Bioprospecting of soil-borne microorganisms and chemical dereplication of their anti-microbial constituents with the aid of UPLC-QTOF-MS and molecular networking approach.

Due to the emergence of drug-resistant microorganisms, the search for broad-spectrum antimicrobial compounds has become extremely crucial. Natural sources like plants and soils have been explored for diverse metabolites with antimicrobial properties. This study aimed to identify microorganisms from agricultural soils exhibiting antimicrobial effects against known human pathogens, and to highlight the chemical space of the responsible compounds through the computational metabolomics-based bioprospecting approach. Herein, bacteria were extracted from soil samples and their antimicrobial potential was measured via the agar well diffusion method. Methanolic extracts from the active bacteria were analyzed using the liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) technique, and the subsequent data was further analyzed through molecular networking approach which aided in identification of potential anti-microbial compounds. Furthermore, 16S rRNA gene sequencing enabled identification of the active bacterial isolates, where isolate 1 and 2 were identified as strains of Bacillus pumilus, whilst isolate 3 was found to be Bacillus subtilis. Interestingly, isolate 3 (Bacillus subtilis) displayed wide-ranging antimicrobial activity against the tested human pathogens. Molecular networking revealed the presence of Diketopiperazine compounds such as cyclo (D-Pro-D-Leu), cyclo (L-Tyr-L-Pro), cyclo (L-Pro-D-Phe), and cyclo (L-Pro-L-Val), alongside Surfactin C, Surfactin B, Pumilacidin E, and Isarrin D in the Bacillus strains as the main anti-microbial compounds. The application of the molecular networking approach represents an innovation in the field of bio-guided bioprospection of microorganisms and has proved to be an effective and feasible towards unearthing potent antimicrobial compounds. Additionally, the (computational metabolomics-based) approach accelerates the discovery of bioactive compounds and isolation of strains which offer a promising avenue for discovering new clinical antimicrobials. Finally, soil microbial flora could serve an alternative source of anti-microbial compounds which can assist in the fight against emergence of multi-drug resistance bacterial pathogens.

The impact of DNA methylation as a factor of Adverse Pregnancy and Birth Outcomes (APBOs): a systematic review protocol.

BACKGROUND: Deoxyribonucleic acid (DNA) methylation is one of the epigenetic modifications that has gained a lot of interest as a factor influencing fetal programming and as a biomarker for adverse pregnancy and birth outcomes (APBOs). Epidemiological studies have demonstrated that DNA methylation can result in adverse pregnancy and birth outcomes (APBOs) including miscarriage, intrauterine growth restriction (IUGR), low birth weight (LBW), sepsis, and preterm birth (PTB), which may later result in diseases in adulthood. However, the mechanism by which DNA methylation influences these APBOs remains unclear. The systematic review will assess the association between global and gene-specific DNA methylation with adverse pregnancy outcomes. METHOD: The Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) 2020 checklist will be followed when conducting this systematic review. To develop the search strategy the PI(E)COS (population, intervention/exposure, comparator/control, outcome, and study designs) framework will be followed. Thus far, the research team has retrieved 4721 from Cochrane Library, PubMed, Web of Sciences, and MEDLINE. Out of these, 584 studies have been screened for eligibility, and approximately 124 studies meet the inclusion criteria. Pending the search results identified from the grey literature. For identification of unpublished studies in journals indexed in electronic databases, Google Scholar will be used. I.M and A.S will separately extract data from the articles and screen them, if there are any disagreements between I.M and A.S, then the L.M will resolve them. The methodological quality and bias risk of the included studies will be evaluated using the Critical Appraisal Skill Programme CASP) checklist. [Formula: see text] and [Formula: see text] alpha = 0.10 statistic will be used for assessing statistical heterogeneity between studies. The Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) approach will be used to assess and grade the overall quality of extracted data. ETHICS AND DISSEMINATION: Ethical approval is not required. The systematic review will assess available literature on possible associations between DNA methylation with adverse pregnancy and birth outcomes (APBOs) including LBW, IUGR, miscarriage, sepsis, and PTB. The findings could help guide future research assessing DNA methylation and other APBOs. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRCRD42022370647.

CO-INFECTION WITH MALARIA AND INTESTINAL PARASITES AND ITS ASSOCIATION WITH ANEMIA IN CHILDREN (ZERO TO TEN YEARS OLD) IN TIKO SUBDIVISION, CAMEROON.

Concomitant infections with malaria and intestinal parasitic infections may be associated with anemia in children (0-10 yr). This study determined the prevalence of co-infection with malaria and intestinal parasitic infections and determined its association with anemia in children (0-10 yr) in Tiko, Cameroon. A hospital-based cross-sectional study was carried out whereby venous blood and stool samples were collected from 377 febrile children. Blood was used to perform a full blood count. Thick and thin blood films were prepared and stained with Giemsa for malaria parasite diagnosis. The formol ether concentration technique was used to analyze the stools. Pearson's chi-square test, Student's t-test, and other statistical analyses were performed. Of the 377 participants, 139 (36.9%) were positive for malaria, 21 (5.6%) had intestinal helminths, 8 (2%) had co-infection, and 79 (21.0%) were anemic. Malaria and anemia were prevalent among the children and were significantly associated (P = 0.025). There was no statistically significant difference (P > 0.05) among age groups. Girls were more often infected with malaria (69, 37.3%), and boys were more often infected with intestinal parasites (13, 7.0%), but there was no statistical association for both malaria and intestinal parasitic infections (IPIs) for both sexes (P > 0.05). Hookworms, Ascaris lumbricoides, and Trichuris trichiura were the intestinal parasites found in this study. There was a significant association between anemia and parasitic co-infection in children (P = 0.003). Malaria and IPIs are prevalent in the Tiko municipality. They play a great role in anemia especially when there is a co-infection. Public education and awareness campaigns are necessary in this municipality.

Factors Related to the Implementation of Preconception Care Recommendations in Selected Districts of Limpopo Province: A Qualitative Study.

Preconception care (PCC) is the provision of biomedical, behavioural, and social health interventions to women and couples before they fall pregnant. The World Health Organization (WHO) developed PCC recommendations in 2013, which were included in the South African maternity care guidelines in 2016. The purpose of PCC is to lessen behaviours and environmental factors leading to maternal ill-health, thus reducing maternal and perinatal mortality rates. OBJECTIVE: To determine the implementation of PCC recommendations at health facilities in the selected districts of Limpopo Province and the associated factors. METHODS: A qualitative exploratory design was used. Nonprobability, purposive sampling was used to sample 29 professional nurses (PNs), and 51 women of childbearing age (WCBA) (19-35 years) from clinics and community health centres (CHCs). Data were collected through in-depth interviews with the professional nurses and focus group discussions with the WCBA. Data analysis was performed through open coding. Measures of trustworthiness were adhered to. Permission to conduct the study was obtained from relevant stakeholders, participation was voluntary and participants signed a consent form prior to data collection. RESULTS: The findings of the study revealed that there was partial implementation of the PCC recommendations in the selected districts of Limpopo Province, PCC provision was dependent on clients' initiation, a knowledge gap regarding PCC recommendations was identified from the professional nurses, and a lack of awareness regarding PCC from the WCBA. CONCLUSION: The preconception period is an important determinant of the pregnancy outcome; therefore, focus should be redirected to the pre-pregnancy period and not only to when the woman is already pregnant. However, to achieve this, professional nurses and WCBA should be empowered regarding PCC and its benefits.

The Epidemiology of Sapovirus in the Etiology, Risk Factors, and Interactions of Enteric Infection and Malnutrition and the Consequences for Child Health and Development Study: Evidence of Protection Following Natural Infection.

BACKGROUND: Sapovirus is one of the principal agents of acute viral enteritis in children. Because it has not been routinely included in diagnostic evaluations, the epidemiology and natural history remain poorly described. METHODS: A birth cohort of 1715 children from 8 countries contributed surveillance samples (n = 35 620) and diarrheal specimens (n = 6868) from 0 to 24 months of age. Sapovirus was detected by quantitative polymerase chain reaction concurrently to other enteropathogens using multiarray cards. Logistic regression was used to identify risk factors, and longitudinal models were employed to estimate incidence rates and evaluate evidence of protective immunity. RESULTS: Sapovirus was detected in 24.7% (n = 1665) of diarrheal stools and 12.8% (n = 4429) of monthly surveillance samples. More than 90% of children were infected and 60% experienced sapovirus diarrhea in the first 2 years of life. Breastfeeding and higher socioeconomic status were associated with reduced incidence of infection and illness. Specimens with sapovirus detected had an increased odds of coinfection with rotavirus (odds ratio [OR], 1.6 [95% confidence interval {CI}, 1.3-2.0]), astrovirus (OR, 1.5 [95% CI, 1.3-1.7]), adenovirus (OR, 1.3 [95% CI, 1.1-1.5]), and Shigella (OR, 1.4 [95% CI, 1.3-1.6]). Prior infection with sapovirus conferred a risk reduction of 22% for subsequent infection (hazard ratio [HR], 0.78 [95% CI, .74-.85]) and 24% for subsequent diarrhea (95% CI, 11.0%-35.0%; HR, 0.76). CONCLUSIONS: Sapovirus is a common cause of early childhood diarrhea. Further research on coinfections is warranted. Evidence of acquired immunity was observed even in the absence of genotype-specific analysis for this pathogen of known genetic diversity.

Burden, Clinical Characteristics, Risk Factors, and Seasonality of Adenovirus 40/41 Diarrhea in Children in Eight Low-Resource Settings.

Background: The application of molecular diagnostics has identified enteric group adenovirus serotypes 40 and 41 as important causes of diarrhea in children. However, many aspects of the epidemiology of adenovirus 40/41 diarrhea have not been described. Methods: We used data from the 8-site Etiology, Risk Factors, and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development Project birth cohort study to describe site- and age-specific incidence, risk factors, clinical characteristics, and seasonality. Results: The incidence of adenovirus 40/41 diarrhea was substantially higher by quantitative polymerase chain reaction than enzyme immunoassay and peaked at ∼30 episodes per 100 child-years in children aged 7-15 months, with substantial variation in incidence between sites. A significant burden was also seen in children 0-6 months of age, higher than other viral etiologies with the exception of rotavirus. Children with adenovirus 40/41 diarrhea were more likely to have a fever than children with norovirus, sapovirus, and astrovirus (adjusted odds ratio [aOR], 1.62; 95% CI, 1.16-2.26) but less likely than children with rotavirus (aOR, 0.66; 95% CI, 0.49-0.91). Exclusive breastfeeding was strongly protective against adenovirus 40/41 diarrhea (hazard ratio, 0.64; 95% CI, 0.48-0.85), but no other risk factors were identified. The seasonality of adenovirus 40/41 diarrhea varied substantially between sites and did not have clear associations with seasonal variations in temperature or rainfall. Conclusions: This study supports the situation of adenovirus 40/41 as a pathogen of substantial importance, especially in infants. Fever was a distinguishing characteristic in comparison to other nonrotavirus viral etiologies, and promotion of exclusive breastfeeding may reduce the high observed burden in the first 6 months of life.

Epidemiology of Campylobacter infections among children of 0-24 months of age in South Africa.

BACKGROUND: Campylobacter spp. are one of the most frequent causes of diarrhoeal disease in humans throughout the world. This study aimed at determining the prevalence and the genotypic distribution of Campylobacter spp. and their association with diarrhoea and child growth in children of less than the age of two in the Limpopo Province of South Africa. METHODS: A total of 4280 diarrheal and non-diarrheal stool samples were collected on a monthly basis from children recruited at birth and followed up to 24 months. All stool samples were screened for the presence Campylobacter antigen using ELISA technique after which CAH 16S primer was used on the positive samples to confirm the presence of Campylobacter. Subsequently, the PCR positive samples were further characterised using species specific primers for Campylobacter jejuni and Campylobacter coli. RESULTS: Campylobacter antigen was detected in 564/4280 (13.2%). Campylobacter was more commonly found in diarrheal stools (20.4%) compared to non-diarrheal stools (12.4%) with a statistically significant difference (χ2 = 7.345; p = 0.006). Throughout the year there were two main peaks of Campylobacter infection one in December- January and the second peak in June. The prevalence of Campylobacter increased with the age of the children up to 11 months after which the prevalence decreased. Out of 564 positive ELISA samples, 257 (45.6%) were confirmed to have 16S rRNA gene for Campylobacter spp. Furthermore, C. jejuni was found to be more prevalent (232/257) than C. coli (25/257) with a prevalence of 90.3% and 9.7%, respectively. Both C. jejuni and C. coli were significantly associated with diarrhea with statistical values of (χ2 = 22.224; p < 0.001) and (χ2 = 81.682; p < 0.001) respectively. Sequences generated from the analysis of hip gene confirmed the PCR positives samples were C. jejuni positive. CONCLUSIONS: This study has delineated a high prevalence of Campylobacter spp. in the study cohort. Moreover, C. jejuni was found to be more prevalent than C. coli both of which were associated with diarrhea. These findings are of clinical and epidemiological significance.

Prevalence and Distribution of Cryptosporidium spp. and Giardia lamblia in Rural and Urban Communities of South Africa

Objective: Enteric diseases remain a serious health problem globally. High prevalence is evident in regions with poor socioeconomic conditions, poor sanitation, and inadequate clean water supply, such as South Africa. Designing an effective strategy, however, requires local knowledge, which can be particularly challenging to acquire in low-and middle-income countries. As the first step in this process, we investigated the prevalence and distribution of protozoan parasites Cryptosporidium and Giardia in the rural and urban gastrointestinal clinics of South Africa. Methods: A cross-sectional study was conducted to assess the prevalence of enteric parasites Cryptosporidium and G. lamblia in rural and urban communities of South Africa. Stool samples were collected from November 2013 to June 2015 from patients with diarrhea (n=227) and without diarrhea (n=257). DNA was extracted and a diagnostic Taqman qPCR assay was performed to detect these protozoan parasites, which was further confirmed by the Sanger sequencing of a few samples. Results: Of the 484 stool specimens collected, 34% (166/484) were positive for either Cryptosporidium spp. or Giardia lamblia parasites, with only 5% containing both parasites (22/484). In both study populations, Cryptosporidium was the most prevalent parasite (overall 25%) followed by Giardia (19%). Conclusion: This study discovered that both Giardia and Cryptosporidium parasites might contribute to diarrheal disease in South Africa and are more prevalent in rural communities. Future studies are needed to identify the source of the infection and design appropriate interventions to reduce the burden of the disease. Amaç: Enterik hastaliklar küresel olarak ciddi bir saglik sorunu olmaya devam etmektedir. Güney Afrika gibi düsük sosyo-ekonomik kosullarin, kötü sanitasyonun ve yetersiz temiz su kaynaklarinin oldugu bölgelerde yüksek prevalans görülmektedir. Ancak etkili bir strateji tasarlamak için, düsük ve orta gelirli ülkelerde edinilmesi özellikle zor olabilecek yerel bir bilgi gerektirmektedir. Bu süreçte biz ilk adim olarak, Güney Afrika'nin kirsal ve kentsel gastrointestinal kliniklerinde protozoan parazitler Cryptosporidium ve Giardia'nin prevalansini ve dagilimini arastirdik. Yöntemler: Güney Afrika'nin kirsal ve kentsel topluluklarinda Cryptosporidium ve G. lamblia enterik parazitlerinin sikligini arastirmak için kesitsel bir çalisma yapildi. Ishali olan (n=227) ve olmayan (n=257) hastalarin Kasim 2013-Haziran 2015 tarihleri arasinda diski örnekleri toplandi. DNA ekstrakte edildi ve bu protozoan parazitleri saptamak için tanisal bir Taqman qPCR tahlili kullanilarak, birkaç örnek Sanger dizilimi ile daha da dogrulandi. Bulgular: Toplanan 484 diski örneginin %34'ü (166/484) Cryptosporidium spp. veya Giardia lamblia parazitleri için pozitifti ve örneklerin sadece %5'i her iki paraziti de içeriyordu (22/484). Her iki çalisma popülasyonunda da Cryptosporidium en yaygin parazitti (toplam %25) ve bunu Giardia (%19) izledi. Sonuç: Bu çalisma, hem Giardia hem de Cryptosporidium parazitlerinin Güney Afrika'daki ishal hastaligina katkida bulunabilecegini ve kirsal topluluklarda daha yaygin oldugunu göstermistir. Hem enfeksiyonun kaynagini belirlemek hem de hastaligin yükünü azaltmak için uygun müdahaleleri tasarlamak için gelecekteki çalismalara ihtiyaç vardir.

Sorption of Fluoride and Bacterial Disinfection Property of Biosynthesized Nanofibrous Cellulose Decorated Ag-MgO-Nanohydroxyapatite Composite for Household Water Treatment.

An innovative and sustainable approach to integrating modified Ag-MgO-nanohydroxyapatite on a nanofibrous cellulose template (CNF-AgMgOnHaP) as a multifunctional adsorbent via a hydrothermal bioreduction route using Citrus paradisi peel extract was developed and examined. The surface morphology and mineralogical properties of CNF-AgMgOnHaP by UV-vis spectroscopy, SEM-EDS, XRD, FTIR, TEM, and BET techniques are reported. Batch fluoride sorption studies and its disinfection potential against common bacteria in surface water were evaluated. The results showed the successful synthesis of a modified multistructural CNF-AgMgOnHaP composite with an improved BET surface area of 160.17 m2/g. The sorption of fluoride by the adsorbent was found to strongly depend on the different sorption conditions with a maximum F- sorption capacity of 8.71 mg/g at 303 K, and pH of 5 with 0.25 g dosage at 10 min contact time (25 ± 3 °C). Equilibrium fluoride sorption onto the CNF-AgMgOnHaP was best described by the Freundlich isotherm model across all the operating temperatures. The overall kinetic results showed that the adsorption mechanisms not only depend on using the pseudo-second-order process but are also governed by the mass transfer of the adsorbate molecules from the external surface onto the pores of the adsorbent. The thermodynamic parameters revealed that the adsorption process of F- onto CNF-AgMgOnHaP was endothermic and spontaneous at the sorbent/solution interface. The synthesized composite also provides some antibacterial activity against common infectious microbes from contaminated drinking water. The overall results suggested that the CNF-AgMgOnHaP nanocomposite possesses the potential for the simultaneous decontamination of pollutants and microbes in drinking water.

Interleukin 10 (IL-10) Production and Seroprevalence of Entamoeba histolytica Infection among HIV-Infected Patients in South Africa.

Infections by the parasite E. histolytica are increasing in HIV-infected individuals. Interleukin (IL-10) plays an important role in maintaining the mucosal barrier. Therefore, the seroprevalence of E. histolytica was investigated in relation to the IL-10 serum concentration among HIV- infected patients. A total of 647 blood samples were collected from asymptomatic HIV-infected patients. The Entamoeba histolytica antigen (GALNAC lectin) and serum antibodies were assessed using specific ELISAs (TECHLAB, Virginia, USA). IL10 blood levels were measured using a commercial ELISA test, and the results were analyzed using parametric and non-parametric statistical tests. The Gal/GALNAC lectin was detected in only 0.5% (3/647) of individuals, and the antibodies against E. histolytica were detected in 65.2% (422/647) of the samples. A significant increase in IL-10 levels was found in 68.1% of patients who were sero-negative for E. histolytica antibodies compared to patients who were sero-positive. There is a high level of exposure to E. histolytica among HIV patients in South Africa, although the prevalence of amoebic liver abscesses might be low. This study revealed that elevated levels of IL-10 might be associated with a reduced risk of amebiasis.

Synthesis, Characterization and In Vitro Antibacterial Evaluation of Pyrenacantha grandiflora Conjugated Silver Nanoparticles.

In the present study, silver nanoparticles (AgNPs) were synthesized using both the chemical and biological methods and conjugated with Pyrenacantha grandiflora extracts. These were then characterized and evaluated for antimicrobial activities against multi-drug resistant pathogens, such as methicillin-resistant Staphylococcus aureus (MRSA), Klebsiella pneumonia, and Escherichia coli. Nanoparticles were analyzed with UV-visible spectrophotometer, transmission electron microscopy (TEM), and energy dispersive X-ray analysis (EDX). Silver nanoparticles, P. grandiflora extracts, and the conjugates were also analyzed with Fourier transform infrared spectroscopy (FTIR). As a result, quasi-sphere-shaped AgNPs with sizes ranging from 5 to 33 nm and spherically shaped AgNPs with sizes ranging from 3 to 25 nm were formed from chemical and biological synthesis, respectively. A well diffusion assay showed that the activity of silver nanoparticles was most improved with acetone extract against all tested bacteria with diameters in the range of 19-24 mm. The lowest MIC value of 0.0063 mg/mL against MRSA was observed when biologically synthesized AgNPs were conjugated with acetone and water extracts. Chemically synthesized silver nanoparticles showed the lowest MIC value of 0.0063 mg/mL against E. coli when conjugated with acetone and methanol extracts. This study indicates that silver nanoparticles conjugated with P. gandiflora tubers extracts exhibit strong antibacterial activities against multi-drug resistant bacterial pathogens. Therefore, biosynthesized conjugates could be utilized as antimicrobial agents for effective disease management due to the synergistic antibacterial activity that was observed.

Molecular epidemiology of Microsporidia among HIV-positive and HIV-negative patients in the Limpopo province, South Africa.

INTRODUCTION: Human microsporidiosis represents an important and rapidly emerging opportunistic disease. The present study investigated the prevalence of microsporidia among HIV positive and HIV negative patients with or without diarrhoea in Vhembe and Mopani Districts in the Limpopo Province. METHODOLOGY: A total of 170 stool samples were collected from these patients and microsporidia species was detected using a Real-Time PCR targeting a conserved region of the small ribosomal subunit rRNA (SSU-rRNA) gene of Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon hellem, and Encephalitozoon cuniculi. RESULTS: Fifty six (32.9%) were positive for microsporidia. The prevalence was higher in HIV negative patients (36.6%) while 24.1% of patients who were HIV positive had microsporidia. Microsporidia was more common among patients aged between 1 and 10 years (52.6%). However among the HIV positive patients, microsporidia prevalence was higher among those that were not taking antiretrovirals (ARVs) compared to those who were on ARVs, (36.6%) and (24.1%), respectively. Microsporidia was also noted to be significantly associated with diarrheal and stomach pains; p = 0.02 and p = 0.048, respectively. Furthermore, microsporidia infections was more prevalent among patients who had animals at home (p = 0.037). CONCLUSIONS: Study has shown a high prevalence of microsporidia among patients attending primary health centers in the Mopani District for the first time. Prevalence of microsporidia was higher among HIV negative and HIV positive patients who were not on ARV treatment. Keeping animals in the household appeared to be a risk of getting infected with microsporidia. Further studies are needed to determine the genetic characteristics of these organisms in the study population.

Enhanced passive surveillance dengue infection among febrile children: Prevalence, co-infections and associated factors in Cameroon.

Dengue virus (DENV) causes a spectrum of diseases ranging from asymptomatic, mild febrile to a life-threatening illness: dengue hemorrhagic fever. The main clinical symptom of dengue is fever, similar to that of malaria. The prevalence of dengue virus infection, alone or in association with other endemic infectious diseases in children in Cameroon is unknown. The aim of this study was to determine the prevalence of dengue, malaria and HIV in children presenting with fever and associated risk factors. Dengue overall prevalence was 20.2%, Malaria cases were 52.7% and HIV cases represented 12.6%. The prevalence of dengue-HIV co-infection was 6.0% and that of Malaria-dengue co-infection was 19.5%. Triple infection prevalence was 4.3%. Dengue virus infection is present in children and HIV-Dengue or Dengue- Malaria co-infections are common. Dengue peak prevalence was between August and October. Sex and age were not associated with dengue and dengue co-infections. However, malaria as well as HIV were significantly associated with dengue (P = 0.001 and 0.028 respectively). The diagnosis of dengue and Malaria should be carried out routinely for better management of fever.

Duration of Postdiarrheal Enteric Pathogen Carriage in Young Children in Low-resource Settings.

BACKGROUND: Prolonged enteropathogen shedding after diarrhea complicates the identification of etiology in subsequent episodes and is an important driver of pathogen transmission. A standardized approach has not been applied to estimate the duration of shedding for a wide range of pathogens. METHODS: We used a multisite birth cohort of children 0-24 months of age from whom diarrheal and monthly nondiarrheal stools were previously tested by quantitative polymerase chain reaction for 29 enteropathogens. We modeled the probability of detection of the etiologic pathogen before and after diarrhea using a log-normal accelerated failure time survival model and estimated the median duration of pathogen carriage as well as differences in subclinical pathogen carriage 60 days after diarrhea onset in comparison to a prediarrhea baseline. RESULTS: We analyzed 3247 etiologic episodes of diarrhea for the 9 pathogens with the highest attributable burdens of diarrhea. The median duration of postdiarrheal carriage varied widely by pathogen, from about 1 week for rotavirus (median, 8.1 days [95% confidence interval {CI}, 6.2-9.6]) to >1 month for Cryptosporidium (39.5 days [95% CI, 30.6-49.0]). The largest increases in subclinical pathogen carriage before and after diarrhea were seen for Cryptosporidium (prevalence difference between 30 days prior and 60 days after diarrhea onset, 0.30 [95% CI, .23-.39]) and Shigella (prevalence difference, 0.21 [95% CI, .16-.27]). CONCLUSIONS: Postdiarrheal shedding was widely variable between pathogens, with strikingly prolonged shedding seen for Cryptosporidium and Shigella. Targeted antimicrobial therapy and vaccination for these pathogens may have a relatively large impact on transmission.

Seasonality of drinking water sources and the impact of drinking water source on enteric infections among children in Limpopo, South Africa.

Enteric infections and water-related illnesses are more frequent during times of relative water abundance, especially in regions that experience bimodal rainfall patterns. However, it is unclear how seasonal changes in water availability and drinking water source types affect enteric infections in young children. This study investigated seasonal shifts in primary drinking water source type and the effect of water source type on enteric pathogen prevalence in stool samples from 404 children below age 5 in rural communities in Limpopo Province, South Africa. From wet to dry season, 4.6% (n = 16) of households switched from a source with a higher risk of contamination to a source with lower risk, with the majority switching to municipal water during the dry season. In contrast, 2.6% (n = 9) of households switched from a source with a lower risk of contamination to a source with higher risk. 74.5% (n = 301) of the total households experienced interruptions in their water supply, regardless of source type. There were no significant differences in enteric pathogen prevalence between drinking water sources. Intermittent municipal water distribution and household water use and storage practices may have a larger impact on enteric infections than water source type. The limited differences in enteric pathogen prevalence in children by water source could also be due to other exposure pathways in addition to drinking water, for example through direct contact and food-borne transmission.

Epidemiology of Shigella infections and diarrhea in the first two years of life using culture-independent diagnostics in 8 low-resource settings.

Culture-independent diagnostics have revealed a larger burden of Shigella among children in low-resource settings than previously recognized. We further characterized the epidemiology of Shigella in the first two years of life in a multisite birth cohort. We tested 41,405 diarrheal and monthly non-diarrheal stools from 1,715 children for Shigella by quantitative PCR. To assess risk factors, clinical factors related to age and culture positivity, and associations with inflammatory biomarkers, we used log-binomial regression with generalized estimating equations. The prevalence of Shigella varied from 4.9%-17.8% in non-diarrheal stools across sites, and the incidence of Shigella-attributable diarrhea was 31.8 cases (95% CI: 29.6, 34.2) per 100 child-years. The sensitivity of culture compared to qPCR was 6.6% and increased to 27.8% in Shigella-attributable dysentery. Shigella diarrhea episodes were more likely to be severe and less likely to be culture positive in younger children. Older age (RR: 1.75, 95% CI: 1.70, 1.81 per 6-month increase in age), unimproved sanitation (RR: 1.15, 95% CI: 1.03, 1.29), low maternal education (<10 years, RR: 1.14, 95% CI: 1.03, 1.26), initiating complementary foods before 3 months (RR: 1.10, 95% CI: 1.01, 1.20), and malnutrition (RR: 0.91, 95% CI: 0.88, 0.95 per unit increase in weight-for-age z-score) were risk factors for Shigella. There was a linear dose-response between Shigella quantity and myeloperoxidase concentrations. The burden of Shigella varied widely across sites, but uniformly increased through the second year of life and was associated with intestinal inflammation. Culture missed most clinically relevant cases of severe diarrhea and dysentery.

Impact of Low-Cost Point-of-Use Water Treatment Technologies on Enteric Infections and Growth among Children in Limpopo, South Africa.

Enteric infections early in life have been associated with poor linear growth among children in low-resource settings. Point-of-use water treatment technologies provide effective and low-cost solutions to reduce exposure to enteropathogens from drinking water, but it is unknown whether the use of these technologies translates to improvements in child growth. We conducted a community-based randomized controlled trial of two water treatment technologies to estimate their effects on child growth in Limpopo, South Africa. We randomized 404 households with a child younger than 3 years to receive a silver-impregnated ceramic water filter, a silver-impregnated ceramic tablet, a safe-storage water container alone, or no intervention, and these households were followed up quarterly for 2 years. We estimated the effects of the interventions on linear and ponderal growth, enteric infections assessed by quantitative molecular diagnostics, and diarrhea prevalence. The silver-impregnated ceramic water filters and tablets consistently achieved approximately 1.2 and 3 log reductions, respectively, in total coliform bacteria in drinking water samples. However, the filters and tablets were not associated with differences in height (height-for-age z-score differences compared with no intervention: 0.06, 95% CI: -0.29, 0.40, and 0.00, 95% CI: -0.35, 0.35, respectively). There were also no effects of the interventions on weight, diarrhea prevalence, or enteric infections. Despite their effectiveness in treating drinking water, the use of the silver-impregnated ceramic water filters and tablets did not reduce enteric infections or improve child growth. More transformative water, sanitation, and hygiene interventions that better prevent enteric infections are likely needed to improve long-term child growth outcomes.

Functionalization and antimicrobial evaluation of ampicillin, penicillin and vancomycin with Pyrenacantha grandiflora Baill and silver nanoparticles.

Some antibiotics have lost their efficacy over common infections and this has led to the search for new antibiotics and chemically altering existing ones for a better control of infectious diseases. In the present study, Pyrenacantha grandiflora tubers extracts were conjugated with ampicillin, penicillin, vancomycin and silver nanoparticles and their antimicrobial activity was evaluated against Escherichia coli, Staphylococcus aureus and Klebsiella Pneumoniae. The reactions were confirmed by formation of new functional groups that were identified by Fourier transmission infrared spectroscopy (FTIR). Minimum inhibitory concentrations were determined using the microdilution assay. Minimum bactericidal concentrations and the fractional inhibition concentration index were also determined. FTIR analysis indicated different functional group associated with conjugation. The activity of ampicillin was improved when conjugated with silver nanoparticles against K. pneumonia and E. coli. Vancomycin showed improvement of activity when conjugated to silver nanoparticles against K. pneumonia. Penicillin was improved by acetone extracts and vancomycin showed to be more effective when conjugated with silver nanoparticles and water extracts. The conjugation of P. grandiflora with penicillin, ampicillin and vancomycin in the presence of silver nanoparticles improved their biological activities. Therefore, the conjugates are medicinally important and can be used to improve the activity of existing antibiotics.

Antimicrobial activity of selected plants against fungal species isolated from South African AIDS patients and their antigonococcal activity.

Background Sexually transmitted infections continue to be a major health concern in sub-Saharan Africa where antimicrobial drugs are becoming ineffective due to increasing resistance. Many healthcare seekers in lower socio-economic settings depend on invasive alien plants administered by traditional health practitioners to treat sexually transmitted infections. Methods Roots of selected plants were analysed for phytoconstituents using standard methods. Both the disc diffusion model and microdilution technique were used to determine the inhibition zone and minimum inhibitory concentration (MIC) of plant extracts against six clinical fungal strains and standard strain of Neisseria gonorrhoea. One-way ANOVA was used to find significant differences. Results Different phytoconstituents such as alkaloids, steroids, cardiac glycosides, terpenes, flavonoids, tannins and saponins were qualitatively detected, depending on plant species. Acetone extracted the highest number of phytoconstituents in Senna didymobotrya, while methanol revealed most from Ricinus communis. Senna didymobotrya showed significant inhibition against Candida glabrata, C. krusei, C. parapsilosis and C. tropicalis. Dichloromethane extract of Catharanthus roseus and methanol extract of S. didymobotrya demonstrated excellent MIC values of 0.03 and 0.08 mg/mL, respectively, against C. glabrata. Catharanthus roseus, Opuntia ficus-indica and Ricinus communis demonstrated moderate to good antigonococcal activity, with all exhibiting more than 63% inhibition. Catharanthus roseus had the best antigonococcal activity with a moderate MIC value of 0.63 mg/mL. Conclusion Some of the plant extracts demonstrated potency towards clinically isolated fungal strains and against N. gonorrhoea, which validate the notion that some of the species need further pharmacological studies for isolation and characterisation of active compounds.

Prevalence and distribution of Entamoeba species in a rural community in northern South Africa.

Amoebiasis occurs worldwide and affects about 20-50 million people annually. Stool samples were collected from patients attending different rural clinics in Northern South Africa in the present study. Microscopic examination was performed for the initial detection of Entamoeba parasites. A multiplex PCR protocol based on the small subunit rRNA gene of E. moshkovskii, E. dispar, and E. histolytica, was used for the differential detection of the three Entamoeba species (collectively referred to as Entamoeba complex). A total of 170 participants were recruited in the study, with a mean age of 35.9 ±â€¯17.8 years and a median of 37.0 years. The prevalence of Entamoeba species was found to be 34.7% and 33% by PCR and microscopy, respectively. E. histolytica had a prevalence of 4.1%, E. dispar 14.7% and E. moshkovskii 15.9%. Of the three species, only E. histolytica was significantly associated with diarrhoea and was more prevalent among HIV patients even in the absence of diarrhoea while the other two were not, although the difference was not significant (p > 0.05). This is the first study in South Africa to describe the prevalence of E. moshkovskii. E. dispar was significantly associated with abdominal pains (p = 0.003). Further studies are needed to clarify the role of E. moshkovskii and E. dispar in abdominal pain and diarrhoea.