OBJECTIVE To identify clinical or clinicopathologic variables that can be used to predict a positive PCR assay result for Anaplasma phagocytophilum infection in equids. ANIMALS 162 equids. PROCEDURES Medical records were reviewed to identify equids that underwent testing for evidence of A phagocytophilum infection by PCR assay between June 1, 2007, and December 31, 2015. For each equid that tested positive (case equid), 2 time-matched equids that tested negative for the organism (control equids) were identified. Data collected included age, sex, breed, geographic location (residence at the time of testing), physical examination findings, and CBC and plasma biochemical analysis results. Potential predictor variables were analyzed by stepwise logistic regression followed by classification and regression tree analysis. Generalized additive models were used to evaluate identified predictors of a positive test result for A phagocytophilum. RESULTS Total lymphocyte count, plasma total bilirubin concentration, plasma sodium concentration, and geographic latitude were linear predictors of a positive PCR assay result for A phagocytophilum. Plasma creatine kinase activity was a nonlinear predictor of a positive result. CONCLUSIONS AND CLINICAL RELEVANCE Assessment of predictors identified in this study may help veterinarians identify equids that could benefit from early treatment for anaplasmosis while definitive test results are pending. This information may also help to prevent unnecessary administration of oxytetracycline to equids that are unlikely to test positive for the disease.

Ranked among the top threats to conservation worldwide, infectious disease is of particular concern for wild canids because domestic dogs (Canis familiaris) may serve as sources and reservoirs of infection. On British Columbia’s largely undeveloped but rapidly changing central and north coasts, little is known about diseases in wolves (Canis lupus) or other wildlife. However, several threats exist for transfer of diseases among unvaccinated dogs and wolves. To gain baseline data on infectious agents in this area, including those with zoonotic potential, we collected blood and stool samples from 107 dogs in 5 remote communities in May and September 2007. Serology revealed that the dogs had been exposed to canine parvovirus, canine distemper virus, Bordetella bronchiseptica, canine respiratory coronavirus, and Leptospira interrogans. No dogs showed evidence of exposure to Ehrlichia canis, Anaplasma phagocytophilum, Borrelia burgdorferi, Dirofilaria immitis, or Cryptococcus gattii. Of 75 stool samples, 31 contained at least 1 parasitic infection, including Taeniid tapeworms, the nematodes Toxocara canis and Toxascaris leonina, and the protozoans Isospora sp., Giardia sp., Cryptosporidium sp., and Sarcocystis sp. This work provides a sound baseline for future monitoring of infectious agents that could affect dogs, sympatric wild canids, other wildlife, and humans.

The present study was conducted to molecular detection of Anaplasma phagocytophilum in infected cows and it is infested ticks in Mosul, Iraq using nested polymerase chain reaction technique (N-PCR) and to investigate the phylogenetic analysis of A. phagocytophilum diagnosed in this study. A total of 50 blood samples (3 ml) were collected from cows in various areas of Mosul city. Additionally, 169 hard ticks were collected from different parts of infected cows. Results revealed that the infection rate of A. phagocytophilum in cows at Mosul city was 72% and in engorged female ticks was 81.2% using N-PCR technique. The infestation rate hard ticks on cows were 46% and four species of hard ticks were microscopically identified and classified includes Hyalomma anatolicum anatolicum (39.64%), Rhipicephalus turanicus (21.89%), Rh. sanguineus (23.07%) and Boophillus annulatus (15.38%) with significantly predominant Hyalomma anatolicum anatolicum. The individual sequencing analysis for five sequences of 16S rRNA gene includes: one extracted from cow blood and four extracted from engorgement female ticks. The sequences of A. phagocytophilum were recorded in the NCBI Genbank under the accession numbers (OR002120.1, OR002121.1, OR002122.1, OR002123.1 and OR002124.1). These sequences were highly identity (100%) to those sequences recorded in the NCBI Genbank such as (MT221233.1, MT221234.1) in Norway, (MK239930.1, MK239931.1) in Southern Korea, (MN170722.1, MN170722.1) in Spain, (MK814411.1, MK814412.1) in South Africa, (LC435049.1, LC435050.1) in Japan and (MH122889.1, OQ727069.1) in Poland, and Malawi respectively. This finding might be advantageous for the future studies and strategically control of this bacterium in the study area.

OBJECTIVE To compare the performance of 5 synthetic peptide–based ELISAs with that of 3 commercially available immunofluorescent assays (IFAs) for serologic diagnosis of anaplasmosis and ehrlichiosis in dogs. SAMPLE A convenience set of 109 serum samples obtained before and at various times after inoculation for 23 dogs that were experimentally infected with Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Ehrlichia chaffeensis, or Ehrlichia ewingii and 1 uninfected control dog in previous studies. PROCEDURES All serum samples were assessed with 5 synthetic peptide–based ELISAs designed to detect antibodies against A phagocytophilum, A platys, E canis, E chaffeensis, and E ewingii and 3 whole organism–based IFAs designed to detect antibodies against A phagocytophilum, E canis, and E chaffeensis. The species-specific seroreactivity, cross-reactivity with the other tick-borne pathogens (TBPs), and diagnostic sensitivity and specificity were calculated for each assay and compared among assays. RESULTS All serum samples obtained from dogs experimentally infected with a TBP yielded positive results on a serologic assay specific for that pathogen. In general, sensitivity was comparable between ELISAs and IFAs and tended to increase with duration after inoculation. Compared with the IFAs, the corresponding ELISAs were highly specific and rarely cross-reacted with antibodies against other TBPs. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that peptide-based ELISAs had enhanced specificity relative to whole organism–based IFAs for detection of antibodies against Anaplasma and Ehrlichia spp, which should facilitate accurate diagnosis and may help detect dogs coinfected with multiple TBPs.

Human granulocytic anaplasmosis (HGA) is caused by the obligate intracellular bacterium Anaplasma (A.) phagocytophilm. Natural killer T (NKT) cells are key players in host defense against various microbial infections. We investigated the role of NKT cells in immune response to A. phagocytophilum infection using NKT-knockout (Jα18-/-) mice. Jα18-/- and wild-type (WT) mice were infected with low-passage A. phagocytophilum and assayed for hepatic histopathology and cytokine production during 7 days post-infection. Compared to WT control, the infected Jα18-/- mice had much less histopathologic lesions and less apoptosis through day 7, and lower concentrations of IFN-γ and IL-12, but not of IL-10. This result suggests that NKT cells are major components in the pathogenesis of HGA.

The antigenic profile of Ehrlichia canis, E risticii, E sennetsu, and E equi was investigated by the use of protein (western) immunoblot technique. Results of analysis of serum from acutely and chronically infected animals indicated that the 4 Ehrlichia species share a unique 25-kD polypeptide in addition to other peptides. Immune sera from dogs inoculated with E canis recognized a wide range of E canis polypeptide antigens, as determined by western blot analysis. A larger number of E sennetsu polypeptides were detected when homologous antiserum and antiserum to E equi were used. The latter antiserum did not recognize antigens of E canis or E risticii. Antisera to E canis, E risticii, and E sennetsu detected E equi antigens. Data indicate that a 25-kD protein is a common antigen among the species of the genus Ehrlichia and that the ascending order of abundance of immunodominant determinants in the 4 species of Ehrlichia studied would be: E risticii leads to E equi leads to E sennetsu leads to E canis. Implications of these findings for diagnosis of ehrlichial infections and prophylaxis are evident.

An intact female tabby Persian cat, 26-month-old, was brought to IPB University Veterinary Teaching Hospital on the 21st of February, 2023, due to vaginal discharge, inappetence, and lethargy. Physical examination revealed a distended abdomen, mammary gland enlargement, and fever (40.1 C). The heart and respiratory rates were 128 beats per minute and 32 times per minute, respectively. The patient body weight was 3.87 kg. The veterinarian palpated masses inside the abdominal cavity and observed obvious purulent and haemorrhagic vaginal discharge. Haematology showed leukopenia and thrombocytopenia, and the eosinophil percentage of white blood cells was increased (eosinophilia). Blood chemistry analysis showed hyperglycaemia. Ultrasonography showed two hyperechoic masses inside two hypoechoic pouches in the queens uterus, suspected as two dead foetuses (no heartbeat). Roentgen revealed abnormalities in the vena cava caudalis, bronchial pattern in the lung, radioopaque masses in the ventral abdomen, radio-opacity changes in the small and large intestines, and abnormalities in the spleen. The patients diagnosis was pyometra, and ovariohysterectomy surgery was performed on the patient. The patient was treated with additional medicines (antibiotics and others) according to the symptoms. The patients symptoms disappeared, except for the fluctuating febrile and pale mucous membrane. A blood smear performed on the 1st of March 2023 revealed Anaplasma phagocytophilum (A. phagocytophilum) in the white blood cells, and immediately doxycycline treatment (10 mg/kg body weight, twice daily) was given. The patient was discharged from the hospital on the 14th of March, 2023, with no febrile or other symptoms reported. This case is the first report of open cervix-type pyometra and anaplasmosis co-occurrence in a Persian cat from IPB University Veterinary Teaching Hospital in Indonesia.