Anaplasma phagocytophilum - the most widespread tick-borne infection in animals in Europe

The bacterium Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila) may cause infection in several animal species including human. The disease in domestic ruminants is also called tick-borne fever (TBF), and has been known for at least 200 years. In Europe, clinical manifestations due to A. phagocytophilum have been recorded in sheep, goat, cattle, horse, dog, cat, roe deer, reindeer and human. However, seropositive and PCR-positive mammalian have been detected in several other species. Investigations indicate that the infection is prevalent in Ixodes ricinus areas in most countries in Europe. A. phagocytophilum infection may cause high fever, cytoplasmatic inclusions in phagocytes and severe neutropenia, but is seldom fatal unless complicated by other infections. Complications may include abortions, and impaired spermatogenesis for several months. However, the most important aspect of the infection at least in sheep is its implication as a predisposing factor for other infections. Factors such as climate, management, other infections, individual conditions etc. are important for the outcome of the infection. A. phagocytophilum may cause persistent infection in several species. Based on the 16S rRNA gene sequences several variants exist. Different variants may exist within the same herd and even simultaneously in the same animal. Variants may behave differently and interact in the mammalian host.     

Resistance to tick-borne fever in young lambs.

Lambs were inoculated with Ehrlichia phagocytophila infected blood at the age of either two or six weeks. The temperature response and degree of parasitaemia were both most pronounced in the lambs inoculated at six weeks old. The half-life of antibodies to E phagocytophila was estimated to be 17.5 days. Lambs inoculated with tick-borne fever (TBF) had lower liveweights in the autumn than TBF-free controls.     

A morphological and molecular study of Anaplasma phagocytophilum transmission events at the time of Ixodes ricinus tick bite

Background

Anaplasma phagocytophilum is the causative agent of human granulocytic anaplasmosis (HGA) in humans and tick-borne fever (TBF) in ruminants. The bacterium invades and replicates in phagocytes, especially in polymorphonuclear granulocytes.

Methods

In the present study, skin biopsies and ticks (Ixodes ricinus) were collected from tick feeding lesions on 38 grazing lambs between two and three weeks after access to pastures. The histopathological changes associated with tick bites and A. phagocytophilum infection, were described. In addition the skin biopsies were examined by immunohistochemistry. Furthermore, samples from blood, skin biopsies and ticks were examined by serology, PCR amplification of msp2 (p44), genotyping of rrs (16S rRNA) variants, and compared with the results obtained from histological and immunohistochemical investigations.

Results

Tick bites were associated with chronic and hyperplastic inflammatory skin lesions in this study. A. phagocytophilum present in skin lesions were mainly associated with neutrophils and macrophages. Bacteria were occasionally observed in the Tunica media and Tunica adventitia of small vessels, but were rarely found in association with endothelial cells. PCR and genotyping of organisms present in blood, ticks and skin biopsies suggested a haematogenous and a local spread of organisms at the tick attachment sites.

Conclusions

The present study describes different aspects of A. phagocytophilum infection at the site of tick bite, and indicates that A. phagocytophilum rarely associates with endothelium during the early pathogenesis of infection.     

A comparative study of clinical manifestations,haematological and serological responses after experimental infection with Anaplasma phagocytophilum in two Norwegian sheep breeds

Background

It has been questioned if the old native Norwegian sheep breed, Old Norse Sheep (also called Norwegian Feral Sheep), normally distributed on coastal areas where ticks are abundant, is more protected against tick-borne infections than other Norwegian breeds due to a continuously high selection pressure on pasture. The aim of the present study was to test this hypothesis in an experimental infection study.

Methods

Five-months-old lambs of two Norwegian sheep breeds, Norwegian White (NW) sheep and Old Norse (ON) sheep, were experimentally infected with a 16S rRNA genetic variant of Anaplasma phagocytophilum (similar to GenBank accession number {"type":"entrez-nucleotide","attrs":{"text":"M73220","term_id":"148293"}}M73220). The experiment was repeated for two subsequent years, 2008 and 2009, with the use of 16 lambs of each breed annually. Ten lambs of each breed were inoculated intravenously each year with 0.4 ml A. phagocytophilum-infected blood containing approximately 0.5 × 10⁶ infected neutrophils/ml. Six lambs of each breed were used as uninfected controls. Half of the primary inoculated lambs in each breed were re-challenged with the same infectious dose at nine (2008) and twelve (2009) weeks after the first challenge. The clinical, haematological and serological responses to A. phagocytophilum infection were compared in the two sheep breeds.

Results

The present study indicates a difference in fever response and infection rate between breeds of Norwegian sheep after experimental infection with A. phagocytophilum.

Conclusion

Although clinical response seems to be less in ON-lambs compared to NW-lambs, further studies including more animals are needed to evaluate if the ON-breed is more protected against tick-borne infections than other Norwegian breeds.     

The effect of two different oxytetracycline treatments in experimental Ehrlichia phagocytophila infected lambs

The effect of 2 different oxytetracycline treatments in acute E. phagocytophila infected lambs was investigated. Twenty 5-month-old lambs of the Dala and Rygja breeds were used. Ten lambs were inoculated intravenously with a stabilate of an ovine E. phagocytophila strain. On the third day of fever, 4 lambs were given long-acting oxytetracycline (Terramycin prolongatum vet, Pfizer) (20 mg/kg) intramuscularly and another 4 lambs were given short-acting oxytetracycline (Terramycin vet, Pfizer) (10 mg/kg) intravenously for 5 consecutive days. The lambs were examined for the presence of Ehrlichia infection by blood smear evaluation, polymerase chain reaction (PCR) and antibody titre against E. equi. One month after the last antibiotic treatment, 250 ml citrate blood from each of these lambs were inoculated into each of 10 susceptible lambs, which were observed during the following 6 weeks. The results indicate that oxytetracycline given in the acute stage of the infection may effectively terminate the development of fever, rickettsemia and weight reduction in E. phagocytophila infected lambs. No difference was observed between the 2 treatment groups. However, at least 3 of 8 antibiotic treated lambs (37.5%) were still infected with granulocytic Ehrlichia 3 months after treatment.     

Persistence of Ehrlichia phagocytophila infection in two age groups of lambs

Tick-borne fever (TBF) is caused by the rickettsiae Ehrlichia phagocytophila and is a common disease in sheep in tick (Ixodes ricinus) infested areas in Norway. Earlier investigations have shown that some sheep could remain infected for several months after the primary infection. In this study, the persistence of E. phagocytophila after experimental infection was investigated in 2 age groups of lambs. Six lambs (1-2 weeks old) and 14 lambs (6-8 months old) were inoculated intravenously with an ovine strain of E. phagocytophila and thereafter examined clinically (including daily body temperature recording) and by haematological and serological (E. equi antibodies) methods for the next 4 months. At the end of this period, the lambs were examined for a TBF infection by blood smear investigation and blood inoculation studies. The infection was demonstrated in 19 (95%) of the 20 lambs.     

Effects of the dose of Ehrlichia phagocytophila on the severity of experimental infections in lambs

Twenty-one lambs were used to investigate whether their response to an infection with Ehrlichia phagocytophila was dose-dependent Four groups of four lambs were infected intravenously with a dilution in physiological saline of E phagocytophila-infected sheep blood containing either 1.3 x 10(5) infected cells, or approximately 43 infected cells, 4.3 infected cells, or 1.3 infected cells (mean values) and four lambs were left uninfected. The incubation period was significantly shorter in the lambs infected with the highest dose of E phagocytophila. However, the clinical and haematological changes observed, and the weekly weight gains of the lambs were independent of the dose of E phagocytophila. As little as one Ephagocytophila infected cell may be enough to transmit the infection.     

Prevalence of gastrointestinal helminths,lungworms and liver fluke in sheep and goats in Norway

The present study describes the occurrence of various gastrointestinal helminths, lungworms and liver flukes in Norwegian sheep and goats as assessed from faecal samples and post mortem examinations performed between 2007 and 2010. Faecal samples for gastrointestinal nematode egg counts were collected from 77 sheep flocks and 30 dairy goat flocks from three geographical regions in Norway. Additionally, thirty-two lambs and 16 adult goats were euthanized for necropsy examination and for identification of adult gastrointestinal nematodes and tapeworms, lungworms and liver flukes. The survey showed that there was a higher mean excretion of trichostrongyle eggs in sheep than in goats at the individual level (392 EPG vs. 154 EPG, p < 0.001). For both host species, the mean prevalence and intensity of excreted trichostrongyle eggs were significantly higher in the southern coastal region compared with the inland and northern regions (p < 0.001). Third stage larvae of Trichostrongylus/Teladorsagia, Haemonchus and Nematodirus type were the most prevalent ones in the coprocultures from sheep, whereas larvae of Trichostrongylus/Teladorsagia and Nematodirus type dominated in goats. The most prevalent gastrointestinal nematode species found at necropsy was Teladorsagia circumcincta (75.0 and 81.2% respectively in sheep and goats), while the largest mean worm burdens were recorded for Haemonchus contortus in sheep (724 ± 623) and T. circumcincta in goats (377 ± 529). Other gastrointestinal nematode species were present at low prevalence or in low numbers.     

Correction to: Population genetic structure of lumpfish along the Norwegian coast: aquaculture implications

The original version of this article unfortunately contained an error where the figure caption for Figures 3 and 4 was mixed.