Transfer of extraintestinal stages of Eimeria vermiformis in the mouse

The oral administration of whole blood and liver or lung homogenates from mice donors inoculated 1-3 days previously with Eimeria vermiformis oocysts produced infection in coccidia-free recipient mice.     

An investigation of sterile immunity against toxoplasmosis in rats

The non-persistent BK strain was examined for its ability to induce sterile immunity in Wistar rats. Groups of 2-9 Wistar rats were inoculated subcutaneously with 5 x 10(4) BK strain tachyzoites per rat. Two months later, 46 rats were dosed by gavage with 2 x 10(1) cysts of the C, ME-49, Prugniaud, C-56, Elg, M-7741 or M3 strains. Another 26 rats were inoculated with 10(3) oocysts of the ME49, M7741, Bear or Hopa-Hopa strains of Toxoplasma gondii. After 2 months, the rats were euthanized and their brains screened microscopically for toxoplasma tissue cysts and bioassayed in mice if negative. As judged by bioassay, the BK strain of Toxoplasma induced statistically significant protection against reinfection only when rats were challenged with cysts of the C and Prugniaud strains or with oocysts of the ME49 strain. Nonetheless, cysts were detected microscopically only in 23% of brains of immunized rats challenged with oocysts of the Bear and Hopa-Hopa strains of Toxoplasma and none of those challenged with tissue cysts of any strain. Tissue cysts were detected in 43 and 48% of non-immunized control rats infected with tissue cysts and oocysts, respectively. The overall infection in control rats (microscopy and bioassay) was 70 and 72% for rats inoculated with cysts and oocysts, respectively. These results are consistent with the divergent results obtained by other investigators with regard to protection after challenge with different complete strains (cyst and oocysts forming) of the parasite, of rats immunized with incomplete strains.     

Experimental Toxoplasma gondii infection in grey seals (Halichoerus grypus)

Laboratory-reared animals were used to assess the susceptibility of seals (Halichoerus grypus) to Toxoplasma gondii infection. Four seals were each orally inoculated with 100 or 10,000 oocysts of T. gondii (VEG strain), and another 4 seals served as negative controls. Occasionally, mild behavioral changes were observed in all inoculated seals but not in control animals. A modified agglutination test revealed the presence of antibodies to T. gondii in sera collected from inoculated seals and mice inoculated as controls. No evidence of the parasite was found on an extensive histological examination of seal tissues, and immunohistochemical staining of tissue sections from inoculated seals revealed a single tissue cyst in only 1 seal. Control mice inoculated with 10 oocysts from the same inoculum given to seals became serologically and histologically positive for T. gondii. Cats that were fed brain or muscle tissue collected from inoculated seals passed T. gondii oocysts in feces. This study demonstrates that T. gondii oocysts can establish viable infection in seals and supports the hypothesis that toxoplasmosis in marine mammals can be acquired from oocysts in surface water runoff and sewer discharge.     

Susceptibility of common voles to experimental toxoplasmosis

Common voles (Microtus arvalis) in groups of nine to 10 animals were inoculated per os with a dose of 1, 10, 1x10(2), 1x10(3), and of the K1 strain of Toxoplasma gondii. All the common voles inoculated with 1 to 1 x 10(3) oocysts remained subclinical and survived. Three of the 10 voles inoculated with 1 x 10(4) oocysts died between days 7 and 12 post inoculation (p.i.). Antibodies were demonstrated in all the infected voles killed on day 60 p.i. The highest antibody titres in voles detected by the dye test (DT) and latex agglutination test (LAT) were 1,024 and 1,280, respectively.     

Survival of Toxoplasma gondii oocysts in Eastern oysters (Crassostrea virginica)

Toxoplasma gondii has recently been recognized to be widely prevalent in the marine environment. It has previously been determined that Eastern oysters (Crassostrea virginica) can remove sporulated T. gondii oocysts from seawater and that oocysts retain their infectivity for mice. This study examined the long-term survival of T. gondii oocysts in oysters and examined how efficient oysters were at removing oocysts from seawater. Oysters in 76-L aquaria (15 oysters per aquarium) were exposed to 1 x 10(6) oocysts for 24 hr and examined at intervals up to 85 days postexposure (PE). Ninety percent (9 of 10) of these oysters were positive on day 1 PE using mouse bioassay. Tissue cysts were observed in 1 of 2 mice fed tissue from oysters exposed 21 days previously. Toxoplasma gondii antibodies were found in 2 of 3 mice fed oysters that had been exposed 85 days previously. In another study, groups of 10 oysters in 76-L aquaria were exposed to 1 x 10(5), 5 x 10(4), or 1 x 10(4) sporulated T. gondii oocysts for 24 hr and then processed for bioassay in mice. All oysters exposed to 1 x 10(5) oocysts were infected, and 60% of oysters exposed to 5 x 10(4) oocysts were positive when fed to mice. The studies with exposure to 1 x 10(4) oocysts were repeated twice, and 10 and 25% of oysters were positive when fed to mice. These studies indicate that T. gondii can survive for several months in oysters and that oysters can readily remove T. gondii oocysts from seawater. Infected filter feeders may serve as a source of T. gondii for marine mammals and possibly humans.     

Examination of attachment and survival of Toxoplasma gondii oocysts on raspberries and blueberries

The consumption of Toxoplasma gondii oocysts on fresh produce may be a means of its transmission to humans. Cats shed T. gondii oocysts, which contaminate produce directly or contaminate water sources for agricultural irrigation and pesticide and fertilizer applications. Cyclospora cayetanensis is a related coccidial parasite, and outbreaks of diarrhea caused by C. cayetanensis have been associated with the ingestion of contaminated raspberries. The oocysts of these coccidians are similar in size and shape, indicating that they may attach to and be retained on produce in a similar manner. In the present study the attachment and survival of T. gondii oocysts on 2 structurally different types of berries were examined. Raspberries and blueberries were inoculated individually with 1.0 x 10(1) to 2.0 x 10(4) oocysts of sporulated T. gondii. Berries inoculated with 2.0 x 10(4) oocysts were stored at 4 C for up to 8 wk. Oocyst viability and recovery were analyzed by feeding processed material to mice. Mice fed T. gondii-inoculated berries stored at 4 C for 8 wk developed acute infections. In other experiments mice fed raspberries inoculated with > or = 1.0 x 10(1) oocysts became infected, whereas only mice fed blueberries inoculated with > or = 1.0 x 10(3) oocysts became infected. This study demonstrates that T. gondii oocysts can adhere to berries and can be recovered by bioassays in mice and that raspberries retain more inoculated oocysts than do blueberries. The results suggest that T. gondii may serve as a model for C. cayetanensis in food safety studies.     

Effect of dietary zinc level on serum carotenoid levels, body and shank pigmentation of chickens after experimental infection with coccidia

Two experiments were conducted to test the effects of a dietary zinc amino acid complex (Zn-AA) and an anticoccidial drug on Eimeria acervulina or Eimeria tenella infections. In each experiment, 288 day-old Three-Yellow-Chickens were used in a 2 x 3 factorial experimental design. Six groups were arranged randomly to receive three levels of Zn-AA (0, 40, or 80 mg/kg) alone or with salinomycin (60 mg/kg). Additionally an uninfected group was set as negative control. At the age of 21 days birds in Exp. 1 were inoculated with 3 x 10(4) sporulated E. acervulina oocysts, while birds in Exp. 2 were inoculated with 1.5 x 10(4) sporulated E. tenella oocysts. In Exp. 1, E. acervulina did not suppress growth performance significantly, but in groups without salinomycin it significantly reduced serum carotenoid levels on day 7 after inoculation and body and shank pigmentation on day 42. Salinomycin medication maintained serum carotenoids and visual colour of inoculated birds, but Zn-AA did not influence these parameters. In Exp. 2, growth performances of infected and uninfected chickens were similar. Infection decreased to only serum carotenoid levels on day 14 after infection, and colour scores on day 42 in the inoculated group without salinomycin and Zn-AA supplementation. The birds that received Zn-AA had significantly higher serum carotenoid levels and colour scores than those that did not. Although supplementation of Zn-AA cannot avoid coccidial damage of caecum, it prevents the reduction of serum carotenoids and pigmentation of Three-Yellow-Chicken infected with E. tenella, but not after infection with E. avervulina. The interactive effects between Zn-AA and salinomycin on growth performance and pigmentation were not significant.     

Suppression of peripheral eosinophilia by the coccidium Eimeria nieschulzi (Apicomplexa: Eimeriidae) in experimentally infected rats

Four groups of 60 rats each were used to examine interspecific interactions between Eimeria nieschulzi and Nippostrongylus brasiliensis. Rats in group 1 served as uninoculated controls. Group 2 rats were each injected subcutaneously with 2.0 X 10(3) L3 larvae of N. brasiliensis. Group 3 rats were each inoculated per os with 2.5 X 10(5) sporulated oocysts of E. nieschulzi. Rats in group 4 were first infected with 2.0 X 10(3) larvae of N. brasiliensis and, at 8 days postinoculation, with 2.5 X 10(5) oocysts of E. nieschulzi. Ten animals from groups 1-3 were sacrificed at 4-day intervals postinoculation and group 4 rats were sacrificed at 4-day intervals beginning after the secondary infection. Blood smears were prepared from each animal to determine differential blood cell counts, bone marrow was examined at the times of peak infection for absolute and relative numbers of eosinophils, portions of the duodenum and jejunum were examined histologically for mast cells, and feces obtained from the cecum and large intestine were examined for ova/gram of feces. Results revealed that relative numbers of peripheral neutrophils and monocytes became elevated during the course of infection for all infected animals, and rats infected with the helminth only also had elevated eosinophil levels. However, rats infected singly with E. nieschulzi, or concurrently with the coccidium and helminth, had peripheral eosinophil levels that were not significantly different from controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection of Toxoplasma gondii by polymerase chain reaction in sera of acutely infected mice

The presence of Toxoplasma gondii DNA was detected in sera of acutely infected mice by polymerase chain reaction. Adult mice were inoculated intraperitoneally with 5 x 10(3) T. gondii RH strain tachyzoites. Five mice were killed every 3 hr from 3 to 21 hr post infection (PI) and every day from 1 to 7 days PI. Toxoplasma gondii DNA was first detected in 60% of the infected mice 18 hr PI and in 100% of the animals 21 hr PI and from 1 to 7 days PI. No mice survived longer than 7 days.     

Feline Toxoplasmosis from Acutely Infected Mice and the Development of Toxoplasma Cysts*

SYNOPSIS. The development of Toxoplasma cysts was studied in mice inoculated with tachyzoites by several routes. After 1–30 days of infection, murine tissues were examined microscopically, and portions or whole carcasses were fed to mice and cats. The feces of the cats were examined for oocyst shedding. Cyst-like structures containing distinct PAS-positive granules were first seen after 3 days of infection with tachyzoites, and became numerous by 6 days. Argyrophilic walls were first seen after 6 days, and became numerous by 16 days of infection with tachyzoites. Prepatent periods to oocyst shedding (PPO) were either “short” (3–10 days) or “long” (19–48 days). The “short” PPO was found only in cats that had ingested mice infected for 3 days or longer, and was related to the development of PAS-positive granules in T. gondii, and to high, 60–100%, oral infectivity rates for cats. The “long” PPO followed the ingestion of mice infected for only 1–2 days, and was related to tachyzoites without distinct PAS-positive granules and low, 32% or less, infectivity for cats. The “long” PPO followed also the ingestion of oocysts and the parenteral inoculation of tachyzoites, bradyzoites, or sporozoites. Using the “short” PPO as a criterion for detecting cysts in tissues, it was shown that (a) numerous cysts developed in mice 5 days after inoculation with tachyzoites, 7–9 days after inoculation with cysts, and 9–10 days after inoculation with oocysts, and (b) cysts developed faster and more frequently in the brain and muscle than in lungs, liver, spleen, and kidneys of mice inoculated with tachyzoites.     

Experimental congenital toxoplasmosis in Wistar and Holtzman rats

Congenital toxoplasmosis was evaluated in Wistar and Holtzman rats using two strains of Toxoplasma gondii isolated in Brazil. Pregnant rats were inoculated by subcutaneous or intraperitoneal routes with 10(6) or 8 x 10(6) tachyzoites of N strain (virulent for mice) and by subcutaneous or oral routes with 10(2) or 1.2 x 10(3) cysts of P strain (avirulent for mice). The tissues of rat pups born from these rats were bioassayed for T. gondii infection. T. gondii was not observed in the pups born from rats inoculated with N strain. In the animals inoculated with P strain, congenital toxoplasmosis occurred in 22.8% (Wistar rats inoculated with 10(2) cysts by the subcutaneous route), 11.4% (Wistar rats inoculated with 10(2) cysts by the oral route), 21.2% (Wistar rats inoculated with 1.2 x 10(3) cysts by the oral route) and 2.9% of fetal infection (Holtzman rats inoculated with 10(2) cysts by the oral route). None of the pups born from chronically infected mother were infected with T. gondii.     

Development of patent infection in immunosuppressed C57Bl/6 mice with a single Cryptosporidium meleagridis oocyst

The ability of Cryptosporidium meleagridis to produce patent infection was studied in adult C57BL/6 mice that were immunosuppressed with dexamethasone phosphate provided in the drinking water at a dosage of 16 microg/ml. Four days after the onset of immunosuppression, mice were orally challenged with 1, 3, 10, or 1,000 C. meleagridis TU1867 oocysts per mouse. The mice were monitored daily for 18 days postinoculation for oocyst shedding. Five of 10 mice given a single oocyst, 4 of 5 mice given 3 oocysts, and all 9 mice given either 10 or 1,000 oocysts became infected and began shedding oocysts 5-7 days after challenge and continued to shed oocysts until the end of the experiment on day 18 postchallenge. Approximately 10(7) oocysts per mouse per day were excreted, regardless of the challenge dose. Neither the noninfected, immunosuppressed nor the inoculated, nonimmunosuppressed control mice shed oocysts. The excreted oocysts were confirmed to be those of C. meleagridis by polymerase chain reaction-restriction fragment length polymorphism analysis. We show that C. meleagridis, originally classified as an avian pathogen but recently found in humans with cryptosporidiosis, can produce patent infection in mice infected with a single oocyst. Moreover, we demonstrate that the immunosuppressed C57BL/6 adult mouse is an ideal host for the propagation of clonal populations of C. meleagridis isolates for laboratory studies.     

Eimerians in Harvest Mice, Reithrodontomys spp., from Mexico, California and New Mexico, and Phenotypic Plasticity in Oocysts of Eimeria arizonensis

ABSTRACT Between May 1979 and August 1991, 48.7% (57/117) of the harvest mice ( Reithrodontomys spp.) examined from 10 localities in Mexico, California and New Mexico had coccidian oocysts in their feces. A total of 46.7% (49/105) of the Reithrodontomys megalotis examined were positive for coccidian oocysts; this included samples from five states in Mexico (47.1%, 8/17), three counties in California (66.7%, 4/6) and two counties in New Mexico (45.1%, 37/82); 66.7% (8/12) of the Reithrodontomys montanus from one county in New Mexico also were infected. Only two coccidian species, Eimeria arizonensis and Eimeria langebarteli , were found in these hosts. Oocysts of E. langebarteli were found only in R. megalotis : in all three infected mice from Madera County, California, in the only mouse from San Bernardino County, California, and in 63% (5/8) of the infected mice from four states in Mexico. Oocysts of E. arizonensis were found in R. megalotis in Mexico, California, and New Mexico and in R. montanus from New Mexico. Sporulated oocysts of E. langebarteli differed slightly from those in previously published reports by having wider oocysts and larger sporocysts. Sporulated oocysts of E. arizonensis were variable in size, with those recovered from R. montanus significantly larger in length and width and sporocyst width than those from R. megalotis . The structure of the oocyst residuum was polymorphic, both within and between host species, and within the same mouse; it could appear as one large globule, two globules, several to many smaller globules, or as a compact mass of many small granules. Oocysts with a variable residuum were larger than those with one globule in all oocyst/sporocyst dimensions. Only 9% (5/57) of the infected mice were discharging oocysts of both eimerians when examined.     

Toxoplasma gondii: protection against colonization of the brain and muscles in a rat model

The objective was to test immune protection against the formation of Toxoplasma gondii tissue cysts in rats. It has been previously shown that 50 T. gondii tissue cysts of strain Me49 are not pathogenic for CF-1 mice, whereas 1 T. gondii tissue cyst of strain M-7741, can be lethal for mice 11-13 days after subcutaneous or oral administration. In the present study, ten rats were fed T. gondii oocysts of strain Me49 and after a further 30 days they were each orally challenged with T. gondii oocysts of strain M-7741. Thirty days after this, they were euthanased and brain and muscle samples inoculated subcutaneously or orally dosed, respectively, to mice for bioassay. None of the mice died, whereas all the mice that were inoculated with brain homogenates or were fed muscle samples from four non-immunized rats that had been inoculated with T. gondii oocysts of strain M-7741, died. These results encourage further research towards achieving vaccinal protection against the formation of T. gondii tissue cysts in meat animals and people.     

Tachyzoite-induced life cycle of Toxoplasma gondii in cats

The tachyzoite-induced cycle of Toxoplasma gondii was studied in 46 cats. Tachyzoites of the M-7741 or Me-49 strain of T. gondii were administered orally to cats by pouring into the mouth or by stomach tube, or by intraintestinal inoculation. Ten weaned cats that had been inoculated with tachyzoites directly in the intestine were killed 1, 3, 6, 9, 12, 15, 18, or 25 days later, and their tissues were studied histologically and bioassayed in mice. Toxoplasma gondii was demonstrable in the blood of 8 cats and in other tissues of all these 10. Four out of five 1- to 8-day-old cats fed tachyzoites by stomach tube became infected with T. gondii, and 1 became ill because of toxoplasmosis. All 19 weaned cats fed tachyzoites (poured into the mouth) became infected, and 6 died of acute toxoplasmosis 9-15 days after being fed T. gondii. Six out of 12 weaned cats fed tachyzoites by stomach tube became infected but were asymptomatic. Overall, 12 out of 26 cats observed for 19 days or more shed oocysts with a prepatent period (pp) of 19 days or more, with the sole exception of 1 cat that shed oocysts with a pp of 5 days. Enteroepithelial stages of T. gondii were not found in any cat before oocysts were shed. Cats shed up to 360 million oocysts in a day, and oocysts were shed for 4-6 days.     

Toxoplasma gondii: differential protection rates by two strains against cyst formation in a rat model

A previous infection with the ME-49 strain of Toxoplasma gondii (of low pathogenicity for mice), protected 17 of 20 rats against formation of brain cysts, following challenge with 10(3) oocysts of the high pathogenicity M3 strain, as determined by bioassay of rat brains in mice. The low pathogenic KSU strain did not afford comparable protection. Protection was further tested in rats that were orally or subcutaneously immunized with cysts or oocysts of the ME-49 strain, and later challenged with 2 x 10(2) cysts or 10(2) oocysts of the highly pathogenic strains M3, M-7741 and C. Protection ranged from 43 to 100%, compared to non immunized control rats and was independent of the stage of ME-49 strain and of the routes used to immunize the rats. The results obtained encourage further investigation into prevention of toxoplasmosis in humans and food animals.     

Extraintestinal sporozoites of chicken Eimeria in chickens and turkeys

Oocysts were found in the feces of chickens (recipients) dosed orally with whole blood, liver, lung, or heart homogenates from chickens and turkeys (donors) inoculated 3 and 4 days previously with a mixture of 3.5 X 10(6) oocysts of chicken Eimeria. No oocysts were found in the feces of recipients given spleen homogenates from these same chickens and turkeys and none were found in the feces of recipients given similar material from uninoculated donors. Intracellular sporazoites were found in the peripheral blood of a turkey inoculated with chicken Eimeria. The results indicate that a small number of sporozoites are capable of invading and surviving for at least 4 days in the peripheral blood of chickens and turkeys.     

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.     

Pathogenicity for several laboratory animals of toxoplasma oocysts originated from naturally infected cats.

The pathogenicity of four strains, O-1, O-2, O-3, and O-4, of Toxoplasma isolated in the form of oocyst from the feces of naturally infected cats was examined for such laboratory animals as mice, rats, rabbits, guinea pigs, and dogs, in comparison with that of the Beverley strain. Suspensions of seven graded doses of oocysts of each strain ranging from 1.0 X 10(-1) to 1.0 X 10(5) were inoculated orally into seven groups of five mice each. The O-1, O-2, and O-3 strains were as pathogenic for mice as the Beverley strain, but the O-4 strain was not so pathogenic as any other strain. Rats, rabbits, guinea pigs, and dogs were inoculated orally with around 1.0 X 10(5) oocysts. The four O strains were not so severly pathogenic for rats and rabbits as to cause death. The O-2 and O-3 strains showed strong pathogenicity for guinea pigs, almost all of which, when inoculated with them, died after manifesting severe clinical symptoms. The pathogenicity of the O-1 and O-2 strains showed essentially the same tendency for dogs as for any other animal. In inoculation with oocysts, as well as with proliferative forms or cysts, the same pathogenicity was not observed in different strains, even if the same species of host animals was used. On the contrary, the same pathogenicity was not always found even in one strain when a different species of animals was used.