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Intervening transcribed spacer region 1 variability in Cyclospora cayetanensis.

Cyclospora cayetanensis is an apicomplexan protozoan parasite which has emerged as an important cause of epidemic and endemic diarrhea. Water-borne as well as food-borne outbreaks have occurred, including a large number of U.S. cases associated with raspberries imported from Guatemala. Molecular markers exist for tracing the epidemiology of many of the bacterial pathogens associated with water-borne or food-borne diarrhea, such as serotyping and pulsed-field electrophoresis. However, there are currently no molecular markers available for C. cayetanensis. The intervening transcribed spacer (ITS) regions between the small- and large-subunit rRNA genes demonstrate much greater sequence variability than the small-subunit rRNA sequence itself and have been useful for the molecular typing of other organisms. Thus, ITS1 variability might allow the identification of different genotypes of C. cayetanensis. In order to determine the degree of ITS1 variability among C. cayetanensis isolates, the ITS1 sequences of C. cayetanensis isolates from a variety of sources, including raspberry-associated cases, cases from Guatemala, and pooled and individual isolates from Peru, were obtained. The ITS1 sequences of all five raspberry-associated isolates were identical, consistent with their origin from a single source. In contrast, one of the two Guatemala isolates and two Peruvian isolates contained multiple ITS1 sequences. These multiple sequences could represent multiple clones from a single clinical source or, more likely, variability of the ITS1 region within the genome of a single clone.

Animals↗

Prevalence of infection with Cryptosporidium parvum and Cyclospora cayetanensis among international travellers.

BACKGROUND: Cryptosporidium parvum and Cyclospora cayetanensis are recognised as possible pathogens of traveller's diarrhoea. AIMS: To identify the prevalence of C parvum and Cyc cayetanensis in travellers returning from developing countries. PATIENTS: Nine hundred and seventy eight stool samples wer taken from 795 patients returning from developing countries. METHODS: Microscopy (iron-haematoxylin stain, SAF concentration, modified acid fast stain) and a commercially available enzyme linked immunosorbent assay (ELISA) kit for the detection of Cryptosporidium antigen in stool. RESULTS: Of the 795 patients in the study, 469 suffered from diarrhoea. Infection with Cyc cayetanensis could be detected in five subjects (1.1%) by acid fast stain, and 13 patients (2.8%) were infected with C parvum. On evaluation, the antigen capture ELISA turned out to be clearly less sensitive for detection of C parvum than microscopy. All patients with either C parvum or Cyc cayetanensis infection suffered from watery diarrhoea. CONCLUSIONS: C parvum and Cyc cayetanensis are not major causes of diarrhoea in international travellers. In cases of persistent watery diarrhoea, however, these pathogens should be taken into account in the differential diagnosis.

Animals↗

Modified detergent Ziehl-Neelsen technique for the staining of Cyclospora cayetanensis.

Cyclospora cayetanensis is a cause of prolonged diarrhoea, mainly in travellers. Laboratory diagnosis may be achieved by a number of methods such as the staining of faecal smears by the modified Ziehl-Neelsen (ZN) technique. Safer methods using this technique have been described for the staining of acid fast bacilli and cryptosporidia by replacing the phenol content of the carbol fuschin stain with various concentrated detergents. In this report the technique was modified slightly using a non-concentrated detergent and applied to the staining of oocysts of C cayetanensis. It was found that oocysts of C cayetanensis do not stain using the modified detergent ZN method when compared with similar preparations containing oocysts of Cryptosporidium spp.

Animals↗

Attempts to establish experimental Cyclospora cayetanensis infection in laboratory animals.

Attempts were made to develop an animal model for Cyclospora cayetanensis to identify a practical laboratory host for studying human cyclosporiasis. Oocysts collected from stool of infected humans in the United States, Haiti, Guatemala, Peru, and Nepal were held in potassium dichromate solution to allow development of sporozoites. The following animal types were inoculated: 9 strains of mice, including adult and neonatal immunocompetent and immune-deficient inbred and outbred strains, rats, sandrats, chickens, ducks, rabbits, jirds, hamsters, ferrets, pigs, dogs, owl monkeys, rhesus monkeys, and cynomolgus monkeys. Most animals were inoculated by gavage, although some of the primates were fed oocysts on food items. The animals were examined for signs of infection, particularly diarrhea, and stool samples were examined for 4-6 wk after inoculation. None of the animals developed patent infections or signs of infection. We conclude that none of the animals tested is susceptible to infection with C. cayetanensis.

Animals↗

Two simultaneous cases of Cyclospora cayatensis enteritis returning from the Dominican Republic.

According to the "International Passenger Survey," published in 1996 by the Office of Trading Standards, 534,000 British people traveled to the Caribbean area (personal communication, ABTA, 1998). The Dominican Republic, the eastern end of a large Caribbean island, has become in recent years one of the most popular destinations for UK holidaymakers as well as for travelers from many other countries. Cyclospora cayatensis has been firmly identified as a cause of gastroenteritis among international travelers,1 including human immunodeficiency virus (HIV)-positive individuals,2 but it has not been previously reported in the literature in British individuals returning from this increasingly popular vacation destination.

Aged↗

Cyclospora in patients with traveller's diarrhea.

Following the description in 1986 of a new enteropathogen called Cyanobacteria-like body (CLB), several outbreaks caused by this microorganism have been described. Since then several research teams have agreed to place CLB in the genus Cyclospora and to establish the epidemiological, pathogenical and clinical features of the illness. This paper describes and assesses those features in 20 travellers diagnosed with CLB and reviews the main literature concerning this enteropathogen. CLB was found in 2.8% of all cases of traveller's diarrhea in our series.

Adult↗

Cyclospora: an enigma worth unraveling.

In part, Cyclospora cayetanensis owes its recognition as an emerging pathogen to the increased use of staining methods for detecting enteric parasites such as Cryptosporidium. First reported in patients in New Guinea in 1977 but thought to be a coccidian parasite of the genus Isospora, C. cayetanensis received little attention until it was again described in 1985 in New York and Peru. In the early 1990s, human infection associated with waterborne transmission of C. cayetanensis was suspected; foodborne transmission was likewise suggested in early studies. The parasite was associated with several disease outbreaks in the United States during 1996 and 1997. This article reviews current knowledge about C. cayetanensis (including its association with waterborne and foodborne transmission), unresolved issues, and research needs.

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Cyclospora cayetanensis among expatriate and indigenous populations of West Java, Indonesia.

From January 1995 through July 1998, we investigated the occurrence of Cyclospora cayetanensis infection associated with gastrointestinal illness or diarrhea in foreign residents and natives of West Java, Indonesia. We found that C. cayetanensis was the main protozoal cause of gastrointestinal illness and diarrhea in adult foreign residents during the wet season. The parasite rarely caused illness in the indigenous population or in children.

Adult↗

Recovery of waterborne oocysts of Cyclospora cayetanensis by Asian freshwater clams (Corbicula fluminea).

Asian freshwater clams (Corbicula fluminea) were exposed for 24 hr in 38 liters of water contaminated with 1.0 x 10(5) Cyclospora cayetanensis oocysts (2.6 x 10(3) oocysts/L). The hemolyph and gill smears of 30 clams were examined by acid-fast stain on days 1, 3, 5, 7, 10, 13, and 18 postexposure (PE). Since no oocysts were detected in the water 24 hr after contamination by the membrane filter-dissolution method, the oocyst retention rate was 4.6 X 10(2) oocysts/clam. The prevalence of oocyst-positive clams significantly decreased (P < 0.01) from 93% to 47% during 13 days PE. None of the clams contained oocysts on day 18 PE; no oocysts were detected in the clam feces. The numbers of oocysts recovered from six clam size classes varied and significantly decreased with smaller clam size (P < 0.01). The lowest prevalence values of oocyst-positive clams, 45% and 34%, were observed in the two lowest size classes: 12.1-14.0 mm and 14.1-16.0 mm, respectively. The prevalence values in the remaining four classes ranged from 84% to 100%. The sampling program demonstrated that the population of 180 clams examined during the study up to 13 day PE could be assessed for C. cayetanensis positivity by random testing of a minimum of 75 clams (42%). When the two lowest clam size classes are eliminated, the population of 114 clams could be assessed by sampling a minimum of 32 clams (28%). The results demonstrate that Corbicula fluminea can recover waterborne oocysts of C. cayetanensis, and could be used as biological indicators of contamination of water with C. cayetanensis oocysts.

Animals↗

Trimethoprim-sulfamethoxazole compared with ciprofloxacin for treatment and prophylaxis of Isospora belli and Cyclospora cayetanensis infection in HIV-infected patients. A randomized, controlled trial.

BACKGROUND: In developing countries, Isospora belli and Cyclospora cayetanensis frequently cause chronic diarrhea in HIV-infected patients. OBJECTIVE: To compare 1 week of trimethoprim-sulfamethoxazole treatment and 1 week of ciprofloxacin treatment in HIV-infected patients with chronic diarrhea caused by I. belli and C. cayetanensis. DESIGN: Randomized, controlled trial. SETTING: HIV clinic in Port-au-Prince, Haiti. PATIENTS: 42 HIV-infected patients with chronic diarrhea due to I. belli (n = 22) or C cayetanensis (n = 20). INTERVENTIONS: Patients were randomly assigned to receive oral trimethoprim-sulfamethoxazole (160 mg or 800 mg) or ciprofloxacin (500 mg) twice daily for 7 days. Patients who responded clinically and microbiologically received prophylaxis for 10 weeks (1 tablet orally, three times per week). MEASUREMENTS: Treatment success was measured by cessation of diarrhea and negative stool examination at day 7. Prophylaxis success was measured by recurrent disease rate. RESULTS: Diarrhea ceased in all 19 patients treated with trimethoprim-sulfamethoxazole. Eighteen of 19 patients had negative results on stool examination at day 7 (95%). Among the 23 patients who received ciprofloxacin, diarrhea ceased in 20 (87% [CI; 66% to 97%]) and 16 had negative results on stool examination at day 7 (70%). By survival analysis, diarrhea from isosporiasis and cyclosporiasis ceased more rapidly with trimethoprim-sulfamethoxazole than with ciprofloxacin. All patients receiving secondary prophylaxis with trimethoprim-sulfamethoxazole remained disease-free, and 15 of 16 patients receiving secondary prophylaxis with ciprofloxacin remained disease-free. CONCLUSIONS: A 1-week course of trimethoprim-sulfamethoxazole is effective in HIV-infected patients with cyclosporiasis or isosporiasis. Although ciprofloxacin is not as effective, it is acceptable for patients who cannot tolerate trimethoprim-sulfamethoxazole.

AIDS-Related Opportunistic Infections↗

Cyclospora species as a cause of diarrhoea in humans.

Within the past 10 years, the coccidian parasite Cyclospora cayetanensis has been clearly recognised as a cause of protracted diarrhoea in humans. Heavy infection results in inflammation and epithelial injury in the duodenum and jejunum. Of the multiple symptoms, diarrhoea may neither be the presenting nor predominant one. Following an incubation period of 12 hours to 11 days, the abrupt onset of watery diarrhoea occurring in a relapsing cyclic pattern is characteristic, lasting from six to eight weeks in immunocompetent patients, but three months in the immunocompromised. Of worldwide distribution, it is endemic in certain regions where it can be associated with rainy seasons. Outbreaks or isolated cases may also occur in developed countries. Contaminated water can transmit the oocysts, and usual control procedures for potable water supplies fail to detect the parasite, which resists chlorination. Diagnosis depends upon laboratory investigations, usually faecal microscopy. For reliable detection of characteristic oocysts in the faeces, special procedures may be necessary. Trimethoprim-sulphamethoxazole is established as the treatment of choice and leads to rapid relief of symptoms, although relapse in immunocompromised patients may require secondary prophylaxis.

Animals↗

Survey for Cyclospora cayetanensis in domestic animals in an endemic area in Haiti.

From January 1997 through July 1998, we examined stool samples from 327 domestic animals, including pigs, cattle, horses, goats, dogs, cats, guinea pigs, chicken, ducks, turkeys, and pigeons in Leogane, Haiti, for the presence of Cyclospora cayetanensis infection. No coccidian oocysts morphologically compatible with C. cayetanensis were detected in any of the animal samples, despite their living in, or near, households with infected individuals. These results suggest that domestic animals are not reservoir hosts for C. cayetanensis and that in this endemic area, humans are the only natural host for this parasite.

Animals↗

[Excessive watery diarrhea and pronounced fatigue due to Cyclospora cayetanensis infection in an HIV infected traveler returning from the tropics].

We report the case of a 46-year-old HIV-1 infected patient who acquired a Cyclopsora cayetanensis infection during travel to southeast Asia. He developed excessive watery diarrhoea and pronounced fatigue, which resolved after treatment with cotrimoxazole. The term Cyclospora cayetanensis was first suggested in 1993 for the infectious agent of diarrhoea in tropical and subtropical regions. The infection is usually self-limiting. However, in HIV-1 infected persons the symptoms are often more severe and protracted. Microbiological diagnosis is performed by light microscopy and at X400 magnification of faeces fixed in SAF medium or 10% formalin preservatives.

AIDS-Related Opportunistic Infections↗

[A new trichromic safranin stain for the detection of Cryptosporidium parvum, Cyclospora cayetanensis, species of Microsporidia and Isospora belli in fecal material].

Cryptosporidium parvum, Isospora belli, Cyclospora cayetanensis and Microsporidia are frequent pathogens in the immunodeficient host, which may cause multiple infections. The above mentioned parasites are found in feces by the application of different specific tintorial techniques. The objective of this work was the development of a stain for the simultaneous detection of these parasites, reducing costs as well as the time taken to make the diagnosis. The safranin-trichrome stain is simple, chip and its results are similar to those of specific tints. All microorganisms are easy to detect and besides being perfectly distinguishable from fungi and faecal elements.

AIDS-Related Opportunistic Infections↗

[Prevalence survey on Cyclospora cayetanensis and Cryptosporidium ssp. in diarrhea cases in Yunnan Province].

OBJECTIVE: To investigate the prevalence and distribution characteristics of Cyclospora cayetanensis and Cryptosporidium ssp. infection in diarrhea cases of Yunnan Province. METHODS: To collect fresh faeces from diarrhea cases in 7 counties/cities, examine the specimens by direct smear with iodine staining and modified acid-fast staining. RESULTS: The infection rate of C. cayetanensis and Cryptosporidium ssp. was 3.97% and 5.29%, respectively. The infection rate of the two pathogenic coccidians was as high as 10.64% and 8.51% in preschool children. C. cayetanensis was found in 3 counties and Cryptosporidium in 6 counties. CONCLUSION: Both C. cayetanensis and Cryptosporidium ssp. are prevalent in Yunnan Province with the latter distributed more widely, and the two pathogens are more prevalent in preschool children.

Adolescent↗

Cyclospora cayetanensis associated with watery diarrhoea in Peruvian patients.

Acid-fast, coccidian-like bodies (Cyclospora cayetanensis) were identified over the last 18 months in the stools of seven Peruvian patients suffering from diarrhoea. The follow-up of two patients revealed a watery, self-limited diarrhoea, which lasted for up to four weeks. The organism was simultaneously identified in the diarrhoeal stools of three members of the same family who drank unchlorinated canal water and in the stools of a duck bred by this family. The organism was not found in the faecal samples of 50 healthy subjects and 10 ducks bred by families without known diarrhoea cases. These findings, albeit preliminary, may be suggesting that besides consumption of untreated water, additional modes of transmission such as contact with domestic animals may be important in this disease. Further studies are needed to assess whether this disease behaves as a zoonotic condition and to ascertain the relative importance of symptom-free subjects in person-to-person transmission of the organism.

Adolescent↗

[3 emerging protozoal infections in The Netherlands: Cyclospora, Dientamoeba, and Microspora infections].

Increased international travelling, an increased number of patients with immunosuppression caused by HIV infection, and renewed interest in known but little studied microorganisms, resulted in a more frequent finding of certain medically important parasites. Three emerging protozoal infection, Cyclospora cayetanensis, Dientamoeba fragilis and Microspora (Enterocytozoon bieneusi and Encephalitizoon) are causative agents of diarrhoea. Encephalitozoon infections are also associated with hepatitis, hepatoconjunctivitis and nephritis. C. cayetanensis infection was diagnosed in 28 patients in the years 1992-1995 in the Academic Medical Centre in Amsterdam, half of these patients returning from a visit to Indonesia. D. fragilis has a prevalence in the Netherlands of 8% among patients with diarrhoea lasting longer than one week referred for parasitological investigation. The prevalence of E. bieneusi in HIV positive patients with diarrhoea was 8-10% in the Academic Medical Centre in the last five years. Infection with Encephalitozoon appears to be endemic in the Netherlands.

Animals↗

Update: outbreaks of Cyclospora cayetanensis infection--United States and Canada, 1996.

Since May 1996, CDC has received reports of clusters and sporadic cases of infection with the parasite Cyclospora cayetanensis that occurred in May and June in the United States and Canada. This report describes preliminary findings of an investigation by the New Jersey Department of Health and Senior Services (NJDHSS) and updates the findings of other ongoing investigations.

Animals↗