Experimental Toxoplasmosis in Rats Induced Orally with Eleven Strains of Toxoplasma gondii of Seven Genotypes: Tissue Tropism, Tissue Cyst Size, Neural Lesions, Tissue Cyst Rupture without Reactivation, and Ocular Lesions.

BACKGROUND: The protozoan parasite Toxoplasma gondii is one of the most widely distributed and successful parasites. Toxoplasma gondii alters rodent behavior such that infected rodents reverse their fear of cat odor, and indeed are attracted rather than repelled by feline urine. The location of the parasite encysted in the brain may influence this behavior. However, most studies are based on the highly susceptible rodent, the mouse. METHODOLOGY/PRINCIPAL FINDINGS: Latent toxoplasmosis was induced in rats (10 rats per T. gondii strains) of the same age, strain, and sex, after oral inoculation with oocysts (natural route and natural stage of infection) of 11 T. gondii strains of seven genotypes. Rats were euthanized at two months post inoculation (p.i.) to investigate whether the parasite genotype affects the distribution, location, tissue cyst size, or lesions. Tissue cysts were enumerated in different regions of the brains, both in histological sections as well in saline homogenates. Tissue cysts were found in all regions of the brain. The tissue cyst density in different brain regions varied extensively between rats with many regions highly infected in some animals. Overall, the colliculus was most highly infected although there was a large amount of variability. The cerebral cortex, thalamus, and cerebellum had higher tissue cyst densities and two strains exhibited tropism for the colliculus and olfactory bulb. Histologically, lesions were confined to the brain and eyes. Tissue cyst rupture was frequent with no clear evidence for reactivation of tachyzoites. Ocular lesions were found in 23 (25%) of 92 rat eyes at two months p.i. The predominant lesion was focal inflammation in the retina. Tissue cysts were seen in the sclera of one and in the optic nerve of two rats. The choroid was not affected. Only tissue cysts, not active tachyzoite infections, were detected. Tissue cysts were seen in histological sections of tongue of 20 rats but not in myocardium and leg muscle. CONCLUSION/SIGNIFICANCE: This study reevaluated in depth the rat model of toxoplasmosis visualizing cyst rupture and clarified many aspects of the biology of the parasite useful for future investigations.

Haematobium eggs detection in human bladder cancer and sporocysts in snail vectors: Seven cases report and a review of the Burkina Faso literature.

Schistosoma haematobium plays a central role in the development of bladder cancer in Burkina Faso. The objective of this study was to determine the presence of S. haematobium in the bladder cancer and in vector snails. For the first time, formalin-fixed tissues embedded in paraffin were analyzed by immunohistochemistry and PCR. Molecular detection has resulted in 7/7 positive bladder cancer. Finally, as the snail vectors were positive. We suggest the use of molecular methods in the snail vectors for the detection of cysts and in cancerous tissues in larger studies.

Eimeria involved in a case of coccidiosis in farmed red-legged partridges (Alectoris rufa) in France: oocyst isolation and gross lesion description after experimental infection.

The aim of the present work was, after a coccidiosis outbreak in a farm rearing red-legged partridges (Alectoris rufa) in Brittany (France), to identify the Eimeria species and describe gross lesions induced by three of them (Eimeria kofoidi, Eimeria caucasica and Eimeria legionensis) after experimental infection. E. kofoidi oocysts measured 19.3 µm × 16.3 µm on average; neither micropyle nor oocyst residuum were present, but one, two or more small polar granules were visible. After inoculation of 300,000 oocysts per partridge, severe gross lesions were observed in the duodenum and jejunum, characterized by thickened oedematous mucosa and lumen filled with thick mucus, gas and sometimes false-membrane due to sloughed epithelium. E. caucasica oocysts were on average 29.8 µm × 19.5 µm in size; no oocyst residuum was observed, but a large granule was well visible. E. caucasica also invaded both the duodenum and jejunum, causing haemorrhagic points on the serosal surface, as well as mucoid duodenitis and catarrhal enteritis when 30,000 oocysts were inoculated per bird. E. legionensis oocysts measured 22.6 µm × 14.9 µm on average; they presented a clear micropyle beneath which one or two granulations were present. E. legionensis mainly invaded the caeca; low mortality was observed at the dosage of 200,000 oocysts per bird. Caecal walls were thickened and caseous material was condensed into off-white cheesy cores. For each species, oocyst shedding started 5 days post inoculation, peaked at 9, 8 and 6 days post inoculation for E. kofoidi, E. caucasica and E. legionensis, respectively, then decreased and persisted until 15 days post inoculation (end of examinations).

Influence of low levels of dietary aflatoxins on Eimeria tenella infections in broilers.

The present study was conducted to evaluate the adverse effects of an interaction between low levels of dietary aflatoxins (AF) and Eimeria tenella infection on broiler chicks. A set of 1-day-old chicks were raised for 35 days in the following groups: a control group, a group fed AF, a group fed AF and inoculated with E. tenella (AF + E.ten), and a group inoculated with E. tenella alone. AF in the contaminated diet were given at 200 ppb starting from the seventh day after hatching while E. tenella was inoculated at a dose of 5 × 10(4) sporulated oocysts per chick at the 14th day after hatching. Worsened performance traits and high mortality were all observed in the treated birds, particularly the AF + E.ten group. Lesion scores and oocyst outputs were not different within groups. Chickens fed with AF had significantly increased serum ALT and ALP activities as well as decreased albumin content. They also showed hepatomegaly, hepatocytic vacuolation and necrosis, an atrophied bursa of Fabricius, and a thymus with tissue depletion. E. tenella-infected broilers displayed a significant reduction in packed cell volume, hemoglobin content and lymphocyte percentage, and showed hemorrhagic typhlitis. The deficits in hepatic function and hematologic parameters as well as the gross pathological, and histopathological changes, were more common and more severe in the group that was exposed to both aflatoxicosis and coccidiosis than in the groups exposed to either treatment alone. Thus, the combination of aflatoxicosis and E. tenella infection may influence the course of coccidial infection due to additive effects.

Investigations and comparative detection of Cryptosporidium species by microscopy, nested PCR and LAMP in water supplies of Ordu, Middle Black Sea, Turkey.

A total of 70 water samples, including tap, river, fountain and well water were collected in the Ordu province, Middle Black Sea, Turkey and investigated for the detection of Cryptosporidium oocysts. The samples were directly screened microscopically for Cryptosporidium oocysts' detection by immunofluorescence test and subsequently DNA was extracted for the molecular detection by loop-mediated isothermal amplification (LAMP) and nested polymerase chain reaction (PCR). Eighteen out of the 70 (25·7%) water samples were found positive for Cryptosporidium spp. by immunofluorescence test and 19 (27·1%) were found positive by LAMP. Nested PCR products were not generated in any of the investigated water samples. A total of 16 randomly selected pellets were spiked with 10 Cryptosporidium oocysts to test the efficiency of the applied method. All the samples were found positive by LAMP for the presence of Cryptosporidium DNA, while the nested PCR assay was positive in only seven (43·75%) out of the 16 examined spiked samples. This is the first report on the occurrence of Cryptosporidium species in environmental and drinking water supplies in the Black Sea area.

Brachylaima aspersae n. sp. (Digenea: Brachylaimidae) infecting farmed snails in NW Spain: morphology, life cycle, pathology, and implications for heliciculture.

The life cycle of Brachylaima aspersae n. sp. (Trematoda: Brachylaimidae) in heliciculture farms is elucidated in light of field and experimental studies. Embryonated asymmetrical eggs (33.3 µm × 20.2 µm) are passed in the faeces of the definitive host, the domestic mouse (Mus musculus), and are ingested by its unique first intermediate host, the helicid snail Helix aspersa aspersa. After hatching, the miracidium develops into a highly branched sporocyst in the connective tissues of the digestive gland. Microcaudate cercariae emerging from this gastropod migrate up the ureter of the second intermediate host, the snails H. a. aspersa and H. a. maxima, and develop into non-encysted metacercariae in the kidney. Following predation of infected snails, the metacercariae develop into adults preferentially in the proximal portion of the duodenum of the definitive host. The strict oioxenic character for the first intermediate host, as well as the cercarial chaetotaxy (3 C(I)V+1 C(I)D, 10 C(II), 5 C(III)V, 14 C(III)L, 2 C(III)D, 16 H, 6 S(I), 6 S(II), 6 S(III), 2 A(I)L+1 A(I)V, 1 A(II)L, 3 ML, 1 P(I)L and 3 P(III)L), the distinct pars prostatica, the variable appearance of testes (rounded to irregular, with smooth or slightly to moderately lobulated margins), the size of eggs, the position of acetabulum (located somewhat posterior to the anterior third of body), and the microhabitat of the adult in the final host allow differentiation of B. aspersae from other well-known species in the genus. Massive infections with sporocysts or metacercariae of this brachylaimid may induce extensive pathological changes in the organs affected. Our results confirm that control of rodents in heliciculture farms is essential to minimize the potential health risks and morbimortality associated with this newly described species.

Involvement of the cytokine MIF in the snail host immune response to the parasite Schistosoma mansoni.

We have identified and characterized a Macrophage Migration Inhibitory Factor (MIF) family member in the Lophotrochozoan invertebrate, Biomphalaria glabrata, the snail intermediate host of the human blood fluke Schistosoma mansoni. In mammals, MIF is a widely expressed pleiotropic cytokine with potent pro-inflammatory properties that controls cell functions such as gene expression, proliferation or apoptosis. Here we show that the MIF protein from B. glabrata (BgMIF) is expressed in circulating immune defense cells (hemocytes) of the snail as well as in the B. glabrata embryonic (Bge) cell line that has hemocyte-like features. Recombinant BgMIF (rBgMIF) induced cell proliferation and inhibited NO-dependent p53-mediated apoptosis in Bge cells. Moreover, knock-down of BgMIF expression in Bge cells interfered with the in vitro encapsulation of S. mansoni sporocysts. Furthermore, the in vivo knock-down of BgMIF prevented the changes in circulating hemocyte populations that occur in response to an infection by S. mansoni miracidia and led to a significant increase in the parasite burden of the snails. These results provide the first functional evidence that a MIF ortholog is involved in an invertebrate immune response towards a parasitic infection and highlight the importance of cytokines in invertebrate-parasite interactions.

[Sarcocystis cruzi (Protozoa: Apicomplexa: Sarcocystiidae) infection in european bison (Bison bonasus) from Bialowieza Forest, Poland]. / Sarcocystis cruzi (Protozoa: Apicomplexa: Sarcocystiidae) u zubra (Bison bonasus) w Puszczy Bialowieskiej.

Samples of oesophagus, diaphragm and heart muscles were taken from one European bison from Bialowieza Forest during seasonal European bison elimination in 2008. Five gram of each muscle was examined by staining small samples in 0.2% aqueous solution of methylene blue. After that they were placed between compressor glasses and examined under a dissecting microscope in order to detect Sarcocystis sp. infection. All sarcocysts observed were counted. The 5-g sample of the heart muscle yielded a total of 756 sarcocysts while that of the diaphragm contained 107 of them and that of the oesophagus--89. All of the sarcocysts were isolated from 1 g of each muscle by using preparation needles (probes). After that the sarcocysts were taken to 0.5% physiological solution and examined under light microscope. The special attention was paid to detection of their cyst wall, which was thin (1 microm up to 1.2 microm) and smooth in all cases. Sometimes villar protrusions were seen on the surface of the cysts. Differences between the size of sarcocysts isolated from different muscle samples were observed. The longest and the slenderest sarcocysts were found in the diaphragm. Slightly smaller in the oesophagus and the smallest ones in the heart muscle tissue. The average size of sarcocysts isolated from diaphragm was 957.6 microm x 112.7 microm. Sarcocysts found in the oesophagus measured 484.1 microm x 194.6 microm and those isolated from the heart muscle attained 305.4 microm x 103.9 microm. All of the sarcocysts isolated from heart, oesophagus and diaphragm muscles were identified Sarcocystis cruzi.