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1.
Ticks Tick Borne Dis ; 15(5): 102351, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38788485

RESUMO

The transplacental transmission of parasites and hemoparasites is crucial for understanding the epidemiology of diseases. This study aimed to assess the prevalence of hemopathogens in bovine fetuses at various gestational periods. Samples were obtained from a slaughterhouse in the state of Minas Gerais, Brazil, and a total of 236 fetuses were collected. DNA extracted from blood samples (145) and organ samples (a pool of brain and spleen) (236) underwent a nested PCR (nPCR) assay to detect Babesia spp., Theileria spp., Trypanosoma vivax, Anaplasma marginale, Anaplasma bovis, Anaplasma phagocytophilum, Ehrlichia minasensis, and hemotropic Mycoplasma spp. Additionally, serological analysis of 145 plasma samples was conducted using the indirect fluorescent antibody test-IFAT to detect IgG against Babesia bovis, Babesia bigemina, A. marginale, and Trypanosoma vivax. The observed prevalence of transplacental transmission was 19.3 %, 6.2 %, 42.7 % and 2.7 %, for A. marginale, B. bigemina, 'Candidatus M. haemobos', and Mycoplasma wenyonii, respectively. The prevalence of A. marginale by gestational trimester was 16 % (13/81) in the second trimester and 23 % (14/60) in the third trimester, with no positive samples in the first trimester. Regarding the species B. bovis and B. bigemina, all evaluated animals tested negative by nPCR, and no serological evidence for B. bovis was found by the IFAT. Babesia bigemina demonstrated an overall seroprevalence of 6.2 % (9/145), with 4.8 % (7/145) in the last trimester and 1.3 % (2/145) in the second trimester of pregnancy. In total, 42.7 % (62/145) of blood samples were positive for 'Candidatus M. haemobos', with 42 % (34/81) in the middle trimester, and 43 % (26/60) in the final trimester of pregnancy. Mycoplasma wenyonni was detected in 2.7 % (4/145) blood samples, all in coinfection with 'C. M. haemobos'. The prevalence by pregnancy trimester was 25 % (1/4) in the first trimester; 1.2 % (1/81) in the second trimester and 3.3 % (2/60) in the third trimester of pregnancy. Hemopathogen DNA was detected in fetus blood samples but not the brain or spleen samples. All the samples were negative for T. vivax, Theileria spp., Anaplasma spp. and Ehrlichia spp. Overall, in this study, approximately 70 % of fetuses were positive for one or more of the studied parasites. No significant associations were observed between pairs of pathogens, except 'C. M. haemobos' and A. marginale.


Assuntos
Doenças dos Bovinos , Mycoplasma , Animais , Brasil/epidemiologia , Bovinos , Feminino , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , Mycoplasma/isolamento & purificação , Gravidez , Prevalência , Babesia/isolamento & purificação , Feto/microbiologia , Feto/parasitologia , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/microbiologia , Theileria/isolamento & purificação , Trypanosoma vivax/isolamento & purificação , Transmissão Vertical de Doenças Infecciosas/veterinária , Anaplasma/isolamento & purificação , Babesiose/epidemiologia , Babesiose/parasitologia , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Ehrlichia/isolamento & purificação
2.
Trop Anim Health Prod ; 56(4): 154, 2024 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-38727782

RESUMO

This study aimed to investigate seven outbreaks of A. marginale infection in two regions of Brazil, affecting taurine, zebu, and crossbred cattle. We assessed the possible causes, treatment measures, and genetic diversity of A. marginale. These outbreaks occurred in two states (Goiás: outbreaks 1-7; Mato Grosso do Sul: outbreak 3), breeds (Holstein, Nellore, and crossbreed), age groups (beef cattle: 18-25 days old and 7-8 months; dairy cattle: 18-25 days old, 13-14 months, and cow after the first birth) and rearing systems (feedlot, pasture, pen in a wood shaving bedding system and compost bedded-pack barns). Metaphylactic or prophylactic treatments varied according to outbreak (imidocarb dipropionate: outbreaks 1-4 and 6; enrofloxacin: outbreaks 5 and 7; diminazene diaceturate: outbreak 5). In outbreaks 6 and 7, the packed cell volume was monitored. In all outbreaks, the practice of needle/syringe sharing was discontinued. For outbreaks 1-3, clinical signs and mortality (range, 4.8-13.3%) occurred 36-45 days after entry into the feedlot. In outbreak 4, A. marginale was diagnosed in 66.2% of the calves (bacteremia, 0-4.5%), with a mortality of 8.6%. Among nursing calves aged 60 days during outbreak 5, 53.8% were infected with A. marginale, with average bacteremia of 2.7% (range, 0-21.3%), and a mortality of 13.8%. In dairy heifers aged 14 months, raised in paddocks lacking vegetation cover and infested with R. microplus, then transitioned to a rotational grazing system also infested with R. microplus, the A. marginale bacteremia ranged from 3.2 to 6.7%, with a mortality of 20%. Before monitoring during outbreak 7, the mortality was 17.9%, but no further deaths were observed after monitoring initiation. In conclusion, possible causes triggering the outbreaks included primary tick infestation, needle/syringe sharing, and stress factors which may have affected the immunological statues of animals in the feedlots. Control measures performed in all outbreaks were effective. The partial msp4 gene sequences of A. marginale generated herein belonged to two haplotypes, but further research would be needed to investigate if this finding has any clinical significance.


Assuntos
Anaplasma marginale , Anaplasmose , Doenças dos Bovinos , Surtos de Doenças , Variação Genética , Animais , Brasil/epidemiologia , Bovinos , Surtos de Doenças/veterinária , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Anaplasma marginale/genética , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Feminino , Criação de Animais Domésticos/métodos , Masculino
3.
Nat Commun ; 15(1): 3988, 2024 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-38734682

RESUMO

Tick-borne bacteria of the genera Ehrlichia and Anaplasma cause several emerging human infectious diseases worldwide. In this study, we conduct an extensive survey for Ehrlichia and Anaplasma infections in the rainforests of the Amazon biome of French Guiana. Through molecular genetics and metagenomics reconstruction, we observe a high indigenous biodiversity of infections circulating among humans, wildlife, and ticks inhabiting these ecosystems. Molecular typing identifies these infections as highly endemic, with a majority of new strains and putative species specific to French Guiana. They are detected in unusual rainforest wild animals, suggesting they have distinctive sylvatic transmission cycles. They also present potential health hazards, as revealed by the detection of Candidatus Anaplasma sparouinense in human red blood cells and that of a new close relative of the human pathogen Ehrlichia ewingii, Candidatus Ehrlichia cajennense, in the tick species that most frequently bite humans in South America. The genome assembly of three new putative species obtained from human, sloth, and tick metagenomes further reveals the presence of major homologs of Ehrlichia and Anaplasma virulence factors. These observations converge to classify health hazards associated with Ehrlichia and Anaplasma infections in the Amazon biome as distinct from those in the Northern Hemisphere.


Assuntos
Anaplasma , Animais Selvagens , Ehrlichia , Filogenia , Floresta Úmida , Carrapatos , Anaplasma/genética , Anaplasma/isolamento & purificação , Anaplasma/patogenicidade , Anaplasma/classificação , Ehrlichia/genética , Ehrlichia/isolamento & purificação , Ehrlichia/classificação , Humanos , Animais , Carrapatos/microbiologia , Animais Selvagens/microbiologia , Anaplasmose/microbiologia , Anaplasmose/epidemiologia , Anaplasmose/transmissão , Guiana Francesa , Ehrlichiose/microbiologia , Ehrlichiose/epidemiologia , Ehrlichiose/veterinária , Ehrlichiose/transmissão , Metagenômica/métodos , Genoma Bacteriano/genética , RNA Ribossômico 16S/genética
4.
Vet Parasitol Reg Stud Reports ; 50: 101011, 2024 05.
Artigo em Inglês | MEDLINE | ID: mdl-38644043

RESUMO

Anaplasmosis and babesiosis are globally distributed arthropod-borne diseases known for causing substantial economic losses due to their high morbidity and mortality rates. This study aims to assess the frequency and epidemiological features associated with the infection of Anaplasma marginale, Babesia bigemina, and Babesia bovis in three Creole cattle breeds (Chino Santandereano (Chino), Casanareño (CAS), and Sanmartinero (SM)) in northeastern Colombia. Between June 2019 and March 2020, a total of 252 Creole cattle were sampled, with Chino, CAS, and SM accounting for 42.8%, 29.5%, and 29.5% of the samples, respectively. Blood samples were subjected to molecular analysis to detect the DNA of A. marginale, B. bigemina, and B. bovis, using species-specific primers. Additionally, Packed Cell Volume (PCV), total serum proteins, and body condition were evaluated. Molecular analyses revealed the presence of B. bigemina, A. marginale, and B. bovis in 83.7% (211/252; 95% CI = 79.1%-88.3%), 59.9% (151/252; 95% CI = 53.8%-66.1%), and 40.9% (103/252; 95% CI = 34.7%-46.9%) of the samples, respectively, with 69% (174/252; 95% CI = 57.8%-80.3%) exhibiting coinfections. Notably, in infected animals, no significant alterations in PCV, total serum proteins, or body condition were observed. Multivariate analyses indicated a statistically significant association between the frequency of A. marginale infection and the breed and season, with a higher frequency in SM during the rainy season (P < 0.05). To our knowledge, this is the first molecular survey that evaluates multiple arthropod-borne pathogens in Colombian Creole breeds. The results revel a high frequency of B. bigemina and A. marginale infections, coupled with a notable frequency of coinfections, all without significant alteration in the PCV, total serum proteins and body conditions. Our findings enhance the understanding of the epidemiological aspects of arthropod-borne pathogens in Colombian Creole breed and contribute to the improvement of sanitary programs for these animals.


Assuntos
Anaplasma marginale , Anaplasmose , Babesia bovis , Babesia , Babesiose , Doenças dos Bovinos , Animais , Bovinos , Colômbia/epidemiologia , Babesiose/epidemiologia , Babesiose/parasitologia , Anaplasma marginale/genética , Anaplasma marginale/isolamento & purificação , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/microbiologia , Babesia/isolamento & purificação , Babesia/genética , Babesia/classificação , Babesia bovis/genética , Babesia bovis/isolamento & purificação , Feminino , Masculino , Prevalência
5.
Ticks Tick Borne Dis ; 15(1): 102254, 2024 01.
Artigo em Inglês | MEDLINE | ID: mdl-37989016

RESUMO

Anaplasmosis, caused by bacteria of the genus Anaplasma, is an important tick-borne disease that causes economic losses to livestock farms in many countries. Even though Anaplasma spp. have been detected in goats and sheep worldwide, few studies investigate the occurrence and genetic identity of these agents in small ruminants from Brazil. Thus, this work aimed to detect and determine the genetic identity of Anaplasma spp. in small ruminants from the Baixo Parnaíba region, state of Maranhão, northeastern Brazil. For this purpose, blood samples were collected from 161 animals (91 goats; 70 sheep) from 4 municipalities in the Baixo Parnaíba region. Sheep and goat serum samples were subjected to recombinant membrane surface protein (MSP5)-based iELISA. Whole blood samples were subject to DNA extraction and molecular diagnosis using PCR assays for Anaplasma spp. targeting msp1ß, msp1α, 16S rRNA and msp4 genes. Positive samples were sequenced and then subjected to Anaplasma marginale msp1α genetic diversity analysis and phylogenetic inferences based on the 16S rRNA and msp4 genes. The serological survey detected the presence of anti-A. marginale IgG antibodies in 18 animals (11.1%): 2.9% (2/70) sheep and 17.4% (16/91) goats. Anaplasma marginale DNA was detected in 2 goats (1.2%) using qPCR based on the msp1ß gene. Two distinct A. marginale msp1α strains, namely α ß and α ß ΓγΓγΓγΓγ were found in the infected goats, each one found in a different animal, both belonging to the H genotype. Phylogenetic analysis based on the 16S rRNA gene showed the sequences positioned in three different clades and grouped with sequences from 'Candidatus Anaplasma boleense', A. platys and A. marginale. Phylogenetic inferences based on the msp4 gene positioned the sequence variants in the A. marginale clade. The present work represents the first molecular detection of sequence variants phylogenetic associated to 'Candidatus Anaplasma boleense' and A. platys and α ß and α ß ΓγΓγΓγΓγ in goats from Brazil.


Assuntos
Anaplasma marginale , Anaplasmose , Doenças das Cabras , Doenças dos Ovinos , Animais , Ovinos , Anaplasma/genética , RNA Ribossômico 16S/genética , Brasil/epidemiologia , Filogenia , Anaplasmose/microbiologia , Ruminantes , Anaplasma marginale/genética , Proteínas de Membrana/genética , Cabras/microbiologia , DNA , Doenças das Cabras/epidemiologia , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/microbiologia
6.
Braz J Microbiol ; 54(3): 2497-2504, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37351788

RESUMO

The coinfections by some microorganisms have been related to severe diseases in humans and animals, where immunosuppressive agents favor opportunistic behavior of other pathogens. A 4-month-old, female mixed-breed dog with a two-week history of inappetence, prostration, emaciation, and respiratory distress was admitted at a veterinary hospital in Brazil. Tachycardia, pale mucous membranes, severe respiratory distress, and a large number of ticks (Rhipicephalus sanguineus s.l.) in different body regions were observed at clinical examination. Hematological examination of dog showed leukocytosis, neutrophilia, mild anemia, and thrombocytopenia, whereas unremarkable values in biochemical tests. Thoracic radiography revealed a pleural effusion image. Blood and the pleural fluid (purulent aspect) samples were subjected to qPCR (16S rRNA and dsb genes) and sequencing, which identified Ehrlichia canis and Anaplasma platys coinfection. An aggregate of coccoid-to-branching or long filamentous microorganisms, surrounded by pyogranulomatous inflammatory reaction was seen at the cytology of the pleural fluid. Bacteriological culture of pleural effusion showed colonies compatible with the genus Nocardia, which revealed gram-positive filamentous organisms with a tendency of fragmentation and were identified as Nocardia otitidiscaviarum in mass spectrometry (MALDI-TOF MS). Therapy of N. otitidiscaviarum isolate using levofloxacin (supported by a previous in vitro susceptibility testing) and doxycycline for E. canis and A. platys resulted in complete resolution of the clinical picture. Here, we report for the first time a triple coinfection by Nocardia otitidiscaviarum, A. platys, and E. canis in a dog with pleural effusion, where debilitating or immunosuppressive conditions induced by A. platys and E. canis coinfection probably contributed to the opportunistic behavior of N. otitidiscaviarum.


Assuntos
Anaplasmose , Coinfecção , Doenças do Cão , Ehrlichiose , Nocardia , Derrame Pleural , Síndrome do Desconforto Respiratório , Humanos , Cães , Feminino , Animais , Lactente , Ehrlichia canis/genética , Anaplasmose/microbiologia , Coinfecção/veterinária , Coinfecção/microbiologia , Ehrlichiose/veterinária , Ehrlichiose/microbiologia , RNA Ribossômico 16S/genética , Nocardia/genética , Derrame Pleural/veterinária , Doenças do Cão/microbiologia
7.
Comp Immunol Microbiol Infect Dis ; 96: 101980, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-37079984

RESUMO

The present study aimed at the molecular detection of Anaplasma spp. in different samples obtained from cattle, goats and free-living Rhipicephalus microplus ticks from Argentina. DNA of members of the Anaplasmataceae family was detected by different PCR assays. The phylogenetic analyses of the obtained partial DNA sequences of the 16 S rDNA gene resulted in the identification of two different Anaplasma spp.: (I) Anaplasma platys-like bacteria (in blood sample from cattle and pools of R. microplus larvae and (II) Candidatus Anaplasma boleense (in blood samples from goats and one pool of R. microplus larvae of R. microplus). Candidatus A. boleense was found in two provinces that belong to different biogeographic regions, which leads to the conclusion that this bacterium may be widely distributed in Argentina. Interestingly, both Anaplasma spp. were found in the same R. microplus population in Chaco province, indicating that these two strains of Anaplasma are circulating in the same tick population. The results of this work represent the first report of the circulation of A. platys-like bacteria and Ca. A. boleense in domestic ruminants and free-living R. microplus ticks in Argentina. Further studies to determine the prevalence of infection, dispersion, clinical impact, transmission routes and cross-reactivity in serological tests of both Anaplasma species are needed.


Assuntos
Anaplasmose , Doenças dos Bovinos , Doenças das Cabras , Rhipicephalus , Animais , Bovinos , Filogenia , Argentina/epidemiologia , Anaplasma/genética , Rhipicephalus/microbiologia , Ruminantes , Cabras/microbiologia , Bactérias , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Doenças das Cabras/epidemiologia , Doenças das Cabras/microbiologia
8.
Res Vet Sci ; 154: 145-150, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36599268

RESUMO

This study aimed to evaluate the frequency of IgG antibodies against A. marginale, the occurrence of this bacterium by qPCR, and the effect of bovine anaplasmosis as a risk factor for clinical cases of retained placenta, mastitis, and abomasal displacement in dairy cattle. For that 179 Holstein cows out of three dairy herds, in the municipality of Sertão, Rio Grande do Sul, Brazil. These cows were on farms that were vulnerable to risk factors that are crucial to susceptibility among these animals to this intracellular hemoparasite. The mean seropositivity for A. marginale from the periods evaluated was 54% on farm A, 69.4% on farm B, and 27.3% on farm C. Molecular diagnosis was performed with qPCR and the mean positivity for A. marginale among the cows on farms A, B, and C in December 2017 was 34.6% (67/179). Infected animals showed clinical cases of retained placenta (6.1%), mastitis (6.1%), and abomasal displacement (0.5%). The association between positivity for anaplasmosis and these clinical cases was assessed through the odds ratio. Our results show that females with a positive qPCR assay for A. marginale had 52.48 times increased probability (OR) to develop clinical cases of retained placenta and mastitis (P < 0.001). These clinical cases negatively impact the productivity of positive females. Thus, implementing preventive and prophylactic control measures to ensure the sanitary quality of the herds is needed to avoid losses due to morbidity and mortality and diminish the economic losses suffered by farmers.


Assuntos
Anaplasmose , Doenças dos Bovinos , Mastite Bovina , Placenta Retida , Feminino , Gravidez , Bovinos , Animais , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Placenta Retida/epidemiologia , Placenta Retida/veterinária , Fatores de Risco , Mastite Bovina/epidemiologia
9.
J Am Vet Med Assoc ; 261(3): 375-383, 2022 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-36563069

RESUMO

OBJECTIVE: To assess exposure to and infection with 3 pathogens (Rickettsia rickettsii, Anaplasma platys, and Ehrlichia canis) vectored by brown dog ticks (Rhipicephalus sanguineus) in sheltered dogs at the western US-Mexico border. ANIMALS: 239 dogs in shelters in San Diego and Imperial counties, US, and Mexicali and Tijuana, Mexico. PROCEDURES: Each dog had blood drawn and basic demographic data collected. PCR was performed to determine active infection with Rickettsia spp, E canis, and A platys. Serology was performed to determine exposure to Rickettsia, Anaplasma, and Ehrlichia species. RESULTS: 2 of 78 (2.6%) dogs sampled in Tijuana were actively infected with R rickettsii. A single brown dog tick collected from a dog in Tijuana was PCR-positive for R rickettsii. Infection with E canis and A platys ranged across shelters from 0% to 27% and 0% to 33%, respectively. Dogs in all 4 locations demonstrated exposure to all 3 pathogens, though Rickettsia and Ehrlichia seropositivity was highest in Mexicali (81% and 49%, respectively) and Anaplasma seropositivity was highest in Tijuana (45%). CLINICAL RELEVANCE: While infection and exposure were highest in sheltered dogs in the southern locations, dogs in all locations demonstrated exposure to all pathogens, demonstrating the potential for emergence and spread of zoonotic pathogens with significant public health consequences in southern California and northern Baja California. In addition, veterinarians and shelter staff should be aware that Ehrlichia or Anaplasma infection may co-occur with Rocky Mountain spotted fever, which is a human health risk.


Assuntos
Anaplasmose , Doenças do Cão , Saúde Única , Rhipicephalus sanguineus , Febre Maculosa das Montanhas Rochosas , Cães , Humanos , Animais , Febre Maculosa das Montanhas Rochosas/epidemiologia , Febre Maculosa das Montanhas Rochosas/veterinária , México/epidemiologia , Rhipicephalus sanguineus/microbiologia , Anaplasma , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Doenças do Cão/epidemiologia
10.
J Dairy Sci ; 105(5): 4421-4433, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35282915

RESUMO

Bovine anaplasmosis causes considerable economic losses in dairy cattle production systems worldwide, ranging from $300 million to $900 million annually. It is commonly detected through rectal temperature, blood smear microscopy, and packed cell volume (PCV). Such methodologies are laborious, costly, and difficult to systematically implement in large-scale operations. The objectives of this study were to evaluate (1) rumination and activity data collected by Hr-Tag sensors (SCR Engineers Ltd.) in heifer calves exposed to anaplasmosis; and (2) the predictive ability of recurrent neural networks in early identification of anaplasmosis. Additionally, we aimed to investigate the effect of time series length before disease diagnosis (5, 7, 10, or 12 consecutive days) on the predictive performance of recurrent neural networks, and how early anaplasmosis disease can be detected in dairy calves (5, 3, and 1 d in advance). Twenty-three heifer calves aged 119 ± 15 (mean ± SD) d and weighing 148 ± 20 kg of body weight were challenged with 2 × 107 erythrocytes infected with UFMG1 strain (GenBank no. EU676176) isolated from Anaplasma marginale. After inoculation, animals were monitored daily by assessing PCV. The lowest PCV value (14 ± 1.8%) and the finding of rickettsia on blood smears were used as a criterion to classify an animal as sick (d 0). Rumination and activity data were collected continuously and automatically at 2-h intervals, using SCR Heatime Hr-Tag collars. Two time series were built including last sequence of -5, -7, -10, or -12 d preceding d 0 or a sequence of 5, 7, 10, or 12 d randomly selected in a window from -50 to -15 d before d 0 to ensure a sequence of days in which PCV was considered normal (32 ± 2.4%). Long short-term memory was used as a predictive approach, and a leave-one-animal-out cross-validation (LOAOCV) was used to assess prediction quality. Anaplasmosis disease reduced 34 and 11% of rumination and activity, respectively. The accuracy, sensitivity, and specificity of long short-term memory in detecting anaplasmosis ranged from 87 to 98%, 83 to 100%, and 83 to 100%, respectively, using rumination data. For activity data, the accuracy, sensitivity, and specificity varied from 70 to 98%, 61 to 100%, and 74 to 100%, respectively. Predictive performance did not improve when combining rumination and activity. The use of longer time-series did not improve the performance of models to predict anaplasmosis. The accuracy and sensitivity in predicting anaplasmosis up to 3 d before clinical diagnosis (d 0) were greater than 80%, confirming the possibility for early identification of anaplasmosis disease. These findings indicate the great potential of wearable sensors in early identification of anaplasmosis diseases. This could positively affect the profitability of dairy farmers and animal welfare.


Assuntos
Anaplasma marginale , Anaplasmose , Doenças dos Bovinos , Anaplasmose/diagnóstico , Anaplasmose/microbiologia , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/microbiologia , Eritrócitos , Feminino , Vacinação/veterinária
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