Tick infestation and prophylaxis of dogs in northeastern Germany: a prospective study.

Ticks transmit various important pathogens to humans and animals, and dogs are frequently exposed to tick infestation. The objective of this study was to examine tick infestation and the characteristics of tick prophylaxis of dogs in the Berlin/Brandenburg area. A total of 441 dogs (392 owners) was examined from March 2010 to April 2011. The dog owners participated in the study for a period of 1-13 months (10.33±2.85; median 11.00). The prevalences of a total of 1728 ticks collected from 251 (57%) of these dogs were: 46.0% Ixodes ricinus, 45.1% Dermacentor reticulatus, 8.8% Ixodes hexagonus, and 0.1% Rhipicephalus sanguineus. The ticks were 75.2% adult females and 24.4% adult males, and 0.4% were nymphs. The average prevalence of apparent infestation of tick-positive dogs was 0.78 ticks/month (median 0.38). Tick infestation was highest in October (5.9±5.8 ticks/dog) and lowest in December (1±0 tick/dog). The highest frequency of infestation was observed during May (117 dogs). The number of ticks found on dogs by owners on a single day varied from one to 70 (median 1). The scutal index indicated that more than 60% of I. ricinus and more than 40% of D. reticulatus had been removed after they had fed for more than 2 days. The heads, necks, chests, and limbs of the dogs were the most common attachment sites. Data for tick prophylaxis with substances licensed for dogs by the Medicinal Products Act (MPA) were available for 124 dogs; a total of 1195 ticks was obtained from these dogs. About two-thirds of the ticks were collected from dogs that were treated incorrectly (n=96) or were not treated (n=60). One third of the ticks were collected from dogs (n=96) that had been treated correctly. Dog-specific characteristics such as length of coat (p=0.011) and body size (p=0.040) played significant roles in tick infestation.

A novel high-resolution melt PCR assay discriminates Anaplasma phagocytophilum and "Candidatus Neoehrlichia mikurensis".

"Candidatus Neoehrlichia mikurensis" (Anaplasmataceae) is an emerging pathogen transmitted by Ixodes ticks. Conventional PCR and the newly developed high-resolution melt PCR were used to detect and discriminate "Candidatus Neoehrlichia mikurensis" and Anaplasma phagocytophilum. Both bacterial species were frequently found in Ixodes ricinus and Ixodes hexagonus but virtually absent from Dermacentor reticulatus. In rodents, "Candidatus N. mikurensis" was significantly more prevalent than A. phagocytophilum, whereas in cats, only A. phagocytophilum was found.

[Tick infestation and tick prophylaxis in dogs in the area of Berlin/Brandenburg--results of a questionnaire study]. / Zeckeninfestation und Zeckenprophylaxe bei Hunden im Raum Berlin/Brandenburg--Ergebnisse einer Fragebogenstudie.

Ticks can transmit different pathogens to humans and animals. Dogs are frequently exposed to tick infestation, which underscores the importance of tick control measures. The objective of this study was to examine the awareness of dog owners regarding tick infestation and tick prophylaxis by a questionnaire survey. During the period from March to December 2010 a total of 616 owners of 670 dogs completed the questionnaire. According to the questionnaire results, 92% of the dogs were previously infested by ticks; 31% of these showed a moderate tick infestation (1-2 ticks a month), almost one in ten dogs was infested by eight or more ticks a month. 17% of the dogs were examined for ticks by the respective owner not at all or only at irregular intervals, 61% of the dogs were examined at least once a day. A tick prophylaxis was performed in 469 dogs (71%). In 353 dogs (53%), registered pharmaceutical products with appropriate label claims were employed. Spot-on products were used most frequently (93%), followed by collars (5%) and sprays (1%).These products were not used as recommended in 56% of the dogs. For further 33% of the dogs, it was not possible to decide if the products were used correctly or not. According to the dog owner statements, tick borne diseases were diagnosed in approximately 2% of the dogs. Dog specific characters, such as coat length, size, age, and walking habits were significant factors influencing the frequency of tick infestation. In summary it can be concluded that nearly every dog in the area of Berlin/Brandenburg is infested by ticks. In the majority of cases the prophylactic and/or therapeutic measures to prevent infestation are not performed correctly.

Molecular biological comparison of different Besnoitia species and stages from different countries.

Besnoitia besnoiti tissue cysts from a recent outbreak in cattle in Germany were characterized with respect to their internal transcribed spacer regions 1, 2, and 18S rDNA gene sequences. These results were compared with own sequences of an Israelian isolate of B. besnoiti and of Besnoitia jellisoni cystozoites stored for years in liquid nitrogen. Furthermore, material was studied that was obtained from white mice (Balb/C) that had been successfully infected by intraperitoneal infection of fresh cystozoites from the German outbreak. All results were then compared and discussed with respect to databank sequences of other Besnoitia species. Comprehensive phylogenetic studies of B. besnoiti isolates from Germany revealed almost identical sequence alignments when compared to previously sequenced B. besnoiti isolates from Israel and Spain. More importantly, phylogenetic analysis revealed two distant clusters of Besnoitia species: the first one includes Besnoitia akodoni, Besnoitia darlingi, and Besnoitia oryctofelisi, while the second cluster includes B. besnoiti, Besnoitia bennetti, Besnoitia tarandi, and the Besnoitia species of rodents (B. jellisoni). The also B. jellisoni named species of the GenBank (AF 076860) must be another one, since our strain derives directly from Frenkel. These findings give strong hints that B. besnoiti has a cycle between rodents and a predator and that cattle and other are only accidental hosts.

Seasonal dynamics of biting midges (Diptera: Ceratopogonidae, Culicoides spp.) on dairy farms of Central Germany during the 2007/2008 epidemic of bluetongue.

The unforeseen outbreak of bluetongue in north-western Europe in August 2006 raised the question, which Culicoides spp. were involved in the transmission of bluetongue virus (BTV). Based on the decision 2007/20/EU of December 2006, a large-scale entomological surveillance programme was initiated in the five affected EU member states including Germany. This paper reports on the entomological findings obtained from March/April 2007 to May 2008 at 15 sampling sites in the federal states of Lower Saxony (eastern region), Mecklenburg-Western Pomerania, Brandenburg and Saxony-Anhalt: The number of captured biting midges in one trap varied from none or few Culicoides during winter (December 2007 to April 2008) to up to more than 12,500 individuals during summer and autumn. Catches of the C. obsoletus group were consistently higher than those of the C. pulicaris group. C. imicola, the principal afro-asiatic vector of BTV, was not detected. High numbers of midges were caught inside the cattle sheds. Eleven pools of biting midges were RT-PCR-positive to BTV-8 including pools of non-engorged midges of the C. obsoletus and of the C. pulicaris groups. The first BTV-genome positive pool of midges was detected in August 2007; the remaining genome-positive pools were detected during October and November 2007.

An appraisal of current and new techniques intended to protect bulls against Culicoides and other haematophagous nematocera: the case of Schmergow, Brandenburg, Germany.

The outbreak of bluetongue (BTV-8) in many parts of north-western Europe led to efforts to curb the spread of the disease, particularly in farms with valuable livestock, as on a stud bull farm in Schmergow, Brandenburg, Germany. In the abundance of the putative BT vectors, Palaearctic Culicoides species, several vector control methods were applied in the hope for a reduction of the target insect populations. Insecticide-impregnated ear tags and regular treatments at 6-week intervals of all bulls with deltamethrin pour on were expected to achieve the desired control of the biting midges. Additionally, insecticide-treated mosquito fences circumventing much of the pens were tried for the first time against Culicoides. Two suction black-light traps (BioGents(R) sentinel traps) helped to monitor the densities of Culicoides and other haematophagous nematocera during the trial period from July to December 2007. Despite all efforts, the densities of Culicoides were not distinctly reduced. Several thousand midges were repeatedly recorded during one-night catches. Examinations of midges and other haematophagous nematocera (Aedes and Anopheles species) revealed high percentages of successful feedings between 10% and 35% for Culicoides and more than 50% for Aedes and Anopheles species. Since all insects were caught inside the pens, the concept of endophily vs exophily or endophagy vs exophagy for some Culicoides species needs to be revised accordingly. Also, stabling of valuable livestock does not reduce the host-vector interface and, hence, the risk of transmission of BT.

Another African disease in Central Europa: Besnoitiosis of cattle. I. Light and electron microscopical study.

The paper reports the first detection of besnoitiosis of cattle in Germany. Just 2 years after the first appearance of the African Bluetongue disease (BTD) of cattle in Central Europe, another African agent of disease has arrived in Germany. While it was proven that the BTD virus was transmitted (after its first appearance) by endemic midges of the genus Culicoides (C. obsoletus, C. pulicaris), nothing is known, how the infectious stages of Besnoitia besnoiti-a member of the so-called cyst-forming coccidia-found their way to a herd in Southern Germany. The infected animals showed all characteristic clinical symptoms of besnoitiosis such as hyposclerodermia, hyperkeratosis, alopecia, and whitish tissue cysts in subcutaneous tissues as well as in the cornea. These cysts had diameters of up to 3 mm and consisted of a dense outer layer (=secondary cyst wall), which surrounded a host cell, that had been enormously enlarged by an inner parasitophorous vacuole containing thousands of 7-9 x 2 mum sized, banana-shaped cyst merozoites (=cystozoites, bradyzoites).Their fine structure was identical to that of published stages of B. besnoiti. During cyst development, the nucleus of the host cell had been hypertrophied and had apparently undergone several divisions, since many flattened, but very large nuclei were seen in light and electron microscopy. Thus, this study proves the arrival of another serious agent of disease of ruminants in Central Europe-a fact which is especially important, since in this species, there is neither information on the way of transmission from animal to animal nor exists concrete information on an efficacious therapy or on the modalities of its import into Germany.

[Dermacentor reticulatus (Fabricius, 1794)--distribution, biology and vector for Babesia canis in Germany]. / Dermacentor reticulatus (Fabricius, 1794)-- Verbreitung, Biologie und Vektor von Babesia canis in Deutschland.

This work deals with occurrence, distribution as well as biology and vectorial capacity of the ornate dog tick (Dermacentor (D.) reticulatus). Until 30 years ago this tick has only been described in a few natural foci in southern Germany. Its distribution has however expanded in the course of the last years. With the exception of northern Germany it is now covering large areas of Germany. This is particularly the case in the Upper Rhine Valley, Saarland and the Mark Brandenburg. However, it is endemic in many other areas of Germany as well. The transformation of agricultural land into fallow land, an increase in host reservoirs and climatic changes are discussed as main contributors to this change. Little is known about the role of this species as a vector for virus, rickettsia, bacteria and protozoa as well as its medical and veterinary importance. D. reticulatus is a vector for Babesia canis canis. It is reported here about a case of autochthonous babesiosis in a dog from Berlin/Brandenburg.

[Pilot study to evaluate the efficiency of insecticide-treated mosquito net fences for the protection of horses against nuisance insects in northern Brandenburg]. / Pilotstudie zur Bewertung der Effizienz insektizid-behandelter Netzzäune zum Schutz von Pferden gegen Weidefliegen im nördlichen Brandenburg.

A fence of black mosquito netting of 100 cm height, pre-treated with 80 mg/m2 of deltamethrin and UV-protected, was used to shelter horses from nuisance and biting insects on pasture in northern Brandenburg. The netting material was attached to the surrounding poles of the existing fences at a height of 15 cm above ground. Three trial groups were selected grazing in spatially separated areas with comparable densities of insect populations. One paddock was completely fenced apart from a wall of 170 cm height and 70 m length. The second pasture had only partial protection with 126 m (13.4%) of fence out of a total perimeter of 942 m. The third pasture served as control. Trap catches outside the fully or partially protected pasture were by at least 60% lower than those recorded for the control pasture. Digital pictures from five different anatomical regions indicated fewer flies on horses kept at the completely or partially protected areas as compared to the control area. The average attack rate in the protected areas amounted to 4.4 and 7.6 flies per horse at the completely or partially protected areas, respectively, as opposed to horses on the control pasture with 172.1 flies. In comparison to the control pasture the horses grazing on the protected areas showed fewer defensive movements, grazing in an undisturbed manner.

A field study to estimate the prevalence of Trypanosoma equiperdum in Mongolian horses.

From May to July 2000, a cross-sectional study was conducted to estimate the prevalence of Trypanosoma equiperdum in the horse population of the central province (Tuv aimag) of Mongolia. On average, four herds were selected from each of the 29 aimag subdivisions (119 herds). From each herd, 10 horses were sampled in proportion to sex and age categories in the respective herds (1190 horses). Sera from 1122 horses were analysed for T. equiperdum antibodies using two serological assays, the complement fixation test (CFT) and the enzyme-linked immunosorbent assay (ELISA). The crude estimate of the CFT and the ELISA seroprevalence was 7.6 and 6.7%, respectively. Concordance between the CFT and ELISA results was high (96%). The highest number of CFT positive animals was detected in one herd in Möngönmorit (6/10), followed by herds in Bayandelger (5/10) and in Bayantsagaan (5/10). Poor body condition was significantly correlated with positive serological status in both CFT and ELISA. A history of abortion appeared to be a risk factor for both CFT and ELISA seropositivity. Blood samples of all horses belonging to herds with at least three (3/10) seropositive animals (CFT and/or ELISA) were analysed by light microscopy and by PCR using a Trypanosoma (Trypanozoon) brucei specific primer pair. No trypanosomes or any other haemoparasites could be detected in Giemsa stained thin blood smears. Eight out of the 130 samples (6.2%) analysed by PCR gave positive signals. Seven out of the eight PCR positive horses were also serologically positive. One PCR (and ELISA) positive stallion from Möngönmorit showed emaciation, scrotal and preputial oedema and an oedematous skin plaque. From the serological and DNA-based results it is concluded, that trypanosome infections occur in horses in the Tuv aimag of Mongolia. Since at present neither serological nor DNA-based tests allow a subspecies specific identification within the subgenus Trypanozoon, no definitive diagnosis can be given for T. equiperdum. Whether the examined herds are infected with T. equiperdum or with T. evansi, the causative agent of surra, remains an open question. However, based on the clinical findings, the negative parasitological results and the concentration of conspicuous seroprevalences in single herds, circumstantial evidence supports the existence of infections with the causative agent of dourine.

Electron microscopic investigations on stages of dog piroplasms cultured in vitro: Asian isolates of Babesia gibsoni and strains of B. canis from France and Hungary.

Stages obtained from two Asian Babesia gibsoni-isolates cultured in vitro were studied by means of transmission electron microscopy and compared to strains of B. canis cultured in vitro. While the developmental stages of the latter preserved their shape in culture, many of the initially small stages of the B. gibsoni strains grew considerably and often looked rather similar to B. canis.