Serological and molecular detection of Toxoplasma gondii in ß. thalassemia patients.

Toxoplasma gondii is a worldwide opportunistic protozoan causing life-threatening infection in immunocompromised patients, while frequently asymptomatic in immunocompetent individuals. The current study aimed to detect T. gondii; serologically and molecularly in ß. thalassemia patients and evaluate the association of infection with some hematological parameters in these patients. Blood samples were collected from 100 ß. thalassemia patients. Serological diagnosis of T. gondii using ELISA for IgG and IgM antibodies was performed. Molecular diagnosis by Real-Time (RE) PCR was performed using specifically designed primers amplifying 389 bp fragments of Toxoplasma genome. 45 patients (45%) had anti-Toxoplasma IgG antibodies with no detectable IgM antibodies while both anti-Toxoplasma IgM and IgG antibodies were noticed in 10 patients (10%). IgM only antibodies were discovered in two cases (2%). The total seropositivity rate among patients was 57%. RE PCR analysis revealed Toxoplasma DNA in 20% out of 100 patients. PCR and serological examination showed slight agreement. A statistically significant relation was observed between the results of IgG and IgM ELISA and PCR for the detection of T. gondii infection among patients with ß. thalassemia. None of the studied risk factors (age, gender, contact with cats, consumption of undercooked meat) or hematological parameters (ESR, anemia degree, ferritin level, type of blood transfusion, spleen status) showed statistically significant association with Toxoplasma infection. It can be concluded that patients with thalassemia have a high risk of infection with T. gondii. RE PCR should be used as a diagnostic method in association with serology especially in immunocompromised patients to increase sensitivity.

Psychological impact of COVID-19 among undergraduate students in Jordan: A cross-sectional study.

Background: Psychopathology could be brought on by an immune system disruption triggered by an infection. Psychiatric residual effects have been noted following prior coronavirus outbreaks. Method: This study is a cross-sectional study that examined the psychopathological influence of the COVID-19 pandemic among survivor students compared to students who have never been diagnosed as positive COVID-19 patients (i.e., healthy students). Further, the study identified the relationship between sociodemographic profiles and mental health. To examine the significance of psychological health differences we used the DASS-21, to measure depression, anxiety, and stress scales. There were seven questions to assess each dimension within the psychological scales. The respondents were selected randomly via online sources and completed the online self-reporting questionnaire between August 23 and October 2, 2021. Results: A total of 750 students completed the online questionnaire, with 677 participants (90.27% of those originally surveyed) included in the final analysis of this study. The mean score of the stress scale for healthy students (M = 16.88, SD = 12.3, N = 537) demonstrated a significantly lower mean compared to COVID-19 survival students (M = 21.7, SD = 11.01, N = 140), t(675) = -4.22, p < 0.001. The mean of the depression scale for healthy students (M = 16.18, SD = 11.94) demonstrated significantly lower mean compared to COVID-19 survival students (M = 20.91, SD = 11. 90), t(675) = -4.18, p < 0.001. The mean of the anxiety scale for healthy students (M = 12.50, SD = 10.72) demonstrated significantly lower mean compared to COVID-19 survival students (M = 19.43, SD = 11. 25), t(675) = -6.74, p < 0.001. Conclusion: University students who were infected with COVID-19 had a traumatic experience, and thus developed psychological symptoms greater than students with no history of this infection.

Molecular diagnosis and subtyping of Blastocystis sp.: Association with clinical, colonoscopic, and histopathological findings.

Introduction: Blastocystis sp. is the most common parasitic infestation in humans. However, its pathogenicity remains controversial. Our aim was to study the prevalence of Blastocystis sp. parasite subtypes in patients with gastrointestinal manifestations referred for colonoscopy and assess possible correlation with clinical, colonoscopic, and histopathological findings. Methodology: One hundred patients with gastrointestinal manifestations referred for colonoscopy were enrolled. Stool samples were collected and examined both microscopically and by real-time quantitative polymerase chain reaction (qPCR) for detection of Blastocystis sp. Subtyping was done for positive samples by qPCR and confirmed by sequencing. Results: qPCR sensitivity far exceeded microscopy in detection of Blastocystis sp. (58% vs. 31%, agreement 38.5%). The most commonly detected subtype was 3 (50%), followed by 2 (32.8%) and 4 (13.8%). Abdominal pain was the most common clinical symptom; inflammation and colitis were the most common abnormal colonoscopic and histopathological findings. The most frequent subtype encountered in those findings was Subtype 3. Conclusions: This study confirmed the importance of using qPCR in diagnosis of Blastocystis sp. An association between abnormal clinical, colonoscopic, and histopathological findings on the one hand, and Blastocystis sp. infestation, especially Subtype 3, on the other hand, is also posed. This necessitates further studies to assess the mechanism of association with pathogenicity.

Comparison of gdh polymerase chain reaction-restriction fragment length polymorphism and tpi assemblage-specific primers for characterization of Giardia intestinalis in children.

Background: Giardia is a diarrheagenic eukaryotic parasite that consists of at least eight morphologically identical but genetically distinct genotypes. Human giardiasis is caused mainly by A and B assemblages. Aim and objectives: The study aimed to compare the performance of gdh polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and tpi assemblage-specific primers in genotyping of G. intestinalis. Materials and Methods: Stool samples of 315 children were microscopically screened for G. intestinalis. Positive samples were genotyped using tpi assemblage-specific primers and gdh semi-nested PCR-RFLP techniques. Results: The prevalence of Giardia was 18.1%. The detected genotypes using tpi and gdh approaches were assemblage A (15.8% vs. 12.7%) and assemblage B (36.8% vs. 74.5%) as single infections and mixed assemblages A and B (47.4% vs. 12.7%). The two approaches showed a moderate agreement (kappa index = 0.413, P < 0.001). PCR-RFLP of gdh gene revealed that sub-assemblages BIII and BIV were equally detected (30.9% each). The remaining samples were equally divided between sub-assemblage AII, mixed BIII and BIV, and mixed AII and BIII (12.7% each). A significant association was detected between the retrieved sub-assemblages and the presence of symptoms. Conclusions: Although both approaches confirmed the predominance of assemblage B, the use of assemblage-specific primers is more effective in elucidating the true picture of mixed assemblage infection.

Pre-and post-treatment evaluation of intestinal inflammation in Giardia and Blastocystis infected children: a community-based study.

Giardia intestinalis and Blastocystis hominis cause frequent infections in children in developing countries. However, the role of intestinal inflammation in their pathogenesis is still poorly understood. Faecal calprotectin (FC) level is used as an indicator of intestinal inflammation and neutrophil migration in the intestinal tract. The present study aimed to evaluate intestinal inflammation by measuring FC level among children infected with either G. intestinalis or B. hominis before and after treatment. Stool samples were collected from 282 children inhabiting a rural area in Egypt and examined microscopically for intestinal parasites. FC level was estimated in a group of children infected with G. intestinalis (n = 12) or B. hominis (n = 12) before and 3 weeks after receiving nitazoxanide (200 mg twice daily for 3 days) and compared to a control group (n = 18) of parasite-free children. Cases of mixed infection were excluded. Nitazoxanide cure rate was 83% in both infections with a remarkable reduction of infection intensity in uncured children. The difference in FC levels between infected children and controls was not statistically significant. Also, the difference between the pre- and post-treatment estimations was not statistically significant. Elevated levels were observed before treatment in three children (two infected with G. intestinalis and one with B. hominis) who displayed normal post-treatment levels. Although G. intestinalis and B. hominis infections appear to cause no remarkable intestinal inflammation, they may induce abnormally elevated FC levels in a subset of children.

Detection of Giardia intestinalis assemblages A and B among children from three villages in the West Delta region, Egypt using assemblage specific primers.

Giardia intestinalis is a common diarrheagenic parasite infecting children globally. It has been classified into eight morphologically identical but genetically distinct genotypes. Human infection is mainly associated with A and B assemblages with variable geographical distribution. The present work aimed to study the epidemiology of assemblages A and B in children inhabiting different areas in Lower Egypt. Stool samples were collected from 315 children and examined microscopically for parasitic infections. Giardia positive samples were genotyped using tpi assemblage specific primers. The prevalence of Giardia was 18.1% among the examined children. Mixed assemblages A and B was more common (47.4%) than single assemblage B (36.8%) or A (15.8%). The distribution of different genotypes was significantly associated with the residence area, animal contact, and handwashing habits. A non-significant association was observed between Giardia assemblages and the clinical manifestations. Assemblage B is the predominant genotype among Egyptian children. The distribution of different Giardia assemblages is strongly associated with the studied area and the habits of its people.

Detection of toxoplasmosis in aborted women in Alexandria, Egypt using ELISA and PCR.

Toxoplasmosis is a worldwide infection that can be acquired through the ingestion of tissue cysts in poorly cooked meat, and/or oblivious intake of sporulated oocysts in cat faeces, and transplacental. The infection in pregnant women is mainly asymptomatic. It produces abortion or congenital infection. The present study aimed to test the utility of polymerase chain reaction (PCR) on placental tissues in comparison to enzyme-linked immunosorbent assay (ELISA) to detect infections with Toxoplasma gondii in aborted women presented to Al-Shatby Maternity Hospital, Alexandria University, Egypt. Specific Toxoplasma gondii IgG and IgM were detected in serum by ELISA. Placental tissues from each participant were subjected to DNA extraction and PCR amplification. It was found that overall seroprevalence was 73%, DNA was detected in placenta tissues by using PCR analysis in 46% of cases. {× 2 (p) 18.124(< 0.001)}. Toxoplasma IgG/IgM by ELISA was positive in 23% of the cases, 20% showed amplified DNA by PCR. Positive IgG without IgM was seen in 27% cases, only 2% of them were positive by PCR. Moreover, positive PCR among positive ELISA IgM aborted women was 21 of the 23 cases. Positive PCR was obtained in three seronegative women. Our results showed that PCR sensitivity was 58.90 specificity 88.89, positive predictive value was 93.48%, and negative predictive value 44.44%. Although ELISA assay is still the gold standard of diagnosis of Toxoplasmosis, other diagnostic modalities are highly required particularly in those ELISA seronegative cases. PCR can be used as a sensitive and precise modality.

Intestinal schistosomiasis among preschool and school-aged children in a rural setting near Alexandria: initiative for elimination.

OBJECTIVE: To assess the status of intestinal schistosomiasis among preschool-aged (PSAC) and school-aged children (SAC) and to compare the efficacy of praziquantel (PZQ) in both groups. METHODS: The study was conducted on 400 children; 103 PSAC and 297 SAC. Diagnosis of Schistosoma mansoni was based on triplicate Kato-Katz thick smears from a single stool sample. To identify the missed cases by Kato-Katz, 120 randomly selected negative cases (38 PSAC and 82 SAC) were screened by real-time PCR. All S. mansoni-positive cases by Kato-Katz were treated by crushed PZQ tablets. Four weeks after treatment, the cure rate was assessed by Kato-Katz smears and real-time PCR. RESULTS: The prevalence of S. mansoni with Kato-Katz was 7.8% among PSAC and 7.4% among SAC. Most of children (63.3%) had light-intensity infection. The cure rate was 100% among PSAC by both techniques, and 91%, and 77.2% among SAC by Kato-Katz and real-time PCR, respectively. In the 120 stool samples screened by real-time PCR, S. mansoni prevalence was 25%; 15.8% and 29.3% were among PSAC and SAC respectively. Treated cases showed a lower range of Ct values than untreated cases. Two melting temperature ranges (Tm = 83-87°C and 89-93°C) were recognised among uncured cases which may point to S. mansoni genetic variability. CONCLUSION: Continuous monitoring and inclusion of PSAC in schistosomiasis control programmes are crucial. Real-time PCR and other molecular tools are recommended for evaluation of the true prevalence, assessment of cure and further studies on genetic diversity.

Molecular Characterization of Giardia intestinalis Detected in Humans and Water Samples in Egypt.

BACKGROUND: Giardia intestinalis is a common cause of gastrointestinal illness especially in children of developing countries. Giardia assemblages A and B are the major human infective genotypes. OBJECTIVE: The present study aimed to investigate the role of water supply in the epidemiology of giardiasis via genotyping G. intestinalis detected in diarrheic children and in water samples in Egyptian rural areas. METHODS: Stool samples of 100 diarrheic children, 40 drinking water samples and 10 raw water samples of canals were examined microscopically for Giardia. DNA was extracted from microscopically positive faecal samples and from all of the collected water samples. Amplification of Giardia tpi gene was performed by a nested PCR using assemblage A- and assemblage B-specific primers. Giardia gdh gene was amplified by a heminested PCR. Giardia genotypes were determined by restriction fragment polymorphism (RFLP) analysis of the amplified products. Sequencing of the amplified products was performed in two faecal and two water samples RESULTS: Giardia intestinalis was detected in 24 children, in none of the drinking water samples and in all canal water samples. Giardia sub-assemblage AII was identified in all stool and raw water samples. The RFLP pattern was confirmed in sequenced samples. CONCLUSION: The presence of the same Giardia sub-assemblage in diarrheic children and in raw water samples shows by molecular evidence the potential for waterborne dissemination of Giardia in Egypt. Further studies are needed to monitor cyst levels and infectivity of the genotype detected in water for risk assessment and management.

Irritable bowel syndrome in Egyptian patients: plausible risk factors and association with intestinal protozoa.

The pathogenesis of irritable bowel syndrome (IBS) is not yet clear. Our study suggested parasitic infection and other plausible risk factors among Egyptian IBS patients. We studied 40 IBS patients diagnosed according to Rome III criteria (Group I), 40 with other gastrointestinal symptoms (Group II) and 40 healthy controls (Group III). Stool samples were examined using direct wet smear, sedimentation technique, trichrome stain and immune-chromatographic tests for Cryptosporidium parvum. IBS patients displayed a significantly greater percentage of Blastocystis hominis infection (45%) than non-IBS patients (20%) and healthy controls (10%). Dientamoeba fragilis was identified in two IBS patients. Detection of B. hominis was independent of demographic characters, IBS subtype, Helicobacter pylori infection or medications, but with a positive association with a history of antibiotic intake with IBS.

Communication with invasive mechanically ventilated patients and the use of alternative devices: integrative review.

BACKGROUND: Communication with patients has always been a major concern in nursing care. Invasive mechanically ventilated (IMV) patients suffer from a communication barrier due to the presence of the endotracheal tube (ETT), which makes them unable to communicate through speech. AIM: The purpose of this review is to examine available evidence regarding existing knowledge, skills, perceptions and barriers to IMV patient communication in order to guide the development of strategies that enhance effective communication with these patients. METHODS: A review of the published literature was conducted between January 2010 and December 2016. RESULTS: The literature support clear and concise communication in all areas of care, especially when patients suddenly become speechless. Invasive mechanically ventilated patients want to be heard, have control over their treatment and contribute to decisions concerning their health. CONCLUSION: There is a need for the establishment of an effective nurse -patient communication strategy, which may include determining the mode of communication used by the patient, waiting and giving time to allow a patient to participate in the communication, confirming the message that was communicated with a patient himself/ herself, and the use of assistive and augmented communication to support comprehension when needed.