Assessment of the quality of multi-institutional data for population-based epidemiological studies: a case study of atoxoplasmosis in Bali mynahs (Leucopsar rothschildi).

To evaluate the potential to conduct population-based epidemiologic studies using multi-institutional data, the quality of data extracted from the MedARKS system was evaluated with regard to atoxoplasmosis in Bali mynahs (Leucopsar rothschildi). Data extracted for 338 Bali mynahs from 34 institutions were analyzed for completeness, correctness, and consistency and subjectively analyzed for accessibility. Data completeness was ascertained by analysis of missing records, tests, and blank fields. Two hundred and sixty four records had animal enclosure information missing, nine records were missing from the parasitology module, and 85 records did not include tests or results. Data correctness was assessed by evaluation of unclear and inaccurate results. From 2,432 parasitology records, 81 tests (3.3%) were not definitive because of uninterpretable entries. Data consistency was assessed by comparing the problem list to positive tests in the parasitology module and listing of buffy coat smears in the clinical pathology module. Overall, six different terms with 28 values were used to denote "positive for Atoxoplasma sp.," and a substantial number of discrepancies were found between problem lists, parasitology modules, and test findings among these data records. In general, this study showed that the data contained in a computer patient record (CPR) system for zoos has great promise for population-level studies if specific areas are addressed, including 1) reduced reliance on free-text data entry, 2) universal use of a standardized vocabulary, 3) use of methods to identify and track individual animals accurately and easily, 4) integration of data checks and maintenance methods, and 5) concerted use of "centralized" animal medical record information.

Morphologic, immunohistochemical, and molecular characterization of a novel Lankesterella protozoan in two White's tree frogs (Litoria caerulea).

Two White's tree frogs (Litoria caerulea) housed at a zoological park died after a short period of lethargy, weight loss, and edema. Detailed postmortem examinations were performed on both frogs, including bacterial cultures and complete histologic examinations. Intracytoplasmatic as well as free protozoan parasites were identified in multiple organs from both frogs. The parasites were identified within erythrocytes, leukocytes, endothelial cells, and hepatocytes. Immunohistochemistry demonstrated a cross-reaction with Toxoplasma gondii antisera. Parasite ultrastructural analysis was performed by transmission electron microscopy. The parasites demonstrated an apical complex containing a conoid, rhoptries, and micronemes, demonstrating it was a member of the phylum Apicomplexa. In addition, the parasites had bipolar paranuclear bodies, organelles that are typical of coccidian sporozoites. The organisms were tentatively identified as members of the genus Lankesterella on the basis of histologic and ultrastructural morphology. A portion of the 18s ribosomal RNA (rRNA) gene was amplified via a polymerase chain reaction, sequenced, and used in a Basic Local Alignment Search Tool search of the GenBank database. The 18s rRNA gene sequence was found to be most similar to gene sequences isolated from Lankesterella organisms (88%). In aggregate, these data support the classification of these protozoa as a novel species of Lankesterella. A causal relationship between frog morbidity and protozoal parasitism was not determined. This is the first report of Lankesterella sp. in White's tree frogs.

Detection of some intestinal protozoa in commercial fresh juices.

Fresh fruit juices are popular, but not always safe. For assessing the likelihood of infection with newly emerging intestinal protozoa, commercial fresh orange, lemon, sugar cane, strawberry, and mango juices were screened by wet mounts, Weber's modified trichrome and modified Ziehl-Neelsen stains. Protozoa viability was done by fluorescein-diacetate/propidium-iodide staining, and infectivity was performed in Swiss albino mice. Results showed that 35.43% were contaminated with one or more of Cryptosporidia, Microsporidia, and Cyclospora, as well as Giardia spp. Strawberry was the most contaminated juice (54.28%), while orange was the slightest (22.86%). Cryptosporidia was the highest contaminant (61.29%), and Cyclospora was the least (14.52%). Microsporidia spp. was the most robust contaminant which retained its viability and infectivity in juices in which it was detected. Moderately acidic strawberry and mango juices and alkaline sugar cane juice pose a possible threat, due to harboring the highest viable and infectious protozoa. Regarding highly acidic juices, viability and infectivity decreased in lemon, yet was not still risk free. Orange juice was comparatively safe, as viability dramatically declined, while infectivity was completely abolished. Hence consumers, especially high risk group, are placed at hazard of contracting intestinal protozoa infections, especially through moderately acidic and alkaline juices.

Gastrointestinal parasites of Howler monkeys (Alouatta palliata) inhabiting the fragmented landscape of the Santa Marta mountain range, Veracruz, Mexico.

In recent years populations of howler monkeys (Alouatta palliata) in southeastern Mexico have decreased substantially due to the transformation and loss of natural habitats. This is especially evident in the Santa Marta mountain range, Veracruz, Mexico where several studies have evaluated the impact of fragmentation on howler monkey populations in order to propose management programs for their conservation. The conditions generated by fragmentation likely change the rates of parasitic infection and could decrease howler survival. In this study, gastrointestinal parasite species richness, prevalence, and egg density of infection were determined in howler groups inhabiting five forest fragments at the Santa Marta mountain range. Two hundred and seventy-eight fresh fecal samples were collected between October 2002 and April 2003. Three parasite species were found during the dry and the wet season in all forest fragments sampled: one unidentified species of Eimeriidae; Trypanoxyuris minutus (Oxyuridae); and Controrchis biliophilus (Dicrocoeliidae). Both the prevalence of T. minutus and infection density for all parasites differed between seasons and fragments (the largest fragment consistently differed from other fragments). Host density, distance to the nearest town, fragment size, fragment shape, and total basal area of food trees explained parasite prevalence, but each species had a different pattern. Although parasite richness was lower, prevalence and density were higher than values reported for howlers in conserved forests. These results suggest that the establishment of biological corridors and animal translocation programs must take into account the parasite ecology of each fragment to avoid higher infection rates and preclude potential consequent mortality.

The impact of health communication and enhanced laboratory-based surveillance on detection of cyclosporiasis outbreaks in California.

We investigated the timing of diagnosis, influence of media information on testing for Cyclospora, and the method used to identify cases during eight cyclosporiasis outbreaks in California in spring of 1997. We found that Internet information, media reports, and enhanced laboratory surveillance improved detection of these outbreaks.

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.

Intestinal parasites in patients with diarrhea and human immunodeficiency virus infection in Zimbabwe.

OBJECTIVES: To determine the prevalence of intestinal parasites and risk factors for infection associated with diarrhea in HIV-infected patients in Harare, Zimbabwe. DESIGN: Prospective observational study. METHODS: Single stool samples were collected from 88 HIV-infected individuals presenting with diarrhea of greater than 1 week duration. Stools were examined for intestinal parasites using modified acid fast stain, fluorescence- labeled monoclonal antibody for Cryptosporidium parvum, as well as a modified trichrome stain and a PCR-based protocol for Enterocytozoon bieneusi. RESULTS: C. parvum was detected in 9% (seven out of 82) of samples evaluated, but no Cyclospora was detected. E. bieneusi was detected in 18% (10 out of 55) of stool by trichrome staining and in 51% (28 out of 55) of stool examined by PCR. Risk factors for E. bieneusi infection were: living in rural areas, consumption of nonpiped water, contact with cow dung and household contact with an individual with diarrhea. CONCLUSION: E. bieneusi infection was common in HIV-infected patients with diarrhea in Zimbabwe and may be acquired through person-to-person and fecal-oral transmission.

Description of Pythonella scleruri n. sp. (Apicomplexa: Eimeriidae) from a brazilian bird rufous-breasted-leaftosser Sclerurus scansor, 1835 (Passeriformes: Furnariidae).

Coccidian oocysts containing 16 sporocysts with 4 sporozoites in each were observed in a faecal sample from Sclerurus scansor collected in the Itatiaia National Park, southeastern region of Brazil. The oocysts are characterized by ellipsoidal shape measuring 42.5 x 32.8 mm, with smooth, thick double-layered wall of a greenish-orange colour. An oocyst residuum of numerous scattered granules among the sporocysts in sporulated ones; 16 round sporocysts, averaging 10.5 x 10 mm each containing four elongated sporozoites; presence of residuum; absence of Stieda body. The presently described coccidian, recorded for the first time in birds, is a new species named P. scleruri.

The genus Hammondia is paraphyletic.

The phylogenetic relationships amongst Hammondia, Neospora and Toxoplasma were investigated by DNA sequence comparisons of the D2/D3 domain of the large subunit ribosomal DNA and the internal transcribed spacer 1. The results obtained allow us to reject the hypothesis that N. caninum and H. heydorni are the same species and show that Hammondia hammondi is probably the sister taxon to Toxoplasma gondii.

Evaluation of five stains in diagnosing human intestinal coccidiosis.

Three hundreds cases over the year 1998 complaining of diarrhoea were examined. The stools were examined by the traditional diagnostic methods and confirmed to be free from intestinal parasites. Cryptosporidium parvum, Isospora belli and Cyclospora cayetanensis were detected by using different types of faecal stains namely modified Ziehl-Neelsen, Kinyoun acid-fast, Auramine-rhodamine, Gomori's trichrome and Giemsa. The number of positive cases were 87 C. parvum (29%), 5 cases L. belli (1.7%) and 12 cases C. cayetanensis (4%). This study showed that the sensitivity of modified Ziehl-Neelsen and Kinyoun Acid-fast were very high (100%) in comparison with the other stains.

Pathogenicity of experimental caryosporosis.

Pathogenicity of the coccidia C. bigenetica and C. simplex was studied in experimentally inoculated pigs, goat kids (untreated and immunosuppressed) and severe combined immunodeficient (SCID) mice. The major pathological changes of caryosporosis were similar in all inoculated animals. In pigs and goat kids, caryosporosis was self-limiting, with clinical responses that included focal swelling and erythema of the muzzle, snout, jaws, cheeks, eyelids, bases of the ears, backs of the necks, scrotum, external genitalia of females, legs and footpads. Histopathological changes were characterized by involvement of the cutaneous mononuclear phagocyte system with an inflammatory exudate containing numerous macrophages, especially around the root sheaths, sensory nervous corpuscles of the hair follicles and surrounding dermal free nerve endings. The tactile hair follicles in the muzzle, snout and upper jaw were most severely changed. In SCID mice, inoculation with C. bigenetica or C. simplex caused a severe, fatal, systemic disease characterized by dissemination of numerous caryosporan developmental stages into the host mononuclear phagocyte system. This study presents evidence that both caryosporan species tested caused similar clinical signs and lesions of dermal coccidiosis in the mammalian secondary hosts.

Besnoitia sp. (Protozoa:Toxoplasmatinae) from Akodon montensis (Rodentia:Cricetidae) in Santa Catarina State, Brazil.

Tissue cysts of Besnoitia sp. were found in muscles and several organs from a naturally infected Akodon montensis captured in the rural area of the municipality of Timbó, Santa Catarina State, in southern Brazil. Indirect fluorescence and enzyme-linked immunosorbent assays carried out with sera from mice chronically infected with Toxoplasma gondii and Besnoitia sp. showed, as expected, a stronger reaction against homologous than heterologous antigens. No cross-protection was observed in mice immunized with T. gondii when challenged with Besnoitia sp. This is the first description of a natural infection of A. montensis by parasites of the genus Besnoitia sp. in Brazil.

Vipera berus and V. ammodytes (Serpentes:Viperidae) represent new host for Caryospora simplex (Apicomplexa:Eimeriidae) in Europe.

During a survey of the coccidian parasites of reptiles, caryosporan oocysts wee found in the faeces of wild and captive European viperid snakes Vipera berus (L.) and V. ammodytes (L.). Thirty two of 37 examined V. berus (86%) and 9 of 17 examined V. ammodytes (53%) specimens were found to be passing caryosporan oocysts. Morphological characters of all caryosporan isolates were identical and fitted well with the description of Caryospora simplex Léger, 1904. Experimental inoculation of severe combined immunodeficient (SCID) mice with seven isolates of C. simplex from V. berus or V. ammodytes confirmed the heteroxenous life cycle pattern, for the first time for isolates of evidently European origin. Caryosporan developmental stages were observed in the connective tissues of the nose, cheeks, ear and scrotum in all inoculated SCID mice. V. berus and V. ammodytes represent new hosts for C. simplex. The present paper represents the first widely based report on coccidian parasites of the genus Caryospora Léger in European viperids. Our findings indicate a wide distribution of C. simplex throughout the range of distribution of snakes of the genus Vipera.

An epizootic of besnoitiosis in goats in Fars province of Iran.

Over 100,000 goats were grazing freely in isolated herds of 100 to 600 goats in a mountainous area of more than 2000 km2 to the north-east of Darab City in Fars Province in southern Iran. Twenty to fifty percent of the goats in this area were infected with besnoitiosis. Sheep, cattle and donkeys grazing in the same area showed no clinical manifestations of the infection. The diagnosis of the infection in the clinically suspected animals was confirmed by histological studies on skin biopsies from the ear tips and carpal and tarsal regions, and the distribution of this protozoon in the tissues and organs of this intermediate host was studied by post-mortem and histological examinations. Skin biopsies from the carpal and tarsal areas of 12% of clinically normal goats from the infected area were also lightly infected with besnoitia cysts.

The morphology and pathology of Besnoitia sp. in reindeer (Rangifer tarandus tarandus).

Four of five reindeer (Rangifer tarandus tarandus) obtained from a Besnoitia sp.- infected herd at the Assiniboine Park Zoo in Winnipeg, Manitoba, Canada, in October 1989, had evidence of mild dermatitis over the articular surfaces of carpal and tarsal joints. Cysts of Besnoitia sp., either surrounded by inflammatory reactions or without evident host response, were present within the dermis, submucosa of the nasal turbinates, periosteum, tendons, testes and hooves. The light microscopic and histochemical features of Besnoitia sp. from reindeer were indistinguishable from those of other Besnoitia spp. described in cattle, rodents and horses. The Besnoitia sp. cysts and organisms from reindeer were unique in that bradyzoite membrane micropores and cytoplasmic enigmatic bodies were not observed. Two cats were fed cysts of Besnoitia sp. but no oocysts were detected in feces for 90 days post-infection.

Caryospora simplex (Apicomplexa: Eimeriidae) from a captive Kaznakov's viper (Vipera kaznakovi).

Caryospora simplex is reported for the first time from the feces of a captive female Kaznakov's viper (Vipera kaznakovi) in Albuquerque, New Mexico (USA). Coccidian meronts and gamonts were observed in the intestinal epithelial cells of another female Kaznakov's viper that died in October 1993.

Coccidian parasites (Apicomplexa) from snakes in the southcentral and southwestern United States: new host and geographic records.

Four hundred thirty-five leptotyphlopid, colubrid, elapid, and viperid snakes were collected from various localities in Arkansas, New Mexico, Oklahoma, and Texas, and their feces were examined for coccidian parasites. Of these, 131 (30%) were passing oocysts or sporocysts of at least 1 coccidian; 88 (67%) of the infected snakes had only 1 species of coccidian when they were examined. Aquatic and semiaquatic snakes accounted for 48% of the infections, whereas strictly terrestrial snakes comprised the other 52%. There was more than a 2-fold difference in prevalence among these 2 groups as 63 of 129 (49%) of the aquatic and semiaquatic snakes versus 68 of 306 (22%) of the terrestrial snakes harbored coccidia. Most terrestrial snakes were infected by species of Caryospora and Sarcocystis that are either facultatively or obligatorily heteroxenous. The aquatic and semiaquatic species most often harbored eimerians. Attempts to transmit some of the Sarcocystis spp. experimentally from Crotalus atrox to Mus musculus, Peromyscus leucopus, Peromyscus maniculatus, or Microtus ochrogaster were unsuccessful. This report documents 27 new host and several distributional records for coccidians from snakes in the southcentral and southwestern United States.

Comparative ultrastructural studies on Besnoitia besnoiti and Besnoitia caprae.

Comparative transmission electron microscopy on Besnoitia besnoiti and on a strain of Besnoitia derived from goats in Kenya revealed that the two organisms differ in their pellicle, micropore, microtubules, nucleus, wall-forming body 1 (W1), amount of lipids and amylopectin. Thus the caprine besnoitia is probably a different organism and the term Besnoitia caprae should continue to be used.

Caprine besnoitiosis: studies on the experimental intermediate hosts and the role of the domestic cat in transmission.

A study was conducted to test the infectivity of bradyzoites of a Besnoitia species infecting goats in Kenya to rats, mice, rabbits, sheep and goats. Only goats developed infection resulting in tissue cyst formation. Eighteen cats were tested for their role in transmission of this Besnoitia species. Ten of the cats were fed on goat tissues with numerous Besnoitia cysts; four cats were orally inoculated with bradyzoites and four others fed on mice and rat carcasses previously inoculated with bradyzoites. None of these cats produced Besnoitia oocysts in their faeces for 30 days.

Isolation of Besnoitia wallacei in Kenya.

The development of Besnoitia wallacei was studied in 13 cats fed on tissues of mice and rats previously infected with B. wallacei. The cats were serially killed between Day 1 and Day 16 of infection, and histological sections from the liver and intestines were examined. Asexual stages were seen in both the small intestines and the liver between Day 6 and Day 16 post-infection. Mature microschizonts in intestinal epithelial cells measured 22.6 microns x 14.7 microns (n = 15). Macroschizonts in intestinal lamina propria measured 66.6 microns x 50.3 microns (n = 25). Those in the liver measured 70.9 microns x 55.0 microns (n = 5). Sexual stages were seen in epithelial cells of the small intestines only.