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PubMed2003

Causes of febrile illnesses after a tick bite in Slovenian children.

BACKGROUND: To establish the etiology in Slovenian children with febrile illnesses occurring after a tick bite. METHODS: Eighty-six febrile patients younger than 15 years referred to our institution in 2001 with a history of a tick bite within 6 weeks before onset of the illness were included in this prospective study. Acute and convalescent serum samples were tested for the presence of antibodies to tick-borne encephalitis virus, Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Ehrlichia chaffeensis, Rickettsia conorii, Babesia microti, Bartonella henselae, Bartonella quintana and Francisella tularensis. Cerebrospinal fluid was investigated in patients in whom meningeal involvement was clinically suspected. Blood and/or cerebrospinal fluid from the patients were cultured in modified Kelly-Pettenkofer medium. PCR was performed to detect ribosomal DNA of A. phagocytophilum and E. chaffeensis. RESULTS: Of 86 patients 33 (38%) were excluded because a well-defined febrile illness not associated with tick bite was established. Tick-borne illness was diagnosed in 28 (53%) of the 53 remaining patients. The most common diagnosis was tick-borne encephalitis (64%), followed by Lyme borreliosis (46%), human monocytic ehrlichiosis and human granulocytic ehrlichiosis (serologic evidence of infection in 9 and 4%, respectively). In 6 (21%) patients there was evidence for infection with more than 1 tick-borne agent. CONCLUSIONS: Tick-borne illness was established in 53% of the patients younger than 15 years presenting with febrile illness occurring within 6 weeks after a tick bite. The most common identified illnesses were tick-borne encephalitis and Lyme borreliosis.

Adolescent↗

PubMed1995

16S rRNA gene sequence of Neorickettsia helminthoeca and its phylogenetic alignment with members of the genus Ehrlichia.

Neorickettsia helminthoeca (tribe Ehrlichieae, family Rickettsiaceae) is the agent of salmon poisoning disease, which affects members of the family Canidae. This bacterium is unusual in that it is the only known obligately intracellular bacterium that is transmitted via a helminth vector. The nucleotide sequence of the N. helminthoeca 16S rRNA gene was determined and compared with the sequences of intracellular bacteria belonging to the alpha subgroup of the Proteobacteria. The N. helminthoeca sequence was most similar to the sequences of two Ehrlichia species, Ehrlichia risticii and Ehrlichia sennetsu (levels of sequence similarity, > 95%). All other members of the tribe Ehrlichieae, including members of the other Ehrlichia species, and the related species Cowdria ruminantium and Anaplasma marginale, were only distantly related phylogenetically (levels of sequence similarity, 84 to 86%). Our results corroborate the results of previous ultrastructural and Western blot (immunoblot) comparisons of N. helminthoeca with other ehrlichial species. The genus Ehrlichia is phylogenetically incoherent and can be separated into three identifiable clusters of species. Each cluster is closely associated with a species classified in another non-Ehrlichia bacterial genus. The close relationships among N. helminthoeca, E. risticii, and E. sennetsu and the striking differences between these organisms and other members of the tribe Ehrlichieae suggest that in the future, these organisms should be treated as members of a new bacterial genus separate from the genus Ehrlichia.

Animals↗

PubMed1998

Ultrastructural differentiation of the genogroups in the genus Ehrlichia.

Ultrastructural characteristics of 15 strains and isolates of ehrlichiae belonging to three genogroups, or clades of genetically related organisms united in the genera Ehrlichia, Cowdria, Anaplasma, Neorickettsia and a strain of Wolbachia pipientis which represents a fourth genogroup in this cluster of species, were studied in continuous cell culture or in vivo: E. canis (Oklahoma strain and VHE isolate), E. muris (AS 145), E. chaffeensis (Arkansas, 91HE17 and Sapulpa), human granulocytic ehrlichiae (HGE)(BDS, 96HE27, 96HE37, #54, #55 and #72), E. equi (MRK), E. sennetsu (Miyayama), E. risticii (HRC-IL). Wolbachia pipientis was studied in the naturally infected Aedes albopictus mosquito cell line Aa23. All organisms were similar in the normal ultrastructure of individual cells and in the ability to form abnormal, pathological ehrlichial cells of the same type irrespective of the species. Normally all ehrlichiae studied in cell culture existed in two morphological forms - reticulate and dense-cored cells, both of which could divide by binary fission. Most alterations were related to their membranes, especially the cell wall. Differences in the structure of intravacuolar microcolonies (morulae) of ehrlichiae and their inter-relations with the host cells allowed differentiation of the genogroups: the E. canis-E. chaffeensis-E. muris genogroup formed large morulae, with many ehrlichiae, often suspended in a fibrillar matrix, and the host cell mitochondria and endoplasmic reticulum usually aggregated near the morulae and were in contact with the morula membrane; the E. phagocytophila-E. equi-HGE group morulae had no fibrillar matrix, no contacts with host cell mitochodria, and they did not aggregate around the morulae; E. sennetsu-E. risticii group usually developed in small individual vacuoles that did not fuse with each other and divided along with the ehrlichiae.

Animals↗

PubMed2004

Ultrastructure and phylogenetic analysis of 'Candidatus Neoehrlichia mikurensis' in the family Anaplasmataceae, isolated from wild rats and found in Ixodes ovatus ticks.

A novel bacterium that infects laboratory rats was isolated from wild Rattus norvegicus rats in Japan. Transmission electron microscopy of the spleen tissue revealed small cocci surrounded by an inner membrane and a thin, rippled outer membrane in a membrane-bound inclusion within the cytoplasm of endothelial cells. Phylogenetic analysis of the 16S rRNA gene sequence of the bacterium found in R. norvegicus rats and Ixodes ovatus ticks in Japan revealed that the organism represents a novel clade in the family Anaplasmataceae, which includes the Schotti variant found in Ixodes ricinus ticks in the Netherlands and the Ehrlichia-like Rattus strain found in R. norvegicus rats from China. The novel clade was confirmed by phylogenetic analysis of groESL sequences found in R. norvegicus rats and Ixodes ovatus ticks in Japan. No serological cross-reactivity was detected between this bacterium and members of the genera Anaplasma, Ehrlichia or Neorickettsia in the family Anaplasmataceae. It is proposed that this new cluster of bacteria should be designated 'Candidatus Neoehrlichia mikurensis'.

Anaplasmataceae↗

PubMed1987

Monoclonal antibodies that distinguish Trypanosoma congolense, T. vivax and T. brucei.

Monoclonal antibodies (MoAbs) were derived against in-vitro-propagated procyclic forms of Trypanosoma congolense, T. vivax, T. brucei brucei and T.b. rhodesiense in order to identify antigens for use in immunodiagnosis of African trypanosomiasis. The antibodies have been tested against procyclic and bloodstream form trypanosomes of 13 T. congolense, six T. vivax six T.b. brucei, four T.b. rhodesiense, five T.b. gambiense and three T. simiae isolates from different geographical areas by indirect immunofluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). The MoAbs raised against T.b. brucei reacted with all the brucei group of trypanosomes but not with T. congolense, T. simiae or T. vivax. Likewise, MoAbs against T. congolense reacted with T. congolense and T. simiae but not with any of the other species, while those against T. vivax reacted with T. vivax only. The antigens recognized by these MoAbs were present in lysates of bloodstream trypanosomes as well as midgut (for T. congolense and T. brucei) and epimastigote forms from infected Glossina morsitans centralis. There was no reactivity of the MoAbs with Theileria parva, Anaplasma marginale, Babesia bigemina or Plasmodium falciparum. These antibodies and the antigens they recognize should, therefore, prove useful in the development of assay systems for immunodiagnosis of African trypanosomiasis.

Animals↗

PubMed1990

Identification of a Theileria mutans-specific antigen for use in an antibody and antigen detection ELISA.

Purified piroplasms of Theileria mutans were used to immunize BALB/c mice to generate monoclonal antibodies (MoAbs). The MoAbs recognized an antigen of a relative molecular mass of 32 kDa in Western blots. This antigen was also recognized by sera from cattle which had recovered naturally from experimental tick-transmission or infections induced by the blood stages of T. mutans. The MoAbs did not react, in indirect immunofluorescence or enzyme-linked immunosorbent assays (ELISA), with the common haemoparasites of cattle, namely, T. parva, T. annulata, Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma congolense, T. vivax or T. brucei. An antigen capture ELISA was established with two of the MoAbs which recognized different epitopes on the 32 kDa molecule. Using this test it was possible to detect circulating antigens or immune complexes in sera collected from cattle during the acute or chronic phases of infection. When the purified 32 kDa protein was used as antigen in a micro-ELISA to detect circulating antibodies in both experimental and field cattle sera, it was found that the titres of antibodies ranged between 1:20 and 1:10,240. Results of this study indicate that the antigen and immune complex capture assays and the antibody detection ELISA can be complementary in the immunodiagnosis of acute and chronic T. mutans infections. Moreover, the tests are useful in the differential diagnosis of the disease and for epidemiological studies.

Animals↗

PubMedDate not supplied

Tick-borne encephalitis virus as a possible cause of optic neuritis in a dog.

A 3-year-old spayed female Siberian Husky was presented due to acute vision loss. Examination revealed bilateral optic neuritis and lymphocytic meningoencephalitis. The serum (1:800) and cerebrospinal fluid (CSF; 1:200) immunoglobulin (Ig)G titers for tick-borne encephalitis virus (TBEV) were elevated as were the serum IgG titer for Anaplasma phagocytophilum (1:640) and serum IgM titer for Toxoplasma gondii (1:20). Intracytoplasmic inclusion bodies such as ehrlichial or anaplasmal morulae were not observed in the CSF or blood smear. The dog was treated with methylprednisone and doxycycline. The left eye regained vision; the right eye remained blind. Anti-inflammatory therapy was stopped on day 18 after diagnosis. Four days later the dog showed evidence of hyperesthesia in the cervical region. Analysis of CSF showed no abnormalities and CSF IgG titers for TBEV and A. phagocytophilum were negative. Funduscopic evidence of active papillitis was absent on day 22 in the left eye and on day 86 in the right eye. On day 243, the dog was presented again with lethargy, ataxia, disorientation and temporary head tilt. The IgG titer for TBEV was again elevated in the CSF (1:800) and in serum (1:400). After interpretation of all findings, we assume that meningoencephalitis and optic neuritis in this patient was caused by TBEV and associated immune-mediated inflammation. In endemic areas, TBEV should be considered as cause of optic neuritis in dogs.

Animals↗

PubMed1994

Taxonomic position of the rickettsiae: current knowledge.

The term rickettsiae initially encompassed all intracellular bacteria. Early rickettsial taxonomy was based on a comparison of a few phenotypic characteristics and recently, molecular studies brought new bases for rickettsial taxonomy. All rickettsial species studied so far belong to the alpha and gamma groups of the Proteobacteria. Ehrlichiae complex groups Cowdria ruminantium, Anaplasma marginale and Wolbachia pipientis and the related parthenogenesis and cytoplasmic incompatibility bacteria, whereas Rochalimaea species group with Bartonella bacilliformis. Rickettsia tsutsugamushi may form an independent lineage, whereas molecular data allow to regroup serologically defined typhus and spotted fever group rickettsiae. The true scale of Rickettsia and Coxiella genera remain to be determined.

Alphaproteobacteria↗

PubMed1996

Comparative sensitivity of two tests for the diagnosis of multiple hemoparasite infection of cattle.

The purpose of this study was to evaluate the efficacy of light microscopy (LM) examination of blood smears and a multiplex polymerase chain reaction (MPCR) assay, in terms of their ability to detect cattle experimentally infected with Babesia bovis, Babesia bigemina, and Anaplasma marginale. Blood samples were collected from 32 intact, 1-2 year old, Holstein bulls, previous to and after simultaneous inoculation of culture-derived or field isolates of B. bovis- and B. bigemina-infected erythrocytes. To establish the triple hemoparasite infection, 16 of the bulls were also inoculated with a calf-derived isolate of A. marginale. The results showed that both tests had 100% specificity. In contrast, the sensitivities of the MPCR assay against the LM test were 93.5% and 70.9%; 96.7% and 100%; and 93.8% and 93.8% for B. bovis, B. bigemina, and A. marginale infection, respectively. The advantages and disadvantages of the MPCR assay to differentially diagnose cattle with multiple hemoparasite infection are discussed.

Anaplasmosis↗

PubMed2000

Culture-derived parasites in vaccination of cattle against tick-borne diseases.

The major economically important tick-borne diseases of cattle are theileriosis, babesiosis, anaplasmosis, and cowdriosis. Culture-derived attenuated schizonts of Theileria annulata have proved to be safe for all types of cattle and they protect against tick-borne theileriosis. T. parva was also successfully grown in vitro; however, inoculation of cattle with allogeneic schizont-infected cells resulted in rejection and destruction of the parasites together with the host cells. The number of schizont-infected cells needed for immunization is greater than for T. annulata theileriosis. Culture-propagated Babesia bovis and B. bigemina were used for large scale vaccination in the field. An avirulent population of Babesia spp. was obtained by in vitro cloning; inoculation of cattle did not induce clinical babesiosis, but produced specific antibodies. Culture-derived exoantigens of Babesia spp. proved to be completely safe for cattle, however, they conferred less protection than live parasites. Cell-cultured Cowdria ruminantium was highly infective for susceptible animals but, attenuated in vitro, could offer a potential source for vaccination. Anaplasma marginale, successfully grown in tick cell culture, may be developed for vaccines. Factors that should be considered in the developing of vaccines against tick-borne diseases include: the protective immune response to the pathogenic parasite developmental stages, virulence, immunological strain differences, and antigenic variations in cattle and in culture.

Anaplasmosis↗

PubMed2000

Integrated molecular diagnosis of Theileria and Babesia species of cattle in Italy.

A reverse line blot hybridization (RLB) test was developed to specifically identify six Theileria spp. (T. annulata, T. parva, T. mutans, T. velifera, T. taurotragi, and T. buffeli/orientalis) and three Babesia spp. (B. bovis, B. bigemina, and B. divergens). No cross reaction was observed with other livestock pathogens (such as Anaplasma marginale, A. centrale, A. ovis, Cowdria ruminantium, Trypanosoma brucei, T. congolense, and T. vivax). This method was used to test bovine blood samples collected in Sicily in April and November, 1998. Preliminary results indicated that T. annulata and T. buffeli/orientalis were the main species observed in cattle blood. Babesia species represented 1.8% and 23.5% in April and November, respectively.

Animals↗

PubMed2003

Ehrlichiae and ehrlichial diseases in china.

The various ticks collected from different areas of China were examined for the existence of ehrlichial agents by polymerase chain reaction (PCR) with genus- or species-specific primers designed on the basis of ehrlichial 16S rRNA genes and sequence analyses. In southern China, E. chaffeensis was detected in Amblyomma testudinarium ticks from infested cattle, Haemaphysalis yeni ticks from hare, and Ixodes ovatus ticks from Muntiacus reevesi. E. canis was identified in Rhipicephalus sanguineus ticks from dogs and Boophilus microplus ticks from goats. A new species of the genus Ehrlichia, closely related to E. chaffeensis, and Anaplasma marginale were found in B. microplus ticks from cattle in Tibet. In northern China, E. chaffeensis was detected in Dermacentor silvarum and I. persulcatus ticks; the granulocytic ehrlichial agents were detected in I. persulcatus ticks from an area where Lyme disease is endemic. Canine ehrlichiosis was found in southern China and E. canis and E. platys were identified in dogs; human ehrlichioses were demonstrated by amplifying the 16S rRNA genes of E. chaffeensis and granulocytic ehrlichial agents from patients' blood specimens. In comparison of 16S rRNA gene sequences, the sequences of E. chaffeensis, E. canis, and E. platys in China were found to be different from that in other countries at certain nucleotide positions. These results reveal that a variety of tick-borne ehrlichial agents and diseases exist in China, and the ehrlichial agents and their tick-vectors are same as or different from that in other countries at species or strain levels.

Animals↗

PubMed1976

Vaccination with Babesia argentina in 5 beef herds in south-eastern Queensland.

Observations on the use of a vaccine containing Babesia argentina in 5 partly susceptible beef herds in south-eastern Queensland were made on 1,029 female breeding cattle over a period of 4 years. Groups averaging about 20 heifers were given 0, 1, 2 or 3 vaccinations. Incidence derived from groups experiencing clinical attacks were 17.9% for unvaccinated cattle and 1.2% for vaccinates. Increasing the number of vaccinations did not appear to increase protection. The one clinical manifestation of infection with B. bigemina was associated with a concurrent reaction to vaccination with Anaplasma centrale. There were no cases of haemolytic anaemia in new-born calves. No severe reactions followed primary vaccination, but 2 revaccinated animals became sick.

Anaplasmosis↗

PubMed1978

Some effects of reduced energy intake on the development of anaplasmosis in Bos indicus cross steers.

Some effects of the plane of nutrition on the development of anaplasmosis in Brahman cross steers were investigated. Batches of 39 and 30 Brahman cross steers, aged approximately 27 months were each divided by stratified randomisation into 4 groups of similar mean PCV and body weight. Two similar experiments, designated A and B were conducted. Groups 1 and 2 were fed a ration of lucerne chaff at the rate of 1 M Cal ME/80 kg live weight/day for 8 weeks aimed to reduce body weight by approximately 5 kg/week. Animals in groups 3 and 4 were fed a ration for the same period aimed to increase body weight by approximately 2 kg/week. Groups 1 and 3 were then inoculated with approximately 10(10) Anaplasma marginale infected erythrocytes and the effects of the subsequent infections during the clinical and recovery phases were examined by measuring humoral antibody response, packed cell volume, parasitaemia and body weight. Groups 2 and 4 were uninfected controls. Anaplasmosis, as measured by three responses, was less severe in the starved animals of group 1. Significant differences in packed cell volume and parasitaemia were detected for short periods between the infected groups 1 and 3. Anaplasmosis caused losses of 6.2% and 5.9% in the mean body weight of group 3 animals in experiments A and B respectively. Most of this loss occurred during the clinical phase of the disease. The disease caused no apparent loss of weight in the infected animals of group 1.

Anaplasmosis↗

PubMed1980

A comparative study of experimental anaplasmosis in Bos indicus and Bos taurus cattle.

Comparative studies were carried out on the susceptibility of 8 Bos taurus and 10 predominantly Bos indicus cross cows aged 48 months to Anaplasma marginale infection. Measurements were carried out on bodyweight, parasitaemia, haematological factors, humoral antibody response and serum transaminase levels. Except for minor differences in reaction Bos indicus and Bos taurus animals were equally susceptible to infecton with the strain of A. marginale employed. Such differences observed were not significant.

Anaplasmosis↗

PubMed1980

Epidemiology of bovine anaplasmosis in beef calves in northern Queensland.

Natural infection of calves with Anaplasma marginale was studied in two endemic areas of tropical north Queensland. Infection, assessed by a micro-complement fixation test, occurred throughout the year but was most frequent just after the summer wet season. Infection was clinically inapparent. Calves vaccinated with A. centrale did not show improved growth rate when compared with non-vaccinated calves. The complement fixation test was shown to be most effective in the detection of recent infections.

Anaplasmosis↗

PubMed1982

Serological studies of babesiosis and anaplasmosis of cattle.

Bos indicus-crossbred calves exposed to natural Babesia bovis infection in wet and dry tropical environments in northern Queensland were tested for antibodies using the indirect haemagglutination (IHA) test. There was evidence of maternal antibodies suggesting endemicity of B. bovis in the 2 environments. Within one year 35.3 to 94.6% of the calves had acquired B. bovis infection. The highest titres and most rapid conversion rates were observed just after the wet season when the tick population was at its peak. Clinical disease was not observed. A third group of Bos indicus-cross cattle of varying ages in a dry tropical environment of north Queensland was examined for antibodies against B. bovis and Anaplasma marginale using the IHA and complement fixation (CF) tests, respectively. B. bovis and A. marginale antibodies were detected on the farm but clinical disease did not occur.

Anaplasmosis↗

PubMed2005

Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil.

BACKGROUND: Infection with Ehrlichia canis causes a highly variable, multisystemic disease in dogs. Nevertheless, many clinicians in Rio de Janeiro, Brazil, use the presence of only thrombocytopenia to make a presumptive diagnosis of E canis infection. OBJECTIVE: The objective of this study was to determine the prevalence of E canis in thrombocytopenic dogs from Rio de Janeiro, Brazil, using polymerase chain reaction (PCR). METHODS: Following DNA extraction of whole blood samples from 226 dogs, PCR assays were done using primers for rickettsial DNA (including Ehrlichia spp, Anaplasma platys and A phagocytophilum) and using E canis-specific primers (16S rRNA gene). Dogs were grouped as thrombocytopenic and nonthrombocytopenic based on platelet counts. The null hypothesis that there was no difference in the prevalence of E canis in these groups was rejected at P<.05. RESULTS: Thirty-six (32.1%) of the thrombocytopenic dogs and 4 (3.5%) of the nonthrombocytopenic dogs were positive for rickettsial gene sequences (P<.0001). Further, 30 (26.8%) of thrombocytopenic dogs and 4 (3.5%) nonthrombocytopenic dogs were positive for E canis-specific gene sequences (P<.0001). CONCLUSIONS: Although the prevalence of E canis infection was higher in thrombocytopenic dogs, less than one third of these dogs had demonstrable E canis infection. Thus, thrombocytopenia is not specific for the detection of E canis infection and should not be used solely to establish a diagnosis of canine ehrlichiosis, even in a geographic area with relatively high disease prevalence.

Animals↗