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1.
Vet Parasitol ; 328: 110188, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38653059

RESUMEN

Canine babesiosis is a rapidly spreading tick-borne disease in Europe, which entails protozoan parasites invading red blood cells. Small extracellular vesicles (EVs) (< 200 nm) were isolated from the serum of 15 healthy and 15 by Babesia canis naturally infected dogs aimed to distinguish EV characteristics and protein profiles. There were no significant differences (P = 0.05) observed in the mean sizes and concentrations of serum EVs between the healthy and canine babesiosis groups. Despite a higher number of Canis lupus proteins detected in EVs from serum of diseased dogs, there were no statistically significant differences (P < 0.05) in the number of protein IDs between the experimental groups. We successfully identified 211 Canis lupus proteins across both experimental groups, of which 147 Canis lupus proteins were validated as being EV-associated. This data set is accessible via the ProteomeXchange PXD047647. EVs isolated from serum of B. canis infected dogs were Cd9+, Cd63+, Cd81+, and Cd82+. Furthermore, 73 Canis lupus proteins were validated as EV-associated and specific for EVs isolated from serum of B. canis-infected dogs. These were predominantly membrane and cytosolic proteins, and innate and adaptive immune system-related proteins, especially those involved in adhesion and proteoglycan mechanisms like integrins. Enrichment was also observed for proteins involved in vascular and cellular responses, including signalling pathways such as VEGF, VEGFR, and the LKB1 network. When only blood-related sites of EV expression were evaluated, the origins of EV proteins were mostly cells of immune system. These were dendritic cells, neutrophils, B cells, monocytes and platelets. In general, proteins were enriched in pathways that collectively regulate various cellular processes, including immune responses, communication, signal transduction, membrane trafficking, and apoptosis. Serum EVs and their protein cargo may have an important role in both the invasion of B. canis and the host's response to the parasitic infection, nevertheless, additional experimental research is warranted. The overall count of identified EV proteins of parasitic origin, meeting cut off criteria of two peptides and 1 % FDR, was relatively low.


Asunto(s)
Babesia , Babesiosis , Enfermedades de los Perros , Vesículas Extracelulares , Proteómica , Animales , Perros , Babesiosis/parasitología , Babesiosis/sangre , Babesia/clasificación , Babesia/aislamiento & purificación , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/sangre , Vesículas Extracelulares/química , Proteómica/métodos , Cromatografía Liquida/veterinaria , Espectrometría de Masas en Tándem/veterinaria , Femenino , Cromatografía Líquida con Espectrometría de Masas
2.
Am J Clin Pathol ; 161(5): 451-462, 2024 May 02.
Artículo en Inglés | MEDLINE | ID: mdl-38113371

RESUMEN

OBJECTIVES: Recent work has demonstrated that automated fluorescence flow cytometry (FLC) is a potential alternative for the detection and quantification of Plasmodium parasites. The objective of this study was to apply this novel FLC method to detect and quantify Babesia parasites in venous blood and compare results to light microscopy and polymerase chain reaction methods. METHODS: An automated hematology/malaria analyzer (XN-31; Sysmex) was used to detect and quantify B microti-infected red blood cells from residual venous blood samples (n = 250: Babesia positive, n = 170; Babesia negative, n = 80). As no instrument software currently exists for Babesia, qualitative and quantitative machine learning (ML) algorithms were developed to facilitate analysis. RESULTS: Performance of the ML models was verified against the XN-31 software using P falciparum-infected samples. When applied to Babesia-infected samples, the qualitative ML model demonstrated an area under the curve (AUC) of 0.956 (sensitivity, 95.9%; specificity, 83.3%) relative to polymerase chain reaction. For valid scattergrams, the qualitive model achieved an AUC of 1.0 (sensitivity and specificity, 100%), while the quantitative model demonstrated an AUC of 0.986 (sensitivity, 94.4%; specificity, 100%). CONCLUSIONS: This investigation demonstrates that Babesia parasites can be detected and quantified directly from venous blood using FLC. Although promising, opportunities remain to improve the general applicability of the method.


Asunto(s)
Babesia , Babesiosis , Eritrocitos , Citometría de Flujo , Citometría de Flujo/métodos , Humanos , Babesiosis/diagnóstico , Babesiosis/sangre , Eritrocitos/parasitología , Babesia/aislamiento & purificación , Babesia/genética , Aprendizaje Automático , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad
3.
Microbiol Spectr ; 10(1): e0241121, 2022 02 23.
Artículo en Inglés | MEDLINE | ID: mdl-35196786

RESUMEN

Equine piroplasmosis (EP) is a serious problem in the horse industry, and controlling EP is critical for international horse trading. EP is caused by two apicomplexan protozoan parasites, Theileria equi and Babesia caballi. Rapid and accurate methods that are suitable for detecting these parasites in the field are crucial to control the infection and spread of EP. In this study, we developed a card to detect antibodies against T. equi and B. caballi based on two colloidal gold immunochromatographic strips according to the principle of the double-antigen sandwich. The proteins equi merozoite antigen 1 (EMA1) and rhoptry protein BC48 are commonly used as diagnostic antigens against T. equi and B. caballi, respectively. On the strip, the purified EMA1 or BC48 protein labeled with colloidal gold was used as the detector, and nitrocellulose membranes were coated with EMA1 or BC48 and the corresponding MAb as the test and control lines, respectively. The protocol takes 10 to 15 min and requires no specialized equipment or chemical reagents, and one test can detect two EP pathogens in one card. Specificity tests confirmed there was no cross-reactivity with sera positive for common equine pathogens. Using a commercial competitive enzyme-linked immunosorbent assay (cELISA) kit for comparison, 476 clinical samples were tested with the card. The coincidence rates were 96.43% and 97.90% for T. equi and B. caballi, respectively. The field trial feedback was uniformly positive, suggesting that this diagnostic tool may be useful for controlling the spread of T. equi and B. caballi. IMPORTANCE Equine piroplasmosis (EP), caused by Theileria equi and Babesia caballi, is an important tick-borne disease of equines that is prevalent in most parts of the world. EP is considered a reportable disease by the World Organization for Animal Health (OIE). The accurate diagnosis and differentiation of T. equi and B. caballi are very important for the prevention, control, and treatment of EP. Therefore, we developed a double-antigen sandwich colloidal gold immunochromatography assay (GICG) to detect T. equi and B. caballi. Two GICG strips were assembled side by side on one card for the detection of T. equi and B. caballi, and the two EP pathogens could be detected in one test. This method was simple, rapid, and specific for the detection of EP; therefore, compared to the previous methods, this method is more suitable for pathogen diagnosis in the field.


Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Babesia/inmunología , Babesiosis/sangre , Enfermedades de los Caballos/sangre , Inmunoensayo/métodos , Theileria/inmunología , Theileriosis/sangre , Animales , Babesia/genética , Babesia/aislamiento & purificación , Babesiosis/diagnóstico , Babesiosis/parasitología , Oro Coloide/química , Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/parasitología , Caballos , Inmunoensayo/instrumentación , Theileria/genética , Theileria/aislamiento & purificación , Theileriosis/diagnóstico , Theileriosis/parasitología
4.
Int J Mol Sci ; 23(3)2022 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-35163517

RESUMEN

Canine babesiosis is an important tick-borne disease worldwide, caused by parasites of the Babesia genus. Although the disease process primarily affects erythrocytes, it may also have multisystemic consequences. The goal of this study was to explore and characterize the serum metabolome, by identifying potential metabolites and metabolic pathways in dogs naturally infected with Babesia canis using liquid and gas chromatography coupled to mass spectrometry. The study included 12 dogs naturally infected with B. canis and 12 healthy dogs. By combining three different analytical platforms using untargeted and targeted approaches, 295 metabolites were detected. The untargeted ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) metabolomics approach identified 64 metabolites, the targeted UHPLC-MS/MS metabolomics approach identified 205 metabolites, and the GC-MS metabolomics approach identified 26 metabolites. Biological functions of differentially abundant metabolites indicate the involvement of various pathways in canine babesiosis including the following: glutathione metabolism; alanine, aspartate, and glutamate metabolism; glyoxylate and dicarboxylate metabolism; cysteine and methionine metabolism; and phenylalanine, tyrosine, and tryptophan biosynthesis. This study confirmed that host-pathogen interactions could be studied by metabolomics to assess chemical changes in the host, such that the differences in serum metabolome between dogs with B. canis infection and healthy dogs can be detected with liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) methods. Our study provides novel insight into pathophysiological mechanisms of B. canis infection.


Asunto(s)
Babesia/patogenicidad , Babesiosis/sangre , Enfermedades de los Perros/parasitología , Metabolómica/métodos , Animales , Estudios de Casos y Controles , Cromatografía Liquida , Enfermedades de los Perros/sangre , Perros , Cromatografía de Gases y Espectrometría de Masas , Interacciones Huésped-Patógeno , Redes y Vías Metabólicas , Espectrometría de Masas en Tándem
5.
Parasit Vectors ; 15(1): 10, 2022 Jan 07.
Artículo en Inglés | MEDLINE | ID: mdl-34991686

RESUMEN

BACKGROUND: Drug resistance and toxic side effects are major challenges in the treatment of babesiosis. As such, new drugs are needed to combat the emergence of drug resistance in Babesia parasites and to develop alternative treatment strategies. A combination of naphthoquine (NQ) and artemisinin is an antimalarial therapy in pharmaceutical markets. The present study repurposed NQ as a drug for the treatment of babesiosis by evaluating the anti-Babesia activity of naphthoquine phosphate (NQP) alone. METHODS: An in vitro growth inhibition assay of NQP was tested on Babesia gibsoni cultures using a SYBR Green I-based fluorescence assay. In addition, the in vivo growth inhibitory effect of NQP was evaluated using BALB/c mice infected with Babesia rodhaini. The parasitemia level and hematocrit values were monitored to determine the therapeutic efficacy of NQP and the clinical improvements in NQP-treated mice. RESULTS: The half maximal inhibitory concentration of NQP against B. gibsoni in vitro was 3.3 ± 0.5 µM. Oral administration of NQP for 5 consecutive days at a dose of 40 mg/kg of body weight resulted in significant inhibition of B. rodhaini growth in mice as compared with that of the control group. All NQP-treated mice survived, whereas the mice in the control group died between days 6 and 9 post-infection. CONCLUSION: This is the first study to evaluate the anti-Babesia activity of NQP in vitro and in vivo. Our findings suggest that NQP is a promising drug for treating Babesia infections, and drug repurposing may provide new treatment strategies for babesiosis.


Asunto(s)
1-Naftilamina/análogos & derivados , Aminoquinolinas/farmacología , Antiprotozoarios/farmacología , Babesia/efectos de los fármacos , Babesiosis/tratamiento farmacológico , 1-Naftilamina/farmacología , 1-Naftilamina/uso terapéutico , Aminoquinolinas/sangre , Aminoquinolinas/uso terapéutico , Animales , Antiprotozoarios/sangre , Antiprotozoarios/uso terapéutico , Babesia/crecimiento & desarrollo , Babesiosis/sangre , Babesiosis/parasitología , Hematócrito , Concentración 50 Inhibidora , Ratones , Ratones Endogámicos BALB C , Parasitemia/parasitología , Distribución Aleatoria
6.
Vet Parasitol ; 300: 109612, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34735844

RESUMEN

Inflammation is a hallmark of the acute Babesia canis infection. Promatrix metalloproteinase (proMMP)-2 and -9 are involved in inflammation, but their levels have not been analyzed in canine babesiosis. We hypothesized that in dogs infected with B. canis, serum proMMP-2 and -9 levels change between presentation and recovery. Degree of the change differs if dogs develop systemic inflammatory response syndrome (SIRS). This study included 24 dogs with an acute B. canis infection, at presentation and after two weeks. We used routine hematology and biochemistry methods, spectrophotometry for the acute-phase proteins, microscopy for parasitemia and zymography for (pro)MMPs. In vitro endothelial cells and leukocyte short-term cultures, and platelet lysates were used to detect specific MMP activity. Statistical analyses included Wilcoxon test for paired samples, Mann-Whitney U test and Spearman's rank correlation. Our results showed that endothelial cells, leukocytes and platelets are the source of proMMP-2 and proMMP-9. Furthermore, both proMMPs were lower at presentation than after recovery (p < 0.001). At presentation, proMMP-9 levels correlated with parasitemia (rho = -0.616, p = 0.009), total leukocyte (rho = 0.704, p < 0.001) and neutrophil counts (rho = 0.741, p < 0.001). Extent of alterations in proMMP-2 levels between presentation and recovery was lower (p = 0.038) in dogs with SIRS than in non-SIRS dogs, while levels of proMMP-9 were comparable between these groups. Our conclusion is that during the acute B. canis infection, low serum levels of proMMP-2 and proMMP-9 at presentation reflect thrombocytopenia and leukopenia. Decreased proMMP-2 level could be associated with SIRS.


Asunto(s)
Babesiosis , Enfermedades de los Perros , Metaloproteinasa 2 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/sangre , Animales , Babesia , Babesiosis/sangre , Enfermedades de los Perros/sangre , Enfermedades de los Perros/parasitología , Perros , Células Endoteliales
7.
Transfusion ; 61(10): 2969-2980, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34368968

RESUMEN

BACKGROUND: Human babesiosis is a zoonotic infection caused by an intraerythrocytic parasite. The highest incidence of babesiosis is in the United States, although cases have been reported in other parts of the world. Due to concerns of transfusion-transmitted babesiosis, the US Food and Drug Administration (FDA) recommended year-round regional testing for Babesia by nucleic acid testing or use of an FDA-approved device for pathogen reduction. A new molecular test, cobas Babesia (Roche Molecular Systems, Inc.), was evaluated for the detection of the four species that cause human disease, Babesia microti, Babesia duncani, Babesia divergens, and Babesia venatorum. STUDY DESIGN AND METHODS: Analytical performance was evaluated followed by clinical studies on whole blood samples from US blood donations collected in a special tube containing a chaotropic reagent that lyses the red cells and preserves nucleic acid. Sensitivity and specificity of the test in individual samples (individual donation testing [IDT]) and in pools of six donations were determined. RESULTS: Based on analytical studies, the claimed limit of detection of cobas Babesia for B. microti is 6.1 infected red blood cells (iRBC)/mL (95% confidence interval [CI]: 5.0, 7.9); B. duncani was 50.2 iRBC/mL (95% CI: 44.2, 58.8); B. divergens was 26.1 (95% CI: 22.3, 31.8); and B. venatorum was 40.0 iRBC/mL (95% CI: 34.1, 48.7). The clinical specificity for IDT was 99.999% (95% CI: 99.996, 100) and 100% (95% CI: 99.987, 100) for pools of six donations. CONCLUSION: cobas Babesia enables donor screening for Babesia species with high sensitivity and specificity.


Asunto(s)
Babesia/aislamiento & purificación , Babesiosis/sangre , Donantes de Sangre , ADN Protozoario/sangre , ARN Protozoario/sangre , Babesia/genética , Babesia microti/genética , Babesia microti/aislamiento & purificación , Babesiosis/diagnóstico , Babesiosis/microbiología , ADN Protozoario/genética , Pruebas Diagnósticas de Rutina , Selección de Donante , Humanos , ARN Protozoario/genética , Sensibilidad y Especificidad , Estados Unidos
8.
Front Immunol ; 12: 623492, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34079537

RESUMEN

Babesia orientalis, a major infectious agent of water buffalo hemolytic babesiosis, is transmitted by Rhipicephalus haemaphysaloides. However, no effective vaccine is available. Essential antigens that are involved in parasite invasion of host red blood cells (RBCs) are potential vaccine candidates. Therefore, the identification and the conduction of functional studies of essential antigens are highly desirable. Here, we evaluated the function of B. orientalis merozoite surface antigen 2c1 (BoMSA-2c1), which belongs to the variable merozoite surface antigen (VMSA) family in B. orientalis. We developed a polyclonal antiserum against the purified recombinant (r)BoMSA-2c1 protein. Immunofluorescence staining results showed that BoMSA-2c1 was expressed only on extracellular merozoites, whereas the antigen was undetectable in intracellular parasites. RBC binding assays suggested that BoMSA-2c1 specifically bound to buffalo erythrocytes. Cytoadherence assays using a eukaryotic expression system in vitro further verified the binding and inhibitory ability of BoMSA-2c1. We found that BoMSA-2c1 with a GPI domain was expressed on the surface of HEK293T cells that bound to water buffalo RBCs, and that the anti-rBoMSA2c1 antibody inhibited this binding. These results indicated that BoMSA-2c1 was involved in mediating initial binding to host erythrocytes of B. orientalis. Identification of the occurrence of binding early in the invasion process may facilitate understanding of the growth characteristics, and may help in formulating strategies for the prevention and control of this parasite.


Asunto(s)
Antígenos de Protozoos/metabolismo , Antígenos de Superficie/metabolismo , Babesia/metabolismo , Babesiosis/parasitología , Adhesión Celular , Eritrocitos/parasitología , Merozoítos/metabolismo , Proteínas Protozoarias/metabolismo , Animales , Antígenos de Protozoos/genética , Antígenos de Superficie/genética , Babesia/genética , Babesia/patogenicidad , Babesiosis/sangre , Búfalos , Eritrocitos/metabolismo , Células HEK293 , Humanos , Merozoítos/genética , Merozoítos/patogenicidad , Proteínas Protozoarias/genética
10.
Vet Rec ; 188(5): e29, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33666974

RESUMEN

BACKGROUND: We aimed to identify prognostic markers and their discriminant score in predicting the lethal outcome of canine Babesia gibsoni. METHODS: Blood samples were collected from 108 client-owned dogs with clinical signs commensurate with babesiosis to analyze haematological, biochemical, haemostatic, antioxidant profile and thiobarbituric acid reactive substance levels. Samples were screened for Babesia infection (microscopic and molecular techniques). Babesiosis-affected dogs were classified into survivors and non-survivors, and 30 healthy dogs were used in the control group. RESULTS: Haemoglobin, thrombocytes, catalase, urea, creatinine, alanine aminotransferase (ALT), alkaline phosphatase, lactate and reticulocytes were highly correlated to survival. Receiver operating characteristics analysis revealed urea, ALT and lactate as specific prognostic markers for the disease. The formula for calculation of discriminant scores (Di) for lethal outcome of the disease was generated with cut-off score 0.141. The scoring system was 79% sensitive and 83% specific in predicting the lethal outcome of the disease. CONCLUSIONS: A scoring system developed from the prognosticating markers may aid in predicting the outcome of Babesia gibsoni infection on the day of presentation itself enabling intensive care for those animals with a cut-off score more than 0.141.


Asunto(s)
Babesia/clasificación , Babesiosis/parasitología , Babesiosis/terapia , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/terapia , Animales , Babesiosis/sangre , Biomarcadores/sangre , Estudios de Casos y Controles , Enfermedades de los Perros/sangre , Perros , Femenino , Masculino , Pronóstico , Resultado del Tratamiento
11.
Parasitol Res ; 120(5): 1915-1919, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33740119

RESUMEN

This study reports the identification and first molecular characterization of Babesia occultans from naturally infected cows in Iran. Microscopic examination showed pyriform trophozoites, and ring-shaped merozoites (>2.5 µm) in Giemsa-stained blood smears obtained from two symptomatic cows in West-Azarbaijan province, Iran. PCR amplification of the partial 18S rRNA gene including the V4 hypervariable region were carried out on DNA extracted from blood samples. BLAST analyses of the partial 18S rRNA (approximately 400 bp) obtained from two cows revealed the presence of B. occultans and the detected sequences were identical to each other. Comparisons of the partial 18S rRNA sequence of the current isolate with other B. occultans sequences from Tunisia, South Africa, Turkey, Pakistan, and China confirmed the relation of the Iranian isolate to the species B. occultans. Sequence analysis of the obtained B. occultans showed 99.5-100% identity to the previously reported isolates. The sequences of B. occultans had 100% identity to a sequence obtained from ticks in Tunisia. This report is the beginning of a path to further research about B. occultans in vectors and reservoirs throughout Iran.


Asunto(s)
Babesia/aislamiento & purificación , Babesiosis/parasitología , Enfermedades de los Bovinos/parasitología , Animales , Babesia/genética , Babesiosis/sangre , Bovinos , ADN Protozoario/genética , Femenino , Irán , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , ARN Ribosómico 18S , Garrapatas/genética
13.
Protein J ; 40(1): 119-130, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33387247

RESUMEN

Babesia microti is a protozoan that mainly parasitizes rodent and human erythrocytes. B. microti infection can result in changes in the expression levels of various proteins in the host serum. To explore the mechanism underlying the regulation of serum proteins by the host during B. microti infection, this study used a data-independent acquisition (DIA) quantitative proteomic approach to perform comprehensive quantitative proteomic analysis on the serum of B. microti-infected mice. We identified and analysed 333 serum proteins during the infectious stage and recovery stage within 30 days of infection by B. microti in mice. Through quantitative analysis, we found 57 proteins differentially expressed in the infection stage and 69 proteins differentially expressed in the recovery stage. Bioinformatics analysis revealed that these differentially expressed proteins were mainly concentrated in organelles, cell parts, and extracellular regions that are mainly involved in immune system, metabolic, and cellular processes. Additionally, the differentially expressed proteins mainly had catalytic activity. Kyoto Encyclopedia of Genes and Genome (KEGG) pathway analysis showed that many of the differentially expressed proteins participate in the complement and coagulation cascade reaction, including complement C3, complement FP, and coagulation factor XII. The results of this study can provide more information for the selection of biomarkers for the early clinical monitoring of babesiosis and help in the treatment of babesiosis.


Asunto(s)
Babesia microti/inmunología , Babesiosis/genética , Proteínas Sanguíneas/genética , Proteínas del Sistema Complemento/genética , Interacciones Huésped-Patógeno/genética , Redes y Vías Metabólicas/genética , Animales , Babesia microti/crecimiento & desarrollo , Babesiosis/sangre , Babesiosis/inmunología , Babesiosis/parasitología , Biomarcadores/sangre , Proteínas Sanguíneas/clasificación , Proteínas Sanguíneas/inmunología , Proteínas del Sistema Complemento/clasificación , Proteínas del Sistema Complemento/inmunología , Factor XII/genética , Factor XII/inmunología , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Ontología de Genes , Interacciones Huésped-Patógeno/inmunología , Redes y Vías Metabólicas/inmunología , Ratones , Ratones Endogámicos BALB C , Anotación de Secuencia Molecular , Análisis de Componente Principal , Proteómica/métodos
14.
J Vet Diagn Invest ; 33(1): 73-79, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33252025

RESUMEN

Detection of bovine Babesia spp. and Anaplasma marginale is based on the reading of Giemsa-stained blood or organ smears, which can have low sensitivity. Our aim was to improve the detection of bovine Babesia spp. and A. marginale by validating a multiplex PCR (mPCR). We used 466 samples of blood and/or organs of animals with signs and presumptive autopsy findings of babesiosis or anaplasmosis. The primers in our mPCR amplified the rap-1a gene region of Babesia bovis and B. bigemina, and the msp-5 region of A. marginale. We used a Bayesian model with a non-informative priori distribution for the prevalence estimate and informative priori distribution for estimation of sensitivity and specificity. The sensitivity and specificity for smear detection of Babesia spp. were 68.6% and 99.1%, and for A. marginale 85.6% and 98.8%, respectively. Sensitivity and specificity for mPCR detection for Babesia spp. were 94.2% and 97.1%, and for A. marginale 95.2% and 92.7%, respectively. Our mPCR had good accuracy in detecting Babesia spp. and A. marginale, and would be a reliable test for veterinarians to choose the correct treatment for each agent.


Asunto(s)
Anaplasma marginale/aislamiento & purificación , Anaplasmosis/diagnóstico , Babesia/aislamiento & purificación , Babesiosis/diagnóstico , Enfermedades de los Bovinos/diagnóstico , Anaplasma marginale/genética , Anaplasmosis/sangre , Animales , Babesia/genética , Babesiosis/sangre , Bovinos , Enfermedades de los Bovinos/sangre , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Sensibilidad y Especificidad , Uruguay
15.
Vet Parasitol ; 289: 109338, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33359970

RESUMEN

Canine babesiosis, a tick-borne haemoprotozoan disease of dogs, is of significance globally due to its rapid spread. A precise confirmatory diagnosis is required to curtail the rapid spread of infection. Our study described the evaluation of recombinant BgSA3 protein based indirect ELISA for sero-diagnosis and sero-surveillance of Babesia gibsoni infection in dogs. A partial BgSA3 gene segment (1921 bp) of B. gibsoni, encoding for recombinant truncated BgSA3 (75 kDa) protein devoid of predicted signal peptide (23 aa) at N-terminus and transmembrane region (20 aa) at C-terminus, was expressed in E. coli using a pET28a(+) vector. The rBgSA3 protein purified under native conditions using Ni-NTA superflow cartridge was confirmed by SDS-PAGE and Western blotting using sera from dogs infected/uninfected with B. gibsoni, and erythrocyte lysate/ plasma from infected/uninfected dogs. The rBgSA3 protein was specific only to B. gibsoni antibodies but did not react with uninfected sera. Further, rBgSA3 protein was evaluated for sero-diagnosis/sero-surveillance using Indirect-ELISA format. There was no cross reactivity to B. vogeli, E. canis, H. canis and D. repens infected dogs serum samples. The diagnostic sensitivity and specificity of rBgSA3 based I-ELISA was found to be 86.4 and 93.1 % respectively, in comparison with cytb based PCR assay. Additionally, rBgSA3-ELISA evaluated using survey serum samples (n = 287), detected 11.85 % samples as positive. In conclusion, B. gibsoni infection, an emerging disease is prevalent in the present study area and the standardized rBgSA3 protein based indirect-ELISA was found to be a specific and sensitive test for large scale sero-diagnosis and sero-surveillance of B. gibsoni infection in dogs.


Asunto(s)
Babesia/aislamiento & purificación , Babesiosis/parasitología , Enfermedades de los Perros/parasitología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Proteínas Protozoarias/sangre , Animales , Antígenos de Protozoos , Babesiosis/sangre , Babesiosis/diagnóstico , Enfermedades de los Perros/diagnóstico , Perros , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Masculino , Vigilancia de la Población , Estudios Seroepidemiológicos
16.
Sci Rep ; 10(1): 18904, 2020 11 03.
Artículo en Inglés | MEDLINE | ID: mdl-33144631

RESUMEN

Canine babesiosis may cause several hematological and biochemical changes, but only limited studies are available regarding the possible differences of changes in animals infected by different Babesia parasites. The study focused on the evaluation of the differences in serum protein electrophoretic pattern between dogs naturally infected with B. gibsoni (17 dogs) and B. canis (40 dogs). The mean values of total proteins, ß1-, ß2- and γ-globulins were in dogs infected with B. gibsoni significantly higher (P < 0.05 and P < 0.001) than in dogs infected with B. canis. The relative concentrations of albumin, α1-, α2-globulins and the A/G ratios were in the B. gibsoni infected dogs significantly lower (P < 0.001), no significant differences were found in the relative concentrations of ß1- and ß2-globulins. Significant differences were found in most of the evaluated parameters when comparing the results in relation to the form of B. canis infection to B. gibsoni infection. Hematological indices showed significant differences between dogs infected with B. gibsoni and the complicated form of B. canis infection. In conclusion, the obtained results suggest differences in the changes of serum protein electrophoretic pattern between dogs infected with both Babesia species and thus, in the response to the infection caused by various Babesia parasites.


Asunto(s)
Babesia/clasificación , Babesiosis/sangre , Electroforesis de las Proteínas Sanguíneas/veterinaria , Proteínas Sanguíneas/análisis , Enfermedades de los Perros/parasitología , alfa-Globulinas/análisis , Animales , beta-Globulinas/análisis , Enfermedades de los Perros/sangre , Perros , Femenino , Masculino , Albúmina Sérica/análisis , gammaglobulinas/análisis
17.
Parasit Vectors ; 13(1): 537, 2020 Oct 29.
Artículo en Inglés | MEDLINE | ID: mdl-33121531

RESUMEN

BACKGROUND: Parasitic infections are among the important causes of death of giant pandas (Ailuropoda melanoleuca) that hamper their survival in the wild. There are about 35 species of parasites which have been identified in giant pandas, but no information is currently available regarding the infection of Babesia in giant pandas. Babesia spp. are common intraerythrocytic parasite in wildlife, transmitted by ixodid ticks, which cause babesiosis. Clinical signs of babesiosis include fever, hemolysis, anemia, jaundice and death. METHODS: A species of Babesia was detected in the blood of a giant panda based on morphology and PCR amplification of the 18S rRNA gene. The phylogenetic relationship of Babesia sp. infecting giant panda was assessed by gene sequence alignment and phylogenetic analysis. RESULTS: Our analysis revealed that the Babesia isolate detected was most similar to an unidentified species of Babesia identified in black bears (Ursus thibetanus japonicus) from Japan (Babesia sp. Iwate, AB586027.1) with a 99.56% sequence similarity, followed by Babesia sp. EBB (AB566229.1, 99.50%) and Babesia sp. Akita (AB566229.1, 99.07%). CONCLUSIONS: To our knowledge, this is the first report of Babesia detected in the giant panda. The results indicate that this Babesia sp. may be a novel species, currently named Babesia sp. strain EBP01.


Asunto(s)
Babesia/clasificación , Babesiosis/parasitología , Filogenia , Ursidae/parasitología , Animales , Babesia/aislamiento & purificación , Babesiosis/sangre , China , Femenino , ARN Ribosómico 18S/genética , Alineación de Secuencia
18.
Rev Bras Parasitol Vet ; 29(3): e012420, 2020 Aug 03.
Artículo en Inglés | MEDLINE | ID: mdl-32756775

RESUMEN

Piroplasm species were analyzed by molecular tools in total 31 blood samples from positive dogs, previously checked by stained slides, stored until DNA extraction between 2016 to 2018 in the laboratory Clinical Analyzes in Niterói, Rio de Janeiro. The piroplasms were identified by PCR, targeting the 18S rRNA gene and sequencing. From the total number of samples only 24 (77.4%) were positive and show adequate nucleotide sequences for interpretation with identity between 93%-100% with Babesia vogeli in compared to the sequences isolated of infected dogs from other states in Brazil deposited on GenBank. Most of dogs infected with B. vogeli had anemia (62.5%) and thrombocytopenia (95.8%). The findings of this study are compatible with previous reports in the literature and highlight B. vogeli as the most incriminated species in canine piroplasmosis in Brazil, and thrombocytopenia the hematological alteration most frequently identified in this infection. It is important to note that this is the first study involving the molecular characterization of piroplasms in the metropolitan region of Rio de Janeiro, based on PCR followed by sequencing.


Asunto(s)
Babesia , Babesiosis , Sangre , Enfermedades de los Perros , Manejo de Especímenes , Animales , Babesia/genética , Babesiosis/sangre , Sangre/parasitología , Análisis Químico de la Sangre , Brasil , Enfermedades de los Perros/sangre , Enfermedades de los Perros/parasitología , Perros , ARN Ribosómico 18S/genética , Manejo de Especímenes/veterinaria
19.
Parasitol Res ; 119(12): 4151-4158, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32715343

RESUMEN

This study was designed to evaluate the effects of Babesia ovis infection on concentrations of some essential acute phase proteins (APPs) including albumin, fibrinogen, serum amyloid A, haptoglobin, and ceruloplasmin as well as total, protein-binding, and lipid-binding sialic acids (TSA, PBSA, and LBSA) and two crucial cytokines including interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α). Some hematological parameters also were evaluated. Furthermore, any probable correlation among the APPs, SAs, IFN-γ, and TNF-α was calculated. A total of 420 Marghoz and Raeini goats with the ages of 1-3 years old from the north and northwest of Iran were examined, and 17 goats confirmed to be infected with B. ovis by both routine microscopic examination of blood films and molecular assays. As the control, 17 healthy goats were included. The results revealed a significant decrease (P < 0.05) in erythrocyte count, hemoglobin level, and pack cell volume as well as a nonsignificant increase in white blood cell count in the diseased animals compared with the control. Additionally, all the APPs, SAs, and cytokines were remarkably higher in the infected animals than the uninfected ones, except for albumin, which was significantly lower. Moreover, a strong and positive correlation was detected among the parameters mentioned above, except for albumin, which was inversely correlated with the other parameters. In conclusion, B. ovis infection is associated with the induction of severe inflammatory reactions in goats, and both SA and APP are significantly involved in the pathophysiology of the disease.


Asunto(s)
Proteínas de Fase Aguda/análisis , Babesiosis/sangre , Citocinas/sangre , Enfermedades de las Cabras/sangre , Enfermedades de las Cabras/parasitología , Animales , Babesia , Biomarcadores/análisis , Biomarcadores/sangre , Recuento de Eritrocitos , Cabras/parasitología , Inflamación/sangre , Inflamación/patología , Interferón gamma/sangre , Irán , Ovinos , Factor de Necrosis Tumoral alfa/sangre
20.
Parasit Vectors ; 13(1): 369, 2020 Jul 22.
Artículo en Inglés | MEDLINE | ID: mdl-32698835

RESUMEN

BACKGROUND: The tick-borne intra-erythrocytic apicomplexan Babesia caballi is one of the etiological agents of equine babesiosis, an economically important disease of equids in most tropical and subtropical areas of the world. Discovering candidate antigens for improved diagnostic tools and vaccines remains needed for controlling equine babesiosis. This study describes the B. caballi sbp4 (Bcsbp4) gene and protein (BcSBP4) and analyzes its antigenicity in infected equids. METHODS: BLAST searches of an uncurated B. caballi assembly genome using the B. bovis SBP4 as a query were carried out, followed by PCR amplification and sequencing of a newly identified BcSBP4. Characterization of this novel gene and protein was performed by bioinformatics analysis, western blots, immunofluorescence (IFA) and an in vitro neutralization test using anti SBP4 peptide antibodies. Antigenicity of recombinant BcSBP4 (rBcSBP4) was tested with sera from field animals (n = 18) using an indirect ELISA (iELISA). RESULTS: Babesia caballi genome searches using B. bovis SBP4 as a query allowed identification of a novel gene termed Bcsbp4. The Bcsbp4 gene encodes for a protein of 30.58 kDa, which is fully conserved among B. caballi isolates from USA and Egypt. Bioinformatics analysis indicates that BcSBP4 contains a signal peptide and lacks additional transmembrane domains. Expression of BcSBP4 in blood stages of B. caballi was confirmed by western blot and IFA using antibodies against synthetic peptides representing putative B-cell epitopes of BcSBP4 predicted by in silico analysis. In vitro neutralization tests using anti-BcSBP4 peptide antibodies showed a marginal, but statistically significant inhibitory effect on the infectivity of B. caballi merozoites in horse red blood cells. Sera from eight B. caballi-infected equids, but none out of ten negative equid control sera, gave a positive signal in an rBcSBP4 based iELISA. CONCLUSIONS: The Bcsbp4 gene is expressed in B. caballi blood stages. The BcSBP4 protein is a potential candidate for developing a novel serological test that could detect B. caballi infection in equids in tropical and subtropical countries worldwide.


Asunto(s)
Babesia , Caballos/parasitología , Proteínas Protozoarias/inmunología , Animales , Anticuerpos Antiprotozoarios/sangre , Babesia/citología , Babesia/genética , Babesia/inmunología , Babesia/metabolismo , Babesiosis/sangre , Babesiosis/diagnóstico , Genes Protozoarios , Enfermedades de los Caballos/diagnóstico , Filogenia , Proteínas Protozoarias/metabolismo , Pruebas Serológicas/métodos
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