Diarrheal Status Research Articles

Whole genome sequence analysis reveals high genomic diversity and potential host-driven adaptations among multidrug-resistant Escherichia coli from pre-weaned dairy calves.

Food-producing animals such as dairy cattle are potential reservoirs of antimicrobial resistance (AMR), with multidrug-resistant (MDR) organisms such as Escherichia coli observed in higher frequency in young calves compared to older cattle. In this study, we characterized the genomes of enteric MDR E. coli from pre-weaned dairy calves with and without diarrhea and evaluated the influence of host-level factors on genomic composition. Whole genome sequence comparative analysis of E. coli (n = 43) revealed substantial genomic diversity that primarily clustered by sequence type and was minimally driven by calf diarrheal disease status (healthy, diarrheic, or recovered), antimicrobial exposure, and dietary zinc supplementation. Diverse AMR genes (ARGs)-including extended-spectrum beta-lactamase genes and quinolone resistance determinants-were identified (n = 40), with unique sets of ARGs co-occurring in gene clusters with large AMR plasmids IncA/C2 and IncFIB(AP001918). Zinc supplementation was not significantly associated with the selection of individual ARGs in E. coli, however analysis of ARG and metal resistance gene pairs identified positive associations between certain aminoglycoside, beta-lactam, sulfonamide, and trimethoprim ARGs with acid, tellurium and mercury resistance genes. Although E. coli in this study lacked the typical virulence factors of diarrheagenic strains, virulence genes overlapping with those in major pathotypes were identified. Among the 103 virulence genes detected, the highest abundance and diversity of genes corresponded to iron acquisition (siderophores and heme uptake). Our findings indicate that the host-level factors evaluated in this study were not key drivers of genomic variability, but that certain accessory genes in enteric MDR E. coli may be enriched. Collectively, this work provides insight into the genomic diversity and host-microbe interface of MDR E. coli from pre-weaned dairy calves.

PREVALENCE OF TOXOCARA CANIS AND ASSESSMENT OF KNOWLEDGE, ATTITUDE, AND PRACTICES OF THE COMMUNITY IN BISHOFTU TOWN, CENTRAL ETHIOPIA

A cross-sectional study was conducted in order to determine the prevalence of Toxocara canis, potential risk factors, and knowledge, attitude, and practices in Bishoftu, central Ethiopia. Fecal samples were taken from 300 randomly selected dogs of different sex, age group, breeds, and diarrheal status. The samples were processed using a flotation technique to detect the eggs of Toxocara canis. The current result revealed an overall prevalence of 32.33% (95% CI: 27.07 - 37.95) of Toxocara canis. The prevalence of the disease was 30.30% (95% CI: 22.61 - 38.90), 31.06% (95% CI: 23.30 - 39.70), and 44.44% (95% CI: 27.94 - 61.90) in young, adult and older dogs, respectively. The prevalence of Toxocara canis was 32.43% (95% CI: 26.77 - 38.50) and 31.71% (95% CI: 18.08 - 48.09) in female and male dogs, respectively. The present findings indicated that there were statistically significant links between the prevalence of Toxocara canis regarding dog breeds and diarrheal status (p < 0.05). The prevalence was higher in German Shepherd breed dogs (56.52%; 95% CI: 34.49 - 76.81) than in local dog breeds (30.32%; 95% CI: 24.97 - 36.11). The prevalence of the disease was higher in diarrheic dogs (46.48%; 95% CI: 34.55 - 58.71) than in non-diarrheic dogs (27.95%; 95% CI: 22.24 - 34.24). The knowledge, attitude, and practice assessment in the present study indicated poor dog management practices and insufficient public awareness. Most dog owners (56.7%; 95% CI: 50.85 - 62.35) did not deworm their dogs regularly, while 90.67% (95% CI: 86.79 - 93.71) fed dogs raw animal products. The present finding indicated that most respondents were not aware of zoonotic canine parasites (83.7%; 95% CI: 81.18 - 89.43) and toxocariasis (80.7%; 95% CI: 75.74 - 84.98). This study demonstrates a higher prevalence of Toxocara canis and lower public awareness about the disease. Therefore, this study recommends strategic deworming of dogs, better dog management, and increased public awareness in order to control the impact of the disease on animal and human health.

Impact of precursor-derived peracetic acid on post-weaning diarrhea, intestinal microbiota, and predicted microbial functional genes in weaned pigs

Post-weaning diarrhea affects piglets in the nursery phase of production, leading to a substantial impact both at the farm and financial levels. The multifactorial etiology of this disease includes housing conditions, pig genetics, microbial composition, and metagenomic assets. Among the common therapeutic approaches, the widely used zinc oxide underwent a European Union ban in 2022 due to its negative environmental impact and correlation to increased antimicrobial resistance. During this study, we have tested two levels of inclusion of the potential antimicrobial alternative peracetic acid, delivered in water via the hydrolysis of the precursors sodium percarbonate and tetraacetylethylenediamine, in comparison to zinc oxide and an untreated control during a 2-week animal study. We assessed the microbial composition and predicted the metagenome, together with performance and physiological parameters, in order to describe the microbial functional role in etiopathology. Both zinc oxide and peracetic acid resulted in amelioration of the diarrheal status by the end of the trial period, with noticeable zinc oxide effects visible from the first week. This was accompanied by improved performance when compared to the first-week figures and a decreased stomach pH in both peracetic acid levels. A significant reduction in both stomach and caecal Proteobacteria was recorded in the zinc oxide group, and a significant reduction of Campylobacter in the stomach was reported for both zinc oxide and one of the peracetic acid concentrations. Among other functional differences, we found that the predicted ortholog for the zonula occludens toxin, a virulence factor present in pathogens like Escherichia coli and Campylobacter jejuni, was less abundant in the stomach of treated pigs compared to the control group. In water, peracetic acid delivered via precursor hydrolysis has the potential to be a valid intervention, an alternative to antimicrobial, to assist the weaning of piglets. Our findings support the view that post-weaning diarrhea is a complex multifactorial disease with an important metagenomic component characterized by the differential abundance of specific predicted orthologs and microbial genera in the stomach and caecum of pigs.

Prevalence And Associated Risk Factors With Entamoeba histolytica Infections In School Children

Entamoeba histolytica is medically important pathogenic parasitic protozoan of human in origin which causes amoebiasis that is common in children of devolving nations in tropical and sub-tropical regions of the world. A total of 100 children, comprising 45 boys and 55 girls, were screened to calculate the prevalence and risk factors of E.histolytica along with their socio-demographic characteristics among school children at Chkadara, Lower Dir, Pakistan. The stool samples were collected and transported to the Parasitology Laboratory, Zoology Separtment, Malakand University for parasitic investigation. Each of the stool samples were processed through floatation and sedimentation procedures and techniques and examined under the microscope first under low and then high power objectives. The protozoan cysts and the helminth eggs were have been investigated and identified through valid keys. The data was analysed through chi-square test via GraphPad version 5. The p vale was considered statistically significant if less than 0.05 at 95% CI. The results indicates that Entamoeba histolytica is a prominent pathogenic protozoan parasite in school children, with a total of 67 cases identified. Boys showed a slightly higher prevalence of this parasite, with 58.8% (n=30/51) cases compared to 58.7% (n=37/63) in boys. Ascaris lumbricoides, is another common intestinal parasite, was observed in a total of 35 cases, (30.7%) with an almost equal distribution among boys (17 cases) and girls (18 cases). Moreover, hookworm species was found in 11 children, with a somewhat higher prevalence in girls 11.1% (7 cases). Giardia spp, although less prevalent, still accounted for 4 cases, equally distributed among boys. E.histolytica was highest in prevalence as 58.7 %( 67/114) followed by Ascaris lumbricoides 30.7 % (n=35/114), then hookworms 9.64 % (n=11/114) while Giardia spp was found in least prevalence 0.87 %( n=1/114). Pattern of infection reveal that most of the students were found to be infected with 2 species of parasites including one is E.histolytica (37%) followed by single infection 20% with E.histolytica while least number of participants were reported to be infected with 3 species of parasites 10%. The overall prevalence was reported as 67%. Age and types of stool were not significantly associated with the prevalence of E.histolytica P>0.05, however all other factors sex, diarrheal status, parent education, parent employment status and economic conditions were significantly related to the prevalence of E.histolytica infection (P<0.05).

Study of the infection rate of Entamoeba histolytica in children in some reference laboratories in Almarj, Libya

This study included about 262 samples collected from the age groups between 1 and 7 years. Some of these children have diarrhoea and their number is about 97, while others have diseases other than diarrhea. These samples were collected by five laboratories in the Libyan city of Al Marj in 2017 where that included (Ibn Zuhr Laboratory, Al-Razi Laboratory for Medical Analysis, Al-Rayada Hospital Laboratory , Almejhr Laboratory, and Tabebk laboratory). This study was conducted to determine the incidence of Entamoeba histolytica infection and the relationship of infection to both gender, age and diarrhoeal status. Of the 262 stool samples examined by routine microscopy, E. histolytica had the highest incidence of intestinal parasites (34.73%), there was no significant relationship between E. histolytica infection to both age and gender. This is what the study concluded.

Gut microbiome variations in Rhinopithecus roxellanae caused by changes in the environment

BackgroundThe snub-nosed monkey (Rhinopithecus roxellanae) is an endangered animal species mainly distributed in China and needs to be protected. Gut microbiome is an important determinant of animal health and population survival as it affects the adaptation of the animals to different foods and environments under kinetic changes of intrinsic and extrinsic factors. Therefore, this study aimed to elucidate gut fecal microbiome profiles of snub-nosed monkeys affected by several extrinsic and intrinsic factors, including raising patterns (captive vs. wild), age, sex, and diarrheal status to provide a reference for making protection strategies.ResultsThe 16S rRNA gene sequencing was firstly used to pre-check clustering of 38 fecal samples from the monkeys including 30 wild and 8 captive (5 healthy and 3 diarrheal) from three Regions of Shennongjia Nature Reserve, Hubei Province, China. Then the 24 samples with high-quality DNA from 18 wild and 6 captive (4 healthy and 2 diarrheal) monkeys were subjected to shotgun metagenomic sequencing to characterize bacterial gut microbial communities. We discovered that the raising pattern (captive and wild) rather than age and sex was the predominant factor attributed to gut microbiome structure and proportionality. Wild monkeys had significantly higher bacterial diversity and lower Bacteroidetes/Firmicutes ratios than captive animals. Moreover, the gut microbiomes in wild healthy monkeys were enriched for the genes involved in fatty acid production, while in captive animals, genes were enriched for vitamin biosynthesis and metabolism and amino acid biosynthesis from carbohydrate intermediates. Additionally, a total of 37 antibiotic resistant genes (ARG) types were detected. Unlike the microbiome diversity, the captive monkeys have a higher diversity of ARG than the wild animals.ConclusionTaken together, we highlight the importance of self-reprogramed metabolism in the snub-nosed monkey gut microbiome to help captive and wild monkeys adapt to different intrinsic and extrinsic environmental change.

Determinants of Mortality Among Severely Malnourished Children in Northern Nigeria.

ObjectivesSevere Acute Malnutrition is a significant cause of mortality in children under the age of 5 years in low-resource settings, including Northern Nigeria. The study aimed to determine the associations between selected risk factors and mortality outcomes in children admitted with SAM in a facility in Katsina State, Northern Nigeria.MethodsA prospective observational cohort of 201 children aged 6 to 59 months who were admitted with severe acute malnutrition (SAM) in stabilization centers in Katsina State, Northern Nigeria between May 18, 2021, and July 20, 2021, (63 days) were assessed followed up. Outcomes were analyzed using Kaplan-Meir analysis to estimate time to death, and Cox proportional-hazard regression model was used to determine predictors of mortality.ResultsThe log-rank test showed significant differences in the probability of death between categories of diarrheal status (log-rank statistic = 9.760, P = .021) and presence of existing disease (comorbidity) (log-rank statistic = 5.338, P = .021). The study identified that severely malnourished children admitted with comorbidities showed significant association with time to event (death) (AHR: 4.109, 95% CI: 1.51, 32.60). The estimated mean time until death was 57.9 days (±3.0) for children without comorbidities and 20.1 (±3.0) days for children with comorbidities. The median survival time was 18 days for children with comorbidities.ConclusionThe presence of comorbidities was significantly associated with mortality. Severely malnourished children with comorbidities had 4 times higher mortality risk than severely malnourished children admitted without comorbidities. Clinicians and health workers should give due emphasis to the early detection and effective management of comorbidities in children with severe acute malnutrition.

Norovirus-specific immunoglobulin A in breast milk for protection against norovirus-associated diarrhea among infants.

BackgroundNorovirus (NV) causes acute gastroenteritis in infants. Humoral and fecal immunoglobulin A (IgA) responses have been correlated with protection against NV; however, the role of breast milk IgA against NV infection and associated diarrhea is still unknown. This study aimed to evaluate the protective role of NV-specific IgA (NV-IgA) in breast milk.MethodsNinety-five breast milk samples collected from mothers enrolled in a 2016–2017 Peruvian birth cohort study were tested for total IgA and NV-IgA by ELISA using GII·4 variants and non-GII·4 genotype virus-like particles (VLPs). Breast milk samples were grouped according to the NV infection and diarrheal status of infants: NV positive with diarrhea (NV+D+, n=18); NV positive without diarrhea (NV+D-, n=37); and NV negative without diarrhea (NV-D-, n=40). The percent positivity and titer of NV-IgA were compared among groups. The cross-reactivity was estimated based on the correlation of ratio between NV-IgA against GII·4 variants and non-GII·4 genotype VLPs.FindingsNV-IgA had high positivity rates against different VLPs, especially against GII (89–100%). The NV+D- group had higher percent positivity (89% vs. 61%, p=0·03) and median titer (1:100 vs 1:50, p=0·03) of NV-IgA than the NV+D+ group against GI·1 VLPs. A relatively high correlation between different GII·4 variants (0·87) and low correlation between genogroups (0·23–0·37) were observed.InterpretationMothers with high positivity rates and titers of NV-IgA in breast milk had NV infected infants with reduced diarrheal symptoms. Antigenic relatedness to the genetic diversity of human norovirus was suggested.Funding National Institute of Allergy and Infectious Diseases, National Institute of Health: 1R01AI108695–01A1 and the Japan Society for the Promotion of Science (Fostering Joint International Research B):19KK0241

Metagenomics reveals impact of geography and acute diarrheal disease on the Central Indian human gut microbiome

ABSTRACT Background The Central Indian gut microbiome remains grossly understudied. Herein, we sought to investigate the burden of antimicrobial resistance and diarrheal diseases, particularly Clostridioides difficile, in rural-agricultural and urban populations in Central India, where there is widespread unregulated antibiotic use. We utilized shotgun metagenomics to comprehensively characterize the bacterial and viral fractions of the gut microbiome and their encoded functions in 105 participants. Results We observed distinct rural-urban differences in bacterial and viral populations, with geography exhibiting a greater influence than diarrheal status. Clostridioides difficile disease was more commonly observed in urban subjects, and their microbiomes were enriched in metabolic pathways relating to the metabolism of industrial compounds and genes encoding resistance to 3rd generation cephalosporins and carbapenems. By linking phages present in the microbiome to their bacterial hosts through CRISPR spacers, phage variation could be directly related to shifts in bacterial populations, with the auxiliary metabolic potential of rural-associated phages enriched for carbon and amino acid energy metabolism. Conclusions We report distinct differences in antimicrobial resistance gene profiles, enrichment of metabolic pathways and phage composition between rural and urban populations, as well as a higher burden of Clostridioides difficile disease in the urban population. Our results reveal that geography is the key driver of variation in urban and rural Indian microbiomes, with acute diarrheal disease, including C. difficile disease exerting a lesser impact. Future studies will be required to understand the potential role of dietary, cultural, and genetic factors in contributing to microbiome differences between rural and urban populations.

Does suboptimal household flooring increase the risk of diarrhoea and intestinal parasite infection in low and middle income endemic settings? A systematic review and meta-analysis protocol

BackgroundWater, sanitation, and hygiene interventions often fail to show long-term impact on diarrhoeal and/or intestinal parasite risk in many low- and middle-income countries. Less attention has been paid to wider contextual factors that may contribute to high levels of contamination in the domestic environment such as household flooring. The purpose of this study will be to assess the association between diarrhoeal and/or intestinal parasite infection status and unimproved/unfinished flooring in low- and middle-income countries.MethodsWe will conduct a comprehensive search of published studies (randomized controlled trials, non-randomized controlled trials, and observational studies) that examined the association between unimproved/unfinished household flooring and diarrhoeal and/or intestinal parasite infection status from January 1, 1980, onwards with no language restriction. The primary outcome will include diarrhoeal and/or intestinal parasite infection status. Databases to be searched include EMBASE, MEDLINE, Web of Science, and Google Scholar. The secondary outcome will be the association between specific pathogens (laboratory confirmed) and unimproved/unfinished household flooring. Independent screening for eligible studies using defined criteria and data extraction will be completed in duplicate and independently. Any discrepancies between the two reviewers will be resolved by consensus and/or arbitration by a third researcher. If data permits, random effects models will be used where appropriate. Subgroup and additional analyses will be conducted to explore the potential sources of heterogeneity (e.g. age group, geographical region) and potential risk of bias of included studies.DiscussionThis review will provide a comprehensive examination of a possible association between suboptimal household flooring and increased risk of enteric pathogen infection, highlight gaps for future research in high risk areas, and inform intervention design for future planned studies in Kenya and/or elsewhere in the region.Systematic review registrationPROSPERO registration number: CRD42019156437

Norovirus-Specific Immunoglobulin A in Breast Milk for Protection Against Norovirus-Associated Diarrhea Among Infants

Background: Norovirus (NV) causes acute gastroenteritis in infants. Humoral and fecal immunoglobulin A (IgA) responses have been correlated with protection against NV; however, the role of breast milk IgA against NV infection and associated diarrhea is still unknown. This study aimed to evaluate the protective role of NV-specific IgA in breast milk. Methods: We analyzed 95 breast milk samples collected from mothers enrolled in a birth cohort study conducted in Peru during 2016–2017. Samples were collected before the first NV infection of infants and tested for total IgA and NV-specific IgA using virus-like particles (VLPs) that represented GII·4 variants and non-GII·4 genotypes. Norovirus infection in infants was assessed by testing their stool samples for NV via real-time PCR. Breast milk samples were grouped according to NV infection and diarrheal status of infants: NV positive with diarrhea (NV+D+, n=18); NV positive without diarrhea (NV+D-, n=37); and NV negative without diarrhea (NV-D-, n=40). The percent positivity and titer of NV-specific IgA (Log10) were compared among groups. Findings: Breast milk IgA had a high percent positivity against different NV VLPs, especially against GII (89–100%). Higher median titer of NV-specific IgA was observed in NV+D- when compared to the NV+D+ group against GII·4 Sydney (NV+D+:500 vs. NV+D-:100, p=0·05) and GI·1 (NV+D+:100 vs. NV+D-:50, p=0·01). Although the median titer of NV IgA against GI·9 was similar, NV+D- had higher distribution of titers when compared to NV+D+ group (NV+D-:100 [interquartile range (IQR):100–500] vs. NV+D+:100 [IQR:50–200], p=0·03). Interpretations: Breast milk IgA highly reacted to different NV VLPs. NV-specific IgA may protect infants from having NV-associated diarrhea, but not from NV infection, thus reducing NV symptoms. Funding Statement: National Institute of Allergy and Infectious Diseases, National Institute of Health: 1R01AI108695–01A1 and the Japan Society for the Promotion of Science (Fostering Joint International Research B): 19KK0241 Declaration of Interests: We declare that we have no conflicts of interest. Ethics Approval Statement: The mothers included in this study were selected from a larger birth cohort study carried out in Peru during 2016-2017. The birth cohort study was approved by the Institutional Review Boards of Asociacion Benefica PRISMA, Universidad Peruana Cayetano Heredia, Johns Hopkins University, University of California-San Francisco. Informed consent were obtained from the mothers or caretakers of the infants. The analysis of archived specimen was approved by the ethics committee of the Tohoku University, Graduate School of Medicine.

Environmental factors associated with diarrhoea prevalence among under-five children in the Mataniko settlements in Honiara, Solomon Islands.

Children aged less than 5 years are often at high risk of diarrhoeal infection. In the Solomon Islands, diarrhoea is the second leading cause of under-five mortality with about one in every 10 children dying from it before reaching 5 years. This study aims to assess environmental factors that are associated with under-five diarrhoea prevalence in the Mataniko informal settlements, in Honiara, Solomon Islands. Three out of the six settlements along the Mataniko River corridor were randomly selected. Caregivers who were taking care of at least one child under 5 years, and had signed a voluntary informed consent form, were included in the study. Instruments employed to collect the study variables were global positioning system technology and a questionnaire. Each child's medical record was used to verify the date of his or her diarrhoeal status. The data were entered and analysed using SPSS (v23). Binary logistic regression was used to measure the strength of association between under-five diarrhoea and the independent variables. A p-value of <0.05 was considered as statistically significant (p<0.05). A total of 205 caregivers with at least one child under 5 years participated in the study. Approximately half (45.9%) of the participants reported that their children (<5 years) had suffered with at least one episode of diarrhoea within the 2 weeks prior to the survey. Of the participants, 73.2% did not own a toilet facility and 61.0% of households were built on low-altitude areas (&le;19 m above sea level), and above half (70.6%) were built near (&le;125 m) the river. The presence of stagnant wastewater, flies, solid waste and water-filled containers near households, plus the distance of under-five households from the river, were found to be directly associated with under-five diarrhoea in the Mataniko informal settlements (p<0.05). Awareness and advocacy programs on environmental hygiene, food hygiene and potential health risks about the river should be ongoing at the community level.

Cryptosporidiosis And Other Intestinal Parasitic Infections And Concomitant Threats Among HIV-Infected Children In Southern Ethiopia Receiving First-Line Antiretroviral Therapy.

BackgroundChildren infected with human immunodeficiency virus (HIV) are at high risk of acquiring intestinal parasitic infections. This study aimed to determine the magnitude of Cryptosporidium and other intestinal parasitic infections and concomitant threats among HIV-infected children.MethodsA hospital-based cross-sectional study was carried out at three antiretroviral therapy clinics in southern Ethiopia from February 2016 to June 2017 in 384 HIV positive children. Socio-demographic and clinical data were collected using structured questionnaires. Direct stool microscopic examination and modified Zeihl–Neelsen staining technique to identify parasites. Chi-square test was conducted to determine the real predictors of the infection. Significant association was considered when p-value <0.05 at 95% CI.ResultsThe overall magnitude of intestinal parasitic infections among the study population was 16.9% (95% CI: 13.0–20.8%). The most predominant parasitic infections were Cryptosporidium spp. (9.6%) and the least was Taenia spp. (0.78%). Diarrheal status (χ2=7.653, df=2, p=0.022) was detected to be the only significant associated variable.ConclusionCryptosporidium infection was found to be the most common intestinal parasitosis among HIV-infected children. Routine screening service for Cryptosporidium and other intestinal parasites is important in the clinical management of HIV-infected children.

Diarrheal status and associated factors in under five years old children in relation to implemented and unimplemented community-led total sanitation and hygiene in Yaya Gulele in 2017.

PurposeThe aim of this study was to assess the diarrheal status and associated factors of under five-years old children among implemented and unimplemented community-led total sanitation and hygiene (CLTSH) in Yaya Gulele district, Ethiopia, in 2017.MethodsCommunity-based comparative cross-sectional study involving 380 households from each implemented and unimplemented CLTSH area was conducted from December 1 to June 30, 2017. Pretested structured questionnaire and a complete observational checklist were used to collect data. Qualitative data were collected via focused group discussions and analyzed manually. Data were checked and entered to Epi-info 3.5.4 and analyzed by SPSS version 20. Bivariate and multivariable logistic regression analyses were computedResultsIn this study, 88% implemented and 66% unimplemented CLTSH had latrine. Of households owning latrine, 85% in implemented and 75% in unimplemented CLTSH utilize latrine properly. In the study area, 12% in implemented and 34% in unimplemented CLTSH area practice open defecation. Two weeks period diarrhea prevalence was 13.4% (95% CI: 10.2-17.3%) in implemented CLTSH and 36.3% (95% CI: 31.7–41.6%) in unimplemented CLTSH. Having two or more children under five years old (adjusted odds ratio [AOR] = 2.33; 95% CI: 1.09–4.96), lack of clean water storage (AOR = 2.36; 95% CI: 1.16–4.80), negative attitude of mothers/caregivers toward diarrhea (AOR = 2.07; 95% CI: 1.06–4.04), presence of feces in the compound (AOR = 1.88; 95% CI: 1.10–3.22), and lack of hand washing facility in the compound (AOR = 2.64; 95% CI: 1.47–4.74) were associated factors of the outcome.ConclusionImplementation of CLTSH is the applicable tool to reduce diarrhea prevalence. Having two or more children under five years old, unclean water storage, negative attitude of mothers toward diarrhea, existence of feces in the compound, and lack of hand washing facility in the compound were associated factors to the diarrheal status of children under five years old. Health workers and local authorities inspire the community to clean water-storage facilities and the environment, to change mothers’ behavior toward diarrhea, as well as construct hand washing facility in the compound.

PCR-ribotype distribution of Clostridium difficile in Irish pigs

Clostridium difficile is an important enteric pathogen in humans causing infections in the healthcare environment and the community. Carriage of C. difficile and C. difficile-related enterocolitis has been reported in piglets worldwide. The aim of this study was to investigate the rates of C. difficile isolation from pigs in Ireland. Faecal samples from piglet litters and sows were collected from six farms in 2015. The sows were non-diarrhoeal at the time of sampling. The diarrhoeal status of the piglets was unknown. C. difficile was isolated from 34/44 (77%) of piglet litter samples and from 33/156 (21%) of sow samples. The isolation rate in sows varied from 3 to 39% and in piglet litters from 72 to 86% depending on farm location. Toxin A and toxin B were present in 99% (66/67) of isolates; and binary toxin in 85% (57/67). Only PCR-ribotypes 078 (88%) and 193 (12%) were identified in piglets. Seven PCR-ribotypes were detected in sow C. difficile isolates: PCR-ribotypes 078 (67%), 050 (12%), 014/020 (6%), 015 (6%), 029 (3%), 035 (3%) and 193 (3%). This study shows that toxigenic C. difficile strains such as PCR-ribotype 078 can be commonly isolated from pigs at different geographical locations in Ireland. Since PCR-ribotype 078 is frequently found in humans in Ireland, this highlights the potential for interspecies transmission.

Effects of mulberry (Morus alba L.) leaf polysaccharides on growth performance, diarrhea, blood parameters, and gut microbiota of early-weanling pigs

This study aimed to investigate the effects of mulberry leaf polysaccharides (MLPs) dietary supplements on the growth performance, diarrhea, blood biochemical parameters, and gut microbiota of early weanling piglets. A total of 150 Duroc–Landrace–Yorkshire crossbred weanling pigs (age: 28±2d; body weight: 9.18±0.46kg), were used in this study. Subjects were randomly divided into five treatment groups with six replicates in each group (n=5 animals per replicate). The dietary treatments were as follows: (1) control treatment (CT): basal diet (BD); (2) low-dose MLPs treatment (LT): 0.3g/kg of MLPs+BD; (3) medium-dose MLPs treatment (MT): 0.6g/kg of MLPs+BD; (4) high-dose MLPs treatment (HT): 0.9g/kg of MLPs+BD; and (5) antibiotic treatment (AT): 0.15g/kg chlortetracycline+BD. The fasting weight of each animal was measured and recorded in the morning at three time points (0, 10, and 21d) in addition to food intake per pigpen. These data were then used to calculate the average daily gain (ADG), average daily feed intake (ADFI), and feed-to-gain (F/G) ratio of each subject. The diarrheal status of each piglet was examined and recorded twice daily (morning and afternoon). At the end of the experiment, blood samples were collected for biochemical analysis, and the contents from the ceacum, colon, and rectum were harvested to identify and quantify relevant gut microbiota. Our results showed that there was no significant differences in the ADG, ADFI, or F/G ratio among MLPs-treated groups (P>0.05), but the ADFI in LT, MT, and HT groups were all higher than those of both the CT and AT groups. Additionally, the F/G ratios in LT, MT, and HT groups were lower than those in the CT and AT groups. The LT, MT and HT groups showed a significant reduction in diarrheal incidence (P<0.05) when compared to the CT and AT groups. Biochemical analysis revealed that blood glucose (GLU) levels in all MLPs-treated groups were significantly (P<0.05) lower than in either the CT or AT groups. Blood urea nitrogen (BUN) levels in the MT and HT groups were significantly (P<0.05) different from those in CT and AT group and T3 levels in HT were significantly (P<0.05) higher than those in either the CT or AT groups. Although T4 levels in MLPs-treated groups were not significantly different from the AT control group, both IGF-1 and GH levels in the MT group were significantly higher than those in either the CT or AT groups (P<0.05). Our microbiota results indicated that the overall effects of LT, MT, and HT in both inhibiting gut Escherichia coli and in promoting gut lactobacilli and bifidobacteria were superior than those of both the CT and AT groups. Notably, the HT group (0.9g/kg MLPs) showed a significantly better inhibitory effect on gut E. coli as well as a significantly better promotional effect on gut-beneficial bacteria when compared to both the CT and AT groups (P<0.05). Collectively, these results suggest that dietary addition of MLPs can improve the ecology of gut microbiota, reduce diarrhea rate, and improve the overall growth performance in early-weanling pigs.

Microbiome data distinguish patients with Clostridium difficile infection and non-C. difficile-associated diarrhea from healthy controls.

ABSTRACTAntibiotic usage is the most commonly cited risk factor for hospital-acquired Clostridium difficile infections (CDI). The increased risk is due to disruption of the indigenous microbiome and a subsequent decrease in colonization resistance by the perturbed bacterial community; however, the specific changes in the microbiome that lead to increased risk are poorly understood. We developed statistical models that incorporated microbiome data with clinical and demographic data to better understand why individuals develop CDI. The 16S rRNA genes were sequenced from the feces of 338 individuals, including cases, diarrheal controls, and nondiarrheal controls. We modeled CDI and diarrheal status using multiple clinical variables, including age, antibiotic use, antacid use, and other known risk factors using logit regression. This base model was compared to models that incorporated microbiome data, using diversity metrics, community types, or specific bacterial populations, to identify characteristics of the microbiome associated with CDI susceptibility or resistance. The addition of microbiome data significantly improved our ability to distinguish CDI status when comparing cases or diarrheal controls to nondiarrheal controls. However, only when we assigned samples to community types was it possible to differentiate cases from diarrheal controls. Several bacterial species within the Ruminococcaceae, Lachnospiraceae, Bacteroides, and Porphyromonadaceae were largely absent in cases and highly associated with nondiarrheal controls. The improved discriminatory ability of our microbiome-based models confirms the theory that factors affecting the microbiome influence CDI.

Intestinal Parasitic Infections in HIV-Infected Patients, Lao People’s Democratic Republic

BackgroundHIV infection is an emerging problem in Laos. We conducted the first prospective study on intestinal parasites, including opportunistic protozoa, in newly diagnosed HIV infected patients, with or without diarrhea. The aims were to describe the spectrum of infections, to determine their prevalence and to assess their associations with diarrhea, CD4 cell count, place of residence and living conditions.MethodologyOne to three stool samples over consecutive days were obtained from 137 patients. The Kato thick smear method, formalin-ethyl concentration and specific stains for coccidia and microsporidia diagnosis were performed on 260 stool samples. Baseline characteristics regarding relevant demographics, place of residence and living conditions, clinical features including diarrhea, were collected using a standardized questionnaire.Principal FindingsThe 137 patients were young (median age: 36 years) and severely immunocompromised (83.9% at WHO stage 3 or 4, median CD4 cell count: 41/mm3). Diarrhea was present in 43.0% of patients. Parasite infection was found in 78.8% of patients, infection with at least two species in 49.6%. Prevalence rates of protozoan and helminth infections were similar (54.7% and 58.4% respectively). Blastocystis sp. was the most frequent protozoa (26.3%). Cryptosporidium sp., Cytoisospora belli and microsporidia, found at low prevalence rates (6.6%, 4.4%, 2.9%, respectively), were described for the first time in Laos. Cryptosporidium sp. was associated with persistent diarrhea. Strongyloides stercoralis was the most prevalent helminth following Opisthorchis viverrini (20.4% and 47.5% respectively). The most immunocompromised patients, as assessed by a CD4 count ≤ 50 cells/mm3, were more likely to be infected with intestinal parasites.Conclusions/SignificanceHIV infection was mainly diagnosed at an advanced stage of immunosuppression in Lao patients. Intestinal parasite infections were highly prevalent regardless of their diarrheal status. Opportunistic infections were reported. Improving the laboratory diagnosis of intestinal parasite infections and the knowledge on their local risk factors is warranted.

Quantitative PCR and culture evaluation for enterotoxigenicEscherichia coli(ETEC) associated diarrhea in volunteers

Recent evidence suggests that the abundance of enteric pathogens in the stool correlates with the presence of clinical diarrhea. We quantified the fecal pathogen after feeding enterotoxigenic Escherichia coli (ETEC) strain H10407 to 30 adult volunteers. Stools were collected daily and examined using qualitative and quantitative (Q) culture. DNA was isolated, and quantitative (Q) PCR targeting the heat-labile toxin (LT) gene was performed. Nine volunteers developed diarrhea. Among 131 stool specimens with complete data, pathogen abundance by QPCR was strongly correlated with Qculture, ρ = 0.61, P < 0.0001. Receiver operating characteristic curve analysis comparing quantitative data against diarrhea status suggested cut-points, based on a maximum Youden Index, of 2.8 × 10(4) LT gene copies and 1.8 × 10(7) CFU. Based on these cut-points, QPCR had a sensitivity and specificity compared with diarrheal status of 0.75 and 0.87, respectively, and an OR of 20.0 (95% CI 5.7-70.2), whereas Qculture had a sensitivity and specificity of 0.73 and 0.91, respectively, and an OR of 28.6 (95% CI 7.7-106.6). Qculture had a sensitivity and specificity of 0.82 and 0.48, respectively and an OR of 4.4 (95% CI 1.2-16.0). The correlation between Qculture and QPCR was highest in diarrheal specimens, and both quantitative methods demonstrated stronger association with diarrhea than qualitative culture.

The effect of New Neonatal Porcine Diarrhoea Syndrome (NNPDS) on average daily gain and mortality in 4 Danish pig herds.

BackgroundThe study evaluated the effect of New Neonatal Porcine Diarrhoea Syndrome (NNPDS) on average daily gain (ADG) and mortality and described the clinical manifestations in four herds suffering from the syndrome. NNPDS is a diarrhoeic syndrome affecting piglets within the first week of life, which is not caused by enterotoxigenic Escherichia coli (ETEC), Clostridium perfringens (C. perfringens) type A/C, Clostridium difficile (C. difficile), rotavirus A, coronavirus, Cystoisospora suis, Strongyloides ransomi, Giardia spp or Cryptosporidium spp.ResultsPiglets were estimated to have a negative ADG of 9 and 14 g when diarrhoeic for 1 day and >1 day respectively. However, if only diarrhoeic on the day of birth, no negative effect on ADG was seen. Piglets originating from severely affected litters were estimated to have a reduced ADG of 38 g. The study did not show an overall effect of diarrhoea on mortality, but herd of origin, sow parity, birth weight, and gender were significantly associated with mortality. In one of the herds, approximately 25% of the diarrhoeic piglets vs. 6% of the non-diarrhoeic piglets died, and 74% of necropsied piglets were diagnosed with enteritis. These findings indicate that the high mortality seen in this herd was due to diarrhoea.ConclusionsNNPDS negatively affected ADG in piglets, and even piglets that were diarrhoeic for one day only experienced a reduction in ADG. However, the study showed that diarrhoea restricted to the day of birth did not affect ADG and suggested this phenomenon to be unrelated to the syndrome. Since the diarrhoeal status of the litter had important effects on ADG, future research on NNPDS probably ought to focus on piglets from severely affected litters.The study showed important dissimilarities in the course of diarrhoea between the herds, and one herd was considerably more affected than the others. Within this herd, NNPDS seemed to be associated with a higher mortality, whereas in general the study did not show lethal effects of NNPDS.