Molecular and serologic evidence of Anaplasma phagocytophilum infection in cats in North America

Anaplasma phagocytophilum DNA was detected in blood of clinically ill cats from Massachusetts (n = 4) and Connecticut (1) by use of polymerase chain reaction assay and DNA sequencing. All 5 cats were allowed outdoors, and Ixodes scapularis were found on 3 cats. Clinical signs of fever, anorexia, and lethargy resolved quickly after treatment with doxycycline or tetracycline. Serum samples from each cat reacted with A. phagocytophilum morulae via an indirect fluorescent antibody assay; positive antibody titers persisted even after 21 to 30 days of treatment with tetracycline. To the authors' knowledge, this is the first report of A. phagocytophilum infection of domestic cats in North America. Results suggest that infection with the organism may be associated with clinical illness in some cats.     

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.     

Microscopic and clinical evidence for Anaplasma (Ehrlichia) phagocytophilum infection in Italian cats

Fifteen cats from Italy with Ehrlichia-like inclusion bodies in their neutrophils were studied. They were diagnosed with Anaplasma (Ehrlichia) phagocytophilum infection on the basis of cytological observation of morulae within 1 to 21 per cent of their neutrophils, clinical signs characteristic of ehrlichiosis and their response to doxycycline. The predominant signs of disease were anorexia, lethargy, hyperaesthesia, muscle and joint pain, lameness, neck rigidity, lymphadenomegaly, gingivitis/periodontitis, conjunctivitis, weight loss, incoordination, pale mucous membranes and hyperglobulinaemia. The cat with inclusions in 21 per cent of its neutrophils had suffered arthralgia, tachypnoea, neck rigidity, vomiting and thrombocytopenia for four months, but recovered promptly after treatment with doxycycline.     

Seroprevalence of Anaplasma phagocytophilum infection in dogs in Germany

To determine the prevalence of Anaplasma phagocytophilum in dogs in Germany serum samples from 1124 dogs that were under suspicion of having anaplasmosis were examined. The samples were tested by an indirect immunofluorescence test (OFT) for antibodies to A. phagocytophilum. The geographical origin of positive cases were analysed with an geographic information system. Antibodies to A. phagocytophilum were found in 563 (50.1%) of the tested dogs. 166 dogs came from Saarland, 161 from North Rhine-Westphalia, 134 from Baden-Wuerttemberg, 33 from Bavaria, 22 from Rhineland-Palatinate, 11 from Hamburg, 10 from Brandenburg, 9 from Lower Saxony, 8 from Hesse and Berlin respectively and 1 from Schleswig-Holstein. Clinical signs and laboratory findings of 26 seropositive dogs were analysed. Those dogs showed a low haematocrit, thrombocytopenia and leucocytoses as well as higher values for alkaline phosphatase, ALAT und bilirubin. The clinical signs were lameness in 13 dogs, lethargy in 5, and uveitis in 3 dogs. Rhinitis and lymphadenopathy was found in 2 dogs and retinal detachment with blindness in 1 dog.     

Seroprevalence of Fasciolosis in Equines of the Black Sea Region,Turkey

This study determined the seroprevalence of fasciolosis in equines (n = 140) by enzyme-linked immunosorbent assay and Western blot. Blood samples were collected from different locations of the Black Sea Region of Turkey. The overall seroprevalence of fasciolosis in equines was 19.3%: 18% of horses (15), 22.6% of donkeys (7), and 19.2% of mules (5). The seroprevalence rates across provinces were as follows: 25% (5/20) in Samsun, 21.9% (7/32) in Tokat, 31.4% (11/35) in Amasya, 7.4% (2/27) in Ordu, and 7.7% (2/26) in Sinop. Three protein bands with molecular weights of 13, 30-40, and 40-65 kDa were determined in antigen clusters in sera from Fasciola spp-infected equine animals.     

Prevalence of Anaplasma phagocytophilum infection and effect on lamb growth

Background

A major challenge in sheep farming during the grazing season along the coast of south-western Norway is tick-borne fever (TBF) caused by the bacteria Anaplasma phagocytophilum that is transmitted by the tick Ixodes ricinus.

Methods

A study was carried out in 2007 and 2008 to examine the prevalence of A. phagocytophilum infection and effect on weaning weight in lambs. The study included 1208 lambs from farms in Sunndal Ram Circle in Møre and Romsdal County in Mid-Norway, where ticks are frequently observed. All lambs were blood sampled and serum was analyzed by an indirect fluorescent antibody assay (IFA) to determine an antibody status (positive or negative) to A. phagocytophilum infection. Weight and weight gain and possible effect of infection were analyzed using ANOVA and the MIXED procedure in SAS.

Results

The overall prevalence of infection with A. phagocytophilum was 55%. A lower weaning weight of 3% (1.34 kg, p < 0.01) was estimated in lambs seropositive to an A. phagocytophilum infection compared to seronegative lambs at an average age of 137 days.

Conclusions

The results show that A. phagocytophilum infection has an effect on lamb weight gain. The study also support previous findings that A. phagocytophilum infection is widespread in areas where ticks are prevalent, even in flocks treated prophylactic with acaricides.     

Molecular detection of Anaplasma phagocytophilum in cattle and Ixodes persulcatus ticks

The tick-borne pathogen, Anaplasma phagocytophilum (A. phagocytophilum), the causative agent of human granulocytic anaplasmosis (HGA), is increasingly becoming a public health concern as an aetiological agent for emerging infectious disease. We found A. phagocytophilum infection in a pooled sample of field-collected Ixodes persulcatus (I. persulcatus) ticks from one district in Hokkaido, Japan. Thus, to further investigate the prevalence in field-collected ticks, we used PCR assays targeting the A. phagocytophilum gene encoding 44 kDa major outer membrane protein (p44) for screening of I. persulcatus ticks and samples from cattle from pastures. Out of the 281 I. persulcatus ticks, 20 (7.1%) were found to harbor A. phagocytophilum DNA. The infection rate for A. phagocytophilum in cattle was 3.4% (42/1251). In future studies, it will be necessary to investigate effects of the infection in order to understand its pathogenesis of A. phagocytophilum in domestic animals.     

Variant -and individual dependent nature of persistent Anaplasma phagocytophilum infection

Background

Anaplasma phagocytophilum is the causative agent of tick-borne fever in ruminants and human granulocytotropic anaplasmosis (HGA). The bacterium is able to survive for several months in immune-competent sheep by modifying important cellular and humoral defence mechanisms. Little is known about how different strains of A. phagocytophilum propagate in their natural hosts during persistent infection.

Methods

Two groups of five lambs were infected with each of two 16S rRNA gene variants of A. phagocytophilum, i.e. 16S variant 1 which is identical to GenBank no {"type":"entrez-nucleotide","attrs":{"text":"M73220","term_id":"148293","term_text":"M73220"}}M73220 and 16S variant 2 which is identical to GenBank no {"type":"entrez-nucleotide","attrs":{"text":"AF336220","term_id":"13325085","term_text":"AF336220"}}AF336220, respectively. The lambs were infected intravenously and followed by blood sampling for six months. A. phagocytophilum infection in the peripheral blood was detected by absolute quantitative real-time PCR.

Results

Both 16S rRNA gene variants of A. phagocytophilum established persistent infection for at least six months and showed cyclic bacteraemias, but variant 1 introduced more frequent periods of bacteraemia and higher number of organisms than 16S rRNA gene variant 2 in the peripheral blood.

Conclusion

Organisms were available from blood more or less constantly during the persistent infection and there were individual differences in cyclic activity of A. phagocytophilum in the infected animals. Two 16S rRNA gene variants of A. phagocytophilum show differences in cyclic activity during persistent infection in lambs.     

Congenital infection with Anaplasma phagocytophilum in a calf in northern Germany

Anaplasma phagocytophilum is a Gram-negative, obligate intracellular tick-transmitted bacterium that replicates in neutrophils. It causes tick-borne fever (TBF) in sheep and cattle, but also elicits febrile disease in humans as well as in other domestic animals such as dogs, horses, and cats. Although increasingly recognized in Europe, the first laboratory-confirmed case of TBF in cattle from Germany has been published only recently. We here present the unusual case of an intrauterine transmission of A. phagocytophilum in a calf from northern Germany. To our knowledge, this is the first report of such an event occurring under field conditions in cattle.     

Molecular and serological evidence for Anaplasma platys and Babesia sp. infection in a dog, imported in Belgium, from Southern Spain

This case report describes a dog suffering from a co-infection with Babesia and Anaplasma parasites. Anaplasma platys was found to be responsible for the anaplasmosis by molecular biology techniques, while microscopical and serological evidence was found for a coexistent babesiosis, although this could not be confirmed by polymerase chain reaction. Moreover, the possible risk of import of exotic pathogens is highlighted.     

Anaplasmosis in Uganda. III. Parasitological and serological evidence of Anaplasma infection in Ugandan goats.

Randomly selected goat sera from north-western, central, and south-western regions of Uganda were analyzed parasitologically and serologically for evidence of anaplasmosis. Prevalence rates of 3.2% by parasitemia, 4.8% by card-agglutination test, and 12.9% by DOT-ELISA combined with western blotting were established. Parasitologically positive samples were consistently serologically positive. Positive samples were all from either the north-western or south-western regions of the country. Goats in these regions graze with cattle and are presumable exposed to the same tick species. There was no evidence of clinical caprine anaplasmosis, whereas bovine anaplasmosis cases are very common. Rhipicephalus evertsi was frequently observed on goats which cograze with cattle.     

Anaplasma phagocytophilum infection in a domestic cat in Finland: Case report

Background

Anaplasmosis is a vectorborne disease caused by the gram-negative bacterium Anaplasma phagocytophilum. This species displays positive tropism to granulocytes and can cause illness in several mammalian species, including cats, dogs, and humans. It is considered as an emerging disease in Europe. The clinical signs are nonspecific and include fever, lethargy, and inappetence. The most typical hematologic abnormality is thrombocytopenia. A tentative diagnosis can be made by detecting intracytoplasmic morulae inside neutrophils. The diagnosis is confirmed by PCR and serology in paired serum samples. A sample for PCR analysis should be taken before treatment. Anaplasmosis is treated with doxycycline.

Case presentation

A feline case of anaplasmosis is presented. The history, clinical presentation, diagnostics, treatment, and follow-up are discussed.

Conclusions

This case indicates that Anaplasma phagocytophilum infects cats in Finland. To provide accurate treatment, anaplasmosis should be listed as a differential diagnosis in cats suffering from acute febrile illness with previous tick exposure.     

Evaluation of sequential coinfection with Anaplasma phagocytophilum and Anaplasma marginale in cattle

OBJECTIVE: To determine whether sequelae of infection differed among single versus double infection with Anaplasma phagocytophilum or Anaplasma marginale, with and without tick salivary extract, in cattle. ANIMALS: Eighteen 13-month old steers. PROCEDURES: Treatment groups of 3 cattle each included A marginale inoculated ID followed on day 35 by A phagocytophilum without tick saliva, A phagocytophilum followed on day 10 by A marginale without tick saliva, A marginale followed on day 35 by A phagocytophilum with tick saliva, A phagocytophilum followed on day 10 by A marginale with tick saliva, tissue culture control injection, and tick saliva control injection. Infection was monitored via clinical observations, CBC, serologic testing, and PCR analysis of blood and tissues. RESULTS: Infected cattle had significantly reduced weight gain. Anemia occurred 25 to 32 days after A marginale infection, which was attenuated by tick saliva. Parasitism was greater if cattle had not previously been inoculated with A phagocytophilum. Nine of the 12 treated cattle had positive results of PCR analysis for A phagocytophilum from at least 1 blood sample. Five tissue samples had positive results of PCR analysis for A phagocytophilum; PCR results for A marginale were positive in spleen, lung, lymph node, heart, and ear skin of infected cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated an important biological interaction between A marginale and A phagocytophilum infection as well as with tick saliva in disease kinetics and severity in cattle, which may be important for interpretation of diagnostic tests and management of disease in areas where both pathogens occur.     

Superinfection occurs in Anaplasma phagocytophilum infected sheep irrespective of infection phase and protection status

Background

Anaplasma phagocytophilum infection in domestic ruminants is widespread in the coastal areas of southern Norway. The bacteria may persist in mammalian hosts. Several genetic variants of A. phagocytophilum exist. In the present study, we investigate whether superinfection occurs in the acute and persistent phase of the infection.

Methods

Five-month-old lambs of the Norwegian Dala breed were experimentally infected with two 16S rRNA gene variants of A. phagocytophilum, i.e. A. phagocytophilum variant 1 (GenBank accession number {"type":"entrez-nucleotide","attrs":{"text":"M73220","term_id":"148293","term_text":"M73220"}}M73220) and variant 2 (GenBank acc. no. {"type":"entrez-nucleotide","attrs":{"text":"AF336220","term_id":"13325085","term_text":"AF336220"}}AF336220). Eighteen lambs were used, two lambs in each group. Eight groups were experimentally inoculated with either variant 1 or 2 on day 0. Six of these groups were then challenged with the other variant on either days 7, 42 or 84, respectively. One group was left uninfected. The occurrence of A. phagocytophilum in blood samples was determined using semi-nested PCR analysis and gene sequencing. Specific antibodies were measured by an indirect immunofluorescence antibody assay (IFA).

Results

A. phagocytophilum variant 1 and 2 differed significantly with regards to clinical reaction and cross-immunity in infected lambs. Both variants were found in the blood after challenge. However, variant 1 was detected most frequently.

Conclusion

The present experiment indicates that superinfection of different genotypes occurs during the acute as well as the persistent phase of an A. phagocytophilum infection, even in lambs protected against the challenged infection.     

Anaplasma phagocytophilum infection (granulocytic anaplasmosis) in a dog from Vancouver Island

A 7-year-old Labrador retriever had nonspecific clinical signs that included lethargy, malaise, and difficult ambulation. The dog was native to Vancouver Island, British Columbia, and had never left this area. Morulae were identified in polymorphonuclear cells. Serologic studies and polymerase chain reaction (PCR) testing confirmed canine anaplasmosis caused by Anaplasma phagocytophilum. The dog recovered after treatment with tetracycline.     

Immunohistological and serological investigations of morbillivirus infection in Black Sea harbour porpoises (Phocoena phocoena)

In the present study the occurrence of morbillivirus infection in harbour porpoises (Phocoena phocoena) from the Black Sea was investigated. Blood and tissue specimens of lung, brain and spleen from 73 stranded or by-caught harbour porpoises derived from the three Black Sea subregions such as Bulgaria, Georgia and Ukraine were collected between 1997 and 1999 and processed for histology, immunohistochemistry and serology. Age determination was performed according to dental growth layers and body length. The age of the investigated population ranged from neonates to a 12-year-old animal. Morbillivirus-specific neutralizing antibodies were detected in 53% of harbour porpoises. Generally, titres were very low and ranged from 20 to 270. There was no correlation between age, geographical origin and titre levels. The most common histological finding (97%) consisted of a mild to severe granulomatous bronchopneumonia due to lung worm infection. There were no changes indicative of a morbillivirus infection. Using immunohistology none of the animals were positive for morbillivirus antigen. However, the serological data are suggestive of a continuously circulating morbillivirus among harbour porpoises from the Black Sea indicating that harbour porpoises may serve as carriers for fatal diseases in susceptible cetacean species.     

Granulocytic anaplasmosis in a horse from Nova Scotia caused by infection with Anaplasma phagocytophilum

A 16-year-old Paint stallion was presented with intermittent fever, inappetance, lethargy, icterus, distal limb edema, and submandibular lymphadenopathy. The horse was native to Nova Scotia and had never left that province. Morulae were detected in granulocytes. Ananaplasma phagocytophilum infection was confirmed by serology and polymerase chain reaction (PCR). The horse responded to treatment with oxytetracycline.