Occult Amebiasis among Pediatric with Enterocolitis in National Referral Hospital in Indonesia.

BACKGROUND: Enterocolitis and gastroenteritis remain major health problems, particularly in children living in developing countries. Intestinal protozoa, such as Entamoeba histolytica, Blastocystis, and Cyclospora, 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 E. histolytica from the nonpathogenic E. dispar and E. moshkovskii. Therefore, this study aimed to identify intestinal parasites, particularly Entamoeba, 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 Blastocystis; 4) copro-antigen assay to detect Cryptosporidium and Giardia; and 5) a polymerase chain reaction (PCR) assay to identify Entamoeba. 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 Blastocystis, E dispar, E. histolytica, E. moshkovskii, Cryptosporidium, and Dientamoeba fragilis, respectively. PCR analysis revealed that 10 samples were positive for Entamoeba infection, eight of which originated from pediatric patients. CONCLUSION: At a national tertiary-referral hospital in Indonesia, Entamoeba infection was most prevalent among pediatric patients with enterocolitis. E. histolytica and E. moshkovskii were the two main species identified by PCR. Therefore, PCR assays and fecal occult-blood tests are recommended in cases of enterocolitis and gastroenteritis.

Unspecific congenital toxoplasmosis in a two-month-old baby.

A two-month-old baby boy diagnosed with unspecific congenital toxoplasmosis was referred by a pediatrician to the Clinical Parasitology referral center at the Faculty of Medicine, Universitas Indonesia. Baby was post-hospitalized in the NICU and required ventilation support for one month. Furthermore, there was history of from various medical conditions, such as intracranial bleeding, convulsion, hypertrophic cardiomyopathy, retinopathy, and renal failure. After two month, there was no significant weight gain, anti-Toxoplasma IgM showed positive results, and anti-Toxoplasma IgM and IgG of the mother were also positive. Baby and mother were successfully treated with pyrimethamine, cotrimoxazole, and folinic acid for one month. At 2 years, there signs of normal motoric, eye, and hearing development with underdeveloped kidneys. Therefore, pre-pregnancy counseling and education aimed at preventing toxoplasmosis during pregnancy should be increased and conducted routinely by health workers or trained cadres to reduce the risk of fetal defects.

Asymptomatic Cryptosporidiosis in Children Living with HIV.

Children living with human immunodeficiency virus (HIV) have an increased risk of opportunistic Cryptosporidium infection. Cryptosporidium usually causes chronic diarrhea that may lead to impaired growth and cognitive function in children. This study aimed to estimate the prevalence of cryptosporidiosis in children, describe its clinical characteristics, and the risk factors. A cross-sectional study involving children aged 6 months to 18 years old with confirmed HIV infection was carried out in Sardjito General Hospital, Yogyakarta. Diagnosis of cryptosporidiosis was made by PCR of 18S rRNA after being screened by microscopic examination. The clinical characteristics and risk factors were obtained from medical records and structured questionnaires. A total of 52 participants were included in the final analysis. The prevalence of cryptosporidiosis was 42.3%. Approximately 68% of the HIV children with cryptosporidiosis were asymptomatic, while those who reported symptoms showed weight loss and diarrhea. Independent risk factors of cryptosporidiosis were diarrhea (AOR 6.5; 95% CI 1.16-36.67), well water as drinking water source (AOR 6.7; 95% CI 1.83-24.93), and drink untreated water (AOR 5.8; 95% CI 1.04-32.64). A high prevalence of asymptomatic cryptosporidiosis was observed among children with HIV infection and PCR screening of Cryptosporidium in high-risk children is advisable.

Treatment of human intestinal cryptosporidiosis: A review of published clinical trials.

The global burden of diarrhea caused by Cryptosporidium parasite is underestimated. In immunocompromised hosts, chronic and severe presentation of intestinal cryptosporidiosis can result in long-term morbidity and high illness costs. The evidence of effective treatments for cryptosporidiosis has been lacking. We reviewed the published clinical trials to bring forward the feasible therapeutic options of human cryptosporidiosis in various populations and settings according to clinical improvement and parasite clearance rates. A total of 42 studies consisting of the use of nitazoxanide, paromomycin, macrolides, somatostatin analogues, letrazuril, albendazole, rifaximin, miltefosine, clofazimine, and colostrum were included in the review. The trials were mostly conducted in small number of individuals infected with human immunodeficiency virus (HIV), and there is inadequate data of controlled trials to suggest the use of these treatment modalities. Nitazoxanide was reported to be highly efficacious only in immunocompetent hosts and was found to be superior to paromomycin in the same group of patients. Macrolides showed no effective results in both clinical and parasitological improvement. Human bovine colostrum should possibly be administered as one of complementary therapeutic modalities along with other antimicrobials to reach optimal parasite eradication. Other trials of therapeutic modalities were terminated due to futility. Currently, available data is intended to aid the development of strategies for improving access to treatments in different clinical settings, as well as to help guide further studies on treatments of human intestinal cryptosporidiosis.

In vitro experiments of Pediculus humanus capitis (Phthiraptera: Pediculidae) resistance to permethrin and 6-paradol in East Jakarta: Detoxification enzyme activity and electron microscopic changes in lice.

BACKGROUND AND AIM: Pediculus humanus capitis, the human head louse, remains a global health problem. This study evaluated the resistance of head lice to permethrin and 6-paradol mediated by in vitro detoxification enzyme activity experiments and to describe physical changes in the lice using scanning electron microscopy (SEM). MATERIALS AND METHODS: The adult stages of P. h. capitis were collected from patients exposed to 1% permethrin and three different concentrations of 6-paradol (0.00005%, 0.0001%, and 0.00015%) using a filter paper diffusion bioassay. Healthy P. h. capitis adults served as the control. The in vitro bioassays were conducted after 10, 20, 30, and 60 min of exposure. The activities of acetylcholinesterase (AChE), glutathione S-transferase (GST), and oxidase were analyzed. Physical changes in the lice were analyzed using SEM. RESULTS: Permethrin and 6-paradol exhibited low toxicity against the lice. At 60 min, 1% permethrin had killed 36.7% of the lice present, while 6-paradol had killed 66.7-86.7%. Permethrin induced significantly elevated AChE, GST, and oxidase activity; 6-paradol also caused significantly elevated AChE, GST, and oxidase activity. Permethrin did not cause any ultrastructural morphological changes on the lice, while 6-paradol severely damaged the head, thorax, respiratory spiracles, and abdomen of the dead lice. CONCLUSION: This in vitro experimental of P. h. capitis is the first study to report P. h. capitis in East Jakarta shows complete resistance to permethrin and 6-paradol, and to describe the associated increase in AChE, GST, and oxidase activity. It was observed that 6-paradol severely damaged the head, thorax, respiratory spiracles, and abdomen of the dead lice.

Toxoplasma gondii SAG2 type III in an atypical presentation of ocular toxoplasmosis in Indonesia.

OBJECTIVE: This study aimed to perform genotyping of Toxoplasma gondii strain or variant causing atypical toxoplasmic uveitis in Indonesian patients. METHODS: Ocular fluid samples originating from 46 uveitis patients with non-specific ocular manifestations were analysed for Toxoplasma infection by PCR of the B1 locus. The clonal type was determined by amplification, sequencing and phylogenetic analysis of SAG2 and GRA6 loci in B1-positive samples. Clinical data were obtained from the medical records. RESULTS: Pan uveitis was the most frequent manifestation (65.2%) and mostly unilateral (76.1%). PCR of the B1 locus identified eight positive subjects (12.5%); six had panuveitis and two of these had diabetes mellitus. Phylogenetic analysis with maximum likelihood of the SAG2 locus in the B1-positive samples resulted in Toxoplasma gondii SAG2 type III allele. No positive result was obtained from the PCR of GRA6 locus. CONCLUSION: Toxoplasma gondii SAG2-type III allele was identified in an atypical presentation of toxoplasmic uveitis in Indonesia.

Blastocystis ST-1 is associated with Irritable Bowel Syndrome-diarrhoea (IBS-D) in Indonesian adolescences.

OBJECTIVE: IBS is a functional gastrointestinal disorder and commonly presents in children and adolescences, presented as diarrhoea, constipation or mixed type. Blastocystis is a common intestinal protozoa found worldwide, which pathogenicity is still controversial. This study aimed to identify the risk factors of IBS, the association between IBS types with Blastocystis subtypes and analyse Blastocystis pathogenicity. DESIGN: A comparative cross-sectional study was conducted among senior high school students. Rome III Criteria for IBS diagnosis, questionnaires on the risk factors of IBS and types of IBS were recorded. Students were further selected and classified into IBS and non-IBS groups to analyse the association between IBS, IBS types with Blastocystis infection and its subtypes. Direct microscopic stool examination to identify single Blastocystis infection was performed, followed by culture in Jones' medium, PCR, sequencing of 18S rRNA and phylogenetic analysis to determine Blastocystis subtype. Data was analysed using SPSS v22.0 and P value <0.05 was considered statistically significant (95% confidence intervals). RESULTS: IBS was found in 30.2% of 454 students, consisted of 33.3% IBS Diarrhoea, 27.7% IBS Mixed, 27.7% IBS Unclassified and 11.1% IBS Constipation. Major risk factors to IBS consisted of family history of recurrent abdominal pain, abuse, bullying and female gender in respective order (OR 3.6-2.1). Blastocystis ST-1 was significantly associated to IBS-D with 2.9 times risk factor. CONCLUSIONS: Blastocystis infection is a risk factor to develop IBS-D type in adolescence; Blastocystis ST-1 can be regarded as a pathogenic subtype.

Effect of Cellulase Enzyme Treatment on Cyst Wall Degradation of Acanthamoeba sp.

AIM: The goal of this study is to know the potential of cellulase in the degradation of cyst wall Acanthamoeba sp. METHODS: Sample of Acanthamoeba sp. obtained from isolate collection of Department of Parasitology FKUI of which two samples come from patient and one sample is from environment. All three samples were cultured using non-nutrient agar (NNA) media and identified by PCR and sequencing. The concentration of cellulase concentration used was 50 U, 100 U, 150 U, 200 U, 250 U, and 300 U with the incubation time used being 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours. Furthermore, treatment results with the most optimum concentration and incubation time were observed by using SEM to see changes in the surface of the walls of the cyst. A cysticidal test was performed to determine the effectiveness cysticidal action of disinfectant solution, cellulase, and the combination of disinfectant solution and cellulase in killing Acanthamoeba sp. cyst assessed by their viability value. RESULTS: The most optimal cellulase concentration in killing Acanthamoeba sp. cysts was 300 U with an incubation time of 24 hours. Percentage of viability of Acanthamoeba sp. which was exposed to a disinfectant solution for 24 hours was 95%, cellulase alone for 24 hours 75%, and the combination of cellulase and disinfectant solution for 24 hours 25%. CONCLUSIONS: Cellulase is capable of degrading Acanthamoeba sp. cyst wall. Optimal cellulase concentration in degrading Acanthamoeba sp. cyst wall is 300 U with an optimal incubation time being 24 hours. The addition of cellulase to the disinfectant solution has the potential to increase the effectiveness of the disinfectant solution because cellulase is capable of degrading the cyst wall allowing the disinfectant solution to enter and kill Acanthamoeba sp. cysts.