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.

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.

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.

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.

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.

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.

Camel (Camelus dromedarius) and sheep (Ovis aries) meat as a source of dog infection with some coccidian parasites.

Experimental infection of dogs with camel (Camelus dromedarius) meat resulted in infection of the dogs with Isospora canis, Hammondia heydorni and Sarcocystis cameli. The dogs fed sheep (Ovis aries) meat passed oocysts of Isospora canis, Isospora ohioensis and sporocyts of Sarcocystis spp. Extraintestinal stages were detected in the intestinal lymph node of a rabbit killed 4 days following inoculation with Isospora ohioensis oocysts. Dogs fed the rabbit (killed 4 days after inoculation with I. ohioensis) passed I. ohioensis oocysts in their faeces 8 days post-infection.

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.

Natural besnoitiosis in a rabbit.

An adult female rabbit was submitted for necropsy after sudden death. Pneumonia and nephritis were diagnosed by gross examination. On histological examination. Besnoitia cysts were seen in the pulmonary interalveolar tissue. The Besnoitia cysts were morphologically normal in appearance, measured 127 to 185 microns in diameter and provoked mild mononuclear inflammatory reaction. To the best of our knowledge there has been no previous report of natural besnoitiosis in rabbits.

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.

Experimental Caryospora bigenetica (Apicomplexa: Eimeriidae) infections in swine (Sus scrofa).

Two Hampshire-Landrace crossbred pigs were found to contain developmental stages of Caryospora bigenetica following oral inoculation with 1 x 10(8) oocysts. One pig was given intramuscular injections of methylprednisolone acetate. Both pigs displayed clinical signs of dermal coccidiosis from 3 to 10 days after inoculation, including swollen jowls and hocks, bilateral ocular discharges, generalized erythema, and lethargy. Meronts and gamonts were observed histologically in numerous tissues including jowl, ear, footpad, tongue, and lung at 10 days postinoculation. The present study is the first report of C. bigenetica infections in swine.

Investigation of transplacental transmission of Caryospora bigenetica (Apicomplexa: Eimeriidae) in mice.

Five pairs of female Swiss-Webster mice were caged with 5 males (2 females/1 male). Eight females were inoculated orally with 2.6 x 10(5) Caryospora bigenetica oocysts either 3 days before mating, 3 days after mating (PMD), 9 PMD, or 16 PMD. The remaining 2 females were inoculated orally with Hanks' balanced salt solution and served as controls. One female from each cage delivered naturally at full term and the second female delivered by cesarean section on postmating day 18. The number of offspring per litter ranged from 7 to 12. One female produced a litter of 3 stillborn and 5 liveborn offspring. Seven of 8 female mice exhibited swollen muzzles and footpads 8 days after inoculation. Caryospora bigenetica was identified in tissues of muzzle, tongue, footpad, uterus, and placenta at necropsy. This is the first report of C. bigenetica in uterus and placenta. Clinical signs and tissue infections were not observed in control mice or in any offspring of the 10 female mice. This study presents evidence that C. bigenetica is not transmitted transplacentally.

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.

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.

Blood parasites of amphibians from Sichuan Province, People's Republic of China.

Two hundred forty-six amphibians from Sichuan Province, People's Republic of China, were examined for blood parasites between April and June 1990. Six trypanosome species were found, 2 of which were not identified because of poor material. Trypanosoma rotatorium (sensu Mayer, 1843) was found in Rana limnocharis and Rana nigromaculata. Trypanosoma chattoni and a T. rotatorium-like species were found in Bufo gargarizans. A sphaeromastigote similar to Trypanosoma tsunezomiyatai was seen in R. limnocharis. The relationship of this parasite to T. chattoni is unclear. Dactylosoma ranarum, Lankesterella minima, and Aegyptianella bacterifera were identified in ranids. Frog erythrocytic virus (FEV) was found in 2 ranids and a bufonid. Differences in shape of FEV and its assumed effect on the host-cell nucleus suggest that different kinds of viruses may be involved.

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.

A coccidian (Apicomplexa: Eimeriidae) with extracytoplasmally located stages in the kidney tubules of golden carp (Carassius auratus gibelio L.) (Cyprinidae).

Numerous coccidian stages were found in the kidney tubules of the golden carp (Carassius auratus gibelio). The merogonial and gamogonial stages were localized extracytoplasmally in the microvillous region of the epithelial cells. The host-parasite interface consisted of i) a large area where the parasite was separated from the host cytoplasm by the parasitophorous vacuole membrane only, and ii) a zone of multiple fusions of the host cell membrane investing the parasite to the neighbouring microvilli. The taxonomic status of the extracytoplasmic stages is not clear, however, their possible appurtenance to Eimeria scardinii, which was frequently found in the kidneys of golden carps in the same population, is discussed.

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.

Rodents are not a source of endogenously-produced, fecally-transmitted Caryospora bigenetica oocysts.

Fifteen Swiss-Webster mice (Mus musculus) and eight cotton rats (Sigmodon hispidus) were inoculated orally with Caryospora bigenetica oocysts. Feces from these animals were collected from 0 to 180 days postinoculation (DPI) and examined for endogenously-produced oocysts using Nomarski microscopy. Oocysts were recovered from mouse feces at 0, 1, 2, 3, 5, 7, 8, 10, and 14 DPI, and from cotton rat feces at 1, 2, and 9 DPI. The recovered oocysts were determined to be from the original inocula due to the presence of thick walls, polar granules, and Stieda and substieda bodies. All animals exhibited clinical signs at 8 DPI. Developmental stages of C. bigenetica were identified in various tissues of seven cotton rats found dead at 9, 10, 11, 12, and 13 DPI. Caryocysts were found in muzzle, tongue, footpad, scrotum, and rectum of mice and cotton rats at 30 DPI. Fecal samples collected from mice on 0, 8, 10, 12, 14, 16, and 18 DPI, and from cotton rats on 0, 9, 11, 13, 15, and 17 DPI were injected subcutaneously into 13 mice. Of the 13 mice, a Caryospora infection was observed only in the mouse inoculated with 0 DPI mouse feces. We propose that endogenously-produced C. bigenetica oocysts are not fecally-transmitted by Swiss-Webster mice or cotton rats.

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.