Genebank accession numbers of sequences of Culicoides species vectors of bluetongue virus in Germany.

The paper offers the genebank accession numbers of Culicoides obsoletus, Culicoides scoticus and Culicoides pulicaris sequences (ITS 1, ITS 2, 18S rRNA) that had been shown to be vectors of the bluetongue virus serotype 8, which was introduced in 2006 into Germany and spread until 2009 all over Central Europe, including parts of England. The numbers are FN 263292 until FN 263323.

A novel Sarcocystis-associated encephalitis and myositis in racing pigeons.

Sarcosporidian cysts in the skeletal muscle of domestic pigeons (Columba livia f. domestica) have previously been attributed to infection with Sarcocystis falcatula, which is shed in the faeces of the opossum (Didelphis virginiana). Here, we describe fatal spontaneous encephalitis and myositis associated with Sarcocystis infections in three flocks of racing pigeons with 47 of 244 animals affected. The clinical course was characterized by depression, mild diarrhoea, torticollis, opisthotonus, paralysis and trembling. Histopathological examination of 13 pigeons revealed generalized severe granulomatous and necrotizing meningoencephalitis and myositis with sarcosporidian cysts. Light and transmission electron microscopy identified cysts in heart and skeletal muscle of 1 to 2 mm in length and 20 to 50 microm in width. These were subdivided into small chambers by fine septae and filled with lancet-shaped cystozoites (7.5 x 1.5 microm) and dividing metrocytes, which is characteristic for Sarcocystis. The cysts had smooth walls and were devoid of protrusions typical of S. falcatula. Polymerase chain reaction amplification and sequencing of the internal transcribed spacer region (ITS-1) and the complete 28S rRNA identified a novel Sarcocystis species with only 51% ITS-1 nucleotide sequence similarity with S. falcatula. A phylogenetic comparison of the 28S rRNA revealed close sequence homologies with Frenkelia microti, Frenkelia glareoli and Sarcocystis neurona. The clinical, histopathological, electron microscopic and genetic data are unlike any previously described protozoan infections in pigeons, suggesting a novel, severe disease due to an as yet undescribed Sarcocystis species.

Transmission of feline calicivirus via the cat flea (Ctenocephalides felis).

In this study, a possible role of the cat flea (Ctenocephalides felis) in transmitting feline calicivirus (FCV) was examined. Fleas were fed via artificial membranes with FCV-spiked bovine blood, free of anti-FCV antibodies. Flea feces were collected daily for 10 days and incubated at room temperature. Infectivity of the feces was tested in vitro using Crandell-Reese Feline Kidney (CRFK) cells. FCV remained infectious for 8 days. These flea feces were also used to oronasally inoculate four specific pathogen-free (SPF) kittens. All kittens were successfully infected as demonstrated by virus isolation from pharyngeal swabs and seroconversion. Two of the cats showed, in addition, clinical signs. Besides the infection of cats with flea feces containing FCV, four SPF kittens were exposed to fleas that were fed with FCV-spiked bovine blood. One of the kittens was successfully infected via this route as demonstrated by virus isolation from pharyngeal swabs and virus isolation. The results of this study show that fleas can spread infectious virus through their feces or by stitch and must be considered a source of infection for uninfected cats.

Light and transmission electron microscopic studies on trophozoites and cyst-like stages of Histomonas meleagridis from cultures.

The present study deals with Berlin strains of Histomonas meleagridis, the specimens of which were cultivated in Dwyer's medium. The light and electron microscopic examination revealed that the cultivated trophozoite stages (reaching about 10 mum in size) appeared more or less spherical, although their surface (covered by a single membrane) showed amoeba-like waves. All stages were uni-nucleated and reproduced by binary fission with an extranuclear spindle apparatus. Some trophozoites appeared ovoid and possessed a single flagellum with a typical microtubular 9 x 2 + 2 arrangement. Furthermore, the latter were characterized by an inner row of typical microtubules (remnant of an axostyle) and a Golgi apparatus (both adjacent to the nucleus), multivesicular structures, hydrogenosomes, and many food vacuoles containing either starch grains or bacteria. Their cytoplasm was densely filled with glycogen granules and ribosomes. Similar stages were also documented in the caeca and cloaca of chicken when being inoculated (via cloaca) with such culture stages. In addition to these typical trophozoites, the cultures contained a low number of 10-mum-sized spherical cyst-like stages with a surrounding amorphous layer. The cytoplasm of some of these cyst-like stages-when studied by electron microscopy-appeared with two membranes or had formed an amorphic, cyst-wall-like layer at their surface, apparently corresponding to their light microscopical appearance. Such stages might be involved in transmission from one host to another and probably have been missed before in microscopical examinations of infected poultry.

Molecular biological features of strains of Histomonas meleagridis.

Berlin strains of Histomonas meleagridis were subcultivated to produce cyst-like stages. These strains were studied for their ITS 1 and 18S rRNA properties and compared with sequences in data banks of other H. meleagridis strains, Dientamoeba fragilis, and some species of the genus Trichomonas and relatives. The Berlin isolates that had previously been shown to be able to develop cyst-like structures (Munsch et al. 2008) represent a significant cluster among the published data of other Histomonas meleagridis isolates and thus the formation of cysts might be a common feature that would open further possibilities of transmission.

Investigations on the life cycle and morphology of Tunga penetrans in Brazil.

In the present study, the life cycle of Tunga penetrans was established in Wistar rats in the laboratory, and the morphology of the resulting developmental stages was studied by means of light and scanning electron microscopy. It was seen that the females enter at a nonfertilized stage through the skin of their hosts. Only there the copulation occurs, while females and males brought together in a Petri dish showed no interest in each other. In any way -- fertilized or not -- the females start about 6 days after penetration and hypertrophy with the ejection of eggs. While fertilized eggs proceed to development, the unfertilized ones remain arrested. The eggs are ovoid and measure about 600 x 320 mum. The larvae hatch from the eggs 1-6 days (mean 3-4) after ejection. Formation of larvae 2 took at least another day, while 4 up to 10 days more were needed until this larva starts pupation (mean 5-7 days). The formation of the adult fleas inside the puparium occurred within 9-15 days (with a maximum hatch at day 12). Adult female fleas having reached the skin of a host start blood sucking within 5 min and prepare to enter the skin. After 24 h, the flea stacked already with two thirds of its body inside the skin. After 40 h, the penetration was completed, and feeding and hypertrophical enlargement started, which was completed on day 6, when eggs became ejected. When studying the morphology of the fleas obtained from different hosts, slight variations were seen, which, however, are not significant for a species separation but may be an indication of the presence of different strains/races or the beginning of such a formation.

Experimental quantification of the feline leukaemia virus in the cat flea (Ctenocephalides felis) and its faeces.

Cat fleas (Ctenocephalides felis) were fed via artificial membranes and infected with the feline leukaemia virus (FeLV) from cell cultures. After removing the fleas from the blood source, the quantity of virus in the flea and its faeces was measured over a defined period of time. The virus was detectable in the fleas for up to 30 h at room temperature and up to 115 h at 4 degrees C. In the faeces, the amount of virus decreased much more slowly--after 2 weeks half of the initial amount of virus could still be detected. Thus the faeces might be a source of further infections, e.g. for the flea larvae or the cat itself.

Field trial of the efficacy of a combination of imidacloprid and permethrin against Tunga penetrans (sand flea, jigger flea) in dogs in Brazil.

In a field trial in Brazil 17 dogs penetrated by females of the jigger flea, Tunga penetrans, were topically treated with a combination of 10% imidacloprid and 50% permethrin (Advantix), while 17 dogs remained untreated. The follow-up controls on days 7, 14, 21 and 28 post-treatment clearly showed that, beginning from day 7, the flea load in treated dogs decreased, so that most of the dogs became free of tungiasis lesions, while in the untreated group the flea load remained high. Since the dogs distribute the flea eggs throughout the village, leading to a high incidence of tungiasis in humans, treatment of dogs probably also decreases the number of cases of tungiasis in the latter.

Effects of a combinations of emodepside and praziquantel on parasites of reptiles and rodents.

A new combination of two anthelmintic compounds containing emodepside and praziquantel (Profender, Bayer AG, Levekusen, Germany) was tested in pet rodents and reptiles. Topical application of the two compounds led to the quick disappearance of nematodes and cestodes from a broad spectrum of hosts including mice, jirds, snakes, anole lizards, turtles, monitor lizards, etc. In reptiles the dosage had to be increased, since the thick outer layer of the epidermis hinders the penetration of the compounds. In animals with an extremely thick epidermis (e.g. monitor lizards, leguans) the new product was applied under the armpits.

Efficacy of a combination of imidacloprid and moxidectin against parasites of reptiles and rodents: case reports.

A new compound containing imidacloprid 10% (w/v) and moxidectin 2.5% (w/v) (Advantage multi, Advocate) was applied as a spot-on treatment to mice experimentally infected with Trichuris muris. Case reports of reptiles found positive for nematode and mite infections following parasitological examination and treated with this compound are also discussed. The results demonstrated that the registered, recommended 2.5% moxidectin concentration for use in dogs was sufficient to eliminate nematodes and mites in reptiles. Infections with nematodes were successfully treated with a single application. Mite infestations in reptiles were eliminated using a treatment repeated on 3 consecutive days.

Determining a diagnostic dose for imidacloprid susceptibility testing of field-collected isolates of cat fleas (Siphonaptera: Pulicidae).

The susceptibility of four laboratory strains of cat fleas, Ctenocephalides felis (Bouche), to imidacloprid was determined by three different laboratories, by using a standardized bioassay protocol. The probit lines generated by the different laboratories were very similar, with LC50 values ranging from 0.32 to 0.81 ppm. Based on these data, a diagnostic dose (DD) of 3 ppm imidacloprid in larval rearing media was provisionally identified for detecting shifts in tolerance, possibly as a consequence of incipient imidacloprid resistance. None of the larvae from the susceptible laboratory strains survived the DD. Eighteen field-collected isolates were evaluated for their susceptibility to imidacloprid and to validate a DD of 3 ppm. Probit lines from 18 field-collected isolates were very similar, with LC50 values ranging from 0.14 to 1.52 ppm. When exposed to the DD, between 3 and 10% of the exposed larvae emerged as adults from only three of the 18 isolates. All other field isolates gave 100% mortality at the DD. Under the criteria established (>5% survivorship at 3 ppm), two isolates would be established on mammalian hosts and more extensive tests conducted to exclude or confirm the presence of resistance. The DD of 3 ppm is robust enough to eliminate most of the susceptible isolates collected until today, yet low enough to identify possible isolates for further testing.

Molecular biological investigations of Brazilian Tunga sp. isolates from man, dogs, cats, pigs and rats.

Twenty-four isolates of Tunga were collected from afflicted humans, dogs, cats, pigs and rats in Brazil. To investigate genetic diversity, a hypervariable section of mitochondrial 16S rDNA was amplified using PCR and subsequently sequenced. In order to compare results with another species of the genus Tunga, three isolates of the recently described Tunga trimamillata were also checked. Whereas eleven isolates (five from cats, three from dogs and three from humans) were of identical sequence, thirteen isolates collected from dogs, humans, pigs and rats showed differences in sequence up to 49%, so that the existence of one or more new species of Tunga may be presumed.

[Humanopathogenic parasites--an unwelcome import]. / Humanpathogene Parasiten warten schon: So entkommt der Urlauber den unliebsamen Begleitern.

Humanopathogenic parasites represent a health risk for travelers to tropical countries. They can be picked up by direct contact, via contaminated food and drink, the bites of blood-sucking insects and leeches, as also via direct penetration through the skin. Not only the bite and sting wounds themselves are unpleasant. The true danger to the victim is represented, in particular, by the bacteria, viruses or parasites that are transmitted in this way. They can give rise to protracted diseases such as hepatitis B, cutaneous or intestinal disorders. The most effective preventive measure is exposure prophylaxis.

Molecular phylogeny of isolates of Ctenocephalides felis and related species based on analysis of ITS1, ITS2 and mitochondrial 16S rDNA sequences and random binding primers.

The phylogenetic relationships among 31 different flea isolates representing seven different species were studied by nucleotide sequence comparison of the internal transcribed spacer 1 (ITS1), internal transcribed spacer 2 (ITS2) and/or mitochondrial 16S ribosomal RNA gene (mt16S-rDNA) to examine the patterns of variation. Results show that all regions are useful in discriminating among flea species. In Ctenocephalides felis and Tunga penetrans, some differences in these gene regions occurred among different isolates within the same species. In the latter case, the differences are in the mt16S-rDNA region, with one isolate showing 48% divergence in nucleotide sequence. The taxonomic implications of this result are unclear at present. The gene regions revealed differences between C. felis isolates only after DNA sequencing the PCR products. Further differentiation among C. felis isolates was obtained using four different random binding primers (decamers) and primers for mammalian aldolase to amplify narrow differences in the genome. Using these primers we were able to discriminate between different C. felis isolates and determine that some of the genetic variation coincided with minor differences in response to the control agent imidacloprid. However, overall findings do not support the existence of subspecies of C. felis.

[Hantavirus-induced acute renal failure. A case report]. / Hantavirus-bedingtes akutes Nierenversagen. Ein Fallbericht.

A non-ischemic acute renal failure may indicate a hantavirus nephropathy, so that appropriate serological examinations should be done. A 19-year-old man had acutely fallen ill with fever, rhinitis, and arthralgia. Clinically, he presented with a polyuric renal failure as well as hypertension, proteinuria, erythro- and leucocyturia. Serologically, IgG- and IgM-hantavirus antibodies were found against the Central European variant of hantavirus Dobrava. At renal biopsy, tubular epithelial swelling, vacuolization, and necrosis as well as interstitial hemorrhages were seen. Electron microscopically, virion particles with a gridlike surface structure as well as with the shape and dimension evident of hantaviruses could be detected in the nucleus and cytoplasm of renal tubular cells. After a clinical course of 3 weeks, the patient was free of complaints and cured with a restitutio ad integrum.

Evidence of horizontal transmission of feline leukemia virus by the cat flea ( Ctenocephalides felis).

The feline leukemia virus (FeLV) is a naturally occurring and widespread retrovirus among domestic cats. The virus is mainly transmitted horizontally through saliva, blood and other body fluids by close contact between cats. Vectors other than cats, e.g. blood-sucking parasites, have not been reported. This study tested the vector potential of the cat flea ( Ctenocephalides felis) for FeLV. In a first feeding, fleas were fed for 24 h with blood from a FeLV-infected cat with persistent viremia. FeLV could be detected in the fleas, as well as in their feces. Fleas were then divided in two populations and fed in a second feeding for 5 h or 24 h with non-infected non-viremic blood. FeLV was again detected in the fleas and their feces. In addition, the two resulting blood samples of the second feeding were subsequently tested for FeLV and both samples were positive for FeLV RNA. The cat flea transmitted the FeLV from one blood sample to another. In a third feeding, the same populations of fleas were fed again with non-infected blood for 5 h or 24 h. This time FeLV was not detected in the fleas, or in the feces or blood samples. Results show that cat fleas are potential vectors for FeLV RNA in vitro and probably also in vivo.