Prevalence and genetic characterization of Cryptosporidium spp. In diarrheic children from Gonbad Kavoos city, Iran

Background: Cryptosporidium is an intestinal protozean parasite causing water-borne and foodborne outbreaks of diarrheal diseases. The present study was per-formed in order to find prevalence and subtypes of Cryptosporidium among children with diarrhea in Gonbad Kavoos City, Northern Iran. Methods: Diarrheic samples were collected from 547 children. The initial parasi-tological diagnosis was made based on detection of oocysts using the modified Ziehl-Neelsen acid-fast staining method. The positive microscopically samples were selected for sequence analysis of partial 60 kDa glycoprotein (gp60) gene. Results: Out of 547 collected samples, 27 (4.94) were positive for Cryptosporid-ium oocysts. Fifteen from 27 positive samples successfully amplified in PCR. Se-quences analysis of gp60 gene in 15 Cryptosporidium isolates revealed that all of them (100) were C. parvum. The results showed three subtypes of IIa subtype family (7 cases) including IIaA16G2R1, IIaA17G1R1, IIaA22G3R1 and one subtype of IId subtype family (8 cases). The most common allele was IId A17G1d (53.3). Conclusion: The predominance of zoonotic subtype families of C. parvum species (IIa, IId) in the present study is in concordance with previous studies in Iran and emphasizes the significance of zoonotic transmission of cryptosporidiosis in the country. © 2015, Tehran University of Medical Sciences (TUMS). All rights Reserved

Discrimination of Entamoeba spp. in children with dysentery

Aim: The present study was performed in order to differentiate E. histolytica and E. dispar in children from Gorgan city, using a PCR method. Background: Differential detection of two morphologically indistinguishable protozoan parasites Entamoeba histolytica and E. dispar has a great clinical and epidemiological importance because of potential invasive pathogenic E. histolytica and non-invasive parasite E. dispar. Patients and methods: One hundred and five dysentery samples were collected from children hospitalized in Taleghani hospital in Gorgan city. The fecal specimens were examined by light microscopy (10X then 40X) to distinguish Entamoeba complex. A single round PCR amplifying partial small-subunit rRNA gene was performed on positive microscopy samples to differentiate E. histolytica/ E. dispar and E. moshkovskii from each other. Results: Twenty-five specimens (23.8%) were positive for Enramoeba complex in direct microscopic examination. PCR using positive controls indicated E. histolytica and E. dispar in two (2/25, 8%) and three (3/25, 12%) samples, respectively. Conclusion: There is a warrant to performing molecular diagnosis for stool examination at least in hospitalized children in order to prevent incorrect reports from laboratories and consequently mistreating by physicians. © 2014 RIGLD, Research Institute for Gastroenterology and Liver Diseases

First Report of Vannellidae Amoebae (Vannella Spp.) Isolated From Biofilm Source

Background: Members of the Vannellidae family are free-living amoebae (FLA) distributed mainly in water and soil sources. The present study reports the first isolation of this genus in the biofilm source from hospital environment in Tehran, Iran.Methods: Biofilm samples were collected from hospital environment. Cultivation was performed in non-nutrient agar covered with a heat-killed Escherichia coli. Cloning of the suspected amoe­bae was done. PCR amplification and Homology analysis using the Basic Local Alignment Search Tool (BLASTn) was performed to search for the most similar reference sequences.Results: Microscopic examination showed numerous fan-shaped amoebae and peculiar cysts differ­ent to the usual shape of typical FLA. Sequence analysis of the PCR- product revealed that the suspected amoebae are highly homologous with Vannella spp. gene (99% identity and 100% query coverage) available in the gene bank database.Conclusion: Although Vannella spp. is not proved to be pathogenic itself, but they are capable of har­boring pathogenic intracellular organisms such as Microsporidian parasites. Thus, identifica­tion of such amoebae can be of clinical importance, as they could lead to transmission of other pathogens to human

Prevalence of Intestinal Parasites in Bakery Workers in Khorramabad, Lorestan Iran

Background: Food contamination may occur through production, processing, distribution and prepara­tion. In Iran especially in Khorramabad, 33° 29' 16" North, 48° 21' 21" East, due to kind of nutrition, cul­ture and economic status of people, bread is a part of the main meal and the consumption of bread is high. In this study, the bakery workers were studied for determining of intestinal parasites prevalence.Methods: The study was carried out during September to November 2010 in Khorramabad. All the 278 baker­ies and the bakery workers including 816 people were studied in a census method and their feces were examined for the presence of parasites by direct wet-mount, Lugol's iodine solution, and formalde­hyde-ether sedimentation, trichrome staining, and single round PCR (For discrimination of Entamoeba spp).Results: Ninety-six (11.9%) stool specimens were positive for different intestinal parasites. Intestinal para­sites included Giardia lamblia 3.7%, Entamoeba coli 5.5%, Blastocystis sp. 2.1%, Entamoeba dispar 0.4%, Hymenolepis nana 0.1%, and Blastocystis sp. 0.1%.Conclusion: In order to reduce the contamination in these persons, some cases such as stool exam every three months with concentration methods, supervision and application of accurate health rules by health ex­perts, training in transmission of parasites are recommended

Molecular Epidemiology of Cryptosporidiosis in Iranian Children, Tehran, Iran

Background: Cryptosporidium is a worldwide protozoan parasite and one of the most common causes of infection and diarrhea in humans and cattle. The aim of the present study was determina­tion of subtypes of Cryptosporidium among children with diarrhea in Tehran by se­quence analysis of the highly polymorphic 60-kDa glycoprotein (GP60) gene.Methods: Fecal samples were collected from 794 diarrheic children. Initial identification of Crypto­spo­ridium was carried out on stool samples by Ziehl-Neelsen acid-fast staining method. DNA was extracted from positive microscopically samples and Cryptosporidium genotypes and subtypes were determined, accordingly."nResults: Out of 794 collected samples, 19 (2.40 %) were positive for Cryptosporidium oocysts. Sequences analysis of GP60 gene showed that 17 (89.47 %) of the positive isolates were Crypto­spori­dium parvum and 2 (10.52 %) were C. hominis. All subtypes of C. parvum isolates belonged to allele families IIa (6/17) and IId (11/17). The most common allele in all 17 isolates belonged to IId A20G1a (41.18%). A22G1 (IF) subtype was detected in two C. hominis isolates of the chil­dren."nConclusion: The predominancy of C. parvum species (specially, IId A20G1a sub­type) in current study underlines the importance of zoonotic Cryptosporidium transmission in Iran

Anthroponotic transmission of Cryptosporidium parvum predominates in countries with poorer sanitation - a systematic review and meta-analysis

Background: Globally cryptosporidiosis is one of the commonest causes of mortality in children under 24 months old and may be associated with important longterm health effects. Whilst most strains of Cryptosporidium parvum are zoonotic, C. parvum IIc is almost certainly anthroponotic. The global distribution of this potentially important emerging infection is not clear. Methods: We conducted a systematic review of papers identifying the subtype distribution of C. parvum infections globally. We searched PubMed and Scopus using the following key terms Cryptospor* AND parvum AND (genotyp* OR subtyp* OR gp60). Studies were eligible for inclusion if they had found C. parvum within their human study population and had subtyped some or all of these samples using standard gp60 subtyping. Pooled analyses of the proportion of strains being of the IIc subtype were determined using StatsDirect. Meta-regression analyses were run to determine any association between the relative prevalence of IIc and Gross Domestic Product, proportion of the population with access to improved drinking water and improved sanitation. Results: From an initial 843 studies, 85 were included in further analysis. Cryptosporidium parvum IIc was found in 43 of these 85 studies. Across all studies the pooled estimate of relative prevalence of IIc was 19.0% (95% CI: 12.9–25.9%), but there was substantial heterogeneity. In a meta-regression analysis, the relative proportion of all C. parvum infections being IIc decreased as the percentage of the population with access to improved sanitation increased and was some 3.4 times higher in those studies focussing on HIV-positive indivduals. Conclusions: The anthroponotic C. parvum IIc predominates primarily in lower-income countries with poor sanitation and in HIV-positive individuals. Given the apparent enhanced post-infectious virulence of the other main anthroponotic species of Cryptosporidium (C. hominis), it is important to learn about the impact of this subtype on human health

Anthroponotic transmission of Cryptosporidium parvum predominates in countries with poorer sanitation - a systematic review and meta-analysis

Background: Globally cryptosporidiosis is one of the commonest causes of mortality in children under 24 months old and may be associated with important longterm health effects. Whilst most strains of Cryptosporidium parvum are zoonotic, C. parvum IIc is almost certainly anthroponotic. The global distribution of this potentially important emerging infection is not clear. Methods: We conducted a systematic review of papers identifying the subtype distribution of C. parvum infections globally. We searched PubMed and Scopus using the following key terms Cryptospor* AND parvum AND (genotyp* OR subtyp* OR gp60). Studies were eligible for inclusion if they had found C. parvum within their human study population and had subtyped some or all of these samples using standard gp60 subtyping. Pooled analyses of the proportion of strains being of the IIc subtype were determined using StatsDirect. Meta-regression analyses were run to determine any association between the relative prevalence of IIc and Gross Domestic Product, proportion of the population with access to improved drinking water and improved sanitation. Results: From an initial 843 studies, 85 were included in further analysis. Cryptosporidium parvum IIc was found in 43 of these 85 studies. Across all studies the pooled estimate of relative prevalence of IIc was 19.0% (95% CI: 12.9–25.9%), but there was substantial heterogeneity. In a meta-regression analysis, the relative proportion of all C. parvum infections being IIc decreased as the percentage of the population with access to improved sanitation increased and was some 3.4 times higher in those studies focussing on HIV-positive indivduals. Conclusions: The anthroponotic C. parvum IIc predominates primarily in lower-income countries with poor sanitation and in HIV-positive individuals. Given the apparent enhanced post-infectious virulence of the other main anthroponotic species of Cryptosporidium (C. hominis), it is important to learn about the impact of this subtype on human health