Cryptosporidium Spp Research Articles

Giardia spp. cysts and Cryptosporidium spp. oocysts in drinking water treatment residues: comparison of recovery methods for quantity assessment

ABSTRACT Water treatment plant (WTP) residues, e.g. sludge and filter backwash water (FBW), may contain pathogenic microorganisms, as Giardia spp. and Cryptosporidium spp. However, recovering protozoa from such matrices lacks a formal and precise protocol, which is imperative to improve research in their detection, removal and inactivation. The latter includes a deeper challenge as some recovery methods may compromise viability. This study applied different recovery methods for G. muris cysts and C. parvum oocysts spiked into settled sludge and FBW obtained from a bench treatment. Procedures in sludge involved direct centrifugation, alkaline and acid flocculation, including purification by immunomagnetic separation (IMS). FBW samples were tested for membrane filtration (MF) and heated Tween® scrapings followed or not by IMS. Propidium iodide (PI) inclusion was used for oocyst viability evaluation prior and after recovery. Results with purified suspensions lead to higher recovery efficiencies (RE) for C. parvum, which was assumed to relate to poor G. muris fluorescence. Analytical quality assessments were carried out with ColorSeed® for the methods that stood out for each matrix and the results indicated lower RE than when organisms from purified suspensions were recovered. Ferric sulphate flocculation and MF, both followed by IMS reached 32.25% and 11.00% RE for Giardia spp. and 19.61% and 2.00% for Cryptosporidium spp., respectively. All of the tested methods affected oocyst viability. These results encourage further research to overcome the matrices complexity explained in this paper and increase RE, taking effects in protozoa viability into consideration.

Use of intestine-related biomarkers for detecting intestinal epithelial damage in neonatal calves with diarrhea.

To evaluate the usefulness of intestinal biomarkers in determining the presence of intestinal epithelial damage in neonatal calves with diarrhea caused by 4 etiologic agents. 40 neonatal calves that were healthy (n = 10) or had diarrhea (30). The study was a cross-sectional study. Results of hematologic analyses and serum concentrations of intestinal fatty acid-binding protein (I-FABP), liver fatty acid-binding protein (L-FABP), trefoil factor 3 (TFF-3), Claudin-3 (CLDN-3), γ-enteric smooth muscle actin (ACTG2), intestinal alkaline phosphatase (IAP), interleukin-8 (IL-8), platelet-activating factor (PAF), and leptin (LP) were compared among calves grouped according to whether they were healthy (control group; G-1) or had diarrhea caused by K99 Escherichia coli (G-2; n = 10), bovine rota- or coronavirus (G-3; 5 each), or Cryptosporidium spp (G-4; 10). Across the 3 time points at which blood samples were obtained and evaluated, the groups of calves with diarrhea generally had markedly higher mean serum concentrations of L-FABP, TFF-3, IAP, IL-8, and LP, compared with the control group. In addition, G-2 also consistently had markedly higher mean serum concentrations of I-FAB and ACTG2 and lower mean serum concentrations of CLDN-3, compared with the control group. Results indicated that degree of intestinal epithelial damage differed among calves grouped by the etiologic agent of diarrhea and that such damage might have been more severe in calves with diarrhea caused by K99 E coli. Additionally, our results indicated that serum concentrations of I-FABP, L-FABP, TFF-3, IAP, IL-8, ACTG2, LP, and CLDN-3 were useful biomarkers of intestinal epithelial damage in calves of the present study.

Investigations from Northern Greece on mussels cultivated in areas proximal to wastewaters discharges, as a potential source for human infection with Giardia and Cryptosporidium

Marine bivalves are usually cultivated in shallow, estuarine waters where there is a high concentration of nutrients. Many micro-pollutants, including the protozoan parasites Giardia duodenalis and Cryptosporidium spp., which also occur in such environments, may be concentrated in shellfish tissues during their feeding process. Shellfish can thus be considered as vehicles for foodborne infections, as they are usually consumed lightly cooked or raw. Therefore, the main objective of this study was to investigate the presence of both parasites in Mediterranean mussels, Mytilus galloprovincialis that are cultivated in Thermaikos Gulf, North Greece, which is fed by four rivers that are contaminated with both protozoa. Moreover, the occurrence of these protozoa was monitored in treated wastewaters from 3 treatment plants that discharge into the gulf. In order to identify potential sources of contamination and to estimate the risk for human infection, an attempt was made to genotype Giardia and Cryptosporidium in positive samples. Immunofluorescence was used for detection and molecular techniques were used for both detection and genotyping of the parasites. In total, 120 mussel samples, coming from 10 farms, were examined for the presence of both protozoa over the 6-month farming period. None of them were found positive by immunofluorescence microscopy for the presence of parasites. Only in 3 mussel samples, PCR targeting the GP60 gene detected Cryptosporidium spp. DNA, but sequencing was not successful. Thirteen out of 18 monthly samples collected from the 3 wastewater treatment plants, revealed the presence of Giardia duodenalis cysts belonging to sub-assemblage AII, at relatively low counts (up to 11.2 cysts/L). Cryptosporidium oocysts (up to 0.9 oocysts/L) were also detected in 4 out of 8 samples, although sequencing was not successful at any of the target genes. At the studied location and under the sampling conditions described, mussels tested were not found to be harboring Giardia cysts and the presence of Cryptosporidium was found only in few cases (by PCR detection only). Our results suggest that the likelihood that mussels from these locations act as vehicles of human infection for Giardia and Cryptosporidium seems low.

Hematopoietic Stem Cell Transplantation for Combined Immunodeficiencies, on Behalf of IEWP-EBMT.

Combined immunodeficiencies (CIDs) are a clinically and genetically heterogeneous group of primary immunodeficiencies (PIDs) that affect T-lymphocyte immunity with abnormal development or function. As compared to severe combined immune deficiencies (SCID), these patients are usually diagnosed later. They display a broad infectious susceptibility; immune dysregulation manifestations and chronic lymphoproliferation are also frequent. These complications and their specific treatments can lead to persistent damage to several organs. Prognosis of CIDs is worse as compared to other PIDs. The curative treatment is usually hematopoietic stem cell transplantation (HSCT), but difficult questions remain regarding the definitive indication of HSCT and its timing; the final decision depends on a conjunction of factors such as immunological parameters, severity of clinical manifestations, and natural history of the disease, when molecular diagnosis is known. CD40L deficiency, a CID caused by mutations in CD40LG gene, well illustrates the dilemma between HSCT vs. long-term supportive treatment. This disease leads to higher risk of developing infections from bacterial and intracellular pathogens, especially Pneumocystis and Cryptosporidium spp. While supportive care allows improved survival during childhood, organ damages may develop with increasing age, mainly chronic lung disease and biliary tract disease (secondary to Cryptosporidium spp. infection) that may evolve later to sclerosing cholangitis, a severe complication associated with increased mortality. Early HSCT before organ damage development is associated with best survival and cure rate, while HSCT remains a risky therapeutic option for older patients, for those with organ damage, especially severe liver disease, and/or for those with limited or no donor availability. Prospective studies are needed to analyze risks of HSCT compared to those of life-long supportive therapy, including quality of life measures.

Occurrence of gastrointestinal protozoans in cats from Londrina, Paraná, Brazil

Protozoans are widely distributed, and several species may parasitize the digestive tracts of cats (Felis catus), and can be transmitted to humans. The present study aimed to evaluate the species and occurrence of gastrointestinal protozoans in cats in the city of Londrina, Paraná, Brazil. A total of 206 cat fecal samples were tested, of which 141 were from shelter animals, and 65 were from pets owned by local people. Samples were processed by parasitological techniques. Coproparasitological techniques (Willis, Faust and Ziehl-Neelsen) were performed for detection of protozoan parasites. Subsequently, all samples were processed by PCR protocols specific to Toxoplasma gondii, Giardia spp., and Cryptosporidium spp. PCR products from positive samples were selected for sequencing. No samples were found to be positive for Cryptosporidium spp. using the Ziehl-Neelsen technique. Using specific PCR protocols, 1/206 (0.48%) samples tested positive for Cryptosporidium spp. After purification, this one positive sample was sequenced, and it demonstrated a 100% identity match to Cryptosporidium muris. Using specific PCR protocols, 13/206 (9.22%) cat fecal samples tested, including 2/65 (3.08%) pet cat fecal samples, were positive for T. gondii. PCR analysis revealed that 37/206 (17.96%) of cat fecal samples were positive for Giardia spp., including 27/141 (19.15%) of shelter cat fecal samples, and 10/65 (15.38%) pet cat fecal samples (p = 0.5124). When sequenced, these positive samples showed a 100% identity match with Giardia duodenalis. This study demonstrated that infections with Cryptosporidium spp., Toxoplasma gondii, and Giardia duodenalis are present in the population of both pet cats and shelter cats in the city of Londrina. This poses a risk to public health, because these parasites have a high zoonotic potential.

Increased Detection of Cryptosporidium and Cyclospora spp. Oocysts in a Major Philippine Watershed Following Rainfall Events

Increased Detection of Cryptosporidium and Cyclospora spp. Oocysts in a Major Philippine Watershed Following Rainfall Events

Opportunistic Intestinal Protozoan Infections and Immunological Status of HIV/AIDS Patients in Jalingo Local Government, Taraba State

The Human Immunodeficiency Virus (HIV) is a virus that attacks the immune system. Opportunistic intestinal protozoans are among the major contributors to the global intestinal parasite load. This study investigated the danger and complication due to intestinal protozoans’ infection among HIV/AIDS patients in Jalingo metropolis. The study also compared the prevalence of opportunistic intestinal protozoa infection and CD⁴⁺ T-Cell count between HIV/AIDS and non HIV patients. The procedure of wet mount technique and faecal concentration methods were employed. The CD⁴⁺ count of patients were determined using the Partee Cyflow Counter, which determines the percentage of CD⁴⁺ cells of individuals. The prevalence of opportunic intestinal protozoan parasites was higher among female positives, 68 (l9.60%) than the male, 50 (16.50%) with no significant difference (p>0.05). The infection based on age groups revealed that the age group 10-15 years had the highest prevalence of 27 (48.21%) while the age group 41-45 years had the least prevalence of 2 (2.63%). The prevalence of intestinal parasites was highly significant among the study participants with CD⁴⁺ count <200 cells/μl. <i>Cryptosporidium spp</i> was significantly associated with HIV positive subjects who had CD⁴⁺ count of less than 200 cell/μl. The prevalence of intestinal parasites was highly significant among the study participants with CD⁴⁺ count <200 cells/ul. The implications of these findings are discussed and solution proposed to curb opportunistic infections among HIV/AIDS patients.

Asymptomatic Cryptosporidium infections in ewes and lambs are a source of environmental contamination with zoonotic genotypes of Cryptosporidium parvum.

Protozoan parasites of the Cryptosporidium genus cause severe cryptosporidiosis in newborn lambs. However, asymptomatic infections also occur frequently in lambs and ewes. In sheep, the most commonly detected Cryptosporidium species are C. ubiquitum, C. xiaoi and C. parvum. Due to a lack of relevant information about such infections in France, we investigated the situation on five dairy sheep farms in the Pyrénées-Atlantiques Department in south-western France in December 2017. Individual fecal samples were collected from 79 female lambs (5–17 days old) and their mothers (72 ewes). Oocysts were screened using Heine staining before and after Bailenger concentrations. Cryptosporidium species identification and genotyping were performed using real-time PCR and gp60 gene sequencing. No cases of clinical cryptosporidiosis were observed in the 79 lambs. Microscopically, Cryptosporidium spp. oocysts were observed in only one lamb on one farm (prevalence 1.3%) and one ewe on another farm (prevalence 1.4%). By contrast, Cryptosporidium spp. DNA was detected in 17 ewes (prevalence ranging from 10.5% to 50% depending on the farm) and in 36 lambs (prevalence ranging from 0% to 77.8% depending on the farm). Only zoonotic Cryptosporidium parvum IId and IIa genotypes were identified when genotyping was possible. Cryptosporidium ubiquitum and C. xiaoi were detected on one and three farms, respectively. We conclude that healthy young lambs and their mothers during the peripartum period could be a source of environmental contamination with oocysts.

Molecular characterization of &lt;i&gt;Cryptosporidium&lt;/i&gt; spp. from migratory ducks around Tokachi subprefecture, Hokkaido, Japan

Cryptosporidium, a waterborne protozoan parasite, has a substantial veterinary and medical impact worldwide. This parasite is more often recognized during waterborne outbreaks because of its resistance to chlorine disinfection, small size making it difficult to inactivate/eliminate through filtration, and presence in many animal species including humans. Migratory waterfowl, in addition to acting as mechanical carriers of Cryptosporidium oocysts, can also serve as natural reservoirs of infection by host-specific Cryptosporidium species. For better understanding of the extent of genetic diversity and inter-relationships among avian isolates of Cryptosporidium, 200 fecal samples of migratory ducks from the Tokachi subprefecture, Hokkaido, Japan were collected and analyzed by nested PCR (N-PCR) at the 18S rRNA gene. N-PCR revealed that 11.5% (23/200) were positive for Cryptosporidium spp. Among all samples, sequence analysis identified that 10% (20/200) were 98–100% identical to Cryptosporidium avian genotype III. On the other hand, 1.5% (3/200) were 99–100% identical to C. baileyi. This is the first molecular study reporting the prevalence of Cryptosporidium in migratory ducks in Japan. Genetic diversity among Cryptosporidium isolates from humans and birds has been reported worldwide. Nevertheless, further studies are important to assess genetic variety and to elucidate the transmission dynamics of Cryptosporidium parasites.

Prevalence and molecular characterization of Cryptosporidium spp. in Père David's deer (Elaphurus davidianus) in Jiangsu, China.

Cryptosporidium is a zoonotic parasite that causes diarrhea in a broad range of animals, including deer. Little is known about the prevalence and genotype of Cryptosporidium spp. in Père David's deer. In this study, 137 fecal samples from Père David's deer were collected between July 2017 and August 2018 in the Dafeng Reserve and analyzed for Cryptosporidium spp. by nested-PCR based on the small subunit ribosomal RNA (SSU rRNA) gene, followed by sequence analyses to determine the species. The 60 kDa glycoprotein (gp60) gene was used to characterize Cryptosporidium spp. Among 137 samples, 2 (1.46%) were positive for Cryptosporidium spp. according to SSU rRNA gene sequencing results. Both samples belonged to the Cryptosporidium deer genotype, with two nucleotide deletions and one nucleotide substitution. The prevalence data and molecular characterization of this study provide basic knowledge for controlling and preventing Cryptosporidium infections in Père David's deer in this area.

Potential sources of infection with selected zoonotic agents in the veterinary work environment - pilot studies.

The problem of occupational biohazards is very important, especially in the field of agriculture and in human and veterinary medicine. The aim of the study was to determine the potential sources of infection in veterinary professionals with selected zoonotic agents, including: Toxoplasma gondii, Giardia duodenalis, Leptospira spp., Cryptosporidium spp. and Coxiella burnetii. A total of 50 air samples from barns, piggeries and veterinary surgeries were examined for the presence of Leptospira spp. and C. burnetii DNA. Serum samples of 86 pigs and 80 cows were tested for the presence of antibodies to Leptospira spp. and to phase I and II C. burnetii antigens. Serum of 70 cats were tested for the presence of antibodies to T. gondii and 65 samples of cat faeces for the presence of T. gondii oocysts. The presence of G. duodenalis and Cryptosporidium spp. were examined in 50 of dog faeces and 50 of bovine faeces samples. DNA of Leptospira spp. was detected in 2 air samples from the piggeries (4%). C. burnetii DNA was not found in any sample. Anti-Leptospira spp. antibodies were detected in 51 (59.3%) of examined pigs. Neither anti-Leptospira spp. nor anti-C. burnetii antibodies were found among samples of bovine serum. Anti-T. gondii antibodies was found in 52 cat serum samples (74.3%). Among samples of cat faeces, no T. gondii oocysts were detected. In one sample of cattle stool (2%), G. duodenalis was detected and in another (2%) - Cryptosporidium spp. G. duodenalis was detected in 7 samples (14%) and Cryptosporidium spp. in 2 samples (2%) of dog faeces. The results of this study demonstrate the potential risk of infection with Leptospira spp. in veterinarians working with pigs. Veterinarians could be also be at risk of infection with T. gondii and G. duodenalis.

Diarrhoeal disease and subsequent risk of death in infants and children residing in low-income and middle-income countries: analysis of the GEMS case-control study and 12-month GEMS-1A follow-on study

SummaryBackgroundThe Global Enteric Multicenter Study (GEMS) was a 3-year case-control study that measured the burden, aetiology, and consequences of moderate-to-severe diarrhoea (MSD) in children aged 0–59 months. GEMS-1A, a 12-month follow-on study, comprised two parallel case-control studies, one assessing MSD and the other less-severe diarrhoea (LSD). In this report, we analyse the risk of death with each diarrhoea type and the specific pathogens associated with fatal outcomes.MethodsGEMS was a prospective, age-stratified, matched case-control study done at seven sites in Africa and Asia. Children aged 0–59 months with MSD seeking care at sentinel health centres were recruited along with one to three randomly selected matched community control children without diarrhoea. In the 12-month GEMS-1A follow-on study, children with LSD and matched controls, in addition to children with MSD and matched controls, were recruited at six of the seven sites; only cases of MSD and controls were enrolled at the seventh site. We compared risk of death during the period between enrolment and one follow-up household visit done about 60 days later (range 50–90 days) in children with MSD and LSD and in their respective controls. Approximately 50 pathogens were detected using, as appropriate, classic bacteriology, immunoassays, gel-based PCR and reverse transcriptase PCR, and quantitative real-time PCR (qPCR). Specimens from a subset of GEMS cases and controls were also tested by a TaqMan Array Card that compartmentalised probe-based qPCR for 32 enteropathogens.Findings223 (2·0%) of 11 108 children with MSD and 43 (0·3%) of 16 369 matched controls died between study enrolment and the follow-up visit at about 60 days (hazard ratio [HR] 8·16, 95% CI 5·69–11·68, p<0·0001). 12 (0·4%) of 2962 children with LSD and seven (0·2%) of 4074 matched controls died during the follow-up period (HR 2·78, 95% CI 0·95–8·11, p=0·061). Risk of death was lower in children with dysenteric MSD than in children with non-dysenteric MSD (HR 0·20, 95% CI 0·05–0·87, p=0·032), and lower in children with LSD than in those with non-dysenteric MSD (HR 0·29, 0·14–0·59, p=0·0006). In children younger than 24 months with MSD, infection with typical enteropathogenic Escherichia coli, enterotoxigenic E coli encoding heat-stable toxin, enteroaggregative E coli, Shigella spp (non-dysentery cases), Aeromonas spp, Cryptosporidium spp, and Entamoeba histolytica increased risk of death. Of 61 deaths in children aged 12–59 months with non-dysenteric MSD, 31 occurred among 942 children qPCR-positive for Shigella spp and 30 deaths occurred in 1384 qPCR-negative children (HR 2·2, 95% CI 1·2–3·9, p=0·0090), showing that Shigella was strongly associated with increased risk of death.InterpretationRisk of death is increased following MSD and, to a lesser extent, LSD. Considering there are approximately three times more cases of LSD than MSD in the population, more deaths are expected among children with LSD than in those with MSD. Because the major attributable LSD-associated and MSD-associated pathogens are the same, implementing vaccines and rapid diagnosis and treatment interventions against these major pathogens are rational investments.FundingBill & Melinda Gates Foundation.

Comparative genomics: how has it advanced our knowledge of cryptosporidiosis epidemiology?

Whole genomic sequencing (WGS) and comparative genomics are increasingly used in the characterization of Cryptosporidium spp. They are facilitated by the establishment of procedures for WGS analysis of clinical specimens without laboratory propagation of pathogens. Results of recent comparative genomics analysis suggest that gene duplication might be associated with broad host ranges of some zoonotic Cryptosporidium species and subtypes, while genetic recombination could be involved in the emergence of virulent subtypes. The availability of WGS data has further facilitated the development of advanced molecular typing tools. The use of these tools together with comparative genomics analyses has begun to improve the investigations of outbreaks in industrialized nations. More WGS data, however, are needed from both industrialized nations and developing countries before we can have in-depth understanding of the population genetics and evolution of Cryptosporidium spp. and genetic determinants of various phenotypic traits in human-pathogenic subtypes.

Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples.

Cryptosporidiosis is an emerging zoonotic disease caused by the worldwide distributed parasitic protozoa Cryptosporidium spp. The host-adapted species Cryptosporidium canis is most frequently found in dogs, although human infections with this species have been described. This study aimed to develop a real-time PCR targeting the HSP70 protein gene for C. canis DNA detection in dog fecal samples collected from two municipalities in the state of São Paulo, Brazil. Furthermore, the occurrence of Cryptosporidium spp. and. C. canis was also determined by nested PCR. Fecal samples from 367 dogs (21 puppies and 346 adults) were purified by water-ether sedimentation. A real-time PCR protocol targeting the HSP70 gene for the species-specific detection of C. canis was developed and compared with nested PCR results. Real-time PCR identified C. canis in 15.3% (58/367) samples. Nested PCR revealed that 10.4% (38/367) of samples were positive for Cryptosporidium spp. All sequenced 18S rRNA amplicons were C. canis. There was a higher prevalence of Cryptosporidium spp. and C. canis in puppies compared to adult dogs. No non-specific amplification was observed in C. canis specific real-time PCR assay.

Endoparasites of Selected Native Non-Domesticated Mammals in the Neotropics (New World Tropics).

In this review, information was summarized on endoparasites found in six non-domesticated neotropical animals. These mammals have the potential to be domesticated. The animals included three rodents, agouti (Dasyprocta leporina), lappe (Agouti paca), and capybara (Hydrochoerus hydrochaeris); a marsupial, manicou (Didelphis marsupialis insularis); and an artiodactyl, the collared peccary (Tayassu tajacu/Peccari tajacu) and a ruminant (the red brocket deer, Mazama americana). While there are many descriptions of the parasites present, the majority of publications failed to note the effect of them on the animals. Most information is available on endoparasites of capybara, while the endoparasites of the red brocket deer were the least reported. The manicou was reported to have had the most number of endoparasites, 44 species of parasites were reported, while there were only 24 endoparasites reported in the lappe. The most common parasites found in these neotropical animals were Paraspidodera uncinata, Strongyloides spp., Eimeria spp., Moniezia benedeni, Trichuris spp., Physocephalus spp., and Giardia spp. A large majority of the studies concluded that these animals were reservoirs for parasites that could affect domesticated livestock. Endoparasites of zoonotic significance were Echinoccocus spp., Trichuris spp., Giardia spp., and Cryptosporidium spp.

Optimization of routine microscopic and molecular detection of parasitic protozoa in SAF-fixed faecal samples in Sweden

Background: Nucleic acid-based methods are increasingly used for screening of gastrointestinal parasites. Microscopy is still used and Swedish routine protocol consists of formalin ethyl-acetate concentration and do not include screening for trophozoites or Cryptosporidium spp. This study aimed to compare detection with the Swedish routine microscopy method to an extended method that includes screening for trophozoites and Cryptosporidium. Furthermore, we also developed a method for DNA recovery from SAF-fixed faecal samples and compared the real-time PCR detection of Giardia intestinalis, Dientamoeba fragilis, Cryptosporidium spp., Entamoeba histolytica and Entamoeba dispar from SAF-fixed and unpreserved faecal samples. PCR results were then compared with microscopy results.Methods: SAF-fixed and unpreserved faecal samples from 1000 patients at the Clinical microbiology laboratory in Region Jönköping County, Sweden, were included. Samples were analysed with routine formalin ethyl-acetate concentration, wet mounts from both concentrated and unconcentrated samples, Ziehl–Neelsen staining on patients with certain symptoms and real-time PCR.Results: We found a significant higher detection rate of parasites with the extended microscopy method compared to the Swedish routine microscopy method when SAF-fixed samples were used. The detection rate with real-time PCR in SAF-fixed samples was equal to the detection rate in unpreserved samples. There was no significant difference in detection comparing extended microscopy and real-time PCR.Conclusion: In conclusion, this study showed that the extended microscopy method increased detection of intestinal protozoa with detection of both trophozoites and Cryptosporidium spp. We also showed that SAF-fixative can be used for detection of parasite-DNA with real-time PCR.

Food protozoa safety assessment and risk in school restaurants in Armenia, Colombia

AbstractThis work assessed the risk of protozoa in 10 school restaurants in Armenia (Quindío, Colombia) by analyzing the presence of Cryptosporidium spp, Giardia duodenalis, Blastocystis, and Cyclospora cayetanensis DNA in the food, water, and living and inert surfaces of school restaurants and in stools of children who ate at these restaurants. Of the 213 food, water, and surface samples, 6.6% were positive using PCR to test DNA for Blastocystis; 3.8% for Cryptosporidium spp; 0.9% for G. duodenalis; and 0% for C. cayetanensis. In 187 stool samples analyzed via microscopy from children who attended the restaurants, 40 (21.4%) were positive for Blastocystis and 21 (11.2%) were positive for Giardia spp. Via PCR, 20 (10.7%) were positive for Cryptosporidium and 0 (0%) for C. cayetanensis. A higher positivity in children's stools for Blastocystis spp was correlated with lower compliance in property conditions and for higher positivity of Giardia spp in children's stool was related to lower knowledge by food manipulators. Inspection scores can identify restaurants with higher risk for protozoa infection.

2541 Cryptosporidial Colonization of a Peptic Ulcer in a Healthy Patient—Random Occurrence or a Risk Factor?

INTRODUCTION: The incidence of Cryptosporidial enteritis (CE) has recently risen in the USA due to water and foodborne outbreaks. It has a self-limited course in healthy hosts. We describe a case of a healthy female with history of gastric bypass surgery who presented with a one-month history of voluminous watery diarrhea and vomiting. This was a rare case of CE diagnosed by endoscopic biopsies of two anastomotic peptic ulcers that were colonized by Cryptosporidium spp. CASE DESCRIPTION/METHODS: 57-year-old female with a history of GERD, iron deficiency anemia, and Roux-en-y gastric bypass complicated by jejunal strictures with revision of the gastro-jejunal anastomosis, presented with a 7-day history of vomiting and crampy epigastric abdominal pain. She recently returned from a two-week vacation to Latin America. Upon arrival, she experienced progressively worsening voluminous watery diarrhea associated with intermittent melanotic stools followed by frequent episodes of non-bilious, non-bloody vomiting along with reduced oral intake. Abdominal exam revealed hyperactive bowel sounds with tenderness to deep palpation of the epigastrium. Her hemoglobin dropped from a baseline of 11.2 g/dL to 9.1 g/dL. CT scan of the abdomen with contrast showed inflammation around the gastro-jejunostomy site. Blood and stool cultures were negative. Upper endoscopy revealed a 1 cm and 2 cm clean-based ulcers with a deep crater at the anastomotic site. Biopsy demonstrated spherical bluish-purple organisms, 2-5 µm in diameter on hematoxylin and eosin, Gomori Methanamine Silver Nitrate and Masson's trichrome stains. They were suggestive of Cryptosporidium, and the patient was diagnosed with CE. Patient's immunodeficiency workup was unremarkable. Patient's symptoms resolved with a three-day course of Nitazoxanide 500 mg twice per day. DISCUSSION: CE is a significant cause of chronic diarrhea in both healthy and immune-compromised individuals, thus stands as a key differential in those presenting with persistent voluminous diarrhea. The modified acid-fast stain carries only 50% sensitivity to this protozoan, and diagnosis is often missed on routine stool ova and parasite tests. Novel diagnostic modalities have proven useful in identifying this protozoan in stool samples. We report the first case documenting Cryptosporidium colonization of anastomotic site peptic ulcers with complete resolution of symptoms following a short course of Nitazoxanide.

DIAGNÓSTICO E IDENTIFICACIÓN DE ESPECIES DE CRYPTOSPORIDIUM EN POBLACIÓN INFANTIL DE AREAS RURALES EN PANAMÁ.

INTRODUCCIÓN: La criptosporidiosis es una enfermedad entérica causada por diversas especies del género Cryptosporidium. Este parásito se asocia con episodios de diarrea aguda, cuadros de malabsorción y efectos negativos en el desarrollo físico y mental, principalmente en niños menores de cinco años. En Panamá se conoce poco sobre la prevalencia de esta infección y las especies involucradas.MATERIALES Y MÉTODOS: Se analizaron 618 muestras de heces de niños(as) menores de cinco años en cinco comunidades del país. Para el diagnóstico de Cryptosporidium se realizó el método de concentración por sedimentación con formol-acetato de etilo y tinción de Kinyoun para el análisis microscópico. Para la identificación de las especies de Cryptosporidium se realizó una PCR-anidada-RFLP del gen SSUrRNA.RESULTADOS: El análisis microscópico mostró una prevalencia de parasitosis intestinal general de 83% y de 1.94% para Cryptosporidium spp en frotis fecales teñidos con tinción de Kinyoun. Mediante una PCR-anidada-RFLP del gen SSUrRNA de Cryptosporidium se logró identificar a Cryptosporidium hominis como la especie más frecuente (62.5%, 5/8). Además, se confirma la presencia de C. meleagridis (25%, 2/8) y C. parvum (C. pestis) (12.5%, 1/8). Adicionalmente, se reporta el hallazgo de Cyclospora cayetanensis en un 2.5% de las muestras procedentes de la comunidad indígena de Emplanada de Chorcha en Panamá.CONCLUSIÓN: En este estudio se describen por primera vez las especies circulantes de Cryptosporidium en niños menores de cinco años procedentes de seis comunidades rurales de Panamá. Aun cuando las prevalencias encontradas para la criptosporidiosis infantil fueron bajas, la presencia de diversas especies de este gregarino representa un llamado de atención a las autoridades de salud para la correcta implementación de medidas de vigilancia, manejo y prevención acordes con el comportamiento epidemiológico de cada especie.

\u201cEnterocytozoon bieneusi and Cryptosporidium: a cross-sectional study conducted throughout Thailand\u201d

BackgroundEnterocytozoon bieneusi and Cryptosporidium spp. are prevalent zoonotic parasites associated with a high burden among children. To date only limited molecular epidemiological data on E. bieneusi and Cryptosporidium spp. in humans living in Thailand has been published.MethodsPCR-based tools were used to detect and characterize E. bieneusi and Cryptosporidium spp. The internal transcribed spacer (ITS) region of the rRNA gene was used to investigate E. bieneusi, and the small subunit (SSU) rRNA gene was used to investigate Cryptosporidium spp., and 697 fecal samples from villagers and school children in rural areas in Thailand were analyzed.ResultsThe infection rates were 2.15% (15/697) for E. bieneusi and 0.14% (1/697) for Cryptosporidium spp. The prevalence of E. bieneusi was significantly high in Loei province. Sequence analysis indicated that the Cryptosporidium isolate was C. parvum. Nine E. bieneusi genotypes were identified, EbpC, Peru12, TMH6, TMH3, TMH7, H, D, and two novel genotypes TMLH1 and TMLH2. E. bieneusi prevalence was significantly higher in male participants than in female participants, and in children aged 3–15 years than in participants aged > 15 years.ConclusionsThe prevalence, genotypes, and zoonotic potential of E. bieneusi were found to vary significantly high even in one country. Transmission routes and key animal carriers of E. bieneusi may be associated with differences in hygiene, sanitation, and cultural behaviors. Further molecular studies including longitudinal studies will be required to unveil epidemiological characteristics of these opportunistic intestinal protozoa in all over the countries.