Escherichia coli.

Enterotoxigenic Escherichia coli (ETEC) constitutes a major cause of diarrhea in young children and animals, particularly in poor regions of the world, as well the traveler’s diarrhea in adult individuals. Type II heat-labile enterotoxin (LT-II) from ETEC can cause profuse watery diarrhea, posing a potential threat to public health and animal husbandry. In the present study, isothermal multiple-self-matching-initiated amplification (IMSA) was established to rapidly detect LT-II producing ETEC. The specificity and sensitivity were assessed, and clinical samples were tested. The established IMSA method had good specificity for the detection of LT-II gene with a limit of detection of 25 CFU/mL, i.e. 2 times higher than that of real-time PCR and other two isothermal amplifications (loop-mediated isothermal amplification, LAMP and cross-primer isothermal amplification, CPA). Meanwhile, in 103 clinical Escherichia coli strains isolated from diarrhea samples, 9 strains with LT-II+ gene were detected (8.73%), corroborating real-time PCR, LAMP and CPA data. Therefore, the IMSA technology applied for the detection of LT-II producing ETEC has a good application prospect for screening clinical samples in primary medical units or common laboratories.

Induction of a long non-coding RNA, lncRNA-Chr1:1226, by Cryptosporidium infection primes intestinal epithelial cells for IFN-γ-mediated host antimicrobial gene transcription

Abstract Cryptosporidium, a protozoan parasite that infects the intestinal epithelium and other mucosal surfaces in animals and humans, is an important opportunistic pathogen in AIDS patients and a common cause of diarrhea in young children in developing countries. Intestinal epithelial cellular defense is key to innate mucosal anti-Cryptosporidium defense but underlying molecular mechanisms are still obscure. Here, we identified several long non-coding RNAs (lncRNAs) that are predominantly expressed in intestinal epithelial cells. Several of such epithelial-enriched lncRNAs, such as lncRNA-Chr1:1226, were upregulated in cells following C. parvum infection. Induction of lncRNA-Chr1:1226 in infected intestinal epithelial cells was controlled by TLR4/NF-κB/Cdc42 signaling and epithelial specific transcription factor Eif3. Induction of lncRNA-Chr1:1226 promoted IFN-γ-mediated epithelial antimicrobial defense, through facilitating STAT1/SWI/SNF-associated chromatin remodeling to promote IFN-γ-mediated transcription of defense genes in intestinal epithelial cells. We observed that IFN-γ-mediated antimicrobial defense was suppressed in neonatal intestinal epithelium. Expression of PRDM1 in the neonatal intestinal epithelium might contribute to suppression of IFN-γ-mediated antimicrobial gene transcription. Furthermore, PRDM1 interacted with lncRNA-Chr1:1226 and PIAS1 to attenuate SWI/SNF-mediated antimicrobial transcription induced by IFN-γ in intestinal epithelium of neonates. Our data demonstrate that lncRNAs, particularly epithelial lncRNAs, may be key regulators in IFN-γ-mediated epithelial defense.

Molecular Genetics for Cryptosporidium

The protozoan parasite Cryptosporidium is a leading cause of severe diarrhea in young children and an important contributor to early childhood mortality. Fully effective drugs and vaccines to treat or prevent cryptosporidiosis are lacking. A main roadblock for their development has the overall poor tractability of this parasite. We established a powerful molecular genetic model to overcome this hurdle. Using this system we can now engineer reporter parasites to derive robust measurements of infection in vitro and in vivo as well asparasite mutants to understand the mechanistic underpinnings of drug sensitivity and resistance in Cryptosporidium. Cryptosporidium is also a fascinating model to understand fundamental parasite biology. How does the parasite's complex sexual lifecycle unfold, and how do parasite, host and commensals interact to shape susceptibility, disease and protection? To address these issues we are pursuing different genetic strategies to identify key factors in the parasite and in the host.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effect of Zinc Supplementation on the Frequency and Consistency of Stool in Children with Acute Diarrhea.

IntroductionAcute diarrhea in young children is a prevalent and distressing pediatric illness. The role of zinc therapy in the improvement of stool consistency and the shortening of the duration of diarrhea is still controversial. The aim of this study is to assess the effect of oral zinc supplementation in acute diarrhea.MethodsChildren of age 28 days till five years presenting in the outpatient department with acute diarrhea were included. Oral zinc supplementation was included in the anti-diarrheal regime of half of the children (n=50); the other half (n=50) were not given zinc. Mean body weight and the frequency and consistency of stool were noted for both groups on Days 1 and 3.ResultsThe zinc group showed a significantly reduced frequency of diarrheal episodes on the third day of intervention (p<.00001). More children in the zinc group had soft to firm stool consistency than in the non-zinc group (p=.01).ConclusionOral zinc supplementation has a promising role in reducing the duration of diarrhea and improving stool consistency in children with acute diarrhea. Oral zinc supplementation should be made a mandatory part of the anti-diarrheal regime for Pakistani children.