Human Intestinal Infections Research Articles

Prevalence and Socio-Economic Factors of Human Intestinal Helminth Infections amongst Primary School Pupils in Bokkos, L.G.A, Plateau State

Intestinal helminth infections remain a significant public health challenge, particularly in low-resource settings where poor sanitation and hygiene practices prevail. This study was conducted in Bokkos Local Government Area of Plateau State to ascertain the prevalence and socio-economic factors of intestinal helminth infections amongst primary school pupils. Cross-sectional descriptive research design and multi-stage sampling technique were used to collect stool samples from 100 pupils, aged 3–13 years. A close-ended pre-tested structured questionnaire was used to obtain responses from 100 respondents. Samples were analysed using sedimentation, direct smear and flotation method. Results revealed a high overall prevalence of intestinal helminth infections 76 (59.9%). The most prevalent helminth specie was Hookworm 27(19.1%), followed by Ascaris lumbricoides 23(17.9%), Trichuris trichiura 15(14.2%) and Strongyloides stercoralis 11(8.7%) respectively. However, there was statistical difference in the prevalence of Specific Intestinal Helminths infection among pupils (P<0.05). There was a statistical difference between sex of pupil and intestinal helminth infections (P<0.05). More females 42(42.0%) were infected than males 34(34.0%). There was a statistical difference between age of pupil and intestinal helminth infections (P<0.05). The highest infection rate, 28(28.0%) was observed in pupils within age group 3 – 6 while the least infection rate 13(13.0%) was observed in the age group 13 and above. There was a statistical difference in the prevalence of intestinal helminths infection among pupils in relation to age (P<0.05). There was a significant association between socio-economic factor and the prevalence of intestinal Helminth infections among Pupils (P<0.05). There was also a significant difference between the risk factors of intestinal helminth infections among pupil (χ2 = 0.109, df = 2, P = 0.021). Conclusively, the high prevalence rate of intestinal helminths among primary school pupils within the study area significantly affects their academic achievement and overall wellbeing. It is therefore recommended that routine deworming programs should be provided by the government with administration of anti-helminthic drugs complemented by monitoring and evaluation mechanisms to assess its effectiveness in reducing infection rates.

Developing Safe Probiotic Solutions for the Treatment of Human Intestinal Infections

In recent years, preparations based on lactic acid bacteria have proven to be effective in the complex therapy of infectious diseases. Despite the large number of positive results of the use of these preparations and their widespread use, there are some risks associated with their use. To minimize these risks, it is necessary to regulate the production, sale, and use of probiotic preparations on an equal basis with medicines. To date, there are no made-in-Kazakhstan probiotic preparations with clinically proven safety and performance in the pharmaceutical market of Kazakhstan. The purpose of this paper was to present preclinical trials of this preparation confirming its safety and medical performance and demonstrate the readiness for clinical trials of the Kazakh probiotic medication AS-Probionorm. The AS-Probionorm probiotic medication developed by us based on an association of specific action lactic acid bacteria (Lactobacillus fermentum 30 and Lactobacillus cellobiosus 36), has high antimicrobial activity against pathogens of intestinal diseases. This study introduces AS-Probionorm as the first locally produced probiotic in Kazakhstan with clinically established safety and effectiveness, marking a significant step forward in the national production of biomedical health solutions.

Human intestinal nematode infections in Sri Lanka: A scoping review.

Sri Lanka, an island located in South Asia, once experienced a notable prevalence of human intestinal nematode infections (HINIs). With the implementation of control programs, infection prevalence was reduced. Detailed information on prevalence, distribution and temporal trends of HINIs is limited. This review aims to explore Sri Lanka's HINI distribution, trends, diagnostics, control and challenges. We reviewed published information on HINIs in Sri Lanka in electronic databases, local journals and grey literature from inception to September 2022. Based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-Scr), a systematic strategy was used for searching, screening, reviewing and data extraction. The screening was initiated with a review of titles and abstracts using specific keywords, followed by a full-text screening based on predefined eligibility criteria. A total of 105 studies were included in the review, with 28 selected for pooled prevalence analysis. The first nationwide survey in 1924 reported a hookworm infection prevalence of 93.1%. By 2017, soil-transmitted helminth (STH) infection prevalence across the island was 0.97% (ascariasis-0.45%, trichuriasis-0.25%, and hookworm infection-0.29%), and the enterobiasis prevalence between 2003 and 2017 ranged from 0% to 42.5%. Strongyloidiasis had been understudied, with a prevalence of 0.1% to 2%. Over the past two decades, the islandwide pooled HINI prevalence was 13.3%. Within specific demographics, it was 6.96% in the general community, 33.4% in plantation sector, and 11.6% in slum communities. During the colonial period, hookworm infection was the commonest HINI, but ascariasis is now more prevalent. The prevailing data relied solely on microscopy, often utilising single stool smears. Mass deworming programs were widely pursued in the first half of the 20th century, initially targeting antenatal women and schoolchildren, and now focusing on specific community groups. National surveys continue monitoring the three main STH infections. The significant reduction in STH prevalence in the country over the past ten decades highlights the effectiveness of public health interventions, particularly mass deworming programs. Despite the success, STH prevalence disparities persist in vulnerable populations like plantation and slum communities, where hygiene and living conditions continue to pose challenges. Reliance on single stool smear microscopy highlights the need for more sensitive diagnostics to better assess infections. Fluctuating enterobiasis prevalence and limited strongyloidiasis data underscore the importance of continued surveillance and targeted interventions for sustained control and eventual elimination. Sri Lanka's experiences and control measures offer valuable insights for low-income countries in South Asia and beyond, particularly in managing HINIs with limited resources.

Diversity of Clostridioides difficile PCR ribotypes isolated from freshwater sediments depends on the isolation method.

Clostridioides difficile is an intestinal pathogen of humans and animals. In community-associated infections, the environment is suggested to play a significant role in overall transmission routes. Although the prevalence of C. difficile in freshwater and soil has been widely studied, little is known about its presence in sediments. In this study, we tested 15 sediment samples collected from various freshwater sources. C. difficile was isolated from all sampled sites, yielding a total of 171 strains grouped into 26 ribotypes, with 001/072 and 014/020 being the most prevalent. Genome sequencing of 37 isolates from 17 PCR ribotypes confirmed the presence of highly related strains in the geographically distant and unlinked water samples. Eight divergent PCR ribotypes from clades C-II and C-III were found in six samples. In each sample, the unbound fraction (supernatant after sediment wash) and bound fraction (sonicated sediment sample) were subjected to enrichment. Sonication was only slightly better than washing in terms of sample positivity (14 positive samples with sonication and 11 with washing). However, sonication substantially increased the diversity of the PCR ribotypes obtained (23 in sonicated samples vs nine in washed samples). In conclusion, sediments are a rich source for investigating the diversity of environmental C. difficile, including isolates from divergent lineages. Selection of the isolation method can significantly impact the diversity of captured PCR ribotypes.IMPORTANCEClostridioides difficile, a pathogenic bacterium that can cause intestinal infections in humans and animals, thrives in the gut but also disperses widely through spores found in the environment. Clinical and environmental strains often overlap with common PCR ribotypes, which are consistently isolated worldwide. Environmental studies have mostly focused on water and soil, but sediments have been very poorly studied. In this study, we investigated the presence of C. difficile in various freshwater sediments and evaluated the effectiveness of two different isolation approaches on positivity rates and strain diversity. C. difficile was found to be highly prevalent in sediments, with an isolation rate of 100%. Sonication proved to be more effective than simple washing for capturing a greater diversity of PCR ribotypes. Overall, this study underscores the widespread presence of C. difficile in freshwater sediments and emphasizes the importance of continued surveillance and monitoring to understand its ecology and transmission dynamics.

First Phase of the Clinical trial of the AS-Probionorm Probiotic Medication for the treatment of Human Intestinal Infections

The effectiveness of probiotic medications in the treatment of infectious diseases of the gastrointestinal tract has been proven in many clinical studies. Despite this, currently, some preparations do not have the appropriate quality and activity against pathogenic microorganisms. The reason for this is the lack of a stage for the selection of antagonists to specific pathogens at the stage of research and medication development. Given the high prevalence of intestinal infections among the population of Kazakhstan and the lack of made-in-Kazakhstan probiotic medications in the pharmaceutical market, there is a need to introduce an effective preparation based on probiotic microorganisms for the treatment of infectious diseases of the gastrointestinal tract. The purpose of this study was to demonstrate the methodology and results of the first phase of clinical trials of this preparation. The study was conducted following the ethical principles of the Helsinki Declaration, the Order of the Acting Minister of Health of the Republic of Kazakhstan, the Order of the Minister of Health of the Republic of Kazakhstan, and the Code of the Republic of Kazakhstan. 20 healthy volunteers of both sexes aged 18 to 50 years with a verified state of health, who had given informed consent, were examined before the start of the study, and the results of clinical measurement of the initial condition were also obtained, which was then compared with daily measurements during the use of the preparation under study and a week after the last administration. As a result of the study, it was concluded that the preparation was well-tolerated without noticeable side effects, which indicates the safety of the medication.

Antimicrobial susceptibility of Salmonella enteritidis isolated in the period from 2020 to 2022 in the Shtip region

Bacteria of the genus Salmonella are widely distributed in nature. They can colonize different hosts and are the most common causes of intestinal infections in humans and animals. The main route of Salmonella transmission to humans is by contaminated food, or undercooked eggs and meat. The aim of this study was to analyze the antimicrobial susceptibility of Salmonella enteritidis, which was the most frequently isolated Salmonella in the Microbiological Laboratory of the Center of Public Health in Shtip. Materials and methods: A total of 1839 samples (12 rectal swabs and 1827 feces), from persons suspect for salmonellosis in the region of Shtip were investigated for the period 2020 to 2022 year. SS (Salmonella Shigella) agar, Rambach agar, and Selenit F media were used for detection of Salmonella spp., and antimicrobial susceptibility testing of the isolated bacteria is performed with disk diffusion method according EUCAST. Results: Salmonella enteritidis is detected in 62 samples (3.37%), isolated strains of Salmonella enteritidis demonstrate high resistance to ampicillin 2 μg (37.09%), and multi-resistant strains (6.45%) were also detected. Conclusion: The appearance of ampicillin and multi-resistant strains of Salmonella enteritidis in the samples analyzed in our study is a sign of concern and an alarm to initiate a change in the algorithm for the treatment of Salmonella enteritidis infections, especially with the use of fluoroquinolones and cephalosporins (of the third generation). It is recommended to introduce a system for the rational use of antimicrobials in human and veterinary health.

Development of Technology for the production of finished forms of Medicinal probiotics

The study aimed to develop a technology for the production of a finished form of medicinal probiotic agents against human intestinal infections based on active strains of lactic acid and propionic acid bacteria with a wide range of biological activity and resistance to antibiotics. From the laboratory collection of lactic acid and propionic acid bacteria isolated from the intestines of healthy people, two associations of bacteria were compiled with antagonism against test cultures of Staphylococcus aureus, Salmonella gallinarum, Mycobacterium B5, Candida albicans, Pasteurella multocida, Bacillus subtilis, Escherichia coli 8739, Klebsiella pneumoniae ATCC 700603 and ATCC BAA 2524, Staphylococcus aureus 3316 and 9, Salmonella enteritidis 35382, and Pseudomonas aeruginosa 835, as well as the ability to produce hydrolytic enzymes amylase and proteinase, B vitamins, and essential amino acids. The resistance of the selected associations of lactic acid and propionic acid bacteria to the used antibiotics has been studied, which will allow for using them, if necessary, in the complex therapy of diseases. Technology for the production of probiotic medication from these associations has been developed. It was found that the most active preparation in terms of bacterial titer and antagonistic activity and the most stable one during storage for 6 months was the liquid preparation obtained by growing association No. 2 (L. plantarum 2v/A-6+L. brevis B-3/A-26+L. acidophilus 27w/60+P. shermanii 8) on nutrient medium No. 1 (De Man, Rogosa and Sharpe agar with CoCl2) using 7% sucrose and 1.5% gelatin as a protector. The liquid preparation from association No. 5 grown on medium No. 1 showed a more complete preservation of production-valuable signs during storage compared to the results of using nutrient medium No. 4, while the use of protector No. 1 was more optimal. To test the stability during the storage of dry preparation forms, an accelerated method was used by warming them up for 15 minutes at 60°C. It was found that after warming up, the best preservation of viable bacterial cells was observed in association No. 2 on nutrient media No. 1 and No. 4, in association No. 5 on medium No. 4 dried with protector No. 2 (7% sucrose and 1.5% gelatin + 7% skim milk powder), while the titer of bacteria was equal to 1.2×109, 3.5×108, and 2.0±0.2×108 colony-forming units/g, respectively. Antagonistic activity in these association variants was observed against all test cultures taken into the study with zones of suppression of their growth ranging from 10 to 24 mm.

Selection of an active association of Probiotic bacteria and the Optimal composition of the Nutrient medium for Cultivation to increase the Therapeutic and Prophylactic effectiveness of a Medicinal probiotic preparation against Intestinal infections

The study aimed to create associations of probiotic bacteria with a wide range of biological activity, resistance to antibiotics, and selection of the nutrient medium composition to increase the effectiveness of probiotics against intestinal infections in humans. The authors found that the tested associations of probiotic bacteria exhibited sufficient antagonistic activity to all test cultures taken in the experiment. Associations No. 2 on media 1 and 4, No. 5 on media 1 and 8, and association No. 9 on media 1 and 8 showed the greatest activity for most test cultures. These associations have a bacterial titer of nx1010colony-forming units/ml, contain proteolytic, amylolytic, and pectolytic enzymes, as well as B vitamins, such as B3, B5, B6, B8, and B12. The authors studied the resistance of selected associations of lactic acid and propionic acid bacteria to the antibiotics used, which allows these associations to be used in complex therapy if necessary. Selected nutrient media and active associations of probiotic bacteria with a wide range of biological activity and antibiotic resistance can be used to create effective targeted medicinal probiotic agents.

The level of discreteness of the signs of the own productivity of repair pigs and the reproductive qualities of sows of different breeding value: criteria for the selection of highly productive animals according to the BLUP index

Products of plant origin, recommended as permanent components of the daily human diet, are rich in vitamins, minerals, plant fibers, as well as various active phytochemicals (polyphenols, flavonoids) and sterols, which have a low calorie content. According to the results of epidemiological and clinical studies, it has been established that the risks of various pathologies are reduced when using a diet that is balanced in terms of nutrients, rich in vegetable fiber and food products of plant origin. The evolution of food technology has led to the appearance on the shelves of Italian supermarkets of products defined as “ready foods” or products – semi-finished products of high quality and safety, which present all the characteristics of freshness, similar to products that have just been harvested. They also include products of vegetable origin, which belong to the fresh-cut category. A limited technological elaboration is applied to this category of products, after which they can be used without further manipulation before consumption. The products of vegetable origin of fresh-cut vegetable have the definition of "potentially dangerous products" due to their possible contamination with pathogenic microorganisms such as Escherichia coli, Salmonella spp. and Listeria monocytogenes. These pathogens very often contaminate fresh-cut vegetable, causing food poisoning in consumers. The main sources of pathogenic contamination by the causative agents of human intestinal infections are the water used for watering plants and exposure to inappropriate temperatures during the storage of plant products. The decisive aspect, without a doubt, remains the sanitary and hygienic characteristics of the product at the time of its consumption. It is fundamental to deepen our knowledge about the transmission, resistance, and growth mechanisms of pathogenic microorganisms in products of the IV range. This will allow the establishment of sampling norms for express diagnostics to reduce the possibility of low-quality products reaching the consumer.

EU requirements regarding hygienic and sanitary aspects of products of vegetable origin of category IV range in Italy

Products of plant origin, recommended as permanent components of the daily human diet, are rich in vitamins, minerals, plant fibers, as well as various active phytochemicals (polyphenols, flavonoids) and sterols, which have a low calorie content. According to the results of epidemiological and clinical studies, it has been established that the risks of various pathologies are reduced when using a diet that is balanced in terms of nutrients, rich in vegetable fiber and food products of plant origin. The evolution of food technology has led to the appearance on the shelves of Italian supermarkets of products defined as “ready foods” or products – semi-finished products of high quality and safety, which present all the characteristics of freshness, similar to products that have just been harvested. They also include products of vegetable origin, which belong to the fresh-cut category. A limited technological elaboration is applied to this category of products, after which they can be used without further manipulation before consumption. The products of vegetable origin of fresh-cut vegetable have the definition of “potentially dangerous products” due to their possible contamination with pathogenic microorganisms such as Escherichia coli, Salmonella spp. and Listeria monocytogenes. These pathogens very often contaminate fresh-cut vegetable, causing food poisoning in consumers. The main sources of pathogenic contamination by the causative agents of human intestinal infections are the water used for watering plants and exposure to inappropriate temperatures during the storage of plant products. The decisive aspect, without a doubt, remains the sanitary and hygienic characteristics of the product at the time of its consumption. It is fundamental to deepen our knowledge about the transmission, resistance, and growth mechanisms of pathogenic microorganisms in products of the IV range. This will allow the establishment of sampling norms for express diagnostics to reduce the possibility of low-quality products reaching the consumer.

Epidemiological characteristics, virulence potential, antimicrobial resistance profiles, and phylogenetic analysis of Aeromonas caviae isolated from extra-intestinal infections.

Aeromonas caviae (A. caviae) is one of the major etiological agents in human intestinal infections reported to be associated with a broad spectrum of extra-intestinal infections with increasing incidence over recent years. Although previous studies have established its significance as a causative agent of both bloodstream and gastrointestinal infections, the characteristics of A. caviae that cause extra-intestinal infections remain unilluminated.In this single-center retrospective study, we investigated epidemiological characteristics, antimicrobial resistance genes and phenotypes, virulence genes, and phyloevolution of 47 clinical A. caviae isolated from patients with extra-intestinal infections from 2017 to 2020. A. caviae strains were identified by biochemical tests and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS), ultimately confirmed to species level by whole-genome sequencing (WGS). Antimicrobial resistance and virulence genes were identified using the Comprehensive Antibiotic Resistance Database (CARD) and the virulence factor database (VFDB), respectively. Phylogenetic analysis of 47 clinical strains was performed by combining with 521 A. caviae strains from NCBI database. A. caviae was an opportunistic pathogen in immunocompromised patients, especially those with underlying hepatobiliary diseases and malignancies. 19 out of 47 isolates were identified as multidrug resistance (MDR) strains. Piperacillin-tazobactam, levofloxacin, gentamicin, amikacin with a resistance rate of less than 10% remained as options to treat extra-intestinal infections. 24 out of 47 isolates exhibited non-susceptibility to cephalosporins and cephamycins, all of which carried β-lactamase gene, including bla MOX, bla PER-3, bla OXA, bla NDM, and bla CphA. Most stains (98%, 46/47) carried at least one of the virulence genes, but extra-intestinal infections had a low mortality rate. Phylogenetic analysis indicated the risk of nosocomial transmission but revealed no outbreak. However, the emergence of MDR and β-lactamase resistance genes in extra-intestinal isolates of A. caviae is becoming an increasing risk to public health and requires attention. This study strengthen our understanding of A.caviae isolated from extra-intestinal infections. It may contribute to the management of extra-intestinal infections as well as the prevention and control of drug resistance.

Molecular genetic characteristics of <i>Vibrio cholerae</i> nono1/nono139 strain, the causative agent of a new case of acute intestinal infection in Rostov-on-Don

During more than 50 years, a heterogeneous population of Vibrio cholerae non-O1/non-O139 (NAGs) exists in Rostov region of Russia, whose representatives periodically cause acute human intestinal infections; the last case was registered in 2018. In 2022, a sick child was again revealed in Rostov, from whom 2 NAG subcultures were isolated. Aim of the study: bioinformatics analysis of whole genome sequences (WGSs) of the 2022 clinical NAG isolates, genetic determinants of pathogenicity factors and relevant products. Materials and methods. Isolation, identification of cultures, as well as antibiotic sensitivity were carried out according to the MUK 4.2.3745-22. SNP analysis of WGSs obtained on the MiSeq Illumina platform were analyzed by means of designed software; BioEdit 7.2.5, Vector NTI Advance 11 package, CARD database were used to identify and analyze genes and relevant deduced products. Results. The isolated cultures with identical phenotypic features, PCR-genotypes and results of SNP analysis, were identified as a non-toxigenic NAG. The agent showed sensitivity to chloramphenicol, ceftriaxone, streptomycin, gentamicin, nalidixic acid, ciprofloxacin, ampicillin, amikacin and intermediate resistance to tetracycline, doxycycline, furazolidon and co-trimoxazole. WGSs lacked CTX and preCTX prophages, pathogenicity island VPI, thermostable and cholix toxin genes, pandemic islands VSP-I and VSP-II. The determinants of the type 6 secretion system (T6SS) were not presented completely, which evidenced in favor of the loss of its functionality. An intact T3SS cluster was revealed, which was adjacent to the nan-nag region of the VPI-2 pathogenicity island in the absence of the proximal and distal parts of the latter. Other determinants of pathogenicity factors were revealed: RTX cluster responsible for synthesis of MARTX cytotoxin, genes for hemolysin HlyA, cytotonic toxin Cef, several serine and metalloproteases. The nucleotide compositions of most determinants differed from the prototypes, but their putative products preserved characteristic active domains. The adhesive activity might be provided by mannose-insensitive MSHA-like pili, since the msh-cluster included all the genes necessary for their production, as well as flagella, as both clusters responsible for their assembling were 99% identical to the prototype. From antibiotic resistance determinants only -lactamase VarG gene was found. Conclusion. The NAG strain examined here contains a sufficient set of intact virulence-associated genetic determinants, expression of which could contribute to the development of the disease.

The accuracy of nucleic acid amplification tests (NAATs) in detecting human intestinal nematode infections: A protocol for a systematic review and meta-analysis.

Human intestinal nematode infections are a global public health issue as they can result in considerable morbidity in infected individuals, mainly in developing countries. These infections continue to go undiagnosed, as they tend to be mainly endemic in resource-poor communities where there is a shortage of experienced laboratory staff and relevant diagnostic technologies. This is further exacerbated by the nature of intermittent shedding of eggs and larvae by these parasites. Diagnostic methods range from simple morphological identification to more specialised high-throughput sequencing technologies. Microscopy-based methods, although simple, are labour-intensive and considerably less sensitive than molecular methods which are rapid and have high levels of accuracy. Molecular methods use nucleic acid amplification (NAA) to amplify the deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) fragments of the parasite to detect and determine its presence using different technologies (NAAT). They have increased the sensitivity of detection and quantitation of intestinal nematode infections, especially in low infection intensity settings. The absence of a gold standard test limits current diagnosis and, in turn, restricts intervention measures and effective control efforts. The objective of this review is to determine the accuracy of NAATs in detecting human intestinal nematode infections using Kato-Katz as the reference test for the most common soil-transmitted helminth (STH) infections and the scotch tape test for enterobiasis and Baermann method for strongyloidiasis. Relevant studies will be identified by searches in electronic databases. Two reviewers will independently screen the literature against eligibility criteria. The methodological quality of studies will then be appraised by two reviewers using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool. Discrepancies will be addressed by a third reviewer. The true positives, false positives, true negatives and false negatives of all the studies will be extracted into contingency tables. In paired forest plots, study-specific sensitivity and specificity with a 95 per cent confidence interval will be displayed. The systematic review of this protocol will report the diagnostic accuracy of currently available NAATs for the detection of human intestinal nematode infections. This will help healthcare providers and administrators determine the diagnostic method to be used in different clinical and preventive settings. Trial registration: PROSPERO registration number for this protocol is CRD42022315730.

Development of hydrogel-forming microneedles for transdermal delivery of albendazole from liquid reservoir

Albendazole (ABZ) is an anthelmintic agent from the benzimidazole group, known as the broad-spectrum antiparasitic drug. ABZ is commonly used to treat human intestinal and systemic infections. Orally administered ABZ tends to have limited efficacy due to its poor solubility. In order to enhance its delivery to the therapeutic target, polyvinyl alcohol-based hydrogel-forming microneedles (HFMs) was developed. HFMs can effectively deliver drugs loaded in the reservoir through the transdermal route with fewer side effects and longer therapeutic duration. In addition, to enhance ABZ's solubility, the drug can be loaded as a liquid reservoir using water-miscible solvents, which will effectively enhance the solubility of ABZ, resulting in higher bioavailability. In this study, HFMs was successfully developed with high swelling abilities, more than 400%. Moreover, the penetration result showed HFMs could penetrate up to 63% into the skin with only a 7.14% of height decrease. The skin integrity test also showed HFMs permeation into the skin, causing no changes in skin integrity after 24 h of application. Incorporated with the liquid reservoir, the ex vivo permeation test showed that the cumulative amount of ABZ permeated through the skin was about 971.23 ± 11.77 µg/cm2. In conclusion, this innovation has a huge potential to overcome the limitations of ABZ in oral preparations and potentially enhance its therapeutic effect through the transdermal route.

Computational Design of a Chimeric Vaccine against Plesiomonas shigelloides Using Pan-Genome and Reverse Vaccinology.

The swift emergence of antibiotic resistance (AR) in bacterial pathogens to make themselves adaptable to changing environments has become an alarming health issue. To prevent AR infection, many ways can be accomplished such as by decreasing the misuse of antibiotics in human and animal medicine. Among these AR bacterial species, Plesiomonas shigelloides is one of the etiological agents of intestinal infection in humans. It is a gram-negative rod-shaped bacterium that is highly resistant to several classes of antibiotics, and no licensed vaccine against the aforementioned pathogen is available. Hence, substantial efforts are required to screen protective antigens from the pathogen whole genome that can be subjected easily to experimental evaluations. Here, we employed a reverse vaccinology (RV) approach to design a multi-antigenic epitopes based vaccine against P. shigelloides. The complete genomes of P. shigelloides were retrieved from the National Center for Biotechnological Information (NCBI) that on average consist of 5226 proteins. The complete proteomes were subjected to different subtractive proteomics filters, and in the results of that analysis, out of total proteins, 2399 were revealed as non-redundant and 2827 as redundant proteins. The non-redundant proteins were further checked for subcellular localization analysis, in which three were localized in the extracellular matrix, eight were outer membrane, and 13 were found in the periplasmic membrane. All surface localized proteins were found to be virulent. Out of a total of 24 virulent proteins, three proteins (flagellar hook protein (FlgE), hypothetical protein, and TonB-dependent hemoglobin/transferrin/lactoferrin family receptor protein) were considered as potential vaccine targets and subjected to epitopes prediction. The predicted epitopes were further examined for antigenicity, toxicity, and solubility. A total of 10 epitopes were selected (GFKESRAEF, VQVPTEAGQ, KINENGVVV, ENKALSQET, QGYASANDE, RLNPTDSRW, TLDYRLNPT, RVTKKQSDK, GEREGKNRP, RDKKTNQPL). The selected epitopes were linked with each other via specific GPGPG linkers in order to design a multi-epitopes vaccine construct, and linked with cholera toxin B subunit adjuvant to make the designed vaccine construct more efficient in terms of antigenicity. The 3D structure of the vaccine construct was modeled ab initio as no appropriate template was available. Furthermore, molecular docking was carried out to check the interaction affinity of the designed vaccine with major histocompatibility complex (MHC-)I (PDB ID: 1L1Y), MHC-II (1KG0), and toll-like receptor 4 ((TLR-4) (PDB: 4G8A). Molecular dynamic simulation was applied to evaluate the dynamic behavior of vaccine-receptor complexes. Lastly, the binding free energies of the vaccine with receptors were estimated by using MMPB/GBSA methods. All of the aforementioned analyses concluded that the designed vaccine molecule as a good candidate to be used in experimental studies to disclose its immune protective efficacy in animal models.

Association of Midgut Bacteria and Their Metabolic Pathways with Zika Infection and Insecticide Resistance in Colombian Aedes aegypti Populations.

Introduction: Aedes aegypti is the vector of several arboviruses such as dengue, Zika, and chikungunya. In 2015–16, Zika virus (ZIKV) had an outbreak in South America associated with prenatal microcephaly and Guillain-Barré syndrome. This mosquito’s viral transmission is influenced by microbiota abundance and diversity and its interactions with the vector. The conditions of cocirculation of these three arboviruses, failure in vector control due to insecticide resistance, limitations in dengue management during the COVID-19 pandemic, and lack of effective treatment or vaccines make it necessary to identify changes in mosquito midgut bacterial composition and predict its functions through the infection. Its study is fundamental because it generates knowledge for surveillance of transmission and the risk of outbreaks of these diseases at the local level. Methods: Midgut bacterial compositions of females of Colombian Ae. aegypti populations were analyzed using DADA2 Pipeline, and their functions were predicted with PICRUSt2 analysis. These analyses were done under the condition of natural ZIKV infection and resistance to lambda–cyhalothrin, alone and in combination. One-step RT-PCR determined the percentage of ZIKV-infected females. We also measured the susceptibility to the pyrethroid lambda–cyhalothrin and evaluated the presence of the V1016I mutation in the sodium channel gene. Results: We found high ZIKV infection rates in Ae. aegypti females from Colombian rural municipalities with deficient water supply, such as Honda with 63.6%. In the face of natural infection with an arbovirus such as Zika, the diversity between an infective and non-infective form was significantly different. Bacteria associated with a state of infection with ZIKV and lambda–cyhalothrin resistance were detected, such as the genus Bacteroides, which was related to functions of pathogenicity, antimicrobial resistance, and bioremediation of insecticides. We hypothesize that it is a vehicle for virus entry, as it is in human intestinal infections. On the other hand, Bello, the only mosquito population classified as susceptible to lambda–cyhalothrin, was associated with bacteria related to mucin degradation functions in the intestine, belonging to the Lachnospiraceae family, with the genus Dorea being increased in ZIKV-infected females. The Serratia genus presented significantly decreased functions related to phenazine production, potentially associated with infection control, and control mechanism functions for host defense and quorum sensing. Additionally, Pseudomonas was the genus principally associated with functions of the degradation of insecticides related to tryptophan metabolism, ABC transporters with a two-component system, efflux pumps, and alginate synthesis. Conclusions: Microbiota composition may be modulated by ZIKV infection and insecticide resistance in Ae. aegypti Colombian populations. The condition of resistance to lambda–cyhalothrin could be inducing a phenome of dysbiosis in field Ae. aegypti affecting the transmission of arboviruses.

Double troubleStrongyloidiasis hyper infection and intestinal tuberculosis co-infection-A rare case report

Strongyloidiasis, a human intestinal parasitic infection is most commonly caused by Strongyloides stercoralis species. Strongyloides Hyperinfection is an accelerated autoinfection occurring in immunocompromised state like tuberculosis, where increased number of infectious filariform larvae are produced. Here, we report a case of 35-year-old male, known case of intestinal tuberculosis, presenting with complaints of pain abdomen, associated with vomiting. Clinical and radiological examination revealed hollow viscous perforation with peritonitis. Surgical ileal resection with anastomosis was performed and histopathological analysis of the specimen revealed eggs and larval and adult forms of Strongyloides stercoralis within crypts and superficial mucosa

Prevalence, virulence genes, and antimicrobial profiles of Escherichia coli O157:H7 isolated from healthy cattle in Tunisia.

Shiga toxin-producing Escherichia coli (STEC) O157:H7 is associated with intestinal infection in humans and is considered an important cause of food-borne diseases. The aim of the study was to assess the incidence of E. coli O157:H7 in fecal samples of healthy cattle collected in slaughterhouses (n = 160) and from five farms (n = 100). E. coli isolates were detected on MacConkey agar. A total of 236 E. coli isolates were recovered from fecal samples of healthy cattle. We used sorbitol MacConkey medium to detect non-sorbitol fermenting colonies. These bacteria were examined for the presence of O157:H7 antigen by latex agglutination. The isolation of E. coli O157:H7 has been confirmed with PCR amplification of rfbEO157 and fliCH7 specific genes for serogroup O157 and with multiplex PCR of stx1, stx2, eaeA, and ehxA. All isolates were examined for their susceptibility to 21 antibiotics by the disc diffusion method. Of the 236 E. coli isolates, 4.2% (10/236) were positive for STEC O157:H7. Shiga toxin gene (stx2) and ehxA were present in 70% of isolates, stx1 and eae were confirmed in 60% of the isolates. Other virulence factors screened (fimH, sfa/focDE, cdt3, traT, iutA, and hlyA) were present among the 10 isolates. All E. coli O157:H7 isolates were sensitive to amoxicillin/clavulanic acid, cefotaxime, cefepime, aztreonam, colistin, and sulfamethoxazole/trimethoprim. All isolates belong to the phylo-group E. This is the first study of the incidence of E. coli O157:H7 in cattle in Tunisia. Our finding proves the existence of STEC O157:H7 in healthy animals producing food for human consumption which could be a source of food-borne disease.

An Engineered λ Phage Enables Enhanced and Strain-Specific Killing of Enterohemorrhagic Escherichia coli.

ABSTRACTBacteriophages (phages) are ideal alternatives to traditional antimicrobial agents in a world where antimicrobial resistance (AMR) is emerging and spreading at an unprecedented speed. In addition, due to their narrow host ranges, phages are also ideal tools to modulate the gut microbiota in which alterations of specific bacterial strains underlie human diseases, while dysbiosis caused by broad-spectrum antibiotics can be harmful. Here, we engineered a lambda phage (Eλ) to target enterohemorrhagic Escherichia coli (EHEC) that causes a severe, sometimes lethal intestinal infection in humans. We enhanced the killing ability of the Eλ phage by incorporating a CRISPR-Cas3 system into the wild-type λ (wtλ) and the specificity by introducing multiple EHEC-targeting CRISPR spacers while knocking out the lytic gene cro. In vitro experiments showed that the Eλ suppressed the growth of EHEC up to 18 h compared with only 6 h with the wtλ; at the multiplicity of infection (MOI) of 10, the Eλ killed the EHEC cells with ~100% efficiency and did not affect the growth of other laboratory- and human-gut isolated E. coli strains. In addition, the EHEC cells did not develop resistance to the Eλ. Mouse experiments further confirmed the enhanced and strain-specific killing of the Eλ to EHEC, while the overall mouse gut microbiota was not disturbed. Our methods can be used to target other genes that are responsible for antibiotic resistance genes and/or human toxins, engineer other phages, and support in vivo application of the engineered phages.IMPORTANCE Pathogenic strains of Escherichia coli are responsible for 0.8 million deaths per year and together ranked the first among all pathogenic species. Here, we obtained, for the first time, an engineered phage, Eλ, that could specifically and efficiently eliminate EHEC, one of the most common and often lethal pathogens that can spread from person to person. We verified the superior performance of the Eλ over the wild-type phage with in vitro and in vivo experiments and showed that the Eλ could suppress EHEC growth to nondetectable levels, fully rescue the EHEC-infected mice, and rescore disturbed mouse gut microbiota. Our results also indicated that the EHEC did not develop resistance to the Eλ, which has been the biggest challenge in phage therapy. We believe our methods can be used to target other pathogenic strains of E. coli and support in vivo application of the engineered phages.

An increase in prevalence of recombinant GII.3[P12] norovirus in sporadic acute diarrhea in children in Nizhny Novgorod, Russia, 2018-2021.

Noroviruses are important etiological agents causing acute intestinal infection in humans. In the last decades, the most common norovirus genotype was GII.4 despite a significant genetic diversity among strains, while the active circulation of noroviruses with other genotypes was observed periodically. This study shows an increase in the detection rate of recombinant GII.3[P12] norovirus in Nizhny Novgorod, Russia, from 6.8% in 2018-2019 to 34.9% in 2020-2021. We performed a phylogenetic analysis based on the nucleotide sequences of noroviruses possessing this genotype obtained in this work, as well as presented in the GenBank database. It has been shown that the circulation of GII.3[P12] noroviruses in the study area was the result of several independent introductions, either directly from the Western Pacific region, or through the Asian part of Russia. The polyphyletic origin, the geographical expansion, and the growth of the epidemic significance of the recombinant GII.3[P12] noroviruses were noted.