A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for Anaplasma/Ehrlichia in Roaming Dogs and Symptomatic Dogs in Trinidad

This study compared two methods to detect cases of canine ehrlichiosis in a field setting. One method was a polymerase chain reaction for the 16S rRNA gene followed by reverse line blot hybridisation with genera and species-specific probes for Anaplasma/Ehrlichia. The second method was an autologous cell culture of peripheral leucocytes isolated from heparinised blood and maintained in a homologous canine serum in Dulbecco’s Modified Eagle medium without antibiotics. The cultures were examined under light microscopy for inclusion bodies after 48 h. Leucocytes were successfully propagated for 20 of the 34 samples submitted for autologous cell culture. Inclusion bodies were observed after cell culture in leucocytes of eight dogs. Two dogs were positive to the Anaplasma/Ehrlichia genera probe and six dogs were positive to the E. canis probe after reverse line blot hybridisation. There was acceptable agreement between reverse line blot hybridisation and cell culture results. Both reverse line blot hybridisation and autologous cell cultures can be used to detect E. canis in subclinical and clinical cases of disease. A definitive diagnosis of E. canis is best achieved by a combination of clinical signs, positive autologous cell culture, and reverse line blot hybridisation results.

Epidemiological status of erhrlichiosis as a high-risk zoonosis in endemic populations

This work was carried out at the animal health veterinary clinic, located on Cuauhtémoc avenue, Colonia La Rivera, in the city of Tuxpan de Rodríguez Cano, Veracruz, Mexico, during the period of October 2019 - March 2020. Aiming to determine the prevalence canine Ehrlichiosis, for which 30 canine E. canina serotypes were identified. For this evaluation, the AB GENOME kit was used, which was based on an indirect ELISA test in canines with apparent symptoms of the disease. The data were processed in the SPSS version 10 statistical package and the differences between treatments were detected by Duncan's test, the level of significance was 95% taking into account the variables of sex, age and race. The results indicated 23% of positive cases of the canines exposed to E. canis and 7 negative cases with a prevalence of 76.66%. According to the results obtained that were based on the association that existed between the presence of antibodies against E. canis, with age, race, sex, it was determined that there is no association between the presence of antibodies against E. canis and none of the variables described above.

Consequences of Canine Ehrlichiosis: Clinical Case Successfully Resolved

This work aims to report the case of a canine that survived erlichiosis with severe consequences of the disease, such as meningitis, hind limb paresis, circle walking, cerebrovascular accident. The onset of paresis was sudden, soon after a hygienic grooming. X-ray examinations ruled out suspected trauma. A cerebrovascular accident is suspected due to vasculitis found by electrocardiogram examination. Traditional treatment for doxycycline-based erlichiosis (200 mg) 1 comp. 2 x day for 24 days was recommended; anti-inflammatory drug Prednisone - IDB for 15 days, antianemic Erythros - 1 comp./day, Leucogen (H) 5 mL (2 x day) immunomodulator for platelet elevation and a peripheral and cerebral vasodilator Revimax - 1 comp./ day were the drugs of choice. The case seems to suggest that the bacterium, through the hematogenous route, reaches the intervetbral discs, paralyzing the pelvic limbs, bladder, anal sphincter and tail, and may even cause neurological disorders and even lead to death of the animal. Since paresis and stroke, as well as hydrocephalus and meningitis, clinical signs were much more atypical and are currently becoming more frequent.

Successful Treatment and Management of Canine Ehrlichiosis-Leishmaniosis-Heartworm Comorbidity

Background: Canine vector borne diseases (CVBD) are common in tropical countries where the climate favors arthropods abundance. Comorbidity with one or more CVBD are reported as clinical complication and worsen prognostic. Canine visceral leishmaniosis (CanL) is an endemic zoonotic disease in Brazil caused by Leishmania infantum, with several restrictions to canine treatment and suggestion of reservoirs euthanasia for disease control. Heart worm (HW) is a helminthic disease caused by Dirofilaria immitis infection in dogs. It is a chronic heart disease, which can lead to death by congestive heart failure. Canine ehrlichiosis (CE) is caused by Ehrlichia canis bacterial infection with a zoonotic potential and fatal to dogs in acute and chronic presentations. Exposed the above, this study aims to describe a successful treatment and management of a dog with CanL, CE, and HW comorbidity. Case: A 3-year-old male uncastrated black Labrador dog, weighing 35 kg, was admitted to the veterinary clinic due to immunochromatographic CanL positive test performed by municipal zoonosis control center active surveillance in August 2014. Clinical exam showed a mild shedding, intermittent eye white/yellow discharge and popliteal lymph nodes enlargement. After positive for CanL, veterinary requested more laboratorial exams. IFAT and ELISA were positive for CanL, blood smear showed presence of microfilaria, and bone marrow cytology showed Ehrlichia spp. morulae and microfilaria. Initial treatment prescribed was oral doxycycline, omeprazole, ranitidine, and domperidone for 30 days, and allopurinol and ivermectin until further recommendation. Additionally, repellent collar, repellent spray and vitamin supplement was indicated. After first month, marbofloxacin for 30 days and three doses of immune stimulant protocol were administrated. After three months of treatment, dog still positive for heartworm, ehrlichiosis, and CanL. Doxycycline protocol was repeated. Dog became consistently negative for all pathogens one year later with persistent thrombocytopenia but without clinical signs, ergo allopurinol and ivermectin were discontinued. After four years of follow up, the animal had an acute pancreatitis and died, with unremarkable total blood count and negative for all pathogens. Discussion: CVBD coinfections are commonly reported as worsen prognostic in endemic regions. The pathogens reported here share a host immunomodulation competence. L. infantum and Ehrlichia spp.downregulates Th1 response, whereas D. immitis increase as Th2 profile. The therapeutic protocol was iniciated by staging CanL. Since the patient had clinical signs, allopurinol was prescribed as a well-established drug for CanL. Marbofloxacin was added due to its high safety drug in clinical improvement of infected dogs with and without renal disease and in vitro effectiveness against L. infantum. Domperidone was used to promote Th1 cytokine profile as INF-γ, IL-2, IL-12, and TNF-α. We used an immunostimulant protocol to favor polarization to the Th1 profile comprised by 30 days of domperidone protocol followed by a vaccine and an immunomodulator. Doxycycline was used successfully for Ehrlichia spp. and HE clearance after two treatment courses and one year of ivermectin every 15 days. The animal presented intermittent coughing episodes on the first treatment course, but no medical intervention was needed besides exercise restriction. Our report shows the successful management of one dog with CanL, CE and HE comorbidity. This success was possible due to early detection and good therapeutic choice.Keywords: canine visceral leishmaniosis, coinfection, Dirofilaria immitis, Erhlichia canis, Leishmania infantum, treatment.

Relationship between ocular abnormalities and hematologic alterations in patients infected naturally by Ehrlichia canis

ABSTRACT: To determine the association between ocular abnormalities and hematologic findings in Ehrlichia canis-infected dogs, 120 dogs suspected of canine ehrlichiosis were studied. All patients were subjected to rapid serologic diagnostic test for Ehrlichia canis, and the diagnosis was confirmed by quantitative PCR. PCR-positive patients underwent ophthalmologic examination and a hemogram and were grouped into patients with thrombocytopenia and anemia (G1), patients with thrombocytopenia (G2), and patients without hematologic disorders (G3). For ophthalmic evaluation, the patients were grouped into those having mild, severe, both mild and severe, and no ocular alterations. All patients presented with uveitis (100%). Severe ocular disorders predominated in G1 and G2 patients. Dogs with anemia and leukopenia showed severe ocular disorders. In dogs with a greater number of leukocytes and copies/µl of the TRAG gene, the intraocular pressure (IOP) tends to decrease and vice versa (r = −0.23, r = −0.26). In conclusion, uveitis is the earliest ocular alteration of canine ehrlichiosis. The severity of ocular abnormalities in patients with ehrlichiosis is associated with thrombocytopenia and may be aggravated by anemia and leukopenia. The hemogram and IOP are useful tools for predicting eye involvement in Ehrlichia canis-infected patients. In regions where Ehrlichia canis is endemic, the use of the tonometer should be routinely in dog general consultation.

Canine Ehrlichiosis in Africa

Canine ehrlichiosis is an infection of canids causing a clinical disease in domestic dogs and asymptomatic infections in wild canids. Currently, the disease assumes a cosmopolitan distribution. This chapter summarises published information on the disease from across Africa. Some studies were able to demonstrate experimental infections in these canids. Different diagnostic methods, cell culture, direct microscopy, serology, hematology, and molecular methods were employed in different studies for detection of ehrlichiosis. Treatment of the disease mainly involved use of oxytetracycline, doxycycline, imidocarb disproportionate, and levamisole. In severe cases, management has involved administration of supportive therapy such as blood transfusion. Generally, though available, the information on different aspects of the disease in the Africa is scant and fragmented. There is still a need to generate more information on the epidemiology, diagnosis, clinical aspects, and treatment of the disease.

Anti-Ehrlichia properties of the dichloromethane fraction of Ageratum conyzoides associated with doxycycline: In vitro study

ABSTRACT: The increasing number of cases of canine ehrlichiosis caused by Ehrlichia canis in hospitals and veterinary clinics has demonstrated the need for a new drug protocol for this disease. Doxycycline is used to treat ehrlichiosis, but the resistance of the microorganism to this treatment protocol, as well as the various side effects to the animals, has become a concern. Several studies have shown a positive interaction with extracts of plants and drugs, which allow for the reduction of the concentration necessary to produce the desired effect, minimizing adverse effects. This study determined the efficiency of the combination of the dichloromethane (DCM) fraction of Ageratum conyzoides L. with anti-Ehrlichia activity and doxycycline by using the checkerboard assay. Plant material was collected in São Luís, northeastern Brazil, followed by extraction in MeOH: H2O (8:2) and partitioning of the DCM fraction. After determining the Minimum Inhibitory Concentration (MIC) of the fraction under study against DH82 cells infected with Ehrlichia canis, it was combined with doxycycline to derive the Fractional Inhibitory Concentration Index (CIF Index). A reduction of 5.83 times the doxycycline minimum inhibitory concentration was observed, showing that this fraction of A. conyzoides composed predominantly by the class of lignans, identified by mass spectrometry notably intensified the activity of doxycycline against E. canis, resulting in a synergistic effect.

Molecular diagnosis and risk factors of canine ehrlichiosis in the municipality of Itabuna-Bahia, Brazil

Ehrlichiosis is an emerging zoonosis worldwide and has had several adverse effects on public health. Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, has the tick Rhipicephalus sanguineus as the vector. The main clinical signs in affected dogs are fever, apathy, anorexia, weight loss, and neurological signs. The diagnosis is made through the association of clinical signs with parasitological, serological, and molecular tests. The aim of this study was to evaluate the occurrence of E. canis infection in dogs from the city of Itabuna-Bahia, as well as to identify the risk factors related to infection. For this, 405 dogs from the Center for Zoonoses Control (CCZ), non-governmental organizations (NGOs), and dogs domiciled and semi-domiciled in the city of Itabuna, southern Bahia, were evaluated. After initial physical evaluation of the dogs, blood samples were collected by venipuncture for subsequent DNA extraction and E. canis testing using the nested Polymerase Chain Reaction (nested-PCR) technique. In addition, an epidemiological questionnaire that included questions related to the animals was administered to the dog owners to identify the risk factors for exposure to the etiological agent and to the vector. Approximately 17% of the dogs in the municipality of Itabuna-Bahia tested positive for E. canis by nested-PCR, a result higher than that found in other studies conducted in the same municipality. Among the factors associated with E. canis infection, contact with other dogs (p = 0.0226) was an important factor for the dissemination of CME, since dogs are reported to be reservoirs of E. canis. Male dogs (p = 0.0016) presented lower risk for E. canis infection. Other studies, however, describe no association between animal gender and infection by E. canis. Preventive measures to reduce exposure to the vector of ehrlichiosis are necessary.

Feline ehrlichiosis – literature review

Ehrlichiosis is a tick-borne infectious disease caused by a bacterium belonging to the Anaplasmataceae family that can infect mammals. In Brazil, canine ehrlichiosis is caused by Ehrlichia canis, and it is endemic in some regions. The bacterium is transmitted by vectors, and especially the Rhipicephalus sanguineus. In cats the disease process is less understood due to scarce research, and lack of standardization of diagnostic tests. Lack of awareness of veterinarians about the importance of the disease in cats is also a contributing factor to scarce information. The available information suggests that the course of the disease in cats is similar to that of the dog. In this article we present a literature review on feline ehrlichiosis describing etiopathogenesis, methods of diagnostic, treatment, and prophylaxis strategies.