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Biomedical subjects

H Mehlhorn

Publications and source records attributed to H Mehlhorn.

At least 37 records · Page 2Linked to original sources

Development of a larval bioassay for susceptibility of cat fleas (Siphonaptera: Pulicidae) to imidacloprid.

Strategies for controlling cat fleas, Ctenocephalidesfelisfelis (Bouché), have undergone dramatic changes in the past 5 yr. With the advent of on-animal treatments with residual activity the potential for the development of insecticide resistance increases. A larval bioassay was developed to determine the baseline susceptibility of field-collected strains of cat fleas to imidacloprid. All four laboratory strains tested showed a similar level of susceptibility to imidacloprid. Advantages of this bioassay are that smaller numbers of fleas are required because flea eggs are collected for the test. Insect growth regulators and other novel insecticides can also be evaluated. Using a discriminating dose, the detection of reduced susceptibility in field strains can be determined with as few as 40 eggs.

Acetone↗

Comparative study on the effects of three insecticides (fipronil, imidacloprid, selamectin) on developmental stages of the cat flea (Ctenocephalides felis Bouché 1835): a light and electron microscopic analysis of in vivo and in vitro experiments.

The effects of three insecticides (fipronil, imidacloprid and selamectin) on developmental stages of cat fleas (Ctenocephalides felis) were studied in vivo, in vitro and by means of light and electron microscopy. The results were documented by video. Adult fleas were attached to the skin of dogs that had been treated 7 days before with one of the three compounds. Furthermore, adult fleas were exposed exclusively to the hair and skin debris of such treated dogs or were placed on filter papers that had been impregnated with one of these three compounds or with the blood of treated dogs. Larval fleas were exposed to hair of treated dogs, to debris obtained by combing treated dogs, to dried blood samples of treated dogs or were placed onto filter papers impregnated with one of the three compounds. In these experiments with adult and larval fleas, it was noted that none of the three insecticides had a repellent effect on adult or larval fleas. Imidacloprid was the only compound that acted exclusively by body contact, and was apparently taken up by adult and larval fleas via the thin, non-sclerotized intersegmental membranes of the flea's body, shown when flea stages were exposed to hairs taken from dogs treated with one of the compounds or placed onto drug-impregnated filter papers. Imidacloprid killed larvae and adult fleas within 1 h, while it took at least 24 h until all adult fleas had died on fipronil- or selamectin-treated dogs, thus allowing longer feeding periods, increasing the risk of transmission of flea-derived diseases. Flea larvae covered with debris from dogs topically treated 7 days before with fipronil, imidacloprid or selamectin died, like the untreated control, within 16-28 h after exposure. This was, however, probably mainly due to a drying effect. Adult and larval fleas exposed to filter papers impregnated with the blood of treated dogs survived longer than 7 days, as did the untreated controls. All three drugs apparently acted on nerves and muscles and thus stopped motility.

Animals↗

In vivo and in vitro effects of imidacloprid on sheep keds (Melophagus ovinus): a light and electron microscopic study.

The effects of imidacloprid (Advantage) on sheep keds (Melophagus ovinus Linne 1758) were studied in vivo and in vitro by means of direct observation (monitored on video tape) and by light and electron microscopy. It was found that: 1. Imidacloprid acted rapidly on all motile stages of the sheep keds. Within 3-4 min after exposure they became immobile and their legs and the abdomen started tetanic trembling movements for 15-30 min, leading to death. 2. The compound is apparently taken up by the body, since it also acted on those sheep keds that had been exclusively exposed to imidacloprid-contaminated filter papers. 3. The compound is available and active for more than 1 month in the wool of sheep; even rainfall does not reduce its efficacy. Body contact between treated mother sheep and their lambs protects them from infestation with these ectoparasites. 4. The compound initiates an ultimately lethal destruction of the ganglia, nerve chords and related muscle fibers, as can be seen in electron micrographs.

Administration, Topical↗

Hammondia heydorni-like oocysts shed by a naturally infected dog and Neospora caninum NC-1 cannot be distinguished.

This study describes transmission experiments using Hammondia heydorni-like oocysts isolated in 1996 from a naturally infected dog. The isolate was designated as H. heydorni-Berlin-1996. Examination of sera from infected intermediate hosts showed immunoblot reactions that resembled patterns observed after Neospora caninum NC-1 infection. Furthermore, N. caninum DNA could be demonstrated in tissue samples (e.g. heart, brain) of experimentally infected intermediate hosts and in oocyst preparations from H. heydorni-Berlin-1996. The isolated oocysts did not induce any detectable disease in any of the inoculated adult intermediate hosts (goats, sheep, gerbils, guinea pigs, multimammate rats, BALB/c mice, SCID mice), even upon immunosuppression. Furthermore, neither histological lesions nor parasite stages could be identified in the tissues of all fetuses recovered from two multimammate rats that had been infected prior to pregnancy. An experiment with one dog fed a second time on infected intermediate host tissue indicated that immunity may prevent repeated oocyst shedding in N. caninum-infected dogs. In addition, the study clearly demonstrates that N. caninum can be readily transmitted by dogs that have ingested exclusively skeletal muscles of infected intermediate hosts. Therefore, the study has consequences for the recommendations for farmers to prevent postnatal transmission of N. caninum to cattle. It indicates that feeding of any tissues of potential intermediate hosts (including sheep, goats, rodents) to final hosts may induce the shedding of oocysts in these hosts and thus pose a risk for post-natal infection of cattle. With respect to oocyst morphology and the infectivity of muscle tissues for final hosts, no differences were seen in comparison with observations made in the past on Isospora bigemina/I. heydorni/H. heydorni. Therefore, earlier studies made on I. bigemina/I. heydorni/H. heydorni have to be re-evaluated critically to determine whether they may have included N. caninum or other protozoan parasites that use dogs as final hosts and have an oocyst morphology resembling that of I. bigemina/I. heydorni/H. heydorni.

Animals↗

Cyclic transmission of Neospora caninum: serological findings in dogs shedding oocysts.

In a previous paper we demonstrated that Hammondia heydorni-like oocysts isolated in 1996 from a naturally infected dog could not be distinguished from the isolate Neospora caninum NC-1. The isolate, designated as H. heydorni-Berlin-1996, was cyclically transmitted using dogs as the final hosts. The present study provides information on the antibody responses of the dogs used for the cyclical transmission of this isolate. The majority of dogs that had shed oocysts showed no sero-conversion with respect to N. caninum tachyzoite surface or immunodominant antigens, either in the indirect fluorescent antibody test or in two Western-blot-based tests. In addition to the examination of responses to immunodominant antigens, we also analysed the antibody reactions of dogs to a high-molecular-weight antigen (152 kDa) in the tachyzoite antigen preparation. The antibodies against this antigen appeared after the dogs had been fed infected intermediate host tissues and shed oocysts. The reaction was observed in dogs between day 35 and day 447 after feeding of intermediate host tissues. Therefore, our study provides initial information on a 152 kDa tachyzoite antigen, which might be a suitable candidate to identify dogs with a history of shedding N. caninum oocysts.

Animals↗

Comparative molecular investigation of Nc5-PCR amplicons from Neospora caninum NC-1 and Hammondia heydorni-Berlin-1996.

The clinical relevance of Neospora caninum as a cyst-forming coccidian parasite is increasingly acknowledged within veterinary medicine, although the pathways of transmission are far from being solved. The parasite is well known for causing diaplacental infections in cows associated with abortion and/or severe damage of the fetus. In addition, it may cause neuromuscular disease in dogs, which thus apparently act as intermediate hosts as well as final hosts. In our previous studies, we have demonstrated that molecular diagnosis of N. caninum infections has a high performance when a highly sensitive polymerase chain reaction (PCR) targeted to the Nc5 region of the parasite is used. The present study indicates that the high sensitivity of the PCR is the consequence of a target dose effect which reflects a high redundancy of Nc5-type sequences within the genome of the parasite. The PCR was shown to amplify a set of DNA molecules exhibiting significant sequence differences. A complex composition of Nc5-type sequences was observed in the parasite isolate N. caninum NC-1 but also in another isolate, designated Hammondia heydorni-Berlin-1996. Investigation of the infection pattern of this parasite in its intermediate and final canine hosts showed it to be indistinguishable from N. caninum NC-1.

Animals↗

Synergistic action of a cyclic depsipeptide and piperazine on nematodes.

The present study describes the synergistic effects of the cyclic depsipeptide BAY 44-4400 and piperazine in the treatment against the nematodes Trichinella spiralis, Heligmosomoides polygyrus, and Heterakis spumosa. The in vitro anthelmintic activity of a combination of the two compounds (1.7 motility units) against T. spiralis larvae was significantly higher than the sum of the individual drug effects (1.3 motility units). With regard to the rate of expulsion of H. polygyrus worms from the intestine of infected mice, an additive effect was observed; piperazine alone exerted an efficacy of 54.4% and BAY 44-4400 alone, one of 44.4%, whereas the combination of these compounds had an efficacy of 97.5%. With regard to the expulsion of H. spumosa worms, the effect of the combination was more than 5 orders of magnitude greater than the sum of the effects of the single compounds, i.e., there was a considerable potentiation of the actions of BAY 44-4400 and piperazine. Moreover, the combination exerted a significantly higher degree of degenerative effects on the intestine and on the nerve chords of H. spumosa as compared with the single compounds.

Animals↗

Neospora caninum: is it really different from Hammondia heydorni or is it a strain of Toxoplasma gondii? An opinion.

The published data concerning Toxoplasma gondii, Hammondia hammondi, H. heydorni and Neospora caninum on one side and between T. gondii on the other were neglected by most authors. As conclusion we are convinced that there are only two valid species: Isospora (Toxoplasma) gondii and Hammondia heydorni. The first includes as a strain H. hammondi and the latter N. caninum. In any case there is absolutely no reason (with respect to general Zoological nomenclature) to create new genera!

Animals↗

Molecular genetic characterization and subcellular localization of Theileria annulata mitochondrial heat-shock protein 70.

A Theileria annulata mitochondrial heat-shock protein of the 70-kDa family (Tamthsp70) was isolated by screening of the cDNA library of a T. annulata-infected bovine lymphoblastoid cell line with an antibody raised against T. annulata schizonts. The Tamthsp70 coding sequence was found to be most closely related to a previously reported mitochondrial hsp70 gene of Eimeria tenella exhibiting a similarity of 67% with mitochondrial hsp70 genes of eukaryotic plants (Pisum sativum, Phaseolus vulgaris) and with dnaK proteins of prokaryotes (Rhizobium meliloti, Agrobacterium tumefaciens). The Tamthsp70 mRNA is expressed within the sporozoite, schizont, and merozoite stages of the parasite, which suggests that it is constitutively transcribed throughout the life cycle. The gene encodes a polypeptide of 681 amino acids and exhibits a mitochondrial targeting sequence and several sequence motifs common to mitochondrial hsp70 and prokaryotic dnaK proteins. The protein level of the Tamthsp70 protein after heat shock decreased slightly during the exposure of infected cells to a temperature of 42 degrees C in comparison with cells cultured at 37 degrees C. By immunofluorescence the protein was located in the area in which the schizonts reside within infected cells. Immunoelectron microscopy showed that the hsp70 protein was predominantly localized in the mitochondria of the parasites. However, it was also found in small amounts in the cytoplasm of the parasite and host cell. This indicates (1) that Tamthsp70 is very probably translated in the parasite cytoplasm and then transported across the mitochondrial membrane into the mitochondrial matrix and (2) that it is transported across the parasite membrane into the host-cell cytoplasm.

Amino Acid Sequence↗

Fly larvae and pupae as vectors for scrapie.

We analysed experimental transmissibility of the scrapie agent by natural vectors. A fly, Sacrophaga carnaria, fed with brains of scrapie-infected hamsters in different developmental stages caused scrapie in hamsters after they ate fly extracts.

Animals↗

Review: Theileria schizonts induce fundamental alterations in their host cells.

The sporozoites of Theileria annulata and T. parva invade bovine leukocytes, where they differentiate into schizonts. The latter can immortalize and induce fundamental changes in their host cells. T. annulata infects mainly major histocompatibility complex class II cells, whereas T. parva preferentially transforms T-lymphocytes, which proliferate continuously without the need for exogenously added growth factors. Most of the cell lines appear to be independent of a growth factor but may express several cytokines that influence the outcome of the disease. The mechanisms underlying this transformation are not well understood. The infected cells show increased activity of casein kinase II and Jun NH2-terminal kinase (JNK), whereas extracellular signal-related kinase 1 and 2 and P38 are not activated. In addition, several transcriptional factors such as NFkB and AP-1 are activated. It has been postulated that parasite proteins either expressed on the surface of the schizonts or secreted into the host cell cytoplasm may interfere with the signal-transduction pathway of the host cells. A possible candidate may the casein kinase II homologue that was identified in schizonts of both T. annulata and T. parva.

Animals↗

Review: the cellular basis of the immunity to and immunopathogenesis of tropical theileriosis.

The intracellular protozoan parasite Theileria annulata causes a severe and often fatal disease of pure and crossbred cattle in tropical and subtropical countries. Animals that recover from the infection are immune against challenge with homologous parasite strains. In the present review we refer to the role of immunocompetent cells and their products in containing the infection or in facilitating the progress of the disease. Parasite-infected host cells produce cytokines, which, depending on their concentration and timing of production, may enhance the establishment of the infection. Thus, cell lines producing high levels of proinflammatory cytokines cause severe postvaccinal reactions when inoculated into cattle. This may be supported by an aberrant non-specific activation of naive T-cells, leading to the production of high levels of gamma-interferon (IFN-y). Under these circumstances development of the specific immune response may be inhibited. At this stage, innate immune reactions are operating to contain the infection. Natural killer cells and macrophages may represent the most important part of this immunity. Antibodies and specific T-lymphocytes, CD4+ T-cells and cytotoxic T-lymphocytes (CTLs), play the most important role in a challenge infection. In this context, CD4+ T-cells produce cytokines required for the clonal expansion of CTLs that kill their target cells in a major histocompatibility complex (MHC) class I-restricted manner. In addition, CD4+ T-cells produce macrophage-activating cytokines such as IFN-gamma. Such activated macrophages produce mediators such as NO, which destroy the intracellular schizonts. Attempts have been directed toward the identification of parasite antigens involved in the induction of immunity. To date, only a limited number of sporozoite and merozoite antigens have been identified and examined for their immunogenicity, and the protection achieved is partial. An effective vaccine must include schizont proteins, notably, those proteins that are secreted into the host cell cytoplasm because these may have access to the MHC class I and II compartments to be presented to CTLs and CD4+ T-cells, respectively. Several schizont proteins have been identified and these are now under investigation.

Animals↗

Effects of imidacloprid on adult and larval stages of the flea Ctenocephalides felis after in vivo and in vitro application: a light- and electron-microscopy study.

The effects of imidacloprid (Advantage) on the larval and adult stages of cat fleas (Ctenocephalides felis) were studied in vivo and in vitro by means of light and electron microscopy. It was found that: 1. The compound acted rapidly on both larval and adult fleas, killing both stages within 20 min of contact. 2. When applied as a spot-on to the skin of dogs, the compound localized in the water-resistant lipid layer of the skin surface and in the hairs but not in the blood. 3. Thus, the compound was not taken up during sucking of the flea but was absorbed via the thin intersegmental membranes, since larval and adult fleas that had only external contact with imidacloprid-impregnated paper or with shaved hairs from imidacloprid-treated dogs showed reactions similar to those shown by fleas sitting on treated skin. 4. The compound led to a continuous blockage of insect-specific nicotinic-acetylcholine receptors (nAChR), causing tetanic muscle cotractions within minutes of exposure. This manifested as intense trembling of the legs and pumping movements of the body. The affected flea stages remained motionless while the nerves and muscles were constantly and irreversibly destroyed due to hyperactivity. The ganglia of the head and thorax and the striated muscles of the flea body and legs were damaged first, whereas the intestinal movements (e.g., visible in larvae) took longer to exhibit damage. In summary, these studies show that imidacloprid kills larval and adult flea stages rapidly via the same mode of action and thus prevents the development of flea populations in human or animal dwellings.

Animals↗

Isolation and characterization of a cDNA clone encoding a thiol proteinase of Sarcocystis muris cyst mero' zoites (Apicomplexa).

A cDNA library of Sarcocystis muris cyst merozoites was screened using a digoxigenin-labeled probe. This probe was derived from a 504-bp polymerase-chain-reaction fragment representing part of a thiol proteinase. Several cDNA clones were isolated, one of which (PH08) consists of a nucleotide sequence of 1694 bp and encodes the complete prepropolypeptide of a cathepsin L-like proteinase. PH08 contains an open reading frame of 394 amino acid (aa) residues with a 46-residue signal sequence, which is followed by a 129-residue propeptide and 219 aa residues of the mature enzyme. Two potential glycosylation sites and a putative polyadenylation signal were also identified. The occurrence of the highly conserved interspersed ERFNIN aa motif, not found in cathepsin B-like proteinases, suggests the classification of the enzyme as a cathepsin L-like proteinase. Results worked out in this study will enable production of the recombinant thiol proteinase of S. muris cyst merozoites necessary for study of the substrate specificity as well as other biochemical parameters of this enzyme.

Amino Acid Sequence↗

Characterization of a genomic region encoding the 32-kDa dense granule antigen of Sarcocystis muris (Apicomplexa).

Two subgenomic libraries constructed from Sarcocystis muris total DNA were screened with a cDNA probe, specific for a 32-kDa protein associated with the dense granules. Two clones reacted positively and were isolated, gDG 32/1 and gDG 32/2. Genomic clone gDG32/1 and part of clone gDG 32/2 have been sequenced. The composite nucleotide sequence of these genomic clones comprises 4.34 kb. It contains a 5' region of 2.14 kb, a first coding region (222 bp, exon I), a noncoding region (608 bp, intron), a second coding region (660 bp, exon II), and a 3' region of 693 bp. The upstream region shows a eukaryotic promoter structure and a consensus sequence for the 5' and 3' splicing sites. Thus the open reading frame (ORF DG32) coding for the 32-kDa protein of the dense granules of S. muris cyst merozoites is interrupted by an intron. To our knowledge, dg32 is the first sarcosporidian mosaic gene to be characterized.

Amino Acid Sequence↗

Redescription of Babesia equi Laveran, 1901 as Theileria equi Mehlhorn, Schein 1998.

The horse-parasitizing species Babesia equi Laveran, 1901 was redescribed as Theileria equi Mehlhorn, Schein 1998 and, thus, transferred from one valid genus to another. This transfer was needed since it turned out that this horse parasite showed the relevant characteristics of theilerians with regard to biological data, morphological features, biochemical properties, and molecular biological relationships.

Animals↗

Amplification of genomic DNA fragments of Sarcocystis muris (Apicomplexa) cyst merozoites encoding a thiol (cysteine) proteinase.

Parasites of the phylum Apicomplexa (Sporozoa) cause diseases such as malaria, toxoplasmosis, or intestinal coccidiosis. Invasive stages possess typical apical organelles such as dense granules that harbor a broad range of polypeptides that are believed to take part in the parasite-host cell interaction. In previous studies a 26-kDa polypeptide of dense granules from Sarcocystis muris cyst merozoites (bradyzoites) was characterized as a thiol (cysteine) proteinase. In this paper a method is demonstrated to amplify DNA fragments from genomic DNA of S. muris cyst merozoites by polymerase chain reaction, which probably code for the 26-kDa antigen.

Amino Acid Sequence↗