Examination Of Stool Samples Research Articles

DOP028 The resolution of fecal calprotectin configurations and their biological functions in gut inflammation

Abstract Background Fecal calprotectin (CP) serves as a worldwide established biomarker used for the diagnosis and management of inflammatory bowel diseases (IBD). However, the biological function of CP in the human gut is still unknown and numerous studies obtained contradictory results. Calprotectin is mainly described to be present as a tetrameric protein complex (S100A8/S100A9)2, while the quaternary protein structure (configuration) of S100A8 and S100A9 in the human gut lumen remains enigmatic.1 In this study we resolved the fecal protein configurations of calprotectin and their biological functions in IBDs. Methods We defined biological functions of S100A8 and S100A9 dimers by antibody-mediated immunoprecipitation of S100A8 and S100A9 from stool obtained from IBD patients coupled with subsequent LC-MS/MS analysis. Next, we analysed human colonic organoids by single-cell RNA sequencing after S100A8 or S100A9 stimulation. Moreover, we orally exposed mice to recombinant human S100A8 and S100A9 protein during experimental gut inflammation and genetically inactivated S100a9 in toxic and genetic mouse models. To identify the protein configurations of S100A8 and S100A9 in the human gut lumen we examined stool samples from IBD patients using size-exclusion chromatography and LC-MS/MS analysis. Results We demonstrate pro-inflammatory actions for S100A8 and S100A9 dimers as oral administration of human S100A8 and S100A9 homodimers, but not CP, worsened experimental enteritis and colitis. In accordance, multi-omic phenotyping revealed that human S100A8 and S100A9 homodimers induce the NLRP1 inflammasome in intestinal epithelium. In turn, genetic deletion of S100a9 ameliorated experimental gut inflammation. S100A8 and S100A9 dimers were frequently detectable in stool from IBD patients from three independent IBD cohorts, but not in healthy individuals. More specifically, fecal S100A9 detection was associated with clinical and endoscopic disease activity in IBD patients with low fecal CP levels (≤150µg/g). Importantly, four fecal CP assays in clinical use were not able to detect S100A8 and S100A9 homodimers. Conclusion Our study is the first to reveal the presence of S100A8 and S100A9 dimers in stool from IBD patients. Moreover, we demonstrate a diagnostic gap for fecal CP assays in clinical use. We establish that fecal S100A9 detection is associated with endoscopic disease activity in IBD patients in which fecal CP fails to identify active disease. We reveal disease-modifying functions for S100A8 and S100A9 homodimers by inducing the NLRP1 inflammasome, while the same effects were not observable for calprotectin. Taken together, our findings may reflect a major advance in the IBD biomarker field. References Jukic, A., Bakiri, L., Wagner, E. F., Tilg, H. & Adolph, T. E. Calprotectin: from biomarker to biological function. Gut 70 , 1978-1988 (2021). https://doi.org/10.1136/gutjnl-2021-324855

Effect Cryptosporidium parvum on Lactoferrin level in children with diarrhea

Cryptosporidium spp. are a group of coccidian parasites of significant pathogenicity to both animals and humans. Cryptosporidiosis is understood to be the underlying factor behind the development of severe and protracted watery diarrhea in individuals diagnosed with acquired immunodeficiency syndrome. This study aimed to diagnosis of infection with the parasite Cryptosporidium parvum in children by microscopic examination of stool samples using the Modified Ziehl-Nelsoon stain and ELISA. As well as detection level of Lactoferrin in samples of patients infected with Cryptosporidium parvum . A total 300 samples children aged 5 months - 5 years, of both sexes, who visited some government hospitals (Tikrit Teaching Hospital, Salah al-Din General Hospital, Baiji General Hospital, Sharqat General Hospital) and some private medical clinics in some areas of Salah al-Din Governorate who suffered from diarrhea and some intestinal symptoms during the period from July 1, 2022 to September 1, 2023. Based on the analysis of 300 stool samples, the present investigation employing MZN revealed an infection rate of 70 (23.33%) positive samples and 230 (76.67%) negative samples. The ELISA technique used in the current study recorded 130 positive samples and a total infection rate of 72.22% with a mean concentration of 2.54±1.13 pg/mL), while the number of negative samples was 50 (27.78%) with a mean concentration of (27.78%) pg/mL out of 90 samples. This study showed increase incidence of infection with cryptosporidium in male than female that were 82(63.08%) and 48(36.92%) respectively, at p-value<0.05. The study observed a significant increase in the average concentration of fecal LF in infected infants (506.125 ± 99.143 pg/mL) compared to the control group (104.183 ± 21.368pg/ml). This study concluded that Modified Ziehl Neelsen stain is the best staining method used in diagnosing infection with the parasite. . Furthermore this study concluded increase level inflammatory immune parameters as lactoferrine in blood serum and stool samples of children infected with the C. parvum parasite

The relationship between Giardia lamblia infection with Ghrelin hormone and Trefoil factor 3 in patients in Salah al-Din Governorate

Giardiasis is considered one of the parasitic diseases affecting humans, caused by a flagellate protozoan known as Giardia lamblia. It is a common gastrointestinal pathogen that can lead to various clinical and other complications. Study aimed to diagnose infection with the G. lamblia in children aged (1-12 years) using a direct wet mount, and to determine Giardia lamblia by targeting the TPI gene using PCR, in addition to evaluating the relationship of infection of Giardia lamblia with (ghrelin and TFF3) using ELISA assay. By examining 238 stool samples for children suffering from diarrhea and some intestinal symptoms visiting some hospitals in Salah al-Din Governorate. 34 (14.2%) positive samples were recorded by microscopic examination. 100 fecal samples using the PCR, showed 60 (60%) positive samples for the G. lamblia. The study revealed a highly significant increase in the level of TFF3 a non-significant decrease in the level ghrelin in the serum of patients with infection. Direct microscopic examination of stool samples It has good sensitivity for detecting G. lamblia parasite, and the TPI gene targeted in PCR technology has high sensitivity and specificity for genotyping detection of the parasite.

Intestinal parasitic infections among children aged 7-14 years in Mizan-Aman city, Southwest Ethiopia: a community-based cross-sectional study.

Intestinal parasitic infections continue to pose a major threat to human health globally, with a particularly high prevalence in developing countries. Soil-borne helminthiasis and schistosomiasis are notably widespread. The objective of the study was to determine the prevalence and contributing factors of intestinal parasites infection among participants aged 7-14 years. Community-wide prevalence study was undertaken from 30 August to 30 September 2021 in Mizan Aman Town. Socio-demographic information was collected using questionnaires. Three of the five kebels were randomly chosen. Households with children aged 7-14 were gathered from the chosen kebels and health post to recruit one eligible subject. Allocation of study subjects to each of the chosen kebels was computed proportionally. Two thick smear of Kato Katz technique was applied to examine stool samples. Data were entered and analyzed using SPSS version 20. To investigate the association between the dependent and independent variables, a logistic regression analysis was conducted. Statistics were considered significant for p-values under 0.05. The overall prevalence of intestinal parasites was 64.6% (215/333). Of these, 51.05% (170/333) were infected with STHs, while 13.5% (45/333) had S. mansoni. T. trichiura was the most prevalent helminth. Infection intensity ranged from light to moderate was observed. Prior information about STHs (aORr:2.022 = CI:1.222-3.340), poor knowledge about STHs (aOR:1.677 = CI:1.057-2.660), unaware of deworming as prevention method of S. mansoni (aOR:2.620:CI:1267-5.418), swimming (aOR:0.448:CI:0.176-0.992) and contact with water (aOR:0.402:CI:0.169-0.957) were significantly associated with the S. mansoni infection. The prevalence of intestinal parasite was high. Heavy infection was not recorded. Beyond mass deworming, the report emphasizes the necessity of ongoing public health interventions to address the high prevalence of these intestinal helminths.

Beyond Poliomyelitis: A 21-Year Study of Non-Polio Enterovirus Genotyping and Its Relevance in Acute Flaccid Paralysis in São Paulo, Brazil.

In the context of the near-global eradication of wild poliovirus, the significance of non-polio enteroviruses (NPEVs) in causing acute flaccid paralysis (AFP) and their impact on public health has gained increased attention. This research, conducted from 2001 to 2021, examined stool samples from 1597 children under 15 years in São Paulo, Brazil, through the AFP/Poliomyelitis Surveillance Program, detecting NPEVs in 6.9% of cases. Among the 100 NPEV-positive strains analyzed, 90 were genotyped through genomic sequencing of the partial VP1 region, revealing a predominance of EV-B species (58.9%), followed by EV-A (27.8%) and EV-C (13.3%). This study identified 31 unique NPEV types, including EV-A71, CVB2, and E11, as the most prevalent, along with the first documented occurrence of CVA19 in Brazil. These findings emphasize the importance of NPEV genotyping in distinguishing AFP from poliomyelitis, enhancing understanding of these viruses' epidemiology. Moreover, it ensures that AFP cases are correctly classified, contributing to the effective surveillance and eradication efforts for poliomyelitis.

Acupuncture and Moxibustion for Inflammatory Bowel Disease: Regulatory Mechanisms Revealed by Microbiome and Metabolomic Analysis.

Acupuncture and moxibustion are widely acknowledged as effective complementary therapies for managing inflammatory bowel disease (IBD) in traditional Chinese medicine. However, the regulatory mechanisms by which these two therapies exert their therapeutic effects in IBD are yet to be fully elucidated. The objective of this study was to investigate the mechanisms of action underlying acupuncture and moxibustion and the regulative differences between them as therapeutic interventions for IBD. Using a dextran sodium sulfate-induced IBD mice model, the effects of the two treatments were evaluated by examination of body weight, stool samples, colon morphology, inflammatory factors, gut microbiota, and metabolites. The results indicated that both acupuncture and moxibustion mitigated body weight reduction; improved the structural characteristics of intestinal tissues; increased levels of anti-inflammatory cytokines including interleukin (IL)-10; and decreased levels of pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-[Formula: see text]), nuclear factor kappa B (NF-[Formula: see text]B), IL-6, IL-1[Formula: see text], and IL-17. Acupuncture and moxibustion had distinct effects on the regulation of the intestinal microbiota and metabolic pathways in IBD mice. Moxibustion regulated a greater number of metabolic pathways than acupuncture, the majority of which were associated with amino acid metabolism, brain signal transmission, energy metabolism, and anti-inflammatory pathways. These findings provide a scientific basis for the differential applications of acupuncture and moxibustion in clinical practice.

Morphological and molecular identification of Hymenolepis spp. in Rattus rattus and children with diarrhea from Upper Egypt.

Hymenolepiasis remains among the most common parasitic zoonoses in developing countries. Little information is available about hymenolepiasis in children in Upper Egypt and rodents' contribution to maintaining the disease's epidemiology. A cross-sectional study was carried out to investigate the occurrence of Hymenolepis spp. in Rattus rattus and children in Asyut Governorate, Egypt. Rodents (n = 100) were randomly trapped from various localities in Asyut Governorate, and stool samples from 120 children were collected from the same localities. Laboratory examination of the collected samples involved investigation of the small intestine of R. rattus for adult worm detection by morphological examination, followed by examination of stool samples of children using direct smear, formol-ether sedimentation technique, and Sheather's sugar flotation technique. Confirmation of Hymenolepis spp. positive samples were performed using polymerase chain reaction targeting the internal transcribed spacer 1 (ITS-1) and restriction fragment length polymorphism (PCR-RFLP). This study revealed the occurrence of Hymenolepis spp. in 45% of the examined R. rattus, comprising 43% positivity for H. diminuta and 2% for mixed infection by H. nana and H. diminuta. Hymenolepis nana was detected in 28.3% of the examined children. PCR-RFLP confirmed these findings, showing 100% sensitivity. Collectively, these findings reveal the potential contribution of R. rattus as an important reservoir for Hymenolepis infection in Upper Egypt. This study concluded that personal education, periodical deworming of children, rodent control, and hygienic measures should be implemented by governmental and nongovernmental organizations to reduce the incidence of infection.

A Study Evaluating Infection with the Parasite Entamoeba Histolytica and Its Relationship to Hemoglobin Level and White Blood Cell Count

This study was conducted to determine the incidence of the histolytic encephalopathy and its relationship to some blood variables (hemoglobin and white blood cells). A number of (120) samples were collected from the general records of Zawia Teaching Hospital from the period (January 2023-December 2023) in the children's department, in addition to examining some samples of shelter cases in the children's department, where we relied on the direct stain method to examine stool samples. The results showed that the most infected age groups are the ages between (6 years - 10 years) at a rate of (19%), in addition to the fact that the most infected gender was females at a rate of (64%), in addition to the fact that it was noted that the infected had a slight deficiency in the hemoglobin percentage, which indicates that the infections were not severe, in addition to the fact that the percentage of white blood cells was (14%), which is considered a high percentage compared to the normal percentage, and the percentage of non-infected indicates that the children's immunity was good and the infection was simple. Keywords: Entamoeba histolytica parasite, infection, relationship, hemoglobin level, white blood cells.

Correlation between Assemblages A and B Giardia lamblia infection with melatonin, ghrelin, Hsp70 and TFF3 biomarkers in patients

Background: Giardiasis is considered one of the parasitic diseases affecting humans, caused by a flagellate protozoan known as Giardia lamblia. It is a common gastrointestinal pathogen that can lead to various clinical and other complications. Objective: Study aimed to diagnose infection with the G. lamblia in children aged (1-12 years) using a direct wet mount, and to determine Assemblages A and B Giardia lamblia by targeting the TPI gene using Nested-PCR, in addition to evaluating the relationship of infection of assemblages with (melatonin, ghrelin, Hsp70 and TFF3) using ELISA assay. Method: By examining 238 stool samples for children suffering from diarrhea and some intestinal symptoms visiting some hospitals in Salah al-Din Governorate. Results: 34 (14.2%) positive samples were recorded by microscopic examination. 100 fecal samples using the Nested-PCR, showed 60 (60%) positive samples for the G. lamblia, including 18 (30%) for the assemblage B, 31 (51.66%) samples for assemblage A, as well as 11 (18.33%) mixed infections of A+B. The study of biomarkers revealed a highly significant increase in the levels of (melatonin, ghrelin, Hsp70 and TFF3) in the serum of patients with mixed A + B infection, while the lowest average concentrations of biomarkers were recorded in children infected with group A. Conclusion: Direct microscopic examination of stool samples It has good sensitivity for detecting G. lamblia parasite, and the TPI gene targeted in Nested-PCR technology has high sensitivity and specificity for genotyping detection of the parasite. Group A is the most common among patient samples. Mixed infections (A+B) have the greatest effect.

Intestinal parasitic infections in Ukrainian war refugee children living in Poland

Introduction: The armed aggression of the Russian Federation against Ukraine in February 2022 forced millions of Ukrainian citizens to flee their country and seek refuge mainly in the European Union. Many of them found shelter in Poland. There are now over one million Ukrainian war refugees living in Poland, including 500,000 children. The attention of Polish medical services has been recently drawn to the previously unknown epidemiological status of infectious and invasive diseases in the paediatric population of Ukrainian refugees. The aim of this study was to assess the prevalence of intestinal parasitic infections in the group of Ukrainian children living in Poland. Materials and methods: The study involved performing parasitological examination of stool samples collected in May 2023 from 37 Ukrainian refugee children aged 3–7 years, who were living in Szczecin (Poland) and were attending a preschool set up to address the needs of the local Ukrainian community. Stool samples were tested by light microscopy using three different methods (direct smear, decantation, flotation) at the Department of Epidemiology and Tropical Medicine at the Military Institute of Medicine – National Research Institute. Results: Intestinal parasites were detected in 3 out of 37 children (8.1% of the study group; there was one case of infection with Ascaris lumbricoides and two cases of infection with Blastocystis spp.). The infected children’s parents or guardians did not observe any diarrhoeas or other gastrointestinal symptoms within 6 months prior to the examination. Conclusions: The presence of intestinal parasites among Ukrainian war refugee children living in Poland justifies the screening for parasitic diseases in this population. The obtained results will allow to take appropriate preventive and therapeutic actions.

Magnitude of anemia and associated factors among pregnant women attending antenatal care in governmental health facilities of Shashemene Town, Oromia region, Ethiopia.

Anemia during pregnancy is a common issue that significantly affects the health of both the mother and her child. Globally, anemia is a major public health concern, affecting both developing and developed countries, with approximately 1.3 billion people affected. Pregnant women are among the most vulnerable to anemia. To assess the magnitude and risk factors of anemia among pregnant women attending antenatal care in Shashemene Town, Oromia, Ethiopia. A facility-based cross-sectional study was conducted among 391 pregnant women in Shashemene Town in April 2022. Data were collected using interviewer-administered questionnaires, along with laboratory examinations of blood and stool samples. The data were entered into EpiData 3.1 and analyzed using the Statistical Package for Social Sciences (SPSS) version 22. Bivariate logistic regression was performed, and variables with a p-value of <0.25 were included in the multivariate logistic regression analysis to identify factors associated with anemia. Adjusted odds ratio (AOR) with 95% CIs were calculated, and a p-value of < 0.05 was considered statistically significant. Finally, the results are presented using narration, descriptive statistics, such as tables, graphs, and charts. The prevalence of anemia was found to be 30.9% (95% CI: 26.4, 35.4%). Factors significantly associated with a reduced risk of anemia included high dietary diversity (AOR = 0.217, 95% CI: 0.105-0.451), no history of excessive menstrual bleeding (AOR = 0.162, 95% CI 0.076-0.345), age 25-34 years (AOR = 0.391, 95% CI 0.173-0.883), and age ≥ 35 years (AOR = 0.068, 95% CI 0.011-0.444). Conversely, a mild upper arm circumference (MUAC) of <23 cm (AOR = 4.939, 95% CI 2.330-10.469), no use of contraceptives (AOR = 4.935, 95% CI 2.207-11.032), and no iron supplementation use (AOR = 3.588, 95% CI 1.794-7.175) were significantly associated with an increased risk of anemia. According to the WHO classification, anemia in this study was found to be a moderate public health issue. High dietary diversity, no previous excessive menstrual bleeding, and age were significantly associated with a reduced risk of anemia, whereas a MUAC of <23 cm, no contraceptive use, and no iron supplementation were significantly associated with an increased risk of anemia. Therefore, promoting diverse diets among pregnant women, providing counseling on the benefits of family planning and iron-folic acid supplements, and improving women's education and empowerment are essential.

Distinct gut microbiota alterations in patients with early‐stage and advanced‐stage hepatocellular carcinoma

AbstractGut microbiota (GM) alterations play a key role in the development of hepatocellular carcinoma (HCC); however, little is known about the changes in GM diversity during the progression of HCC. Thus, we analyzed the differences in the fecal microbiota of patients with early‐stage and advanced‐stage HCC. This cross‐sectional study examined stool samples from adults with early‐stage HCC (n = 31) and advanced‐stage HCC (n = 44). The taxonomic composition of the GM was determined by 16S ribosomal RNA gene sequencing of stool samples. There was no significant difference in the alpha‐diversity of the GM between groups; however, beta‐diversity was significantly different between the early‐stage and advanced‐stage groups. Further analysis indicated that the genus Veillonella, family Enterobacteriaceae, order Enterobactriales, and class Gammaproteobacteria were more abundant in the advanced‐stage group than the early‐stage group. Patients with early‐stage and advanced‐stage HCC exhibit different patterns of GM diversity. Furthermore, specific taxa—including Veillonella, Enterobacteriaceae, Enterobacteriales, and Gammaproteobacteria—are enriched in patients with advanced‐stage HCC.

Multi-infection parasitic nematodes in deer at Waru Farm Land, Ciampea, Bogor Regency

Deer is a protected wild animal in Indonesia, and currently, deer are often kept and used as part of amusement rides in recreational areas. This study aimed to identify the incidence of worm infections in deer kept in the Waru Farm Land recreation area in Tegalwaru, Ciampea Bogor. Nine deer were kept by Waru Farm Land, consisting of spotted deer and Timor deer. Samples were collected immediately after the deer had defaecated. Stool sample examinations included the flotation test, McMaster’s test, and multilevel filtration. The examination results showed that one deer had multiple parasitic nematode infections. Examination revealed strong and trichurid-type eggs. The degree of infection showed a very mild value, namely less than 50 EPG, so it can be concluded that worm infection in deer at Waru Farm Land is very low. Treatment by administering anthelmintics to deer that were positively infected with worms should be given, as well as preventing infection in other deer.

High-throughput detection of parasites and ova in stool using the fully automatic digital feces analyzer, orienter model fa280

BackgroundIntestinal parasitic infections can harm health by causing malnutrition, anemia, impaired growth and cognitive development, and alterations in microbiota composition and immune responses. Therefore, it is crucial to examine stool samples to diagnose parasitic infections. However, the traditional microscopic detection method is time-consuming, labor-intensive, and dependent on the expertise and training of microscopists. Hence, there is a need for a low-complexity, high-throughput, and cost-effective alternative to labor-intensive microscopic examinations.MethodsThis study aimed to compare the performance of a fully automatic digital feces analyzer, Orienter Model FA280 (People’s Republic of China) with that of the formalin-ethyl acetate concentration technique (FECT). We assessed and compared the agreement between the FA280 and the FECT for parasite detection and species identification in stool samples. The first part of the study analyzed 200 fresh stool samples for parasite detection using the FECT and FA280. With the FA280, the automatic feces analyzer performed the testing, and the digital microscope images were uploaded and automatically evaluated using an artificial intelligence (AI) program. Additionally, a skilled medical technologist conducted a user audit of the FA280 findings. The second set of samples comprised 800 preserved stool samples (preserved in 10% formalin). These samples were examined for parasites using the FECT and FA280 with a user audit.ResultsFor the first set of stool samples, there was no statistically significant difference in the pairwise agreements between the FECT and the FA280 with a user audit (exact binomial test, P = 1). However, there were statistically significant differences between the pairwise agreements for the FECT and the FA280 with the AI report (McNemar’s test, P < 0.001). The agreement for the species identification of parasites between the FA280 with AI report and FECT showed fair agreement (overall agreement = 75.5%, kappa [κ] = 0.367, 95% CI 0.248–0.486). On the other hand, the user audit for the FA280 and FECT showed perfect agreement (overall agreement = 100%, κ = 1.00, 95% CI 1.00–1.00). For the second set of samples, the FECT detected significantly more positive samples for parasites than the FA280 with a user audit (McNemar’s test, P < 0.001). The disparity in results may be attributed to the FECT using significantly larger stool samples than those used by the FA280. The larger sample size used by the FECT potentially contributed to the higher parasite detection rate. Regarding species identification, there was strong agreement between the FECT and the FA280 with a user audit for helminths (κ = 0.857, 95% CI 0.82–0.894). Similarly, there was perfect agreement for the species identification of protozoa between the FECT and the FA280 with user audit (κ = 1.00, 95% CI 1.00–1.00).ConclusionsAlthough the FA280 has advantages in terms of simplicity, shorter performance time, and reduced contamination in the laboratory, there are some limitations to consider. These include a higher cost per sample testing and a lower sensitivity compared to the FECT. However, the FA280 enables rapid, convenient, and safe stool examination of parasitic infections.Graphical

Prevalence of Soil-Transmitted Helminth Infections and Associated Risk Factors Among School Children in Dembecha Town, Ethiopia.

Soil-transmitted helminth (STH) infections are among the most common infections that affect the poorest and most deprived communities. In most developing countries, children aged 5 to 15 years are at risk for chronic helminth infection and associated morbidity. This study aimed to determine the prevalence of STH infections and associated risk factors among three government elementary schools in Dembecha town, Ethiopia. We conducted a school-based cross-sectional study involving 316 participants between November 2019 and March 2020. A systematic random sampling method was used to select study participants from the study schools. Data related to the sociodemographic characteristics of the study participants and risk factors for STH infections were collected using a pretested questionnaire survey. Parasitological examinations of stool samples were performed using the formal-ether concentration method. Study participants aged 5 to 15 years were enrolled in this study. The overall prevalence of STH infection was 21.5% (68/316). Ascaris lumbricoides ranked highest, with a prevalence of 11.4%, followed by hookworms 7.3%, Trichuris trichiura 1.9%, and Strongloides stercoralis 0.9%. Age groups of 10-15 years (AOR =3.109; 95% CI: 1.033, 9.350), residence in Kebele 2 (AOR =2.990; 95% CI: 1.082, 8.264), illiterate mothers (AOR =4.689; 95% CI: 1.410, 15.59), and a family size of 4-6 (AOR =3.286; 95% CI: 1.299, 8.313) were significantly associated with STH infections. The prevalence of STH infections remains an important health issue for study participants. Therefore, school deworming programs twice a year are crucially needed until the prevalence falls below the level of public health importance.

Genetic map of the isolated Cryptosporidium parasite from children with diarrheal in the city of Mosul

The current study was conducted between the beginning of October 2022 and the end of March 2023 to identify the prevalence of Cryptosporidium parivum infection among children under five years using microscopic and molecular methods. The study recorded 43(37.71%) infected children diagnosed using the Modified Zeihl Neelsen Stain and Gimsa stain. On the other hand, a molecular examination of stool samples using PCR showed that 25(21.92%) children out of 115 collected stool samples were infected with this parasite. DNA Sequencing was performed for specimens diagnosed with Cryptosporidium parasite, and the new isolates were registered for the first time in Nineveh governorate in NCBI under the accession number 757376. Keywords: Cryptosporidium disease, Genetic map.

Blastocystis sp. and Toxocara spp. Coinfection in Patients with Clinically Manifested Skin Allergy

In recent years, more and more publications reflect the relationship between clinically apparent allergy and infection with Blastocystis spp. or Toxocara spp. Our main goal is to examine allergy patients infected with Blastocystis sp. for the presence of anti-Toxocara IgG antibodies, intending to detect co-infection and its influence on the allergic condition of patients. For a period of 3 years (2020 – 2022), 1011 patients hos¬pitalized in the Allergology Clinic at the University Hospital, Pleven, were examined for intestinal protozoa and helminths. Fresh stool samples for microscopic parasitological examination were used. From persons infected with Blastocystis sp. venous blood was obtained for the determination of specific anti-Toxocara antibodies. Of the patients with allergic symptoms examined for intestinal helminths and protozoa, 40 were infected with Blastocystis sp. Out of them, seven (17.5%) were positive for anti-Toxocara IgG antibodies on the ELISA test, and Western blot confirms the diagnosis. All individuals with Blastocystis / Toxocara co-infection were established against the background of chronic urticaria, but no difference was found in the severity of clinical manifestations in comparison with those with only blastocystosis and those without evidence of parasitic infection. The data of our study, give us reason to consider that the establishment of parasitic infections in persons with skin allergic manifestations is not unusual, but in general, it is very difficult to determine whether they are the cause of the allergic condition or are a finding in the course of diagnostic clarification.

Gut butyrate-producers confer post-infarction cardiac protection

The gut microbiome and its metabolites are increasingly implicated in several cardiovascular diseases, but their role in human myocardial infarction (MI) injury responses have yet to be established. To address this, we examined stool samples from 77 ST-elevation MI (STEMI) patients using 16 S V3-V4 next-generation sequencing, metagenomics and machine learning. Our analysis identified an enriched population of butyrate-producing bacteria. These findings were then validated using a controlled ischemia/reperfusion model using eight nonhuman primates. To elucidate mechanisms, we inoculated gnotobiotic mice with these bacteria and found that they can produce beta-hydroxybutyrate, supporting cardiac function post-MI. This was further confirmed using HMGCS2-deficient mice which lack endogenous ketogenesis and have poor outcomes after MI. Inoculation increased plasma ketone levels and provided significant improvements in cardiac function post-MI. Together, this demonstrates a previously unknown role of gut butyrate-producers in the post-MI response.

Anti-Cryptosporidium oocysts polyclonal antibodies for cryptosporidiosis diagnosis and protection

Cryptosporidiosis is an intestinal infection that is triggered by the protozoan parasite Cryptosporidium spp. Cryptosporidium oocysts can spread from one host to another either through direct contact with infected hosts' faeces or through indirect means (consumption of contaminated water or food). Significant numbers of oocysts are produced as a result of the rapid growth of the parasite within the infected hosts. For proper care of cryptosporidiosis, a laboratory diagnosis is necessary. Therefore, this study aimed to produce anti-Cryptosporidium parvum (C. parvum) oocyst immunoglobulin (Ig)G polyclonal antibodies (pAbs). The produced pAbs were used in the detection of C. parvum oocysts antigens in stool and serum samples of infected calves. Moreover, pAbs were tested in protection of balb-c male mice from cryptosporidiosis infection. C. parvum oocysts were used in the preparation of antigens to be used in the immunization of New Zealand white rabbits. pAb was purified by ammonium sulphate precipitation method, caprylic acid purification method and diethylaminoethyl (DEAE) anion exchange chromatographic method. Sandwich enzyme-linked immunosorbent assay (ELISA) (using prepared pAb) scored higher sensitivity (85% and 95% for serum and stool samples) than that (80%) of microscopic examination of stool samples. Moreover, pAb significantly reduced the oocysts shedding, decreased inflammatory cytokines and enhanced the loss in the body weight of protected animals. The prepared pAb succeeded in the diagnosis of cryptosporidiosis in calves with high sensitivity either in the serum or stool samples. Our results indicated the usefulness of using the prepared pAb in protection against cryptosporidiosis.

The Association Between Vitamin D Levels and Intestinal Parasites

Immune system is known to be affected by vitamin D deficiency. In this study, we investigated whether there was a significant association between vitamin D levels and intestinal parasites. A total of 239 patients treated at the internal medicine outpatient clinics with gastrointestinal complaints were included in this study. Demographic characteristics of the patients were recorded. Examination of stool samples were repeated three times within 10 days. Cellophane band method, native-lugol, sedimentation and modified Kinyoun's acid fast stain methods were used for the detection of parasites in the stool samples. The samples were prepared, stained and examined under a microscope. Vitamin D levels were examined in the blood samples collected from the patients. Patients having vitamin D level under 20 ng/ml were diagnosed with vitamin D deficiency. Binary logistic regression analysis showed that serum vitamin D level was significant informative variable for only Iodamoeba and Cyclospora. No association between vitamin D level and the presence of other intestinal parasites was found. The risk of Iodamoeba positivity was 2.54 times higher in the patients with a serum vitamin D level under 20 ng/ml ($$p&lt;0.01$$). Similarly, the risk of Cyclospora positivity was 2.44 times higher in the patients with a serum vitamin D level under 20 ng/ml compared to those with a serum vitamin D level over 20 ng/ml ($$p&lt;0.01$$). The risk for positivity of Iodamoeba and Cyclospora that are among the intestinal protozoa is high for people with vitamin D deficiency.