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Activity of buparvaquone against Theileria cervi in white-tailed deer.

Buparvaquone, a naphthoquinone with known efficacy against Theileria parva parva in cattle, was tested for activity against Theileria cervi piroplasms in both an in vitro culture system and in vivo in experimentally infected white-tailed deer. The in vitro data showed a significant decrease in the incorporation of 3H-hypoxanthine by infected red blood cells treated with buparvaquone when compared to that seen with imidocarb and chloroquine treatment. In both intact and splenectomized deer treated with buparvaquone (2.5 mg kg-1) a gradual decrease in piroplasm parasitaemia was observed following treatment. However, in the splenectomized deer, parasitaemia levels returned to near pretreatment values after approximately 2 weeks.

Animals↗

Responses of peripheral blood mononuclear cells in calves infected with Theileria sergenti.

The role of peripheral blood mononuclear cell (PBMC) in Theileria sergenti-infected calves was studied by various in vitro assay systems. Proliferation of T cells in mixed lymphocyte protozoa culture (MLPC) increased with parasitemia, and the addition of monoclonal antibodies against T. sergenti merozoites in this MLPC enhanced the response. However, the addition of antibody-positive autologous serum resulted in the suppression of the response. Cell-mediated cytotoxicity of PBMC increased after peak parasitemia. This cytotoxicity increased on co-cultivation of PBMC with T. sergenti merozoites, but the addition of autologous serum suppressed the response.

Animals↗

Immunoprophylaxis against Theileria annulata with protein from plasma membrane of infected lymphoblasts.

Cross-bred (Bos taurus male x Bos indicus female) calves were protected against the homologous sporozoite-induced challenge of Theileria annulata when immunised with protein from the plasma membrane of macroschizont-infected lymphoblasts of allogeneic origin. However, such protection was parasite-strain specific with the plasma membrane originating from lymphoblasts infected and transformed by the same isolate of T. annulata sporozoites. No protection ensued when the infected lymphoblasts and sporozoites were from immunologically different isolates of T. annulata. There was enhanced proliferation of cells and evidence for lymphokine, originating from antigen-sensitised lymphocytes, demonstrable as macrophage migration inhibition factor in peripheral blood lymphocytes of the calves immunised by plasma membrane protein and challenged by sporozoites of homologous origin.

Animals↗

A simple field diagnostic smear test for bovine besnoitiosis.

A fast and inexpensive skin biopsy smear was used for confirming suspected clinical cases of bovine besnoitiosis. The technique was based on the demonstration of Besnoitia besnoiti bradyzoites (cystic stages), which appeared stumpy, each organism 6.2 microns by 3.1 microns in size, or banana-shaped, 7.7 microns by 1.5 micron in affected skin smears. A more rapid non-surgical technique, scleral conjunctival scraping, revealed similar bradyzoites, thus enhancing the diagnostic value of conjunctival cysts in more chronic infections. The technique will aid prompt management decisions to contain suspected outbreaks in herds not routinely served by tissue-processor-equipped diagnostic laboratories.

Animals↗

Clinical neosporosis in a 4-week-old Hereford calf.

Most reports of neosporosis associated with abortion in cattle are in dairy cattle and infected calves do not survive beyond 7 days of age. This paper reports neosporosis in a 4-week-old Hereford calf. The calf was full term and appeared clinically normal at birth. At 2 weeks of age, the calf had weakened and was unable to nurse unassisted. The calf was killed at 4 weeks because of paralysis. The primary lesions were in the brain and were associated with Neospora caninum tachyzoites. The diagnosis was confirmed by immunohistochemical staining with anti-Neospora rabbit serum.

Animals↗

Canine neosporosis in South Africa.

Neospora caninum was identified in tissues of one neonatal and two adolescent dogs by a specific immunohistochemical technique during a retrospective study. The histological lesions included multifocal polymyositis, multifocal parasitic myocarditis, interstitial pneumonia, severe multifocal encephalitis and myelitis with perivascular cuffing. This report extends the known range of the parasite to the African continent.

Animals↗

Cell surface interaction of the protozoan Gregarina with concanavalin A beads - implications for models of gregarine gliding.

Con A Sepharose beads can be translocated over the surface of the protozoan Gregarina and in (forward) moving gregarines, the bead may be moved backwards relative to the substrate. The speed of bead movement is not constant over the surface of the cell, but has a maximum value in the central region of the deutomerite. The mass of the individual beads used in this study was about the same order of magnitude as the mass of a gregarine, i.e. considerable force must be generated at the gregarine cell surface. The implications of these experiments to models of gregarine locomotion are considered. The close similarities between this system and flagellar surface motility of Clamydomonas studied by particle movement and gliding motility are discussed.

Animals↗

Demonstration of actin in the protozoon--Gregarina.

Actin was identified in the motile trophozoites of Gregarina by indirect immunofluorescence microscopy. Ultrastructurally actin was found by immunogold labelling to be associated with the internal cytomembranes of the ectoplasmic folds and the general ectoplasmic region below the folds. Western blotting with antibodies to actin identified polypeptides with molecular weights around 43kD and 98kD in non-ionic detergent extracted trophozoites. The possibility of two distinct forms of cytoplasmic actin (43kD and 98kD) was considered.

Actins↗

Synthesis and in vitro activities of ferrocenic aminohydroxynaphthoquinones against Toxoplasma gondii and Plasmodium falciparum.

Fourteen ferrocenyl aminohydroxynaphthoquinones, analogues of atovaquone, were synthesized from the hydroxynaphthoquinone core. These novel atovaquone derivatives were tested for their in vitro activity against two apicomplexan parasites of medical importance, Toxoplasma gondii and Plasmodium falciparum, including resistant strains to atovaquone (T. gondii) and chloroquine (P. falciparum). Three of these ferrocenic atovaquone derivatives composed of the hydroxynaphthoquinone core plus an amino-ferrocenic group and an aliphatic chain with 6-8 carbon atoms were found to be significantly active against T. gondii. Moreover, these novel compounds were also effective against the atovaquone-resistant strain of T. gondii (Ato(R)).

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Feline cytauxzoonosis.

Cytauxzoon felis is a protozoan hemoparasite of wild and domestic cats. In domestic cats, it causes severe clinical disease with high mortality.

Animals↗

Ultrastructural study of developmental stages of Mattesia dispora (Neogregarinorida: Lipotrophidae), a parasite of the flour moth Ephestia kuehniella (Lepidoptera).

The ultrastructure of merozoites, gamonts and oocysts of the neogregarine Mattesia dispora and their development in larvae of the flour moth Ephestia kuehniella were studied by electron microscopy. The apical complex of free macronuclear merozoites was very distinct in micrographs of sections, the polar rings being especially prominent. Two gamonts associated in head-to-head syzygy and the apical complexes served as the contact point during pairing. At this stage the rhoptries became reduced and the conoid widened. The gamonts had a foam-like appearance in the light microscope. Paired gamonts formed an envelope and developed into a gametocyst, within which the gamonts were separated by a distinct border. Four gametes and two residual cells developed inside the gametocyst. The gametes were covered with a single membrane. The gametes fused in pairs to form two spherical zygotes, each covered by two membranes and with one large nucleus. The external layer appeared more undulated than the inner one. A single membrane covered each residual cell. Walls were formed around both zygotes to produce two oocysts. Each mature oocyst was lemon-shaped with polar plugs and eight peripheral sporozoites, which had a pellicle similar to that of the merozoites, lay beneath the thick oocyst wall.

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Vector-borne parasites decrease host mobility: a field test of freeze or flee behaviour of willow ptarmigan.

Transmission mode has been suggested to be a strong predictor of virulence. According to theory, the transmission of vector-borne parasites should be less dependent on host mobility than directly transmitted parasites. This could select for increased exploitation of host resources in parasites transmitted by vectors, which may be manifested as higher virulence. Here, we test the prediction that there is an association between transmission mode and the effect on host mobility by comparing parasite infection levels and mobility in willow ptarmigan (Lagopus lagopus L.). We examined the endoparasite infracommunities of individual hosts to obtain annual, quantitative data on four vector-transmitted species (Leucocytozoon lovati, Trypanosoma avium, Haemoproteus mansoni and microfilaria), two directly transmitted species (Trichostrongylus tenuis and Eimeria sp.) and two species with indirect life cycles (Hymenolepis microps and Parionella urogalli). We then used observed variations in freeze-or-flee responses of individual willow ptarmigan to assess whether parasite intensities were related to scored freezing responses. From a field data set covering a period of 9 years from a single area, we found that stronger freezing responses were associated with higher intensities of vector-borne parasites, especially with higher intensities of the haemosporidian L. lovati. Freezing responses were not associated with parasites transmitted in other ways. Thus, high intensities of vector-borne parasites tended to reduce host movements, while parasites with other transmission modes did not.

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Sexual comparisons in immune ability, survival and parasite intensity in two damselfly species.

Recent evolutionary studies have suggested that females have a more robust immune system than males. Using two damselfly species (Hetaerina americana and Argia tezpi), we tested if females produced higher immune responses (as phenoloxidase and hydrolytic enzymes), had a higher survival (using a nylon implant inserted in the abdomen and measuring survival after 24h) and fewer parasites (gregarines and water mites) than males. We also tested whether immune differences should emerge in different body areas (thorax vs. abdomen) within each sex with the prediction that only females will differ with the abdomen having a higher immune response than their thorax since the former area, for ecological and physiological reasons, may be a target zone for increased immune investment. Animals were adults of approximately the same age. In both species, females were more immunocompetent than males, but only in H. americana females were immune responses greater in the abdomen than in the thorax. However, there were no differences in survival and parasite intensity or the probability of being parasitised between the sexes in either of the two species. Thus, this study lends partial support to the principle that females are better at defending than males despite the null difference in parasitism and survival.

Abdomen↗

Use of ribosomal DNA sequence data to characterize and detect a neogregarine pathogen of Solenopsis invicta (Hymenoptera: Formicidae).

A neogregarine parasite of the red imported fire ant, Solenopsis invicta, was discovered recently in Florida and tentatively placed in the Mattesia genus based on morphological characterization. S. invicta infected with this Mattesia species exhibited a characteristic yellowing of the cuticle which was designated Mattesia "yellow-head disease" (YHD). The 18S rRNA gene sequence from Mattesia YHD was elucidated and compared with the neogregarine pathogens, Mattesia geminata and Ophriocystis elektroscirrha. The sequence data support the previous conclusion that Mattesia YHD is a new species that infects S. invicta. Furthermore, high sequence identity between Mattesia YHD, M. geminata (95.7%), and O. elektroscirrha (86.2%) correctly place the YHD organism in the Mattesia genus and Neogregarinorida order. Oligonucleotide primer pairs were designed to unique areas of the 18S rRNA genes of Mattesia YHD and S. invicta. Multiplex PCR resulted in sensitive and specific detection of Mattesia YHD infection of S. invicta.

Animals↗

Evaluation of an innate immune reaction to parasites in earthworms.

Encapsulation is an essential process of the invertebrate immune system and includes the prophenoloxidase (proPO) cascade. We present an assay for evaluating this immune response, now newly adapted to earthworms. Coelomic fluid is withdrawn and coelomocytes are stained with l-Dopa. We studied assay repeatability and the correlation between number of PO-active cells and infection level of the parasitic protozoan Monocystis sp. in the earthworm Lumbricus terrestris. Our study showed high assay repeatability although the expected negative relationship between PO-active coelomocytes and parasite load was not observed; yet a suggestion toward a positive relationship was detected. This finding is contrary to previous assumptions that presume coelomocyte concentrations to be the independent variable determining parasite load.

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Effect of Gregarina sp. parasitism on the susceptibility of Blattella germanica to some control agents.

Gregarines are enteric parasites of invertebrates but little is known about the negative effects of this parasitism on host species. The present study evaluates the influence of the parasitism of Gregarina sp. on the survival of Blattella germanica and methods for elimination of gregarine infection in laboratory rearing systems. Insects were dissected and the infection was detected in 80% of a sample of 50 adults. Diseased cockroaches had swollen abdomens, slower movement at high incidences of the protozoan, and short antennas. Dead cockroaches showed darkened body and putrid smell, indicating septicaemia. Infected insects were more susceptible than healthy cockroaches when treated with Metarhizium anisopliae and triflumuron.

Animals↗

Detection of Mattesia oryzaephili (Neogregarinorida: Lipotrophidae) in grain beetle laboratory colonies with an enzyme-linked immunosorbent assay.

An indirect sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of the neogregarine Mattesia oryzaephili was developed with monoclonal antibodies. It was used to screen laboratory colonies of Oryzaephilus surinamensis, Cryptolestes ferrugineus, C. pusillus, and C. turcicus from the United States, Canada, and Australia. All of the colonies except C. turcicus had larvae that tested positive with the percent of positives ranging from 0.2 to 83.9, but only colonies that tested positive had reported population declines. This assay will make possible epizootiological studies to assess the impact of M. oryzaephili on pest populations.

Animals↗

Differential diagnosis of Perkinsus species by polymerase chain reaction-restriction fragment length polymorphism assay.

Perkinsosis is an infection of marine molluscs caused by the protistan parasites of the genus Perkinsus, which has been classified by the OIE as a disease that warrants notification. In the present study, we have applied a molecular genetic approach to develop an optional method for the specific identification of Perkinsus species. A species-specific polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay of the rRNA ITS region was developed to identify and distinguish among Perkinsus species. A taxonomic key was established that allows successful identification of Perkinsus species using a single restriction enzyme (Rsa I) to discriminate P. chesapeaki and P. marinus or by a combination of two endonucleases (Rsa I plus Hinf I) to discriminate P. olseni and P. mediterraneus. In order to validate the RFLP assay, the PCR products were cloned and sequenced, and its phylogenetic affinity was determined. Phylogenetic analysis confirmed the specific identification carried out by RFLPs. Herein is the first report of P. olseni in Manila clams from the NW Adriatic Sea (Italy), which we identified by employing this method. The PCR-RFLP assay herein described may be useful to provide accurate, rapid and inexpensive identification of Perkinsus species, and may aid in ongoing epizooetiological studies and diseases control programmes.

Animals↗