ABSTRACT
BACKGROUND: Bovine babesiosis is caused by infection with the protozoal parasite Babesia bovis, which is transmitted by Rhipicephalus (Boophilus) spp. It can cause mortality rates up to 90% in immunologically naive Bos taurus cattle. In south Texas, R. (B.) microplus is known to infest nilgai antelope (Boselaphus tragocamelus); however, their susceptibility to infection with B. bovis and their role in the transmission of the parasite remain unknown. In this study, we challenged nilgai antelope with B. bovis and evaluated their susceptibility to infection. METHODS: Nilgai were needle inoculated with ≈108 B. bovis-parasitized erythrocytes (merozoites) or a homogenate of B. bovis-infected larval ticks (sporozoite) delivered intravenously. Bos taurus beef calves were inoculated in parallel, as this strain of B. bovis is lethal to cattle. Temperature and hematocrit were monitored daily over the course of each study, and whole blood was collected for molecular [polymerase chain reaction (PCR)] and serological [indirect enzyme-linked immunosorbent assay (ELISA)] diagnostic evaluation. Histological sections of nilgai cerebral tissue were examined for evidence of infection. Recipient bovine calves were sub-inoculated with blood from nilgai challenged with either stage of the parasite, and they were monitored for clinical signs of infection and evaluated by a PCR diagnostic assay. Red blood cells (RBCs) from prechallenged nilgai and B. taurus beef cattle were cultured with an in vitro B. bovis merozoite culture to examine colonization of the RBCs by the parasite. RESULTS: Nilgai did not display clinical signs of infection upon inoculation with either the merozoite or sporozoite stage of B. bovis. All nilgai were PCR-negative for the parasite, and they did not develop antibodies to B. bovis. No evidence of infection was detected in histological sections of nilgai tissues, and in vitro culture analysis indicated that the nilgai RBCs were not colonized by B. bovis merozoites. Cattle subinoculated with blood from challenged nilgai did not display clinical signs of infection, and they were PCR-negative up to 45 days after transfer. CONCLUSIONS: Nilgai do not appear to be susceptible to infection with a strain of B. bovis that is lethal to cattle. Tick control on these alternative hosts remains a critical priority, especially given their potential to disseminate ticks over long distances.
Subject(s)
Antelopes , Babesia bovis , Babesiosis , Animals , Babesia bovis/genetics , Babesia bovis/pathogenicity , Babesia bovis/isolation & purification , Babesia bovis/immunology , Babesiosis/parasitology , Cattle , Antelopes/parasitology , Cattle Diseases/parasitology , Erythrocytes/parasitology , Texas , Virulence , Rhipicephalus/parasitology , Female , Polymerase Chain ReactionABSTRACT
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.
Subject(s)
Anaplasma marginale , Anaplasmosis , Babesia bovis , Babesia , Babesiosis , Cattle Diseases , Animals , Cattle , Colombia/epidemiology , Babesiosis/epidemiology , Babesiosis/parasitology , Anaplasma marginale/genetics , Anaplasma marginale/isolation & purification , Anaplasmosis/epidemiology , Anaplasmosis/microbiology , Cattle Diseases/epidemiology , Cattle Diseases/parasitology , Cattle Diseases/microbiology , Babesia/isolation & purification , Babesia/genetics , Babesia/classification , Babesia bovis/genetics , Babesia bovis/isolation & purification , Female , Male , PrevalenceABSTRACT
Babesiosis is a tick-borne disease with global impact caused by parasites of the phylum Apicomplexa, genus Babesia. Typically, acute bovine babesiosis (BB) is characterized by fever, anemia, hemoglobinuria, and high mortality. Surviving animals remain persistently infected and become reservoirs for parasite transmission. Bovids in China can be infected by one or more Babesia species endemic to the country, including B. bovis, B. bigemina, B. orientalis, B. ovata, B. major, B. motasi, B. U sp. Kashi and B. venatorum. The latter may pose a zoonotic risk. Occurrence of this wide diversity of Babesia species in China may be due to a combination of favorable ecological factors, such as the presence of multiple tick vectors, including Rhipicephalus and Hyalomma, the coexistence of susceptible bovid species, such as domestic cattle, yaks, and water buffalo, and the lack of efficient measures of tick control. BB is currently widespread in several regions of the country and a limiting factor for cattle production. While some areas appear to have enzootic stability, others have considerable cattle mortality. Research is needed to devise solutions to the challenges posed by uncontrolled BB. Critical research gaps include risk assessment for cattle residing in endemic areas, understanding factors involved in endemic stability, evaluation of parasite diversity and pathogenicity of regional Babesia species, and estimation of whether and how BB should be controlled in China. Research should allow the design of comprehensive interventions to improve cattle production, diminish the risk of human infections, and increase the availability of affordable animal protein for human consumption in China and worldwide. In this review, we describe the current state of BB with reference to the diversity of hosts, vectors, and parasite species in China. We also discuss the unique risks and knowledge gaps that should be taken into consideration for future Babesia research and control strategies.
Subject(s)
Babesiosis/epidemiology , Cattle Diseases/parasitology , Rhipicephalus/parasitology , Tick-Borne Diseases/prevention & control , Animals , Babesia bovis/isolation & purification , Babesia bovis/pathogenicity , Babesiosis/transmission , Buffaloes/parasitology , Cattle , Cattle Diseases/epidemiology , Cattle Diseases/transmission , China/epidemiology , DNA, Protozoan/genetics , Tick-Borne Diseases/epidemiology , Tick-Borne Diseases/parasitologyABSTRACT
Bovine babesiosis is economically the most important arthropod-borne disease of cattle worldwide. The most significant damage caused by bovine babesiosis is attributed to Babesia bovis due to its higher pathogenicity. This study aimed to develop a real-time PCR method followed by HRM (high-resolution melting) analysis for the simultaneous detection of B. bovis and B. bigemina, enabling a semi-quantitative analysis of Babesia levels using a single-tube reaction. The HRM was compared with real-time PCR using species-specific hydrolysis probes. The HRM analysis allowed to differentiate both Babesia species and was sensitive in the detection and differentiation of 10% for each Babesia species in the sample. Our results suggest the use of this method to estimate the prevalence of infections by B. bovis or B. bigemina as an alternative to the methods of absolute quantification by real-time PCR since it neither requires precise estimates of the number of DNA loads nor the construction of calibration curves. The simultaneous detection of the two Babesia species can be used to characterise the infection levels in cattle populations from different geographical regions, allowing a better control of these diseases.
Subject(s)
Babesia , Real-Time Polymerase Chain Reaction/methods , Animals , Babesia/genetics , Babesia/isolation & purification , Babesia bovis/genetics , Babesia bovis/isolation & purification , Babesiosis/parasitology , Cattle , Cattle Diseases/parasitology , DNA, Protozoan/genetics , Tick-Borne Diseases/parasitologyABSTRACT
Babesia bovis is a known causative agent of bovine babesiosis and is widely distributed across China. Rapid detection and accurate identification of B. bovis is essential for follow-up management and epidemiological investigations. In this study, a cross-priming amplification combined with vertical flow (CPA-VF) assay was developed. The detection limit of the CPA-VF assay targeting the 18S rRNA gene was 320 fg per reaction at 61 °C for 60 min. No cross-reactions were observed with other piroplasms infective to cattle. Furthermore, 36 blood samples from experimentally-infected animals were accurately assessed using the CPA-VF assay. The performance of the CPA-VF assay was compared with the results of conventional PCR for 219 blood samples from the field. Our results demonstrate that the CPA-VF assay is a practical and effective diagnostic tool for bovine babesiosis caused by B. bovis infection.
Subject(s)
Babesia bovis/isolation & purification , Babesiosis/diagnosis , Cattle Diseases/diagnosis , Nucleic Acid Amplification Techniques/veterinary , Animals , Babesiosis/parasitology , Cattle , Cattle Diseases/parasitology , Cross-Priming , Nucleic Acid Amplification Techniques/methods , RNA, Protozoan/analysis , RNA, Ribosomal, 18S/analysis , Sensitivity and SpecificityABSTRACT
This aim of the present study was to estimate the prevalence of haemopathogens in cattle in Beni Hamidene locality, district of Constantine (Νortheastern Algeria). Between June and October 2014, 169 bovines from 25 farms were included in this survey, 32 (18.9%) among them were suspected of piroplasmosis and/or anaplasmosis. Infection prevalences were estimated by microscopic examination of Giemsa-stained blood smears and blood samples from all included cattle (n = 169). Animals were infected by Theileria annulata (65/169; 38.46%), Anaplasma marginale (22/169; 13%) and Babesia bovis (5/169; 3%). Two co-infection patterns were found: Theileria annulata/Anaplasma marginale (7.69%) and Theileria annulata/Babesia bovis (1.18%). Only one farm had no cattle infected by any of the haemopathogens. There was a signification difference of T. annulata infection prevalence according to age category (p =.04). These results emphasised mainly the presence of bovine tropical theileriosis in northeastern, Beni Hamidene locality, province of Constantine, Algeria.
Subject(s)
Anaplasmosis/epidemiology , Babesiosis/epidemiology , Cattle Diseases/epidemiology , Coinfection/veterinary , Theileriasis/epidemiology , Algeria/epidemiology , Anaplasma marginale/isolation & purification , Anaplasmosis/microbiology , Animals , Babesia bovis/isolation & purification , Babesiosis/parasitology , Cattle , Cattle Diseases/microbiology , Cattle Diseases/parasitology , Coinfection/epidemiology , Coinfection/microbiology , Coinfection/parasitology , Cross-Sectional Studies , Female , Male , Prevalence , Theileria annulata/isolation & purification , Theileriasis/parasitologyABSTRACT
BACKGROUND: The tick vector Rhipicephalus microplus which transmits Babesia spp. and rickettsial pathogens has not been reported in Kenya since 1998. More recently, the pathogenic Babesia bovis has been detected in cattle blood DNA. The status of R. microplus in Kenya remains unknown. This study employed morphological and molecular tools to characterize R. microplus originating from Kenya and assess the genetic relationships between Kenyan and other African R. microplus genotypes. METHODS: Ticks were collected in south-eastern Kenya (Kwale County) from cattle and characterized to investigate the existence of R. microplus. Genetic and phylogenetic relationships between the Kenyan and other annotated R. microplus reference sequences was investigated by analysis of the cytochrome c oxidase subunit 1 (cox1) gene. To further characterize Kenyan ticks, we generated low coverage whole genome sequences of two R. microplus, one R. decoloratus and R. appendiculatus. A B. bovis specific TaqMan probe qPCR assay was used to detect B. bovis in gDNA from R. microplus ticks. RESULTS: Occurrence of R. microplus was confirmed in Kwale County, Kenya. The Kenyan R. microplus cox1 sequences showed very high pairwise identities (> 99%) and clustered very closely with reference African R. microplus sequences. We found a low genetic variation and lack of geographical sub-structuring among the African cox1 sequences of R. microplus. Four complete mitochondrial (mt) genomes for two R. microplus, one R. decoloratus and one R. appendiculatus were assembled from next generation sequence data. The mitochondrial genome sequences of the two Kenyan R. microplus ticks clustered closely with reference genome sequences from Brazil, USA, Cambodia and India forming R. microplus Clade A. No B. bovis was detected in the Kwale R. microplus DNA. CONCLUSIONS: These findings confirm the presence of R. microplus in Kenya and suggest that R. microplus Clade A is prevalent in cattle in sub-Saharan Africa. These and other recent findings of widespread occurrence of R. microplus in Africa provide a strong justification for urgent surveillance to determine and monitor the spread of R. microplus and vector competence of Boophilus ticks for B. bovis in Africa, with the ultimate goal of strategic control.
Subject(s)
Babesia bovis/isolation & purification , Rhipicephalus , Tick Infestations/veterinary , Animals , Arachnid Vectors/genetics , Arachnid Vectors/parasitology , Babesia bovis/genetics , Cattle , Cattle Diseases/epidemiology , Disease Vectors , Electron Transport Complex IV/genetics , Epidemiological Monitoring , Genes, Protozoan , Genome, Mitochondrial , Kenya/epidemiology , Pathology, Molecular/methods , Phylogeny , Rhipicephalus/genetics , Rhipicephalus/parasitologyABSTRACT
Babesia bovis and Babesia bigemina are tick-transmitted piroplasms that cause severe damage to the livestock industry in tropical regions of the world. Recent studies demonstrated differences in infection levels of these haemoparasites among bovine breeds and variation between individual cows regarding resistance to these diseases. This study aimed to estimate the repeatability and correlations between B. bovis and B. bigemina using two cattle breeding systems, an individual system (IS) and a collective paddock system (CPS). All animals were Holstein breed, and the levels of B. bovis and B. bigemina in blood samples were estimated by quantitative polymerase chain reaction (qPCR). The estimated correlations for the B. bigemina and B. bovis DNA copy number for IS and CPS were moderate and high, respectively, whereas repeatability estimates for both systems and both Babesia species were moderate. Although we cannot infer that the type of rearing system directly influenced the correlation and repeatability coefficients, it appears that the bovine parasitemia burden may be dependent on (or determine) the parasitemia burden on ticks because the bovines remained in the same place for a longer time in both systems. Thus, the babesiosis infection levels of the ticks may have been uniform, a phenomenon that also ensures greater uniformity in cattle infection. This factor may have favored the occurrence of infected ticks leading to higher repeatability estimates and correlations. Our study confirms high variability in resistance/susceptibility between breeds, and the high correlations found may be linked to this characteristic and the most intensive breeding type of dairy cattle. Besides, under the present study conditions, the estimated correlations suggest that measuring an infection level of one Babesia species can predict the level of infection of the other.
Subject(s)
Babesia bovis , Babesia , Babesiosis/epidemiology , Cattle Diseases , Cattle/parasitology , Animals , Babesia/isolation & purification , Babesia bovis/isolation & purification , Breeding , Cattle Diseases/epidemiology , Cattle Diseases/parasitology , DNA, Protozoan/isolation & purification , Dairying , ParasitemiaABSTRACT
Babesia bovis parasites present a serious and significant health concern for the beef and dairy industries in many parts of the world. Difficulties associated with the current diagnostic techniques include the following: they are prone to human error (microscopy) or expensive and time-consuming (polymerase chain reaction) to perform. Little is known about the biochemical changes in blood that are associated with Babesia infections. The discovery of new biomarkers will lead to improved diagnostic outcomes for the cattle industry. Vibrational spectroscopic technologies can record a chemical snapshot of the entire organism and the surrounding cell thereby providing a phenotype of the organism and the host infected cell. Here, we demonstrate the applicability of vibrational spectroscopic imaging techniques including Atomic Force Microscopy Infrared (AFM-IR) and confocal Raman microscopy to discover new biomarkers for B. bovis infections. Furthermore, we applied Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) to detect B. bovis in red blood cells (RBCs). Based on changes in the IR spectral bands, with ATR-FTIR in combination with Partial Least Squares-Discriminant Analysis we were able to discriminate infected samples from controls with a sensitivity and specificity of 92.0% and 91.7%, respectively, in less than 2 min, excluding sample extraction and preparation. The proposed method utilized a lysis approach to remove hemoglobin from the suspension of infected and uninfected cells, which significantly increased the sensitivity and specificity compared to measurements performed on intact infected red blood cells (intact infected RBC, 77.3% and 79.2%). This work represents a holistic spectroscopic study from the level of the single infected RBC using AFM-IR and confocal Raman to the detection of the parasite in a cell population using ATR-FTIR for a babesiosis diagnostic.
Subject(s)
Babesia bovis/chemistry , Babesiosis/diagnosis , Cattle Diseases/diagnosis , Spectrophotometry, Infrared/methods , Spectrum Analysis, Raman/methods , Animals , Babesia bovis/isolation & purification , Babesiosis/parasitology , Biomarkers/chemistry , Cattle , Cattle Diseases/parasitology , Discriminant Analysis , Erythrocytes/parasitology , Least-Squares Analysis , Microscopy, Atomic Force , Microscopy, ConfocalABSTRACT
Anaplasmosis and babesiosis are tick-borne diseases widely disseminated in cattle herds in many parts of the world. These diseases represent important causes of death and economic losses in several countries, including Brazil, and are characterized by hemolytic disease and anemia. Animals of all ages may be affected. Although transplacental infections are known to occur, abortion, stillbirth and neonatal death directly associated with Anaplasma marginale and especially Babesia spp. infections have rarely been documented in cattle. The objective of the present study is to describe the pathological and molecular findings of two cases of bovine abortion, two cases of stillbirth and two cases of neonatal death associated with intrauterine anaplasmosis and/or babesiosis in southern Brazil. All cases occurred in beef farms in the state of Rio Grande do Sul, between 2017 and 2019. Angus and crossbred calves were affected. At the necropsy, the main gross lesions observed included different degrees of splenomegaly, enlarged and yellow liver, thick and grumous bile, pallor or jaundice of mucous membranes and carcass, and dark kidneys. Four calves also presented cherry-pink discoloration of the central nervous system. Cytological slides enabled the observation of intraerythrocytic organisms consistent with Babesia bovis (3/6) and A. marginale (2/6). Through PCR assays, it was possible to detect three cases of Babesia sp. infection alone, and one case of Anaplasma sp. infection alone. Co-infections with Anaplasma sp. and Babesia sp. were detected in two cases. These findings reaffirm that anaplasmosis and babesiosis should be considered as an important differential diagnosis of fetal loss, stillbirth and neonatal death in cattle in areas where these diseases occur.
Subject(s)
Abortion, Veterinary/pathology , Anaplasma/isolation & purification , Anaplasmosis/microbiology , Babesia bovis/isolation & purification , Babesiosis/parasitology , Cattle Diseases/pathology , Stillbirth/veterinary , Abortion, Veterinary/microbiology , Anaplasmosis/pathology , Animals , Babesiosis/pathology , Brazil , Cattle , Cattle Diseases/microbiology , HumansABSTRACT
The objective of this study was to instrument a serological assay for the epidemiological diagnosis of bovine babesiosis in Mexico, using the Babesia bigemina recombinant protein RAP-1 (rRAP-1α) as antigen. rRAP-1α, r12d3 and rGP45 were the three recombinant antigens initially tested. Based on the highest titres obtained in the indirect ELISA (iELISA) with the positive control serum, using similar antigen concentrations, rRAP-1α was selected for further use. The diagnostic sensitivity and specificity rates estimated for the iELISA with rRAP-1α as antigen were 89.9% and 86.5%, respectively, while for the Indirect Fluorescent Antibody Test (IFAT), the gold standard assay, the sensitivity was 86.66% and the specificity was 95%. The ĸ agreement value determined was 0.52, indicating a moderate agreement between the iELISA and IFAT assays. The instrumented iELISA with rRAP-1α as antigen shows an excellent specificity rate and an acceptable sensitivity that allows for the detection of antibodies to B. bigemina in cattle naturally exposed to the vector tick Rhipicephalus microplus. By using the iELISA-rRAP-1α, along with an iELISA with recombinant Merozoite Surface Antigen (rMSA-1) for antibody determination against Babesia bovis in the serum samples collected from cattle at 'La Posta' experimental station in Mexico, a seroprevalence of 20.3% was estimated for B. bigemina and 19.4% for B. bovis, while 36.89% of samples were positive for both Babesia species. The iELISA test promises to be a safe and low-cost type of diagnosis available to cattle producers in Mexico and would facilitate the definition of herd immunity status to implement measures of control adapted for the prevention of bovine babesiosis outbreaks.
Subject(s)
Antigens, Protozoan/immunology , Babesia/immunology , Babesiosis/epidemiology , Cattle Diseases/epidemiology , Fluorescent Antibody Technique, Indirect/veterinary , Rhipicephalus/parasitology , Animals , Babesia/isolation & purification , Babesia bovis/immunology , Babesia bovis/isolation & purification , Babesiosis/diagnosis , Babesiosis/parasitology , Cattle , Cattle Diseases/diagnosis , Cattle Diseases/parasitology , Enzyme-Linked Immunosorbent Assay/veterinary , Mexico/epidemiology , Recombinant Proteins , Seroepidemiologic StudiesABSTRACT
Bovine babesiosis is a tick-transmitted haemoparasitic disease caused by Babesia bovis and B. bigemina affecting cattle of tropical and subtropical regions around the world. Pathogens are transmitted by the tick vector Rhipicephalus microplus displaying a widespread distribution in northeastern Argentina. The disease is characterized by significant animal morbidity and mortality resulting in considerable economic loss. In this study, B. bovis and B. bigemina infection was investigated in a cattle herd of 150 adult bovines of pure Braford breed raised in a tick-hyperendemic field using molecular and serum antibody tests. A highly sensitive nested polymerase chain reaction (nPCR) assay targeting a species-specific region of the apocytochrome b gene resulted in direct B. bovis and B. bigemina detection in 27.3% and 54.7% of bovines, respectively. A recently developed immunochromatographic strip test (ICT) based on recombinant forms of spherical body protein 4 and the C-terminal region of rhoptry-associated protein 1 showed that 71.3% and 89.3% of bovines were seropositive for B. bovis and B. bigemina, respectively. The mixed infection rate as observed by direct (19.3%) and indirect detection (65.3%) coincided with those expected, respectively. Importantly, four months after sampling, nine bovines of the studied herd showed clinical signs of bovine babesiosis of which six animals eventually died. Microscopic detection of infected erythrocytes in Giemsa-stained blood smears confirmed B. bovis infection. Our study demonstrates that although animals showed a relatively high and very high rate of immunity against infection with B. bovis (71.3%) and B. bigemina (89.3%) parasites, respectively, clinical cases and fatalities due to the infection with B. bovis were observed. It is proposed that the most adequate control measure in the studied epidemiological situation is to vaccinate animals to prevent losses and/or an outbreak of bovine babesiosis.
Subject(s)
Babesia/isolation & purification , Babesiosis/epidemiology , Cattle Diseases/epidemiology , Rhipicephalus/parasitology , Animals , Argentina/epidemiology , Babesia/genetics , Babesia/immunology , Babesia bovis/genetics , Babesia bovis/immunology , Babesia bovis/isolation & purification , Babesiosis/parasitology , Cattle , Cattle Diseases/parasitology , Chromatography, Affinity/veterinary , Female , Male , Polymerase Chain Reaction/veterinary , Sensitivity and Specificity , Species SpecificityABSTRACT
The water buffalo industry is a vital part of the Philippine livestock economy and is an essential contributor to the developing local dairy industry. Although relatively less susceptible to diseases, water buffaloes can still be infected and can act as reservoirs of tick-borne pathogens (TBPs). However, limited information is available regarding the prevalence of tick-borne infections in water buffaloes in the Philippines. This study was conducted to identify TBPs harbored by water buffaloes and to characterize these pathogens molecularly. One hundred water buffalo blood samples collected from three areas in Bohol, Visayas region, Philippines were screened for various TBPs using pathogen-specific PCR assays. TBPs were detected in 46% of the samples (39% singly infected, 7% coinfected). The pathogens detected were Anaplasma marginale (29%), Babesia bovis (21%), and B. bigemina (3%). None of the blood samples were positive for Theileria annulata, T. orientalis, and B. ovata. A. marginale infection rates were significantly higher (37.5%) among water buffaloes aged ≤6 years (P = 0.046) than those >6 years old (18.2%) and was detected only in Bulgarian Murrah (36.1%) and US Murrah (25.9%) breeds. Phylogenetic analyses revealed that groEL sequences of A. marginale were 100% identical with isolates from the Philippines (Batangas and Cebu) and China. Two B. bigemina RAP-1a gene sequences were identical to each other and were homologous with previous isolates from Thailand, Indonesia, Uruguay, and the Philippines. Moreover, four B. bovis SBP-2 partial sequences obtained in this study had 92.4-99.7% identities. This study is the first molecular detection and characterization of A. marginale, B. bigemina and B. bovis in water buffaloes in the Visayas region, and the first molecular confirmation of B. bovis infection in water buffaloes in the country. The findings presented in this study may serve as baseline data for crafting effective tick-borne disease surveillance and prevention programs in Bohol and in the Philippines.
Subject(s)
Anaplasmosis/epidemiology , Babesiosis/epidemiology , Buffaloes/microbiology , Phylogeny , Theileriasis/epidemiology , Tick-Borne Diseases/veterinary , Anaplasma marginale/genetics , Anaplasma marginale/isolation & purification , Animals , Babesia/genetics , Babesia/isolation & purification , Babesia bovis/genetics , Babesia bovis/isolation & purification , Buffaloes/parasitology , DNA, Protozoan/genetics , Female , Genetic Variation , Philippines/epidemiology , Polymerase Chain Reaction , Theileria annulata/genetics , Theileria annulata/isolation & purification , Tick-Borne Diseases/microbiology , Tick-Borne Diseases/parasitology , Ticks/microbiology , Ticks/parasitologyABSTRACT
The diseases caused by hemoprotozoan parasites in cattle often result in economic losses. In Sri Lanka, previous studies found that the up-country wet zone, which is located in central Sri Lanka, was characterized by a high rate of Theileria orientalis and a low rate of Theileria annulata compared with the dry zone. In this study, DNA samples were prepared from the blood of 121 cattle in Galle, a coastal district located in low-country wet zone in Sri Lanka, and were PCR-screened for Babesia bovis, Babesia bigemina, T. annulata, T. orientalis, and Trypanosoma theileri. All the parasite species, except B. bovis, were detected among the surveyed cattle. The animals had a high rate of T. orientalis (100%) and a low rate of T. annulata (1.6%), as in the up-country wet zone. Babesia bigemina and Tr. theileri were detected in 19.0% and 20.6% of the animals, respectively, and their infection rates were higher in the animals reared in extensive management systems (32.8% and 27.9%, respectively) than in those managed in intensive/semi-intensive systems (5.0% and 13.3%, respectively). Genotypic analyses found that the T. orientalis mpsp type 5 was predominant similar to up-country wet zone, and that Tr. theileri consisted of seven catl genotypes, including two new genotypes (IL and IM) and four previously detected genotypes (IA, IB, II, and IK). These findings suggest that the hemoprotozoan infection profiles are largely conserved within the wet zone, despite differences in the geography, cattle breeds, and management practices between the up-country and low-country wet zones.
Subject(s)
Babesia bovis/isolation & purification , Cattle Diseases/epidemiology , Theileria/isolation & purification , Trypanosoma/isolation & purification , Trypanosomiasis/veterinary , Animals , Babesia bovis/genetics , Babesiosis/epidemiology , Cattle/parasitology , Cattle Diseases/parasitology , Climate , DNA, Protozoan/genetics , Genotype , Geography , Phylogeny , Polymerase Chain Reaction , Sri Lanka/epidemiology , Theileria/genetics , Theileriasis/epidemiology , Trypanosoma/genetics , Trypanosomiasis/epidemiologyABSTRACT
Tick-borne diseases (TBDs) have a large impact on animal health and the livelihood of livestock owners, particularly in developing countries. Although climatic and ecological conditions in Pakistan may favour the transmission of tick-borne pathogens (TBPs), only a few systematic studies have been carried out on TBPs and the diseases that they cause in this country. To improve our understanding of the distribution of TBPs, 3,807 ticks were collected from ruminants (n = 369) on 108 livestock farms (semi-arid zone = 36, arid zone = 72) in Punjab Province. After morphological identification ticks were pooled into 405 pools (Hyalomma anatolicum = 300, Rhipicephalus microplus = 89, Hyalomma dromedarii = 9, Rhipicephalus turanicus = 7) based on their species, locality of collection, and the host. DNA from each pool was screened by a Reverse Line Blot (RLB) hybridization assay for the presence of Anaplasma, Ehrlichia, Rickettsia, Babesia, and Theileria species. DNA from at least one TBP was found in 142 (35.1%) pools. Among the positive pools, 91 (64.1%) had a mixed infection with two or more TBPs, whereas 51 (35.9%) pools were infected with a single TBP. The detected pathogens not only included species that were known to be endemic in Pakistan, such as Theileria annulata (6.7%), Theileria orientalis (3.5%), Anaplasma marginale (5.7%), Anaplasma centrale (2.7%), Anaplasma ovis (1.5%), Babesia bigemina (0.7%), and Babesia bovis (0.2%), but also several TBPs that had not been reported to occur in Pakistan before. This included Ehrlichia minasensis (3.2%), an Anaplasma platys-like organism (1.2%), Babesia occultans (0.2%), and Rickettsia massiliae (0.2%), as well as two previously uncharacterized species: Ehrlichia sp. Multan (18.0%) and Anaplasma sp. (BL099-6) (2.22%). The pathogenicity of these novel species remains to be examined. This study shows that a much broader spectrum of TBPs is present in Pakistan than previously thought, including several zoonotic pathogens.
Subject(s)
Cattle Diseases/epidemiology , Livestock , Tick-Borne Diseases/veterinary , Ticks/microbiology , Ticks/parasitology , Anaplasma/genetics , Anaplasma/isolation & purification , Animals , Babesia/genetics , Babesia bovis/genetics , Babesia bovis/isolation & purification , Cattle , Cattle Diseases/microbiology , Cattle Diseases/parasitology , DNA, Bacterial/genetics , DNA, Protozoan/genetics , Ehrlichia/genetics , Ehrlichia/isolation & purification , Female , Livestock/microbiology , Livestock/parasitology , Pakistan/epidemiology , Polymerase Chain Reaction/veterinary , Rickettsia/genetics , Rickettsia/isolation & purification , Theileria annulata/genetics , Theileria annulata/isolation & purification , Tick-Borne Diseases/epidemiology , Tick-Borne Diseases/microbiology , Tick-Borne Diseases/parasitologyABSTRACT
BACKGROUND: Babesia bigemina and B. bovis are two economically important hemoparasites affecting both cattle and buffaloes involved in dairy and beef production. In Colombia, although some parasitological and serological studies suggest an endemicity of these pathogens in areas under 1000 m, little is known about its molecular prevalence in different host. The objective of this study was to estimate the prevalence and molecular traits of these parasites in cattle and buffaloes from two Colombian regions. METHODS: Between 2014 and 2016, a three-point longitudinal survey was designed in farms from Caribbean and Orinoquia regions to evaluate the molecular prevalence of B. bigemina and B. bovis using a nested PCR (n-PCR) targeting hypothetical protein (hyp) and rhoptry-associated protein (RAP-1) genes, respectively. A total of 1432 cattle, 152 buffalo and 1439 Rhipicephalus microplus samples were analyzed. Moreover, phylogenetic relationship of isolates was analyzed using the 18S rRNA gene. RESULTS: A molecular prevalence of 31.6% (24.2% for B. bigemina and 14.4% for B. bovis), 23.6% (6.5% for B. bigemina and 17.7% for B. bovis) and 4.3% (3.5% for B. bigemina and 1.0% for B. bovis) was observed in cattle, buffaloes and Rhipicephalus microplus, respectively. Higher values of infection were observed during the wet season and late wet season; nevertheless, other variables such as age, production type, sex, breed and babesiosis control were also significantly associated with infection. Prevalence analysis showed that B. bovis infection was higher in cattle that coexist with buffaloes, when compared to those which did not. For each species, phylogenetic analyses revealed a high genetic diversity of isolates without clusters related to the isolation source. CONCLUSIONS: To our knowledge, this is the first longitudinal survey that evaluates through molecular methods, the infection of B. bigemina and B. bovis in two important livestock regions from Colombia. This study reveals that the prevalence of infection by Babesia spp., in cattle and buffaloes are modulated by seasonal variations, host factors and vector traits. Our results provide new insights on the epidemiological aspects of infection of Babesia spp., in cattle and buffaloes, which must be taken into consideration when babesiosis control programs are implemented in the study area.
Subject(s)
Babesia/isolation & purification , Babesiosis/epidemiology , Buffaloes/parasitology , Cattle Diseases/epidemiology , Animals , Babesia/genetics , Babesia bovis/genetics , Babesia bovis/isolation & purification , Babesiosis/parasitology , Cattle , Cattle Diseases/parasitology , Colombia/epidemiology , Female , Genetic Variation , Longitudinal Studies , Male , Phenotype , Phylogeny , Polymerase Chain Reaction/veterinary , PrevalenceABSTRACT
Bovine babesiosis is a serious threat to the cattle industry. We prepared blood DNA samples from 13 cattle with clinical babesiosis from the Badulla (n = 8), Jaffna (n = 3), and Kilinochchi (n = 2) districts in Sri Lanka. These DNA samples tested positive in PCR assays specific for Babesiabovis (n = 9), Babesia bigemina (n = 9), and Babesiaovata (n = 1). Twelve cattle were positive for B. bovis and/or B. bigemina One cow was negative for the tested Babesia species but was positive for Babesia on microscopic examination; the phylogenetic positions of 18S rRNA and cytochrome oxidase subunit III gene sequences suggested that the cow was infected with Babesia sp. Mymensingh, which was recently reported from a healthy cow in Bangladesh. We then developed a novel Babesia sp. Mymensingh-specific PCR assay and obtained positive results for one other sample. Analysis of gene sequences from the cow with positive B. ovata-specific PCR results demonstrated that the animal was infected not with B. ovata but with Babesia sp. Hue-1, which was recently reported from asymptomatic cattle in Vietnam. The virulence of Babesia sp. Hue-1 is unclear, as the cow was coinfected with B. bovis and B. bigemina However, Babesia sp. Mymensingh probably causes severe clinical babesiosis, as it was the sole Babesia species detected in a clinical case. The present study revealed the presence of two bovine Babesia species not previously reported in Sri Lanka, plus the first case of severe bovine babesiosis caused by a Babesia species other than B. bovis, B. bigemina, and Babesiadivergens.
Subject(s)
Babesia/genetics , Babesia/isolation & purification , Babesiosis/microbiology , Cattle Diseases/microbiology , Animals , Babesia/classification , Babesia/cytology , Babesia bovis/genetics , Babesia bovis/isolation & purification , Babesiosis/epidemiology , Babesiosis/pathology , Babesiosis/physiopathology , Cattle , Cattle Diseases/pathology , Cattle Diseases/physiopathology , DNA, Protozoan/genetics , Female , Phylogeny , Polymerase Chain Reaction/veterinary , Protozoan Proteins/genetics , RNA, Ribosomal, 18S/genetics , Sequence Analysis, DNA/veterinary , Sri Lanka/epidemiologyABSTRACT
Water buffaloes (Bubalus bubalis) are raised in tropical and subtropical regions of the world, and act as hosts of Babesia bovis parasites and the tick vector Rhipicephalus microplus. As no clinical cases of B. bovis-infection have been reported, we hypothesized that, unlike bovines, water buffaloes respond asymptomatically to an acute infection. To test this hypothesis, we inoculated two groups of 24-month-old Mediterranean breed water buffaloes with 108 erythrocytes infected with two Argentine B. bovis isolates: BboM2P (nâ¯=â¯5) or BboS2P (nâ¯=â¯5). These strains displayed mild (BboM2P) or high (BboS2P) pathogenicity in Bos taurus calves of the same age (nâ¯=â¯5 and nâ¯=â¯1, respectively), when tested in parallel. In water buffaloes, no changes in body temperature were observed with both strains, and no hematocrit changes were detected in BboM2P-inoculated animals. In contrast, in the BboS2P-inoculated water buffalo group significant but relatively minor reductions in haematocrit values were noted compared to the infected bovine. The parasitemia attained in water buffaloes was considerably lower than in bovines and could only be detected by nested PCR, or indirectly via serology, whereas in most bovines, it could also be detected in Giemsa-stained smears under the light microscope. Our results show that water buffaloes present no or significantly mitigated clinical symptoms to B. bovis infections and suggest that they are able to substantially reduce and/or eliminate B. bovis parasites from circulation by an efficient innate immune mechanism.
Subject(s)
Babesia bovis/isolation & purification , Babesiosis/parasitology , Cattle Diseases/diagnosis , Parasitemia/diagnosis , Animals , Babesia bovis/genetics , Babesia bovis/immunology , Babesia bovis/pathogenicity , Babesiosis/diagnosis , Babesiosis/immunology , Buffaloes/immunology , Buffaloes/parasitology , Cattle , Cattle Diseases/blood , Cattle Diseases/parasitology , DNA, Protozoan/genetics , Enzyme-Linked Immunosorbent Assay/veterinary , Erythrocytes/parasitology , Hematocrit , Immunity, Innate , Male , Parasitemia/epidemiology , Polymerase Chain Reaction/veterinary , Rhipicephalus/microbiology , Serologic TestsABSTRACT
Little is known about the occurrence of haemoparasites in cattle in communal grazing areas of Mungwi District of Northern Province, Zambia. Clinical signs and post mortem lesions are pathognomonic of mixed tick-borne infections especially babesiosis, anaplasmosis and East Coast fever. The main objective of this study was to screen selected communal herds of cattle for tick-borne haemoparasites, and identify the tick vectors associated with the high cattle mortalities due to suspected tick-borne diseases in the local breeds of cattle grazing along the banks of the Chambeshi River in Mungwi District, Northern Province, Zambia. A total of 299 cattle blood samples were collected from July to September 2010 from Kapamba (nâ¯=â¯50), Chifulo (nâ¯=â¯102), Chisanga (nâ¯=â¯38), Kowa (nâ¯=â¯95) and Mungwi central (nâ¯=â¯14) in the Mungwi District. A total of 5288 ticks were also collected from the sampled cattle from April to July 2011. DNA was extracted from the cattle blood and the hypervariable region of the parasite small subunit rRNA gene was amplified and subjected to the reverse line blot (RLB) hybridization assay. The results of the RLB assay revealed the presence of tick-borne haemoparasites in 259 (86.6%) cattle blood samples occurring either as single (11.0%) or mixed (75.6%) infections. The most prevalent species present were the benign Theileria mutans (54.5%) and T. velifera (51.5%). Anaplasma marginale (25.7%), Babesia bovis (7.7%) and B. bigemina (3.3%) DNA were also detected in the samples. Only one sample (from Kapamba) tested positive for the presence of T. parva. This was an unexpected finding; also because the tick vector, Rhipicephalus appendiculatus, was identified on animals from Kowa (14.0%), Chisanga (8.5%), Chifulo (6.0%) and Kapamba (1.4%). One sample (from Kapamba) tested positive for the presence of Ehrlichia ruminantium even though Amblyomma variegatum ticks were identified from 52.9% of the sampled animals from all study areas. There was significant positive association between T. mutans and T. velifera (pâ¯<â¯0.001) infections, and between A. marginale and B. bovis (pâ¯=â¯0.005). The presence of R. microplus tick vectors on cattle was significantly associated with B. bovis (odds ratio, ORâ¯=â¯28.4, pâ¯<â¯0.001) and A. marginale (ORâ¯=â¯42.0, pâ¯<â¯0.001) infections, while A. variegatum presence was significantly associated with T. mutans (ORâ¯=â¯213.0, pâ¯<â¯0.001) and T. velifera (ORâ¯=â¯459.0, pâ¯<â¯0.001) infections. Rhipicephalus decoloratus was significantly associated with B. bigemina (ORâ¯=â¯21.6, pâ¯=â¯0.004) and A. marginale (ORâ¯=â¯28.5, pâ¯<â¯0.001). Multivariable analysis showed a significant association between location and tick-borne pathogen status for A. marginale (pâ¯<â¯0.001),â¯T. mutans (pâ¯=â¯0.004),â¯T. velifera (pâ¯=â¯0.003) and T. taurotragi (pâ¯=â¯0.005). The results of our study suggest that the cause of cattle mortalities in Mungwi during the winter outbreaks is mainly due to A. marginale, B. bovis and B. bigemina infections. This was confirmed by the clinical manifestation of the disease in the affected cattle and the tick species identified on the animals. The relatively low prevalence of T. parva, B. bigemina, B. bovis and E. ruminantium could indicate the existence of endemic instability with a pool of susceptible cattle and the occurrence of disease outbreaks.
Subject(s)
Cattle Diseases/epidemiology , Tick-Borne Diseases/veterinary , Ticks/parasitology , Anaplasma/genetics , Anaplasma/isolation & purification , Anaplasma marginale/genetics , Anaplasma marginale/isolation & purification , Anaplasmosis/blood , Anaplasmosis/epidemiology , Anaplasmosis/microbiology , Anaplasmosis/mortality , Animals , Babesia/genetics , Babesia/isolation & purification , Babesia bovis/genetics , Babesia bovis/isolation & purification , Babesiosis/blood , Babesiosis/epidemiology , Babesiosis/mortality , Babesiosis/parasitology , Cattle , Cattle Diseases/parasitology , DNA, Bacterial/genetics , DNA, Protozoan/genetics , Ehrlichia ruminantium/isolation & purification , Heartwater Disease/blood , Heartwater Disease/epidemiology , Heartwater Disease/microbiology , Humans , Theileria/genetics , Theileria/isolation & purification , Theileriasis/blood , Theileriasis/epidemiology , Theileriasis/parasitology , Tick-Borne Diseases/epidemiology , Tick-Borne Diseases/microbiology , Tick-Borne Diseases/parasitology , Zambia/epidemiologyABSTRACT
Babesiosis, theileriosis, anaplasmosis, and heartwater are tick-borne diseases (TBD) that threaten livestock production in sub-Saharan Africa including Benin. This country has been faced with an invasion of Rhipicephalus microplus, a major vector for babesiosis, theileriosis, and anaplasmosis over the last decade. Yet, data on TBD and the impact of the invasive ticks are lacking, making risk level evaluation and disease control arduous. In this study, epidemiological features of Babesia bovis, B. bigemina, Theileria spp., Anaplasma marginale and Ehrlichia ruminantium infections in Benin cattle were investigated in R. microplus-invaded and non-invaded areas. Detection of pathogens was based on species-specific PCR assays and resulting data were used to identify risk factors. Genetic diversity and phylogenies were then evaluated using several markers. Out of 207 samples examined, 170 (82.1%), 109 (52.7%), 42 (20.3%) 24 (11.6%) and 1 (0.5%) were positive for T. mutans, A. marginale, B. bigemina, B. bovis and E. ruminantium, respectively. Animal gender (for B. bovis), exposure to R. microplus (for B. bigemina and A. marginale), animal age (for B. bigemina and A. marginale) and cattle breed and/or antiprotozoal treatment (for T. mutants) significantly modulated pathogen occurrence. In addition, R. microplus exposure was significantly related to co-infection patterns and cases of clinical theileriosis and/or anaplasmosis were recorded among cattle highly exposed to the tick. In the genetic characterization, Theileria spp. and E. ruminantium sequences were conserved. Babesia spp. and A. marginale, however, showed high sequence polymorphisms that indicate the presence of several strains and may be linked to R. microplus invasion. Taken together, these results ascertain the endemicity of tick-borne infections in Benin and suggest that the characteristics of Babesia spp. and A. marginale infections in R. microplus-invaded and non-invaded areas are different.