Subtypes and phylogenetic analysis of Blastocystis sp. isolates from West Ismailia, Egypt.

In Egypt, Blastocystis sp. is not yet on the diagnostic list of parasitology reports, and information about its subtypes (STs) is scarce. This study investigated its prevalence and its STs/alleles, performed phylogenetic analysis, and considered the distribution of risk factors associated with Blastocystis sp. infections in West Ismailia, Ismailia governorate. Sociodemographic data, exposure factors, and previous parasitic infection status were recorded for symptomatic and asymptomatic individuals. Microscopy, polymerase chain reaction, sequencing, and phylogenetic analysis for Blastocystis sp. isolated from fecal samples were performed. Eighty Blastocystis sp.-infected individuals (15.3%) were examined. The age of the individuals ranged between 0.60 and 85.0 (mean 17.10 ± 15.70), the male/female ratio was 33/47, and the asymptomatic/symptomatic ratio was 55/25. The findings demonstrate clear evidence of direct contact with animals, poor water quality, and previous parasitic infections. Eleven samples yielded three Blastocystis STs (ST1: allele 4, ST2: alleles 9 and 12, and ST3: allele 34), with ST3 (45.5%) representing the most common subtype. Phylogenetic analysis with a robust bootstrap revealed three distinct clades for isolates of each subtype. This study updates the epidemiological knowledge of the distribution of Blastocystis sp. STs in Egypt and expands the current understanding of the prevalence, risk factor frequencies, and genetic diversity of this protist in the studied area.

Free-living amoebae and other neglected protistan pathogens: Health emergency signals?

Protistan parasites have an undisputed global health impact. However, outside of a few key exceptions, e.g. the agent of malaria, most of these infectious agents are neglected as important health threats. The Symposium entitled "Free-living amoebae and neglected pathogenic protozoa: health emergency signals?" held at the European Congress of Protistology in Rome, July 2019, brought together researchers addressing scientific and clinical questions about some of these fascinating organisms. Topics presented included the molecular basis of pathogenicity in Acanthamoeba; genomics of Naegleria fowleri; and epidemiology of poorly diagnosed enteric protistan species, including Giardia, Cryptosporidium, Blastocystis, Dientamoeba. The Symposium aim was to excite the audience about the opportunities and challenges of research in these underexplored organisms and to underline the public health implications of currently under-appreciated protistan infections. The major take home message is that any knowledge that we gain about these organisms will allow us to better address them, in terms of monitoring and treatment, as sources of future health emergencies.

Blastocystis hominis transmission by non-potable water: a case report in Italy.

In the reported case, a 41-year-old Italian man came to the clinician's observation reporting cramps, bloating and watery diarrhoea a few days after drinking water indicated as unpotable from a fountain in a farm area. The medical suspicion was directed at both gluten intolerance and enteric infection, eventually of waterborne origin. Gluten intolerance was investigated by intestinal biopsy and excluded, while stool analyses ruled out infective bacteriological or viral agents and parasites. Subsequently, a persistent eosinophilia was revealed and a parasitological analysis was again suggested, planning for a more sensitive molecular method. Therefore, a multiplex-PCR of enteric protozoa species DNA was performed on an intestinal biopsy and faecal samples revealing only Blastocystis hominis protozoa, subsequently typed as subtype 1 by RFLP-PCR method. B. hominis is an anaerobic protozoa found in the human and animal intestinal tract, recently associated with a pathogenic role characterized by chronic development. Since blastocystosis has been demonstrated as a waterborne infection, a sample of water matrix was analysed, revealing the B. hominis subtype 1 DNA inside. A probable water transmission of Blastocystis infection has been demonstrated in this case report. Only a probiotic treatment based on Saccharomyces boulardii was administered to the patient and this apparently resolved the infection. In summary, the case described here is a chronic blastocystosis of possible waterborne origin, controlled by assuming a yeast treatment.

The relationship between nucleoside triphosphate hydrolase (NTPase) isoform and Toxoplasma strain virulence in rat and human toxoplasmosis.

Avirulent strains of Toxoplasma gondii possess only the nucleoside triphosphate hydrolase II (NTPaseII) isoform, whilst virulent strains possess both NTPaseI and NTPaseII. To determine if it is possible to identify the infective strain type (virulent or avirulent) in T. gondii infections by serological methods, we developed isoform-specific peptide ELISAs from the NTPaseI and NTPaseII antigens of T. gondii. When rats were immunized with either recombinant NTPaseI or NTPaseII, the ELISA could differentially identify antibody reactivity to each NTPase isoform. This ELISA was then used to test six groups of rats that were infected with either one of three virulent (RH, P or Ent) or three avirulent (Me49, C or TPR) strains of T. gondii. No differential antibody reactivity was detected by either whole recNTPase ELISA or peptide ELISA in the sera of rats, whether infected by virulent or avirulent strains of T. gondii. We also studied a panel of human sera from patients infected with known laboratory strains of T. gondii or naturally infected patients where the parasite was isolated and its virulence determined in mice. Differential reactivity to whole recNTPase isoforms was detected in some human sera, but this reactivity was not detected by the isoform-specific peptide ELISAs. Although the NTPase peptides do exhibit differential antibody reactivity, this is not correlated with the virulence status of the infecting strain.

Toxoplasmosis in pregnancy is still an open subject.

OBJECTIVE: To assess the consequences of a systematic screening for toxoplasma infection in pregnant women in a potentially high risk population. METHODS: We have investigated all consecutive women with likely toxoplasma seroconversion in a referral center setting. Data were obtained from 68 women for whom an acute infection during pregnancy was considered likely or definite. They were all treated with antibiotics in the first instance and offered, if in the first or second trimester, amniocentesis for detection of vertical transmission (PCR and IFAT). Third trimester seroconversions and positive cases after amniocentesis were offered more aggressive antibiotic treatment. RESULTS: Five fetuses/neonates (7%) were found to be infected. Four of them were diagnosed prenatally at amniocentesis, two women decided for termination of pregnancy, two were treated and gave birth to seronegative, normally developing children. One case was found postnatally, after a third trimester conversion; this developed into hydrocephalus with neurologic impairment. DISCUSSION: Prenatal screening and antibiotic treatment of mothers infected with toxoplasmosis showed good feasibility in our infection-susceptible population. However, there were some weak points; for example, the high number of invasive procedures and the questionable prevention of mother to child transmission in the second to third trimester.