Diagnostic Tools for the Identification of Babesia sp. in Persistently Infected Cattle

Bovine babesiosis is a tick-borne disease of cattle caused by the protozoan parasites of the genus Babesia. Babesia bovis, Babesia bigemina and Babesia divergens are considered by International health authorities (OIE) as the principal species of Babesia that cause bovine babesiosis. Animals that recover from a babesial primo infection may remain as persistent carriers with no clinical signs of disease and can be the source of infection for ticks that are able to acquire Babesia parasites from infected cattle and to transmit Babesia parasites to susceptible cattle. Several procedures that have been developed for parasite detection and diagnosis of this infectious carrier state constitute the basis for this review: A brief description of the direct microscopic detection of Babesia-infected erytrocytes; PCR-based diagnostic assays, which are very sensitive particularly in detecting Babesia in carrier cattle; in-vitro culture methods, used to demonstrate presence of carrier infections of Babesia sp.; animal inoculation, particularly for B. divergens isolation are discussed. Alternatively, persistently infected animals can be tested for specific antibabesial antibodies by using indirect serological assays. Serological procedures are not necessarily consistent in identifying persistently infected animals and have the disadvantage of presenting with cross reactions between antibodies to Babesia sp.

Download Full-text

Assessing the Immunochromatographic Test Strip for Serological Detection of Bovine Babesiosis in Uganda

Microorganisms ◽

10.3390/microorganisms8081110 ◽

2020 ◽

Vol 8 (8) ◽

pp. 1110 ◽

Cited By ~ 2

Author(s):

Dickson Stuart Tayebwa ◽

Amany Magdy Beshbishy ◽

Gaber El-Saber Batiha ◽

Mariam Komugisha ◽

Byaruhanga Joseph ◽

...

Keyword(s):

Test Strip ◽

Spherical Body ◽

Babesia Bovis ◽

Diagnostic Tools ◽

Immunochromatographic Test ◽

Babesia Bigemina ◽

Bovine Babesiosis ◽

Body Protein ◽

Eastern Uganda ◽

Good Agreement

In Uganda, bovine babesiosis continues to cause losses to the livestock industry because of shortages of cheap, quick, and reliable diagnostic tools to guide prescription measures. In this study, the presence of antibodies to Babesia bigemina and Babesia bovis in 401 bovine blood samples obtained from eastern and central areas of Uganda were detected using enzyme-linked immunosorbent assays (ELISAs) and immunochromatographic test strips (ICTs). The ELISA and ICT test used targeted the B. bigemina C-terminal rhoptry-associated protein (RAP-1/CT17) and B. bovis spherical body protein-4 (SPB-4). Using ELISA, single-ICT and dual-ICT, positive samples for B. bovis were detected in 25 (6.2%), 17 (4.3%), and 14 (3.7%) samples respectively, and positive samples for B. bigemina were detected in 34 (8.4%), 27 (6.7%), and 25 (6.2%), respectively. Additionally, a total of 13 animals (3.2%) had a mixed infection. The correlation between ELISA and single-ICT strips results revealed slight agreement with kappa values ranging from 0.088 to 0.191 between both methods, while the comparison between dual-ICT and single-ICT results showed very good agreement with kappa values >0.80. This study documented the seroprevalence of bovine babesiosis in central and eastern Uganda, and showed that ICT could, after further optimization, be a useful rapid diagnostic test for the diagnosis of bovine babesiosis in field settings.

Download Full-text

First Report on Molecular Identification of Babesia bigemina in a Nervous Signs Evident Naturally Infected Holstein-Friesian Cattle from Central India

Indian Journal of Animal Research ◽

10.18805/ijar.b-4306 ◽

2021 ◽

Author(s):

V. Agrawal ◽

G. Das ◽

A. Jaiswal ◽

A.K. Jayraw ◽

G.P. Jatav ◽

...

Keyword(s):

Blood Sample ◽

Genomic Dna ◽

Blood Smear ◽

Clinical Signs ◽

Central India ◽

Babesia Bovis ◽

Thin Blood Smear ◽

Babesia Bigemina ◽

Bovine Babesiosis ◽

Holstein Friesian

Background: Bovine babesiosis caused by an intraerythrocytic apicomplexan protozoon responsible for the most prevalent and costly tick borne diseases (TBD’s) of cattle throughout the globe. Cerebral babesiosis of bovine is fatal and mainly caused by Babesia bovis. To the knowledge of author, there is no confirm molecular report of Babeisa bigemina caused cerebral babesiosis in cattle. Therefore, authors want to report Babesia bigemina caused cerebral babesiosis on record. Methods: In the year 2015, a Holstein-Friesian cow aged 3 years and weighing approximately 300 kg, was attended at Jabalpur, (M.P.) with the clinical signs of high rise in temperature (104°F), recumbency, severe dysponea, peculiar sound during open mouth breathing, pale color of eye conjunctiva and mucous membrane of vagina, convulsions, sever anaemia, paddling of legs at frequent interval. After preparation of peripheral thin blood smear from animal at the site of collection and fixation with methanol, blood sample brought to Department of Veterinary Parasitology, College of Veterinary Science and A.H, Jabalpur and stained by standard protocol for Giemsa staining. Genomic DNA was isolated from the collected blood sample using QIAamp® DNA blood mini kit following the manufacturer’s recommendations and PCR was performed. Conclusion: The thin blood smear examination revealed the presence of Babesia parasite. The species of Babesia was confirmed by molecular amplification of genomic DNA as B. bigemina. This might be the first confirmed report of cerebral babesiosis caused by B. bigemina from Central India.

Download Full-text

Bovine babesiosis: Cattle protected in the field with a frozen vaccine containing Babesia bovis and Babesia bigemina cultured in vitro with a serum-free medium

Parasitology International ◽

10.1016/j.parint.2017.11.004 ◽

2018 ◽

Vol 67 (2) ◽

pp. 190-195 ◽

Cited By ~ 6

Author(s):

Carmen Rojas-Martínez ◽

Roger Iván Rodríguez-Vivas ◽

Julio Vicente Figueroa Millán ◽

Carlos Ramón Bautista-Garfias ◽

Roberto Omar Castañeda-Arriola ◽

...

Keyword(s):

Babesia Bovis ◽

Serum Free Medium ◽

Free Medium ◽

Babesia Bigemina ◽

Bovine Babesiosis ◽

Serum Free

Download Full-text

Applying Machine Learning to Predict the Exportome of Bovine and Canine Babesia Species That Cause Babesiosis

Pathogens ◽

10.3390/pathogens10060660 ◽

2021 ◽

Vol 10 (6) ◽

pp. 660

Author(s):

Stephen J. Goodswen ◽

Paul J. Kennedy ◽

John T. Ellis

Keyword(s):

Machine Learning ◽

Severe Disease ◽

Economic Loss ◽

Babesia Bovis ◽

Training Data ◽

Babesia Bigemina ◽

Bovine Babesiosis ◽

Canine Babesiosis ◽

Clinically Significant ◽

Significant Disease

Babesia infection of red blood cells can cause a severe disease called babesiosis in susceptible hosts. Bovine babesiosis causes global economic loss to the beef and dairy cattle industries, and canine babesiosis is considered a clinically significant disease. Potential therapeutic targets against bovine and canine babesiosis include members of the exportome, i.e., those proteins exported from the parasite into the host red blood cell. We developed three machine learning-derived methods (two novel and one adapted) to predict for every known Babesia bovis, Babesia bigemina, and Babesia canis protein the probability of being an exportome member. Two well-studied apicomplexan-related species, Plasmodium falciparum and Toxoplasma gondii, with extensive experimental evidence on their exportome or excreted/secreted proteins were used as important benchmarks for the three methods. Based on 10-fold cross validation and multiple train–validation–test splits of training data, we expect that over 90% of the predicted probabilities accurately provide a secretory or non-secretory indicator. Only laboratory testing can verify that predicted high exportome membership probabilities are creditable exportome indicators. However, the presented methods at least provide those proteins most worthy of laboratory validation and will ultimately save time and money.

Download Full-text

Assessment of Draxxin® (tulathromycin) as an inhibitor of in vitro growth of Babesia bovis, Babesia bigemina and Theileria equi

International Journal for Parasitology Drugs and Drug Resistance ◽

10.1016/j.ijpddr.2018.04.004 ◽

2018 ◽

Vol 8 (2) ◽

pp. 265-270 ◽

Cited By ~ 10

Author(s):

Marta G. Silva ◽

Nicolas F. Villarino ◽

Donald P. Knowles ◽

Carlos E. Suarez

Keyword(s):

Babesia Bovis ◽

In Vitro Growth ◽

Babesia Bigemina ◽

Theileria Equi

Download Full-text

Epidemiology and genotyping of Anaplasma marginale and co-infection with piroplasms and other Anaplasmataceae in cattle and buffaloes from Egypt

Parasites & Vectors ◽

10.1186/s13071-020-04372-z ◽

2020 ◽

Vol 13 (1) ◽

Author(s):

Amira AL-Hosary ◽

Cristian Răileanu ◽

Oliver Tauchmann ◽

Susanne Fischer ◽

Ard M. Nijhof ◽

...

Keyword(s):

Blood Smear ◽

Anaplasma Marginale ◽

Carrier State ◽

Babesia Bovis ◽

Diagnostic Tools ◽

Enzyme Linked Immunosorbent Assay ◽

Babesia Bigemina ◽

Infection Level ◽

Significant Difference ◽

One Year

Abstract Background Anaplasma marginale is an obligate intracellular bacterium and the main cause of bovine anaplasmosis in tropical and subtropical regions. In Egypt, data regarding the prevalence of A. marginale in ruminant hosts and of the circulating genotypes is lacking. This study therefore aimed to (i) investigate the presence, epidemiology and genotypes of A. marginale in cattle and buffaloes in Egypt, (ii) to evaluate suitable diagnostic tools and (iii) to identify co-infections of A. marginale with other selected tick-borne pathogens. Methods Blood samples were collected from 394 animals (309 cattle and 85 buffaloes) from three different areas in Egypt. For the detection of A. marginale infection, several tests were compared for their sensitivity and specificity: blood smear analysis, enzyme-linked immunosorbent assay (ELISA), PCR, real-time PCR and reverse line blot (RLB) assay. Co-infections with A. marginale, piroplasms and other Anaplasmataceae were surveyed by RLB while A. marginale genotypes were identified by amplifying and sequencing the partial msp1α gene. Results Anaplasma marginale DNA was amplified by qPCR in 68.3% of cattle and 29.4% of buffaloes. RLB showed infection with A. marginale in 50.2% of cattle and 42.5% of buffaloes. Blood smear analysis detected this agent in 16.2% of cattle and 2.4% of buffaloes. ELISA showed specific antibodies against A. marginale in 54.9% of cattle. Anaplasma marginale was associated, in cattle and buffaloes, with several tick-borne pathogens (Theileria annulata, Babesia bovis, Babesia bigemina, Babesia occultans and Anaplasma platys). A significant difference of A. marginale infection level was noticed in cattle, where animals between 3–5-years-old had a higher prevalence (79.2%) compared to those older than 5 years (36.4%) and younger than 3 years (59.7%) and one year (64.5%), respectively (P = 0.002281). Microsatellite analysis identified 15 different genotypes. Conclusions The epidemiological findings revealed high prevalence of A. marginale in cattle and buffaloes in all the investigated areas. The circulation of diverse genotypes was observed, most of these A. marginale genotypes being specific for Egypt. The qPCR assay was confirmed to be the most sensitive tool for detection of A. marginale in cattle and buffaloes even in the carrier state, highlighting the importance of using suitable diagnostic tests.

Download Full-text

Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi

10.21203/rs.3.rs-58018/v1 ◽

2020 ◽

Author(s):

Carlos Suarez ◽

Marta G. Silva ◽

Reginaldo G. Bastos ◽

J. Stone Doggett ◽

Michael K. Riscoe ◽

...

Keyword(s):

Mononuclear Cells ◽

Babesia Bovis ◽

Host Cells ◽

Bovine Babesiosis ◽

Theileria Equi ◽

Peripheral Blood Mononuclear ◽

Equine Piroplasmosis ◽

Apicomplexan Parasites ◽

And Control

Abstract Background: The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. Endochin-like quinolones (ELQ)-300 and ELQ-316 proved safe and efficacious against related apicomplexans, such as Plasmodium spp., and ELQ-316 was also effective against B. microti, without showing toxicity in mammals.Methods: Inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. Percentage of parasitized erythrocytes was measured by flow cytometry. Effect of the ELQ drugs on the viability of actively replicating horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by ELISA. Results: We calculated IC50 ranging from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 at 72 hr among all cultured parasites tested. None of the parasites tested were able to replicate in cultures in the presence of the ELQs-300 and ELQ-316 at IC100, which range from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 hours. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions: ELQ-300 and ELQ-316 have a significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

Download Full-text

RON2, a novel gene in Babesia bigemina, contains conserved, immunodominant B-cell epitopes that induce antibodies that block merozoite invasion

Parasitology ◽

10.1017/s0031182019001161 ◽

2019 ◽

Vol 146 (13) ◽

pp. 1646-1654 ◽

Cited By ~ 1

Author(s):

Juan Mosqueda ◽

Mario Hidalgo-Ruiz ◽

Diana Alexandra Calvo-Olvera ◽

Diego Josimar Hernandez-Silva ◽

Massaro Wilson Ueti ◽

...

Keyword(s):

B Cell ◽

Single Copy ◽

Inhibitory Effect ◽

Babesia Bigemina ◽

Bovine Babesiosis ◽

Serum Samples ◽

Merozoite Invasion ◽

B Cell Epitopes ◽

Specific Antibodies

AbstractBovine babesiosis is the most important protozoan disease transmitted by ticks. In Plasmodium falciparum, another Apicomplexa protozoan, the interaction of rhoptry neck protein 2 (RON2) with apical membrane antigen-1 (AMA-1) has been described to have a key role in the invasion process. To date, RON2 has not been described in Babesia bigemina, the causal agent of bovine babesiosis in the Americas. In this work, we found a ron2 gene in the B. bigemina genome. RON2 encodes a protein that is 1351 amino acids long, has an identity of 64% (98% coverage) with RON2 of B. bovis and contains the CLAG domain, a conserved domain in Apicomplexa. B. bigemina ron2 is a single copy gene and it is transcribed and expressed in blood stages as determined by RT-PCR, Western blot, and confocal microscopy. Serum samples from B. bigemina-infected bovines were screened for the presence of RON2-specific antibodies, showing the recognition of conserved B-cell epitopes. Importantly, in vitro neutralization assays showed an inhibitory effect of RON2-specific antibodies on the red blood cell invasion by B. bigemina. Therefore, RON2 is a novel antigen in B. bigemina and contains conserved B-cell epitopes, which induce antibodies that inhibit merozoite invasion.

Download Full-text