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Infection & Chemotherapy May 2024Enterocolitis and gastroenteritis remain major health problems, particularly in children living in developing countries. Intestinal protozoa, such as , and are...
BACKGROUND
Enterocolitis and gastroenteritis remain major health problems, particularly in children living in developing countries. Intestinal protozoa, such as , and are frequently associated with these conditions. Amebic colitis can cause serious complications, including fulminant necrotizing colitis, toxic megacolon, extraintestinal amebiasis, and stunting in children. The diagnosis of amoebiasis is challenging, relying on microscopic examination, which cannot distinguish from the nonpathogenic and Therefore, this study aimed to identify intestinal parasites, particularly , their prevalence, and the clinical characteristics of patients admitted for enterocolitis and gastroenteritis at a tertiary-referral hospital.
MATERIAL AND METHODS
A cross-sectional, retrospective study was conducted at a national, tertiary-referral government hospital, in Jakarta. Of the 111 retrieved medical records from hospitalized patients with enterocolitis and gastroenteritis, 54 (48.6%) fecal samples were still available in the laboratory storage and referred to the parasitology laboratory. All fecal samples underwent the following tests: 1) direct stool examination, after staining with 1% Lugol's solution, and using the water-ether concentration method; 2) modified acid-fast staining for coccidian parasites; 3) Jones' culture medium to detect ; 4) copro-antigen assay to detect and and 5) a polymerase chain reaction (PCR) assay to identify . Clinical and demographic data were obtained from the medical records.
RESULTS
Largely, patients (44.1%) were from the cohort of young children ≤5 years old, followed by adults aged 19-60 years old (24.3%). Both cohorts exhibited polyparasitism. Intestinal parasites were detected in 17 out of the 54 samples (31.4%). These included 6 (11.1%), 2 (3.7%),5 (9.2%), 3 (5.5%), 2 (3.7%), and 1 (1.8%) samples that were positive for , , , , and respectively. PCR analysis revealed that 10 samples were positive for infection, eight of which originated from pediatric patients.
CONCLUSION
At a national tertiary-referral hospital in Indonesia, infection was most prevalent among pediatric patients with enterocolitis. and were the two main species identified by PCR. Therefore, PCR assays and fecal occult-blood tests are recommended in cases of enterocolitis and gastroenteritis.
PubMed: 38859717
DOI: 10.3947/ic.2023.0099 -
Parasitology Jan 2020The presence of bacterial DNA in Dientamoeba fragilis DNA extracts from culture poses a substantial challenge to sequencing the D. fragilis genome. However, elimination...
The presence of bacterial DNA in Dientamoeba fragilis DNA extracts from culture poses a substantial challenge to sequencing the D. fragilis genome. However, elimination of bacteria from D. fragilis cultures has proven difficult in the past, presumably due to its dependence on some unknown prokaryote/s. This study explored options for removal of bacteria from D. fragilis cultures and for the generation of genome sequence data from D. fragilis. DNA was extracted from human faecal samples and xenic D. fragilis cultures. Extracts were subjected to 16S ribosomal DNA bacterial diversity profiling. Xenic D. fragilis cultures were then subject to antibiotic treatment regimens that systematically removed bacterial species depending on their membrane structure (Gram-positive or Gram-negative) and aerobic requirements. The impact of these treatments on cultures was assessed by 16S amplicon sequencing. Prior to antibiotic treatment, the cultures were dominated by Gram-negative bacteria. Addition of meropenem to cultures eliminated anaerobic Gram-negative bacteria, but it also led to protozoan death after 5 days incubation. The seeding of meropenem resistant Klebsiella pneumoniae strain KPC-2 into cultures before treatment by meropenem prevented death of D. fragilis cells beyond this 5 day period, suggesting that one or more species of Gram-negative bacteria may be an essential nutritional requirement for D. fragilis. Gram-positive cells were completely eliminated using vancomycin without affecting trophozoite growth. Finally, this study shows that genome sequencing of D. fragilis is feasible following bacterial elimination from cultures as the result of the major advances occurring in bioinformatics. We provide evidence on this fact by successfully sequencing the D. fragilis 28S large ribosomal DNA subunit gene using culture-derived DNA.
Topics: Anti-Bacterial Agents; Bacteria; Bacterial Physiological Phenomena; Culture Techniques; Dientamoeba; Genetic Variation; Genome, Protozoan; RNA, Ribosomal, 16S; RNA, Ribosomal, 28S
PubMed: 31452478
DOI: 10.1017/S0031182019001173 -
Parasitology Research Jun 2023Intestinal parasitic infections are a global health problem that causes morbidity and mortality, especially in children living in rural areas. In this study, stool...
Intestinal parasitic infections are a global health problem that causes morbidity and mortality, especially in children living in rural areas. In this study, stool samples of pediatric patients with gastrointestinal complaints were examined by conventional and molecular methods to determine the prevalence of intestinal parasites. A total of 100 pediatric patients with gastrointestinal complaints and 50 healthy children were included in the study. Stool samples were collected from each child and examined by direct microscopic examination (native-Lugol method), formol-ethyl acetate concentration technique, Kinyoun's acid-fast staining, and Wheatley trichrome staining methods. Real-time PCR was used for the detection of Blastocystis spp. and D. fragilis in the stool samples. Sanger sequencing was used to identify Blastocystis spp. subtypes. One or more intestinal parasites were found in 12% (n = 100) of the patient group and 1% (n = 50) of the control group using conventional techniques. By using real-time PCR, Blastocystis spp. was discovered in 14% (14/100) of the patient group and 8% (4/50) of the control group. There was no significant difference in the frequency of Blastocystis spp. between the two groups. The most prevalent Blastocystis subtype was ST1 and the most frequent allele was a2 among the samples successfully amplified and sequenced. D. fragilis was detected in 17% (17/100) of the patient group and 8% (4/50) of the control group by real-time PCR. The prevalence of D. fragilis was not significantly different between the patient and control groups, as well. Blastocystis spp. and D. fragilis were found in high prevalence in pediatric patients with gastrointestinal complaints in this study. Although the role of these protists as a pathogen in humans is still controversial, it is supposed to the presence of the parasites are associated with gastrointestinal disorders such as diarrhea, abdominal pain, nausea, and vomiting. More case-control studies are needed to understand the pathogenic or commensal role of these parasites on the intestinal microbiota, especially in both patients with gastrointestinal disorders and healthy individuals.
Topics: Animals; Humans; Child; Parasites; Blastocystis Infections; Feces; Intestinal Diseases, Parasitic; Blastocystis; Gastrointestinal Diseases; Prevalence
PubMed: 37036521
DOI: 10.1007/s00436-023-07836-0 -
Enfermedades Infecciosas Y... Oct 2022Dientamoeba (D.) fragilis is a common intestinal protozoan with an unresolved clinical significance. The association between D. fragilis and the etiology of...
INTRODUCTION
Dientamoeba (D.) fragilis is a common intestinal protozoan with an unresolved clinical significance. The association between D. fragilis and the etiology of gastrointestinal symptoms in children is unclear. Metronidazole is often used for treatment. The aims of this study are to clarify the clinical relevance of D. fragilis in children with gastrointestinal symptoms, and to determine the clinical and microbiological efficacy of metronidazole in D. fragilis-infected children with gastrointestinal complaints.
METHODS
A prospective case-control study was performed from October 2017 to February 2019. A total of 106 individuals aged 1-17 were included. Out of the 106; 59 showed gastrointestinal symptoms (case group), and 47 were without gastrointestinal symptoms (control group). We excluded 2 patients from the case group. D. fragilis was diagnosed by real-time PCR in stool samples. A 10-day course of oral Metronidazole was prescribed in D. fragilis positive children with GI symptoms. Clinical data before and after the treatment as well as peripheral eosinophilia in previous blood samples, were recorded.
RESULTS
Of the 104 participants, D. fragilis was found in 17 (29.8%) children from the case group, whereas in the control group the parasite was detected in 11 patients (23.4%) with an odds ratio (OR) of 1.39 (IC 95% 0.53-3.75, p=0.46). The most prevalent clinical manifestation was abdominal pain (46/57, 80.7%). Seventeen cases with a positive PCR received anti-parasitic treatment according to the established protocol, although during the collection period we received only 11 stool samples to perform the post-treatment follow-up. The PCR of the D. fragilis remained positive in 3 patients (3/11, 27.27%). Despite achieving the eradication of the parasite, 4/8 patients (50%) continued with digestive symptoms.
CONCLUSIONS
According to our study there were no differences between the D. fragilis infection in children with or without gastrointestinal symptoms. No relation was found between the clinical and microbiological responses after said D. fragilis treatment. Therefore, we conclude that it is not justified to look specifically for D fragilis in pediatric patients with abdominal symptoms.
Topics: Case-Control Studies; Child; Dientamoeba; Dientamoebiasis; Humans; Metronidazole; Real-Time Polymerase Chain Reaction
PubMed: 36195407
DOI: 10.1016/j.eimce.2022.03.013 -
Turkiye Parazitolojii Dergisi May 2022Parasitic infections emerge as a significant health problem, especially in underdeveloped and developing countries. Epidemiological data play an important role in taking...
OBJECTIVE
Parasitic infections emerge as a significant health problem, especially in underdeveloped and developing countries. Epidemiological data play an important role in taking effective measures against parasitic diseases.
METHODS
Clinical samples (stool, blood, bone marrow and tissue samples, etc.) that were sent to Hacettepe University Hospitals Parasitology Laboratory between 2014 and 2019 were analyzed retrospectively.
RESULTS
The positivity rates of the parasites detected in this study are as follows; sp. (71.6%), (13.3%), (4.7%), spp. (1.9%), (1.8%) and spp. (0.3%). In this study, four of the patients were found to be positive for spp. and two patients for and four patients for spp. cysts and/or trophozoites examined by Trichrome staining in our study were not detected within six years.
CONCLUSION
According to this data and in the light of the results obtained from different regions of our country, it will be possible to properly direct the necessary strategies for the diagnosis, treatment of parasitic infections and the implementation of preventive measures.
Topics: Animals; Dientamoeba; Faculty; Feces; Giardia lamblia; Humans; Intestinal Diseases, Parasitic; Prevalence; Retrospective Studies
PubMed: 35604188
DOI: 10.4274/tpd.galenos.2022.84429 -
Microbial Pathogenesis Feb 2020Calprotectin is a protein that is mostly released from neutrophils, monocytes, macrophages and submucosal epithelial cells. Fecal calprotectin (f-CP) is a marker of...
Calprotectin is a protein that is mostly released from neutrophils, monocytes, macrophages and submucosal epithelial cells. Fecal calprotectin (f-CP) is a marker of intestinal inflammation. There are some discussions about the pathogenicity of D. fragilis in the gastrointestinal tract. In this study, we investigated whether f-CP level is a factor supporting the pathogenicity of D. fragilis. The f-CP levels were evaluated in patients with only D. fragilis positive in comparison with healthy controls. Moreover, the levels of f-CP were investigated in fecal samples of D. fragilis negative patients with gastrointestinal complaints. The fecal samples were collected from three groups. Three groups of fecal samples were examined directly microscopy, trichrome staining, cultivation, enzyme immunoassay (EIA) and real-time PCR assay. In the first group (Group 1, n = 34), patient stool samples with gastrointestinal symptoms (without other pathogens) found only with D. fragilis were included. In the second group (Group 2, n = 31), there were patients' stool samples with gastrointestinal symptoms that D. fragilis was negative (but there may be other pathogenic agents). In the control group (Group 3, n = 23), we used fecal samples collected from healthy volunteers without any infection or gastrointestinal complaints. The collected fecal samples were stored at -20 °C until analysis. Levels of f-CP were determined by using human calprotectin ELISA kits. Total of 88 patients were enrolled in three different groups. We obtained f-CP levels as follows: 33.40 ng/mg protein in the group 1, 15.99 ng/mg protein in the group 2 and 1.54 ng/mg protein in the group 3. Statistically significant difference in f-CP levels of the group 1 and the group 2 were obtained when compared with healthy controls (p < 0.0001). However, the f-CP levels of the group 1 were not significantly different from the group 2 (p > 0.99). In conclusion, increased levels of f-CP are shown as a marker of an inflammatory disease of the lower gastrointestinal tract in infected humans. There is continues controversy about the pathogenicity of D. fragilis in symptomatic and asymptomatic patients. The findings of this study contribute to the ongoing debate about the pathogenicity of D. fragilis. In our study, the potential pathogenicity of D. fragilis is associated with increased f-CP concentrations with parasite detection in the fecal samples and therefore we assume that the parasite is not only a harmless commensal. In summary, higher levels of f-CP found in D. fragilis positive patients suggest the importance of researches that support the pathogenicity of indicated parasite.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers; Child; Child, Preschool; Dientamoeba; Dientamoebiasis; Disease Susceptibility; Feces; Female; Humans; Leukocyte L1 Antigen Complex; Male; Middle Aged; Prognosis; Sensitivity and Specificity; Symptom Assessment; Young Adult
PubMed: 31730996
DOI: 10.1016/j.micpath.2019.103868 -
International Journal of Obesity (2005) Jan 2022Obesity is a chronic disease whose pathogenesis has been related to changes in the intestinal microbiota. Yet, the role of protozoa and other unicellular eukaryotic...
BACKGROUND
Obesity is a chronic disease whose pathogenesis has been related to changes in the intestinal microbiota. Yet, the role of protozoa and other unicellular eukaryotic parasites in this microenvironment is still largely unknown. Their presence within the gut ecosystem in obese subjects warrants further study, as well as their influence on the host metabolism and comorbidities.
METHODS
Herein, a single center, cross-sectional study of 104 obese individuals was performed to assess the presence of six intestinal unicellular parasites in stool using a commercially available kit, and to evaluate its relationship with the presence of abdominal symptoms, metabolic comorbidities, variations in body composition and nutritional deficiencies.
RESULTS
The overall parasitic colonization rate was 51%, with Blastocystis sp., identified as the most frequent (44.2%), followed by Dientamoeba fragilis (11.5%) and Giardia intestinalis (8.7%), and significantly related to the consumption of ecological fruits and vegetables. Contrary to what previous studies pointed out, colonization with parasites species was significantly associated with fewer abdominal symptoms and depositions per day. The presence of parasites did not correlate with any nutritional deficiencies nor differences in body composition, while it did with significant lower HOMA-IR levels and a lower trend towards metabolic syndrome.
CONCLUSION
Obese subjects frequently harbor unicellular enteric parasites, apparently without clinical nor nutritional harm. This evidence suggests that carrying these microorganisms, from an endocrinological perspective, has a beneficial effect, especially on insulin resistance and possibly on the development of related comorbidities.
Topics: Adult; Animals; Blastocystis; Cross-Sectional Studies; Female; Gastrointestinal Tract; Humans; Male; Obesity, Morbid; Parasites; Spain
PubMed: 34650200
DOI: 10.1038/s41366-021-00980-6 -
International Journal For Parasitology.... Dec 2019Dientamoeba fragilis is a trichomonad parasite of the human intestine that is found worldwide. However, the biological cycle and transmission of this parasite have yet...
Dientamoeba fragilis is a trichomonad parasite of the human intestine that is found worldwide. However, the biological cycle and transmission of this parasite have yet to be elucidated. Although its pathogenic capacity has been questioned, there is increasing evidence that clinical manifestations vary greatly. Different therapeutic options with antiparasitic drugs are currently available; however, very few studies have compared the effectiveness of these drugs. In the present longitudinal study, we evaluate 13,983 copro-parasitological studies using light microscopy of stools, during 2013-2015, in Terrassa, Barcelona (Spain). A total of 1150 (8.2%) presented D. fragilis. Of these, 739 episodes were finally analyzed: those that involved a follow-up parasitology test up to 3 months later, corresponding to 586 patients with gastrointestinal symptoms (53% under 15 years of age). Coinfection by Blastocystis hominis was present in 33.6% of the subjects. Our aim was to compare therapeutic responses to different antiparasitic drugs and the factors associated with the persistence of D. fragilis post-treatment. Gender, age, and other intestinal parasitic coinfections were not associated with parasite persistence following treatment. Metronidazole was the therapeutic option in most cases, followed by paromomycin: 65.4% and 17.5% respectively. Paromomycin was found to be more effective at eradicating parasitic infection than metronidazole (81.8% vs. 65.4%; p = 0.007), except in children under six years of age (p = 0.538). Although Dientamoeba fragilis mainly produces mild clinical manifestations, the high burden of infection means we require better understanding of its epidemiological cycle and pathogenicity, as well as adequate therapeutic guidelines in order to adapt medical care and policies to respond to this health problem.
Topics: Adolescent; Adult; Antiprotozoal Agents; Child; Dientamoeba; Dientamoebiasis; Feces; Female; Humans; Longitudinal Studies; Male; Metronidazole; Middle Aged; Paromomycin; Spain; Treatment Outcome; Young Adult
PubMed: 31759244
DOI: 10.1016/j.ijpddr.2019.10.005 -
Journal of Pediatric Gastroenterology... Feb 2022
Topics: Child; Dientamoeba; Feces; Humans; Microbiota
PubMed: 34338236
DOI: 10.1097/MPG.0000000000003267 -
Turkiye Parazitolojii Dergisi Aug 2021() is a flagellated protozoan with an amoeba-like morphology, located in the gastrointestinal tract. The hypothesis was that the parasite was transported by () eggs....
OBJECTIVE
() is a flagellated protozoan with an amoeba-like morphology, located in the gastrointestinal tract. The hypothesis was that the parasite was transported by () eggs. This study aimed to determine the association of and with the genotypes of the identified strain of . Results of trichrome staining were compared with those of polymerase chain reaction (PCR), which is widely used in the diagnosis of .
METHODS
A total of 391 samples were obtained. The stool and cellophane slide samples were sent together to the Parasitology Department Laboratory, Faculty of Medicine, Aydın Adnan Menderes University, between 1 October 2017 and 1 October 2018. Stool samples of all patients with (n=74) and without (n=74) infection were used. All samples were examined for the presence of by trichrome staining and PCR. The 18S ribosomal RNA region of isolates was sequenced. Demographic characteristics and clinical findings of the patients were evaluated.
RESULTS
was detected in 42 (28.37%) of 148 samples; 28 (66.6%) of them were detected in patients with infection. The coexistence of two parasites was significant (p<0.05). All isolates sequenced were genotype 1. No significant relationship was found between the presence of parasites and clinical findings, living area and gender (p>0.05).
CONCLUSION
is frequently associated with , so the presence of should be also considered in affected patients. The use of high-sensitivity molecular methods such as PCR is important in preventing false results.
AMAÇ
(), amip benzeri morfolojiye sahip, gastrointestinal yerleşimli, kamçılı bir protozoondur. Parazitin () yumurtalarıyla taşındığı hipotezi kabul görmektedir. Çalışmamızda ve birlikteliğini incelemek, bulunan ’lerin genotiplerini belirlemek ve tanısında yaygın olarak kullanılan trikrom boyama ile polimeraz zincir reaksiyon (PZR) yöntemlerini karşılaştırmak amaçlanmıştır.
YÖNTEMLER
Çalışmamızda Aydın Adnan Menderes Üniversitesi Tıp Fakültesi, Parazitoloji Laboratuvarı’na 1 Ekim 2017-1 Ekim 2018 tarihleri arasında dışkı ve selofan lam örneği birlikte gönderilmiş toplam 391 olgu örneği incelenmiştir. Selofanlı lam örneklerinde saptanan tüm gönüllü olguların (74 olgu) dışkı örneği ile negatif 74 olgunun dışkı örneği çalışılmıştır. Tüm dışkılar trikrom boyama ve PZR yöntemleri ile varlığı açısından incelenmiştir. Saptanan izolatlarının 18S ribozomal RNA bölgesi sekanslanmıştır. Olguların demografik özellikleri ve kliniği değerlendirilmiştir.
BULGULAR
Toplam 148 olgunun 42’sinde (%28,37) saptanmıştır. pozitif olan 42 olgunun %66,6’sını pozitif olgular oluşturmuş ve iki parazitin birlikteliği anlamlı bulunmuştur (p<0,05). Sekanslanan tüm izolatlar genotip 1 olarak saptanmıştır. Klinik bulgular, yaşanılan bölge ve cinsiyet ile parazit varlığı arasında anlamlı bir ilişki saptanamamıştır (p>0,05).
SONUÇ
Araştırmamızda ’in sıklıkla ile birliktelik gösterdiği ve bu olgularda varlığına ayrıca dikkat edilmesi gerektiği vurgulanmıştır. Yanlış sonuçları engellemede, yüksek duyarlılığa sahip PZR gibi yöntemlerin önemi bir kez daha görülmüştür.
Topics: Animals; Dientamoeba; Dientamoebiasis; Enterobiasis; Enterobius; Feces; Humans; Polymerase Chain Reaction
PubMed: 34346875
DOI: 10.4274/tpd.galenos.2021.7257