HALLAZGO DE MÓRULAS COMPATIBLES CON ANAPLASMA PLATYS EN UN CANINO DE LA CIUDAD DE ESPERANZA

Anaplasma platys is a Gram-negative, intracellular obligate bacterium that is transmitted by Rhipicephalus sanguineus ticks and has been identified in most of the country. It is the causative agent of canine infectious cyclic thrombocytopenia. The objective of this work is to report the finding of morulae compatible with A. platys in the blood smear of a 10-year-old male Weimaraner canine who was treated at the Hospital de Salud Animal of the Facultad de Ciencias Veterinarias of the Universidad Nacional del Litoral, in the city of Esperanza. The case could not be followed up, because the patient died a few days after his first consultation. The detection of obligate intracellular pathogens in blood smears has low sensitivity and specificity, therefore confirmation of the causative agent must be performed using PCR. Due to the severity of the condition presented in this case, it is necessary to differentiate the presence of Ehrliquia canis and confirm the finding of A. platys using PCR. Current scientific evidence reveals that the zoonotic potential of A. platys is very low or nil.

Vector-Borne Pathogens in Ticks and Fleas of Client-Owned Dogs in Metro Manila, Philippines

Rhipicephalus sanguineus s.l. and Ctenocephalides felis are considered the most prevalent ectoparasites of dogs in the Philippines. Vector-borne pathogens (VBPs) in these ectoparasites pose health risks to humans and animals. This study aimed to confirm the morphological and molecular identity of tick and flea species parasitising dogs in Metro Manila (Philippines) and molecularly investigate the possible presence of Bartonella spp., Rickettsia spp., Ehrlichia canis, and Anaplasma platys DNA. Ticks (n = 58) and fleas (n = 52) on dogs from three veterinary clinics in Metro Manila were collected and identified morphologically and molecularly via amplification and sequencing of cytochrome c oxidase I (cox1). Aliquots of ectoparasite DNA underwent real-time polymerase chain reaction (qPCR) screening for VBPs. All ticks were R. linnaei (formerly R. sanguineus s.l. “tropical lineage”), while all fleas were C. felis from clade 6 of the tropical II cluster/“Cairns” clade known from Australia. DNA of B. clarridgeiae was detected in 10% of fleas. DNA of R. felis was detected in 10% of fleas and in 3.8% of ticks. DNA of E. canis and A. platys was not detected. This study confirmed the presence of ticks and fleas as frequent ectoparasites on dogs and VBP presence emphasises the importance of preventative actions for animal health and welfare.

First report on molecular prevalence and identification of Anaplasma platys in dogs in Khon Kaen, Thailand

Background and Aim: Anaplasma platys is a blood parasite that infects platelets, causing thrombocytopenia. Rhipicephalus sanguineus ticks are believed to transmit A. platys. To identify A. platys, nested polymerase chain reaction (PCR) has proven to be an effective diagnostic tool. In this study, the molecular prevalence of A. platys infection in dogs was investigated for the 1st time in the Khon Kaen region of Thailand. The association between risk factors and A. platys infection was also evaluated. Materials and Methods: A total of 130 blood samples were collected from dogs in Khon Kaen, Thailand. DNA from the samples was extracted and nested PCR was applied for molecular analysis. Platelet count and packed cell volume (PCV) levels were measured. Platelet counts were categorized into four grades: Non-thrombocytopenia (platelets >200,000 cells/μL), mild thrombocytopenia (platelets 150,000-200,000 cells/μL), moderate thrombocytopenia (platelets 100,000-150,000 cells/μL), and severe thrombocytopenia (platelets <100,000 cells/μL). Four categories for PCV levels of >37%, 30-37%, 20-29%, and <20% were defined as no anemia, mild anemia, moderate anemia, and severe anemia, respectively. DNA sequencing was analyzed using BTSeq™ (Barcode-Tagged Sequencing; CELEMICS, Seoul, South Korea) for similarity index. Results: Among the 130 samples, 9 (6.9%) were positive for A. platys infection. There was an association between low platelet count and infection (p<0.05). PCV level was also associated with A. platys infection (p<0.05). DNA sequencing results of the nine positive samples showed similarity to known sequences of A. platys with 99.36-100% nucleotide identity. These results suggested low genetic diversity in A. platys infecting dogs in the Khon Kaen area. Conclusion: By amplifying 16S rRNA, A. platys infection was detected in the blood of Thai dogs. Further work should be performed to identify risk factors potentially associated with A. platys infection in dogs in Khon Kaen. Other related factors should also be considered, such as location and breeding, as well as the environmental characteristics of each locality. In addition, sampling a larger number of animals may reveal predictors for the positivity of A. platys in dogs in this region.

Morphological, molecular and MALDI-TOF MS identification of ticks and tick-associated pathogens in Vietnam

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been reported as a promising and reliable tool for arthropod identification, including the identification of alcohol-preserved ticks based on extracted leg protein spectra. In this study, the legs of 361 ticks collected in Vietnam, including 251 Rhiphicephalus sanguineus s.l, 99 Rhipicephalus (Boophilus) microplus, two Amblyomma varanensis, seven Dermacentor auratus, one Dermacentor compactus, and one Amblyomma sp. were submitted for MALDI-TOF MS analyses. Spectral analysis showed intra-species reproducibility and inter-species specificity and the spectra of 329 (91%) specimens were of excellent quality. The blind test of 310 spectra remaining after updating the database with 19 spectra revealed that all were correctly identified with log score values (LSV) ranging from 1.7 to 2.396 with a mean of 1.982 ± 0.142 and a median of 1.971. The DNA of several microorganisms including Anaplasma platys, Anaplasma phagocytophilum, Anaplasma marginale, Ehrlichia rustica, Babesia vogeli, Theileria sinensis, and Theileria orientalis were detected in 25 ticks. Co-infection by A. phagocytophilum and T. sinensis was found in one Rh. (B) microplus.

Clinical variations observed among the main hemoparasitosis caused by Rhipicephalus sanguineus in dogs

Hemoparasitosis are diseases of great importance in the practice of a small animal’s veterinarian, due to their high number of cases in the clinical routine of these species, and the wide occurrence of their vectors. Related to hemoparasitosis in dogs, the tick Rhipicephalus sanguineus has the greatest importance due to parasitizing dogs, perpetuating hemoparasitosis in this host group. The present work describes a literature review about the three main hemoparasitosis in the routine of a veterinarian in Brazil, ehrlichiosis, caused by Ehrlichia canis, babesiosis caused by the protozoa Babesia canis and Babesia gibsoni, and anaplasmosis caused by Anaplasma platys, comparing them regarding the differences between clinical signs and laboratory alterations, such as the different types of anemia caused by these conditions that culminate in clinical signs related to them. It also aims to describe the similarity in the diagnostic techniques used and infection treatments, which in all cases is used tetracycline, the most commonly used is doxycycline, and highlight the need for further studies about this topic.

HEMOPARASITOSE EM UMA CANINA- RELATO DE CASO

Introdução: A hemoparasitose é uma patologia de alta casuística na rotina clínica veterinária, desencadeada por bactérias, protozoários, helmintos, possuindo como principal transmissor o carrapato em períodos do ano de maior incidência de calor e umidade, tendo em vista o potencial zoonótico. A babesiose é causada pelos protozoários Babesia Canis e Babesia gibsoni transmitida pelo carrapato Rhipicephalus sanguineus, esses parasitas causam a hemólise das hemácias. Já a anaplasmose é oriunda de bactérias intracelulares Anaplasma platys ou Anaplasma phagocytophilum transmitidas pelo carrapato Rhipicephalus sanguineus, causando trombocitopenia. A rangeliose é ocasionada por um piroplasma Rangelia vitalli, transmitida por carrapatos Rhipicephalus sanguineus e Amblyomma aerolatum, se replicando nos eritrócitos. Objetivo: Relatar um caso clínico de uma canina. Relato de caso: canina castrada, sem raça definida, com 1 ano e dois meses, pesando 11,4 Kg, sendo atendida em uma clínica veterinária particular em Caxias do Sul/RS, apresentando apatia, hematoquezia, anorexia, prostração, epistaxe, sangramento em pontas de orelhas, mucosas hipocoradas com petéquias e febre. Durante o atendimento foi realizada coleta sanguínea para avaliação hematológica, esfregaço sanguíneo periférico da orelha, bioquímica sérica (creatinina, fosfatase alcalina, ureia, transaminase pirúvica) e urinálise, apresentando alterações, tais como anemia regenerativa, leucocitose por neutrofilia com desvio a esquerda, linfocitose, trombocitopenia, na urinálise houve presença de hemoglobina, leucócitos, proteinúria. No exame de esfregaço de sangue periférico apresentou-se negativo para hemoparasitas. O diagnóstico definitivo foi através da Reação em Cadeia da Polimerase (PCR), na qual evidenciou os agentes patogênicos de babesiose, rangeliose e anaplasmose. O tratamento instituído foi doxiciclina (5 mg/kg/q.12h durante 28 dias), azitromicina (10 mg/Kg/q.24h durante 10 dias), diaceturato de diminazeno (0,5ml/2Kg) repetindo em 14 dias a dose com prévia aplicação de atropina (0,044 mg/Kg), suplemento vitamínico mineral aminoácido (1 ml/q.24h). O animal se manteve bem clinicamente durante o tratamento, obtendo cura da enfermidade. Conclusão: Mesmo com o diagnóstico negativo de esfregaço sanguíneo periférico, com a suspeita clínica do animal foi solicitado PCR para confirmar a patologia, assim foi de suma importância a pesquisa de hemoparasitas pelos sinais clínicos do canino, juntamente do tratamento precoce e assertivo.

Molecular detection and risk factors for Anaplasma platys infection in dogs from Egypt

Background Anaplasma platys is a tick-borne bacterium which infects blood platelets of dogs, causing canine cyclic thrombocytopenia. The disease is distributed worldwide, particularly in the tropics and subtropics, but information on the epidemiology of A. platys infection in dogs is fragmentary in many countries, including Egypt. In this study, we investigated the prevalence and risk factors associated with A. platys infection in dogs from Egypt. Methods A conventional PCR targeting a fragment of the 16S rRNA gene of A. platys was used to screen 500 dogs from five North Egyptian governorates. DNA sequencing and phylogenetic analysis were performed for one of the positive samples. Results The overall prevalence of A. platys in the studied dogs was 6.4%. Females of the German shepherd breed without veterinary care had higher odds for A. platys positivity. High tick infestation and lack of anti-tick treatment were also identified as risk factors for A. platys infection. Phylogenetic analysis revealed that the sequence obtained herein was closely related to sequences from Egypt, South Africa and Uruguay. Conclusions This is the first large-scale epidemiological study of A. platys in Egypt, where female German shepherd dogs without veterinary care, as well as dogs with high tick infestation and without anti-tick treatment are at a higher risk of infection. Graphical abstract

Molecular detection of Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos, Trypanosoma evansi and first evidence of Theileria sinensis-associated bovine anaemia in crossbred Kedah-Kelantan x Brahman cattle

Background Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos and Trypanosoma evansi, and determine their phylogenetic relationships and haemato-biochemical abnormalities in naturally infected cattle. Methods Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of Anaplasma and Mycoplasma species, MPSP genes of T. orientalis and T. sinensis, MSP4 gene of A. marginale, 16SrRNA gene of Candidatus Mycoplasma haemobos, and RoTat1.2 VSG gene of Trypanosoma evansi, in sixty-one (61) clinically ill Kedah-Kelantan x Brahman cattle in Pahang, Malaysia. Results A total of 44 (72.13%) cattle were infected with more than one blood pathogen. Theileria species was the blood pathogen with the highest molecular detection rate (72.13, 95% CI 59.83–81.81%). Nucleotide blast analyses of all sequences demonstrated high degree of molecular similarity (98–100%) in comparison with their respective reference sequences. Analysis of 18SrRNA gene sequences of Theileria species and 16SrRNA gene sequences of Anaplasma species revealed Theileria sinensis and Anaplasma platys respectively as additional species detected in these cattle. MPSP-PCR analysis was conducted for further confirmation of T. sinensis. The blood picture of eight infected cattle groups revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. High mean erythrocyte fragility values were common in infected cattle groups. Anaemia of the macrocytic normochromic type and spherocytes were observed in the T. evansi and Anaplasma platys + Theileria sinensis double species co-infected cattle group. Normocytic normochromic anaemia was observed in the T. sinensis infected cattle group. Significant (p < 0.05) increases in serum liver and kidney parameters, total protein, globulin, total and unconjugated bilirubin and decreased albumin values were observed in the T. evansi infected cattle when compared to clinically healthy cattle. Conclusion We present the first evidence of Theileria sinensis-associated bovine anaemia (TSABA) in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of A. platys, there is an urgent need for appropriate preventive and control measures against these blood pathogens.

Aspectos epidemiológicos de hemoparasitose em gatos domésticos (Felis catus) da região Amazônica Mato-Grossense

Gatos domésticos (Felis catus) correm o risco de serem acometidos por agentes transmitidos por carrapatos, mesmo sendo menos susceptíveis a infestações desses vetores. Dentre esses agentes, Anaplasma platys vem chamando atenção por suas características patogênicas que podem se tornar ainda mais severas, quando associada a outros agentes bacterianos. O objetivo desta pesquisa foi avaliar de modo retrospectivo279 laudos de exames para pesquisa de hemoparasitoses em gatos domésticos, realizados na região Amazônica Mato-grossense entre os anos de 2016 e 2018. A associação com as variáveis independentes (raça, sexo, idade e estação do ano)foi realizada através do teste Exato de Fisher com nível de confiança de 95%, a razão de prevalência (RP) foi calculada através da regressão de Poisson, todas as análises foram realizadas pelo programa SPSS versão 2.0. Dentre os laudos analisados, foram detectadas apenas formas parasitárias com características de Anaplasma platys, este agente foi detectado em 41 (14,7%) dos 279 laudos de gatos domésticos analisados neste estudo.Destaca-se maior prevalência da infecção em filhotes, contudo não houve distinção de prevalência entre períodos de estiagem e chuvosos, assim como nas variáveis raça e sexo não foi observada diferença significativa.