Molecular detection of Anaplasma species in dogs in Colombia

Abstract Anaplasma platys and A. phagocytophilum are tick-borne pathogens that parasitize platelets and neutrophils, respectively, of humans and animals. The former is the etiological agent of canine cyclic thrombocytopenia, while the latter is that of canine granulocytic anaplasmosis. This work involved the detection and identification of Anaplasma species in blood samples from dogs in Colombia, using molecular techniques. Between December 2008 and April 2009, blood samples were drawn from the cephalic vein of 91 dogs in the central-western region of Colombia (cities of Bogota, Villavicencio and Bucaramanga) and stored in tubes containing EDTA. These samples were used in 16S rRNA-Anaplasma spp. nPCR and the preparation of blood smears. One (1.1%) of the 91 sampled dogs showed inclusions suggestive of Anaplasmataceae agents in the cytoplasm of platelets. Based on PCR followed by sequencing and phylogenetic analysis, A. platys and Anaplasma sp. closed related to A. phagocytophilum were detected in two and one dog, respectively. Interestingly, all the samples were negative for specific msp-2-A. phagocytophilum real-time qPCR, suggesting the circulation of an Anaplasma species phylogenetically related to A. phagocytophilum in dogs in the aforementioned region. Hence, Anaplasma spp. circulates among dogs in Colombia, albeit with low frequency. To the best of authors' knowledge, this is the first molecular detection of Anaplasma spp. in dogs in Colombia.

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Molecular detection of feline arthropod-borne pathogens in cats in Cuiabá, state of Mato Grosso, central-western region of Brazil

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612013000300011 ◽

2013 ◽

Vol 22 (3) ◽

pp. 385-390 ◽

Cited By ~ 25

Author(s):

Natasha Gandolfi Miceli ◽

Fernando Antonio Gavioli ◽

Luiz Ricardo Gonçalves ◽

Marcos Rogério André ◽

Valéria Régia Franco Sousa ◽

...

Keyword(s):

Molecular Detection ◽

Western Region ◽

Mato Grosso ◽

Animal Shelters ◽

Blood Samples ◽

Cytauxzoon Felis ◽

The City ◽

Veterinary Hospital

Hemotrophic mycoplasmas (hemoplasmas), Bartonellasp., Hepatozoon sp. and Cytauxzoon felis are prominent pathogens that circulate between cats and invertebrate hosts. The present study aimed to detect the presence of DNA from hemoplasmas,Bartonella sp., Hepatozoon sp. andCytauxzoon felis, and then confirm it by means of sequencing, in blood samples from cats in Cuiabá, MT, Brazil. From February 2009 to February 2011, blood samples with added EDTA were collected from 163 cats that were being housed in four different animal shelters in the city of Cuiabá, state of Mato Grosso, Brazil and from 15 cats that were admitted to the veterinary hospital of the Federal University of Mato Grosso (UFMT). Out of the 178 cats sampled, 15 (8.4%) were positive for hemoplasmas: four (2.2%) forMycoplasma haemofelis, 12 (6.7%) for ‘Candidatus M. haemominutum’ and one (0.5%) for ‘Candidatus M. turicensis’. One cat (0.5%), a patient that was attended at the veterinary hospital, was coinfected with M. haemofelis, ‘Candidatus M. haemominutum’ and ‘Candidatus M. turicensis’, based on sequencing confirmation. Four cats were positive for Bartonella spp.: three (1.7%) for B. henselae and one (0.5%) for B. clarridgeiae. None of the animals showedCytauxzoon sp. or Hepatozoon sp. DNA in their blood samples. This study showed that cats housed in animal shelters in the city of Cuiabá, state of Mato Grosso, are exposed to hemoplasmas andBartonella species.

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First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum from a Clinical Case of Canine Granulocytic Anaplasmosis in Japan

Japanese Journal of Infectious Diseases ◽

10.7883/yoken.jjid.2017.558 ◽

2018 ◽

Vol 71 (4) ◽

pp. 302-305 ◽

Cited By ~ 3

Author(s):

Yuichi Fukui ◽

Seigo Ohkawa ◽

Hisashi Inokuma

Keyword(s):

Phylogenetic Analysis ◽

Clinical Case ◽

Molecular Detection ◽

Granulocytic Anaplasmosis ◽

Canine Granulocytic Anaplasmosis

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Molecular Detection and Differentiation of Theileria lestoquardi, T. ovis and T. annulata in Blood of Goats and Ticks in Kermanshah Province, Iran

Journal of Arthropod-Borne Diseases ◽

10.18502/jad.v13i3.1539 ◽

2019 ◽

Author(s):

Mozhgan Rahmani-Varmale ◽

Mousa Tavassoli ◽

Bijan Esmaeilnejad

Keyword(s):

Nested Pcr ◽

Molecular Detection ◽

Ixodid Ticks ◽

Hyalomma Anatolicum ◽

Blood Samples ◽

Theileria Lestoquardi ◽

Blood Smears ◽

Pcr Rflp ◽

Pcr Products ◽

Theileria Spp

Background: This study was carried out to identify Theileria spp. infections in goats and ticksin Kermanshah Province, western Iran from May–Sep 2015. Methods: For differentiation of different Theileria spp. both blood and tick samples were examined by nested PCR-RFLP. Results: Light microscopy of blood smears revealed Theileria spp. infection in 22 (5.5%), while 68 (17%) of blood samples were positive using nested PCR. Out of 68 positive samples, 85.3% (58/68) and 11.7% (8/68) were respectively positive for Theileria ovis and T. lestoquardi. Mixed infection was detected in 3% (2/68) cases. Overall, 420 ixodid ticks belong to seven different hard ticks species were collected from goats. Rhipicephalus turanicus 112 (26.7%), R. sanguineus 95 (22.6%), R. bursa, 91(21.7%), Hyalomma anatolicum, 55(13.1%), H. excavatum 27(6.4%), H. marginatum, 22(5.3%) and Dermacentor marginatus, 18(4.2%) were the main tick species infesting goats. The PCR products obtained from ticks were subjected to the differentiation of Theileria species. Respectively, 2 and 8 pools of H. marginatum and R. turanicus salivary glands were infected with T. ovis and T. lestoquardi. In addition, T. annulata and T. lestoquardi infection weredetected in three pools of H. anatolicum. Conclusion: This is the first report of goats and collected ticks to Theileria spp infection in Iran. The results suggest that T. ovis has a higher prevalence than T. lestoquardi. It is also postulated H. marginatum, R. turanicus and H. anatolicum might play an important role in the field as a vector of Theileria spp in this area.

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First report of Cystoisospora belli parasitemia in a patient with acquired immunodeficiency syndrome

Acta Parasitologica ◽

10.1515/ap-2016-0023 ◽

2016 ◽

Vol 61 (1) ◽

Cited By ~ 2

Author(s):

Jorge Néstor Velásquez ◽

Cecilia Alicia di Risio ◽

Cristina Beatriz Etchart ◽

Agustín Víctor Chertcoff ◽

Mónica Gabriela Nigro ◽

...

Keyword(s):

Lamina Propria ◽

Acquired Immunodeficiency Syndrome ◽

Chronic Diarrhea ◽

Tissue Level ◽

Molecular Techniques ◽

Blood Samples ◽

Blood Smears ◽

Immunodeficiency Virus ◽

Acquired Immunodeficiency ◽

Immunodeficiency Syndrome

AbstractCystoisospora belli in patients with the acquired immunodeficiency syndrome (AIDS) has been described as cause of chronic diarrhea and disseminated cystoisosporosis. Diagnosis of intestinal cystoisosporosis can be achieved at the tissue level in the villus epithelium of the small bowel. Disseminated cystoisosporosis is diagnosed by microscopy identification of unizoite tissue cysts in the lamina propria of the intestine. We report a case of disseminated cystoisosporosis in a human immunodeficiency virus (HIV)-infected patient with detection of parasitemia. We studied a 39-year old patient with AIDS and chronic diarrhea by analysis of stool and duodenal biopsy samples. Blood samples were also collected and examined by light microscopy and molecular techniques for C. belli DNA detection. The unizoite tissue cyst stages were present in the lamina propria, with unsporulated oocysts in feces. Zoites were present in blood smears and DNA of C. belli was detected in blood samples. Our study identified a new stage in the life cycle of C. belli. Detection of parasitemia is a novel and noninvasive tool for diagnosis of disseminated cystoisosporosis.

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Serological, Hematologic, and PCR Studies of Cattle in an Area of Switzerland in Which Tick-Borne Fever (Caused by Ehrlichia phagocytophila) Is Endemic

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.5.3.325-327.1998 ◽

1998 ◽

Vol 5 (3) ◽

pp. 325-327 ◽

Cited By ~ 9

Author(s):

Nicola Pusterla ◽

Jeannine Berger Pusterla ◽

Ueli Braun ◽

Hans Lutz

Keyword(s):

Seasonal Variations ◽

Nested Pcr ◽

Molecular Detection ◽

Clinical Signs ◽

Pcr Amplification ◽

Blood Samples ◽

Ehrlichia Phagocytophila ◽

Blood Smears ◽

Antibody Titers ◽

First Time

ABSTRACT The purpose of this study was to examine the seasonal variations in seroprevalence to Ehrlichia phagocytophila in cattle pastured during the summer months in an area where tick-borne fever is endemic. The study was performed during a 1-year period from April 1996 to March 1997 and involved 34 cows, 22 pregnant heifers, and 14 calves. Blood samples, collected from all 70 cattle once a month, were used to determine serum immunoglobulin G titers by indirect immunofluorescence. In addition, blood smears were examined for Ehrlichiaorganisms, and PCR amplification was performed for the molecular detection of E. phagocytophila. Prior to the pasture period, the seroprevalence was 16%. Two weeks after the start of pasturing, it was 43%, after which it progressively increased and reached a maximum of 63% in September. The seroprevalence progressively decreased after the end of pasturing to a low of 23%. The variation in antibody titers was similar to that of seroprevalence.E. phagocytophila organisms were detected in blood smears of 7 animals and by nested PCR in 12. Only four cows, which were on the pastures of endemicity for the first time, had clinical signs of ehrlichiosis. This study demonstrated marked seasonal variations in seroprevalence and in serum titers of antibody to E. phagocytophila in cattle. The incidence of clinical signs of ehrlichiosis was increased in cattle grazing on the pastures of endemicity for the first time.

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Molecular detection of Trypanosoma evansi (Kinetoplastida: Trypanosomatidae) in procyonids (Carnivora: Procyonidae) in Eastern Amazon, Brazil

Ciência Rural ◽

10.1590/0103-8478cr20150679 ◽

2016 ◽

Vol 46 (4) ◽

pp. 663-668 ◽

Cited By ~ 2

Author(s):

Paulo Cesar Magalhães-Matos ◽

Rafaelle Cunha-Santos ◽

Paulo Geovani Silva Sousa ◽

Francisco Dantas Sampaio-Júnior ◽

Flávia de Nazaré Leite Barros ◽

...

Keyword(s):

Whole Blood ◽

Molecular Detection ◽

Natural Infection ◽

Trypanosoma Evansi ◽

Free Living ◽

Blood Samples ◽

Blood Smears ◽

In Captivity ◽

Whole Blood Samples ◽

Wild Life

ABSTRACT: The present study aimed to diagnose the natural infection of captive and free-living procyonids with Trypanosoma evansi in the states of Amapá and Pará, Brazil. From February 2012 to August 2013, whole blood samples and blood smears were obtained from 45 free-living procyonids and from nine procyonids kept in captivity in wild life refuges and zoobotanical parks in the states of Amapá and Pará. Whole blood samples were collected and kept at -20ºC for the detection of T. evansi DNA by PCR using the RoTat 1.2 forward and RoTat 1.2 reverse primers. In addition, the blood smears were processed and examined for the presence of trypomastigote forms of T. evansi. T. evansi DNA was detected in 18.52% (10/54) of the procyonids, namely, in captive crab-eating raccoons and captive and free-living coatis in Pará State. No trypomastigote forms were observed in the blood smears. DNA from T. evansi was detected in P. cancrivorus and N. nasua in Pará State, being this the first such report in P. cancrivorus.

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Molecular detection of Ehrlichia canis and Anaplasma platys in dogs from municipality of Belém, State of Pará, Brazil

Ciência Rural ◽

10.1590/0103-8478cr20190414 ◽

2019 ◽

Vol 49 (12) ◽

Author(s):

Verucia Maria Dias Brandão ◽

Pedro Henrique Marques Barrozo ◽

Luciane Oeiras Sousa ◽

Rafaelle Cunha dos Santos ◽

Katiane Schwanke ◽

...

Keyword(s):

Nested Pcr ◽

Molecular Detection ◽

Ehrlichia Canis ◽

Anaplasma Platys ◽

Blood Samples ◽

Stray Dogs ◽

Significant Difference ◽

The City

ABSTRACT: The occurrence of diseases transmitted by ticks in dogs is very frequent in Brazil, among these diseases we can highlight the ehrlichiosis and anaplasmosis, which are caused by Ehrlichia canis and Anaplasma platys, respectively. The objective of this study was to survey the occurrence of these pathogens in blood samples from domiciled and stray dogs from the city of Belém, Pará. Two hundred and seventy six dogs were sampled for convenience, and the DNA extracted from the blood of these animals was submitted to nested-PCR for research of E. canis and A. platys. E. canis DNA was detected in 39.4% (109/276) and A. platys DNA in 23.1% (64/276) of the samples, there was a statistically significant difference between the frequency of these agents (P<0.0001), and there was coinfection in 13.4% (37/276) of animals. The frequency of detection of these parasites was higher in stray dogs than in those domiciled for both E. canis (OR=2.84) and A. platys (OR=10.5). Considering the results, it was possible to conclude that E. canis and A. platys are present in the studied population, with stray dogs being more affected by these parasites.

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Canine Babesiosis in Northwestern India: Molecular Detection and Assessment of Risk Factors

BioMed Research International ◽

10.1155/2014/741785 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5 ◽

Cited By ~ 13

Author(s):

Amritpal Singh ◽

Harkirat Singh ◽

N. K. Singh ◽

N. D. Singh ◽

S. S. Rath

Keyword(s):

Risk Factors ◽

Molecular Detection ◽

Winter Season ◽

Canine Babesiosis ◽

Babesia Gibsoni ◽

Blood Samples ◽

Molecular Assays ◽

Northwestern India ◽

Blood Smears ◽

Thin Blood Smears

In the current study, a total of 214 blood samples from dogs in and around Ludhiana, Punjab (India), suspected for canine babesiosis were examined with conventional and molecular assays. Examination of Giemsa-stained peripheral thin blood smears revealed an overall prevalence of 7.47% (16/214) for canine babesiosis encompassing 0.93% (2/214) of largeBabesiaand 6.54% (14/214) ofBabesia gibsoni. However, molecular diagnosis revealed 15.42% (33/214) samples positive forB. gibsoniinfection as evident by the presence of 671 bp amplicon. The results of multivariate analysis showed that the prevalence ofB. gibsoniwas associated with various risk factors, namely, age (P<0.001; OR: 0.398; CI 95%: 0.080–1.799), sex (P=0.022; OR: 0.849; CI 95%: 0.403–1.791), breed of host (P=0.371; OR: 3.345; CI 95%: 1.045–10.710), and season (P=0.230; OR: 2.143; CI 95%: 0.788–5.830). The prevalence ofB. gibsoniwas higher in summer as compared to winter season and in younger dogs, while breed and sex of the host were not significantly associated with the occurrence of the disease.

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Analytical validation of a real-time hydrolysis probe PCR assay for quantifying Plasmodium falciparum parasites in experimentally infected human adults

Malaria Journal ◽

10.1186/s12936-021-03717-y ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Claire Y. T. Wang ◽

Emma L. Ballard ◽

Zuleima Pava ◽

Louise Marquart ◽

Jane Gaydon ◽

...

Keyword(s):

Clinical Trials ◽

Plasmodium Falciparum ◽

18S Rrna ◽

Drug Efficacy ◽

18S Rrna Gene ◽

Molecular Techniques ◽

Qpcr Assay ◽

Rrna Gene ◽

Analytical Sensitivity ◽

Blood Samples

Abstract Background Volunteer infection studies have become a standard model for evaluating drug efficacy against Plasmodium infections. Molecular techniques such as qPCR are used in these studies due to their ability to provide robust and accurate estimates of parasitaemia at increased sensitivity compared to microscopy. The validity and reliability of assays need to be ensured when used to evaluate the efficacy of candidate drugs in clinical trials. Methods A previously described 18S rRNA gene qPCR assay for quantifying Plasmodium falciparum in blood samples was evaluated. Assay performance characteristics including analytical sensitivity, reportable range, precision, accuracy and specificity were assessed using experimental data and data compiled from phase 1 volunteer infection studies conducted between 2013 and 2019. Guidelines for validation of laboratory-developed molecular assays were followed. Results The reportable range was 1.50 to 6.50 log10 parasites/mL with a limit of detection of 2.045 log10 parasites/mL of whole blood based on a parasite diluted standard series over this range. The assay was highly reproducible with minimal intra-assay (SD = 0.456 quantification cycle (Cq) units [0.137 log10 parasites/mL] over 21 replicates) and inter-assay (SD = 0.604 Cq units [0.182 log10 parasites/mL] over 786 qPCR runs) variability. Through an external quality assurance program, the QIMR assay was shown to generate accurate results (quantitative bias + 0.019 log10 parasites/mL against nominal values). Specificity was 100% after assessing 164 parasite-free human blood samples. Conclusions The 18S rRNA gene qPCR assay is specific and highly reproducible and can provide reliable and accurate parasite quantification. The assay is considered fit for use in evaluating drug efficacy in malaria clinical trials.

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